Using design of experiments to optimize derivatization with methyl chloroformate for quantitative analysis of the aqueous phase from hydrothermal liquefaction of biomass.

PubMed

Madsen, René Bjerregaard; Jensen, Mads Mørk; Mørup, Anders Juul; Houlberg, Kasper; Christensen, Per Sigaard; Klemmer, Maika; Becker, Jacob; Iversen, Bo Brummerstedt; Glasius, Marianne

2016-03-01

Hydrothermal liquefaction is a promising technique for the production of bio-oil. The process produces an oil phase, a gas phase, a solid residue, and an aqueous phase. Gas chromatography coupled with mass spectrometry is used to analyze the complex aqueous phase. Especially small organic acids and nitrogen-containing compounds are of interest. The efficient derivatization reagent methyl chloroformate was used to make analysis of the complex aqueous phase from hydrothermal liquefaction of dried distillers grains with solubles possible. A circumscribed central composite design was used to optimize the responses of both derivatized and nonderivatized analytes, which included small organic acids, pyrazines, phenol, and cyclic ketones. Response surface methodology was used to visualize significant factors and identify optimized derivatization conditions (volumes of methyl chloroformate, NaOH solution, methanol, and pyridine). Twenty-nine analytes of small organic acids, pyrazines, phenol, and cyclic ketones were quantified. An additional three analytes were pseudoquantified with use of standards with similar mass spectra. Calibration curves with high correlation coefficients were obtained, in most cases R (2)  > 0.991. Method validation was evaluated with repeatability, and spike recoveries of all 29 analytes were obtained. The 32 analytes were quantified in samples from the commissioning of a continuous flow reactor and in samples from recirculation experiments involving the aqueous phase. The results indicated when the steady-state condition of the flow reactor was obtained and the effects of recirculation. The validated method will be especially useful for investigations of the effect of small organic acids on the hydrothermal liquefaction process.

Determination of D- and L-amino acids produced by cyanobacteria using gas chromatography on Chirasil-Val after derivatization with pentafluoropropyl chloroformate.

PubMed

Zahradnícková, Helena; Husek, Petr; Simek, Petr; Hartvich, Petr; Marsálek, Blahoslav; Holoubek, Ivan

2007-08-01

A rapid and simple method was developed for the determination of free amino acids (AAs) released from cyanobacteria. The procedure involves trapping of AAs from the centrifuged cyanobacterial culture fluid on a cation-exchange resin, their release together with the resin by direct treatment with the reaction medium, followed by immediate derivatization with a corresponding chloroformate. The extractive alkylation transfers the analytes into an organic phase, an aliquot of which is subjected to GC analysis. Identification and quantification of AAs was performed by GC/MS and GC/FID, respectively, using propyl chloroformate (PCF) as the derivatization reagent. For chiral analysis, the cyanobacteria extracts were treated with 2,2,3,3,3-pentafluoropropyl chloroformate (PFPCF) to create more volatile analytes. Separation of the AA enantiomers was accomplished on a Chirasil-Val capillary column and the D/L enantiomeric ratios were determined. AAs of cyanobacteria are considered to be important for the assessment of energy flow in an aquatic food web, nutrition value of cyanobacteria in a food web and for cell-cell communication within cyanobacteria. The highest levels of AAs were found in the summer period at the beginning of the season (July). In the September and October samples, the amount of AAs was lower, the number of D-AAs decreased and the D/L ratio was higher than in the July sample. Based on the obtained results it can be assumed that young populations excrete AAs in higher concentrations and a different composition compared to actively growing populations.

Analysis of ammonium nitrate headspace by on-fiber solid phase microextraction derivatization with gas chromatography mass spectrometry.

PubMed

Lubrano, Adam L; Andrews, Benjamin; Hammond, Mark; Collins, Greg E; Rose-Pehrsson, Susan

2016-01-15

A novel analytical method has been developed for the quantitation of trace levels of ammonia in the headspace of ammonium nitrate (AN) using derivatized solid phase microextraction (SPME) fibers with gas chromatography mass spectrometry (GC-MS). Ammonia is difficult to detect via direct injection into a GC-MS because of its low molecular weight and extreme polarity. To circumvent this issue, ammonia was derivatized directly onto a SPME fiber by the reaction of butyl chloroformate coated fibers with the ammonia to form butyl carbamate. A derivatized externally sampled internal standard (dESIS) method based upon the reactivity of diethylamine with unreacted butyl chloroformate on the SPME fiber to form butyl diethylcarbamate was established for the reproducible quantification of ammonia concentration. Both of these compounds are easily detectable and separable via GC-MS. The optimized method was then used to quantitate the vapor concentration of ammonia in the headspace of two commonly used improvised explosive device (IED) materials, ammonium nitrate fuel oil (ANFO) and ammonium nitrate aluminum powder (Ammonal), as well as identify the presence of additional fuel components within the headspace. Published by Elsevier B.V.

A rapid novel derivatization of amphetamine and methamphetamine using 2,2,2-trichloroethyl chloroformate for gas chromatography electron ionization and chemical ionization mass spectrometric analysis.

PubMed

Dasgupta, A; Spies, J

1998-05-01

Amphetamine and methamphetamine are commonly abused central nervous system stimulants. We describe a rapid new derivatization of amphetamine and methamphetamine using 2,2,2-trichloroethyl chloroformate for gas chromatography-mass spectrometric analysis. Amphetamine and methamphetamine, along with N-propyl amphetamine (internal standard), were extracted from urine using 1-chlorobutane. The derivatization with 2,2,2-trichloroethyl chloroformate can be achieved at room temperature in 10 minutes. The electron ionization mass spectrum of amphetamine 2,2,2-trichloroethyl carbamate showed two weak molecular ions at m/z 309 and 311, but showed diagnostic strong peaks at m/z 218, 220, and 222. In contrast, chemical ionization of the mass spectrum of amphetamine 2,2,2-trichloroethyl carbamate showed strong (M + 1) ions at m/z 310 and 312 and other strong diagnostic peaks at m/z 274 and 276. The major advantages of this derivative are the presence of a diagnostic cluster of peaks due to the isotopic effect of three chlorine atoms (isotopes 35 and 37) in the derivatized molecule and the relative ease of its preparation. We also observed strong molecular ions for derivatized methamphetamine in the chemical ionization mass spectrum, but the molecular ions were very weak in the electron ionization mass spectrum. We used the scan mode of mass spectrometry in all analyses. When using a urine standard containing 1,000 ng/mL of amphetamine (a 7.4-micromol/L concentration) and methamphetamine (a 6.7-micromol/L concentration), the within-run precisions were 4.8% for amphetamine and 3.6% for methamphetamine. The corresponding between-run precisions were 5.3% for amphetamine and 6.7% for methamphetamine. The assay was linear for amphetamine and methamphetamine concentrations of 250 to 5,000 ng/mL (amphetamine, 1.9-37.0 micromol/L; methamphetamine, 1.7-33.6 micromol/L). The detection limit was 100 ng/mL (amphetamine, 0.74 micromol/L; methamphetamine, 0.67 micromol/L) using the scan mode of electron ionization mass spectrometry. We observed good a correlation between the concentrations of amphetamine and methamphetamine in five urine specimens positive for amphetamines using the more conventional pentafluoropropionyl derivative and our new derivative using 2,2,2-trichloroethyl chloroformate.

Determination of octylphenol and nonylphenol in aqueous sample using simultaneous derivatization and dispersive liquid-liquid microextraction followed by gas chromatography-mass spectrometry.

PubMed

Luo, Shusheng; Fang, Ling; Wang, Xiaowei; Liu, Hongtao; Ouyang, Gangfeng; Lan, Chongyu; Luan, Tiangang

2010-10-22

A simple and fast sample preparation method for the determination of nonylphenol (NP) and octylphenol (OP) in aqueous samples by simultaneous derivatization and dispersive liquid-liquid microextraction (DLLME) was investigated using gas chromatography-mass spectrometry (GC/MS). In this method, a combined dispersant/derivatization catalyst (methanol/pyridine mixture) was firstly added to an aqueous sample, following which a derivatization reagent/extraction solvent (methyl chloroformate/chloroform) was rapidly injected to combine in situ derivatization and extraction in a single step. After centrifuging, the sedimented phase containing the analytes was injected into the GC port by autosampler for analysis. Several parameters, such as extraction solvent, dispersant solvent, amount of derivatization reagent, derivatization and extraction time, pH, and ionic strength were optimized to obtain higher sensitivity for the detection of NP and OP. Under the optimized conditions, good linearity was observed in the range of 0.1-1000 μg L⁻¹ and 0.01-100 μg L⁻¹ with the limits of detection (LOD) of 0.03 μg L⁻¹ and 0.002 μg L⁻¹ for NP and OP, respectively. Water samples collected from the Pearl River were analyzed with the proposed method, the concentrations of NP and OP were found to be 2.40 ± 0.16 μg L⁻¹ and 0.037 ± 0.001 μg L⁻¹, respectively. The relative recoveries of the water samples spiked with different concentrations of NP and OP were in the range of 88.3-106.7%. Compared with SPME and SPE, the proposed method can be successfully applied to the rapid and convenient determination of NP and OP in aqueous samples. Copyright © 2010 Elsevier B.V. All rights reserved.

In vitro Antioxidant and Antiproliferative Activities of Various Solvent Fractions from Clerodendrum viscosum Leaves.

PubMed

Shendge, Anil Khushalrao; Basu, Tapasree; Chaudhuri, Dipankar; Panja, Sourav; Mandal, Nripendranath

2017-07-01

Free radicals such as reactive oxygen and nitrogen species, generated in the body, play an important role in the fulfillment of various physiological functions but their imbalance in the body lead to cellular injury and various clinical disorders such as cancer, neurodegenaration, and inflammation. The objective of this study is to fight this problem, natural antioxidant from plants can be considered as possible protective agents against various diseases such as cancer which might also modify the redox microenvironment to reduce the genetic instability. This study was designed to evaluate the antioxidant and antiproliferative potential of Clerodendrum viscosum fractions against various carcinomas. In this present study, 70% methanolic extract of C. viscosum leaves have been fractionated to obtain hexane, chloroform, ethyl acetate, butanol, and water fractions, which were tested for their antioxidant and anticancer properties. It was observed that chloroform and ethyl acetate fractions showed good free radical scavenging properties as well as inhibited the proliferation of human lung cancer (A459), breast (MCF-7), and brain (U87) cells. Moreover, they arrested the cell cycle at G2/M phase of breast and brain cancer. These inhibitory effects were further confirmed by bromodeoxyuridine uptake imaging. Phytochemical investigations further indicate the presence of tannic acid, quercetin, ellagic caid, gallic acid, reserpine, and methyl gallate which might be the reason for these fractions' antioxidant and antiproliferative activities. Clerodendrum viscosum leaf chloroform and Clerodendrum viscosum leaf ethyl acetate fractions from C. viscosum showed good reactive oxygen species and reactive nitrogen species scavenging potential. Both the fractions arrested cell cycle at G2/M phase in MCF-7 and U87 cells which lead to induce apoptosis. Crude extract of Clerodendrum viscosum leaves was fractionated using different solventsAmong them, chloroform and ethyl acetate fractions exhibited excellent free radical scavenging propertiesThe same fractions inhibited the proliferation of human lung cancer (A459), breast (MCF-7), and brain (U87) cellsChloroform and ethyl acetate fractions arrested the cell cycle at G2/M phase of breast and brain cancerPhytochemical investigations further indicate the presence of several bioactive principles present in them. Abbreviations used: CVLME: Clerodendrum viscosum leaf methanolic extract; CVLH: Clerodendrum viscosum leaf hexane; CVLC: Clerodendrum viscosum leaf chloroform; CVLE: Clerodendrum viscosum leaf ethyl acetate; CVLB: Clerodendrum viscosum leaf butanol; CVLW: Clerodendrum viscosum leaf water; BrdU: Bromodeoxyuridine; WST-1: Water soluble tetrazolium salt.

Separation of chemical groups from bio-oil aqueous phase via sequential organic solvent extraction

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ren, Shoujie; Ye, Philip; Borole, Abhijeet P

Bio-oil aqueous phase contains a considerable amount of furans, alcohols, ketones, aldehydes and phenolics besides the major components of organic acids and anhydrosugars. The complexity of bio-oil aqueous phase limits its efficient utilization. To improve the efficiency of bio-oil biorefinery, this study focused on the separation of chemical groups from bio-oil aqueous phase via sequential organic solvent extractions. Due to their high recoverability and low solubility in water, four solvents (hexane, petroleum ether, chloroform, and ethyl acetate) with different polarities were evaluated, and the optimum process conditions for chemical extraction were determined. Chloroform had high extraction efficiency for furans, phenolics,more » and ketones. In addition to these chemical groups, ethyl acetate had high extraction efficiency for organic acids. The sequential extraction by using chloroform followed by ethyl acetate rendered that 62.2 wt.% of original furans, ketones, alcohols, and phenolics were extracted to chloroform, over 62 wt.% acetic acid was extracted to ethyl acetate, resulting in a high concentration of levoglucosan (~53.0 wt.%) in the final aqueous phase. Chemicals separated via the sequential extraction could be used as feedstocks in biorefinery using processes such as catalytic upgrading of furans and phenolics to hydrocarbons, fermentation of levoglucosan to produce alcohols and diols, and hydrogen production from organic acids via microbial electrolysis.« less

Separation of chemical groups from bio-oil aqueous phase via sequential organic solvent extraction

DOE PAGES

Ren, Shoujie; Ye, Philip; Borole, Abhijeet P

2017-01-05

Bio-oil aqueous phase contains a considerable amount of furans, alcohols, ketones, aldehydes and phenolics besides the major components of organic acids and anhydrosugars. The complexity of bio-oil aqueous phase limits its efficient utilization. To improve the efficiency of bio-oil biorefinery, this study focused on the separation of chemical groups from bio-oil aqueous phase via sequential organic solvent extractions. Due to their high recoverability and low solubility in water, four solvents (hexane, petroleum ether, chloroform, and ethyl acetate) with different polarities were evaluated, and the optimum process conditions for chemical extraction were determined. Chloroform had high extraction efficiency for furans, phenolics,more » and ketones. In addition to these chemical groups, ethyl acetate had high extraction efficiency for organic acids. The sequential extraction by using chloroform followed by ethyl acetate rendered that 62.2 wt.% of original furans, ketones, alcohols, and phenolics were extracted to chloroform, over 62 wt.% acetic acid was extracted to ethyl acetate, resulting in a high concentration of levoglucosan (~53.0 wt.%) in the final aqueous phase. Chemicals separated via the sequential extraction could be used as feedstocks in biorefinery using processes such as catalytic upgrading of furans and phenolics to hydrocarbons, fermentation of levoglucosan to produce alcohols and diols, and hydrogen production from organic acids via microbial electrolysis.« less

Multiresidue screening of endocrine-disrupting chemicals and pharmaceuticals in aqueous samples by multi-stir bar sorptive extraction-single desorption-capillary gas chromatography/mass spectrometry.

PubMed

Van Hoeck, Els; Canale, Francesca; Cordero, Chiara; Compernolle, Sien; Bicchi, Carlo; Sandra, Pat

2009-02-01

A multiresidue method for screening endocrine-disrupting chemicals (EDCs) and pharmaceuticals in aqueous samples is presented. Four 10-mL aliquots of water were taken for stir bar sorptive extraction (SBSE) and they were treated in the following way. In sample one, in situ derivatization was performed with acetic acid anhydride to improve the extraction efficiencies and chromatographic analysis of phenolic compounds. For the same reasons, aliquot two was treated with ethyl chloroformate to improve amine and acid extraction and analysis, and aliquot three with tetraethylborate to enhance organotin compound extraction and analysis. Methanol was added to sample four to stop adsorption of apolar solutes on the wall. After SBSE, the four stir bars, together with a plug of glass wool impregnated with bis(trimethylylsilyl)trifluoroacetamide (BSTFA) to derivatize hydroxyl functionalities, were introduced into the same thermal desorption tube, heat-desorbed, and analyzed simultaneously by capillary GC/MS. The figures of merit of the method were evaluated with an EDC model mixture. In scan-mode MS, the limits of detection (LODs) were in the range 1-500 ng/L, while the LODs dropped by a factor of 50-100 when ion monitoring MS was applied to the targets. The performance of the method was illustrated by analysing some real-world water samples.

Analytical platform for metabolome analysis of microbial cells using methyl chloroformate derivatization followed by gas chromatography-mass spectrometry.

PubMed

Smart, Kathleen F; Aggio, Raphael B M; Van Houtte, Jeremy R; Villas-Bôas, Silas G

2010-09-01

This protocol describes an analytical platform for the analysis of intra- and extracellular metabolites of microbial cells (yeast, filamentous fungi and bacteria) using gas chromatography-mass spectrometry (GC-MS). The protocol is subdivided into sampling, sample preparation, chemical derivatization of metabolites, GC-MS analysis and data processing and analysis. This protocol uses two robust quenching methods for microbial cultures, the first of which, cold glycerol-saline quenching, causes reduced leakage of intracellular metabolites, thus allowing a more reliable separation of intra- and extracellular metabolites with simultaneous stopping of cell metabolism. The second, fast filtration, is specifically designed for quenching filamentous micro-organisms. These sampling techniques are combined with an easy sample-preparation procedure and a fast chemical derivatization reaction using methyl chloroformate. This reaction takes place at room temperature, in aqueous medium, and is less prone to matrix effect compared with other derivatizations. This protocol takes an average of 10 d to complete and enables the simultaneous analysis of hundreds of metabolites from the central carbon metabolism (amino and nonamino organic acids, phosphorylated organic acids and fatty acid intermediates) using an in-house MS library and a data analysis pipeline consisting of two free software programs (Automated Mass Deconvolution and Identification System (AMDIS) and R).

Amino acid analysis in physiological samples by GC-MS with propyl chloroformate derivatization and iTRAQ-LC-MS/MS.

PubMed

Dettmer, Katja; Stevens, Axel P; Fagerer, Stephan R; Kaspar, Hannelore; Oefner, Peter J

2012-01-01

Two mass spectrometry-based methods for the quantitative analysis of free amino acids are described. The first method uses propyl chloroformate/propanol derivatization and gas chromatography-quadrupole mass spectrometry (GC-qMS) analysis in single-ion monitoring mode. Derivatization is carried out directly in aqueous samples, thereby allowing automation of the entire procedure, including addition of reagents, extraction, and injection into the GC-MS. The method delivers the quantification of 26 amino acids. The isobaric tagging for relative and absolute quantification (iTRAQ) method employs the labeling of amino acids with isobaric iTRAQ tags. The tags contain two different cleavable reporter ions, one for the sample and one for the standard, which are detected by fragmentation in a tandem mass spectrometer. Reversed-phase liquid chromatography of the labeled amino acids is performed prior to mass spectrometric analysis to separate isobaric amino acids. The commercial iTRAQ kit allows for the analysis of 42 physiological amino acids with a respective isotope-labeled standard for each of these 42 amino acids.

In vitro Antioxidant and Antiproliferative Activities of Various Solvent Fractions from Clerodendrum viscosum Leaves

PubMed Central

Shendge, Anil Khushalrao; Basu, Tapasree; Chaudhuri, Dipankar; Panja, Sourav; Mandal, Nripendranath

2017-01-01

Background: Free radicals such as reactive oxygen and nitrogen species, generated in the body, play an important role in the fulfillment of various physiological functions but their imbalance in the body lead to cellular injury and various clinical disorders such as cancer, neurodegenaration, and inflammation. Objective: The objective of this study is to fight this problem, natural antioxidant from plants can be considered as possible protective agents against various diseases such as cancer which might also modify the redox microenvironment to reduce the genetic instability. This study was designed to evaluate the antioxidant and antiproliferative potential of Clerodendrum viscosum fractions against various carcinomas. Materials and Methods: In this present study, 70% methanolic extract of C. viscosum leaves have been fractionated to obtain hexane, chloroform, ethyl acetate, butanol, and water fractions, which were tested for their antioxidant and anticancer properties. Results: It was observed that chloroform and ethyl acetate fractions showed good free radical scavenging properties as well as inhibited the proliferation of human lung cancer (A459), breast (MCF-7), and brain (U87) cells. Moreover, they arrested the cell cycle at G2/M phase of breast and brain cancer. These inhibitory effects were further confirmed by bromodeoxyuridine uptake imaging. Phytochemical investigations further indicate the presence of tannic acid, quercetin, ellagic caid, gallic acid, reserpine, and methyl gallate which might be the reason for these fractions’ antioxidant and antiproliferative activities. Conclusion: Clerodendrum viscosum leaf chloroform and Clerodendrum viscosum leaf ethyl acetate fractions from C. viscosum showed good reactive oxygen species and reactive nitrogen species scavenging potential. Both the fractions arrested cell cycle at G2/M phase in MCF-7 and U87 cells which lead to induce apoptosis. SUMMARY Crude extract of Clerodendrum viscosum leaves was fractionated using different solventsAmong them, chloroform and ethyl acetate fractions exhibited excellent free radical scavenging propertiesThe same fractions inhibited the proliferation of human lung cancer (A459), breast (MCF-7), and brain (U87) cellsChloroform and ethyl acetate fractions arrested the cell cycle at G2/M phase of breast and brain cancerPhytochemical investigations further indicate the presence of several bioactive principles present in them. Abbreviations used: CVLME: Clerodendrum viscosum leaf methanolic extract; CVLH: Clerodendrum viscosum leaf hexane; CVLC: Clerodendrum viscosum leaf chloroform; CVLE: Clerodendrum viscosum leaf ethyl acetate; CVLB: Clerodendrum viscosum leaf butanol; CVLW: Clerodendrum viscosum leaf water; BrdU: Bromodeoxyuridine; WST-1: Water soluble tetrazolium salt. PMID:28808404

Evaluation of larvicidal and nymphicidal potential of plant extracts against Anopheles subpictus Grassi, Culex tritaeniorhynchus Giles and Aphis gossypii Glover.

PubMed

Bagavan, A; Kamaraj, C; Rahuman, A Abdul; Elango, G; Zahir, A Abduz; Pandiyan, G

2009-04-01

The acetone, chloroform, ethyl acetate, hexane and methanol extracts of peel and leaf extracts of Citrus sinensis, Ocimum canum, Ocimum sanctum and Rhinacanthus nasutus were tested against fourth instar larvae of malaria vector, Anopheles subpictus Grassi, Japanese encephalitis vector, Culex tritaeniorhynchus Giles (Diptera: Culicidae) and feeding deterrence to nymphs of cotton pest, Aphis gossypii Glover (Homoptera: Aphididae). The larval and nymph mortality were observed after 24 h of exposure. All extracts showed moderate larvicidal and nymphicidal effects; however, the highest mortality was found in peel chloroform extract of C. sinensis, leaf ethyl acetate extracts of O. canum and O. sanctum and leaf chloroform extract of R. nasutus against the larvae of A. subpictus (LC(50) = 58.25, 88.15, 21.67 and 40.46 ppm; LC(90) = 298.31, 528.70, 98.34 and 267.20 ppm), peel methanol extract of C. sinensis, leaf methanol extract of O. canum, ethyl acetate extracts of O. sanctum and R. nasutus against the larvae of C. tritaeniorhynchus (LC(50) = 38.15, 72.40, 109.12 and 39.32 ppm; LC(90) = 184.67, 268.93, 646.62 and 176.39 ppm), peel hexane extract of C. sinensis, leaf methanol extracts of O. canum and R. nasutus and leaf ethyl acetate extract of O. sanctum against the nymph of A. gossypii (LC(50) = 162.89, 80.99, 73.27 and 130.19 ppm; LC(90) = 595.40, 293.33, 338.74 and 450.90 ppm), respectively. These results suggest that the peel methanol extracts of C. sinensis and O. canum, ethyl acetate leaf extract of O. sanctum and leaf chloroform and ethyl acetate extract of R. nasutus have the potential to be used as an ideal eco-friendly approach for the control of the A. subpictus, C. tritaeniorhynchus and A. gossypii.

Alpha-glucosidase Inhibitory and Antioxidant Potential of Antidiabetic Herb Alternanthera sessilis: Comparative Analyses of Leaf and Callus Solvent Fractions.

PubMed

Chai, Tsun-Thai; Khoo, Chee-Siong; Tee, Chong-Siang; Wong, Fai-Chu

2016-01-01

Alternanthera sessilis is a medicinal herb which is consumed as vegetable and used as traditional remedies of various ailments in Asia and Africa. This study aimed to investigate the antiglucosidase and antioxidant activity of solvent fractions of A. sessilis leaf and callus. Leaf and callus methanol extracts were fractionated to produce hexane, chloroform, ethyl acetate, butanol, and water fractions. Antiglucosidase and 1,1-diphenyl-2-picrylhydrazyl scavenging activities as well as total phenolic (TP), total flavonoid (TF), and total coumarin (TC) contents were evaluated. Lineweaver-Burk plot analysis was performed on leaf and callus fractions with the strongest antiglucosidase activity. Leaf ethyl acetate fraction (LEF) had the strongest antiglucosidase (EC 50 0.55 mg/mL) and radical scavenging (EC 50 10.81 μg/mL) activity among leaf fractions. Callus ethyl acetate fraction (CEF) and chloroform fraction had the highest antiglucosidase (EC 50 0.25 mg/mL) and radical scavenging (EC 50 34.12 μg/mL) activity, respectively, among callus fractions. LEF and CEF were identified as noncompetitive and competitive α-glucosidase inhibitors, respectively. LEF and CEF had greater antiglucosidase activity than acarbose. Leaf fractions had higher phytochemical contents than callus fractions. LEF had the highest TP, TF, and TC contents. Antiglucosidase and antioxidant activities of leaf fractions correlated with phytochemical contents. LEF had potent antiglucosidase activity and concurrent antioxidant activity. CEF had the highest antiglucosidase activity among all fractions. Callus culture is a promising tool for enhancing production of potent α-glucosidase inhibitors. Leaf ethyl acetate fraction (LEF) had the strongest antiglucosidase (EC 50 0.55 mg/mL) and radical scavenging (EC 50 10.81 μg/mL) activity among leaf fractionsCallus ethyl acetate fraction (CEF) and chloroform fraction had the highest antiglucosidase (EC 50 0.25 mg/mL) and radical scavenging (EC 50 34.12 μg/mL) activity, respectively, among callus fractionsLEF and CEF were identified as noncompetitive and competitive á-glucosidase inhibitors, respectivelyAntiglucosidase and antioxidant activities of leaf fractions correlated with phytochemical contents. Abbreviations used: LHF: Leaf hexane fraction, LCF: Leaf chloroform fraction, LEF: Leaf ethyl acetate fraction, LBF: Leaf butanol fraction, LWF: Leaf water fraction, CHF: Callus hexane fraction, CCF: Callus chloroform fraction, CEF: Callus ethyl acetate fraction, CBF: Callus butanol fraction, CWF: Callus water fraction, TP: Total phenolic, TF: Total flavonoid, TC: Total coumarin.

Human Cytochrome P450 Enzyme Modulation by Gymnema sylvestre: A Predictive Safety Evaluation by LC-MS/MS.

PubMed

Rammohan, Bera; Samit, Karmakar; Chinmoy, Das; Arup, Saha; Amit, Kundu; Ratul, Sarkar; Sanmoy, Karmakar; Dipan, Adhikari; Tuhinadri, Sen

2016-07-01

Traditionally GS is used to treat diabetes mellitus. Drug-herb interaction of GS via cytochrome P450 enzyme system by substrate cocktail method using HLM has not been reported. To evaluate the in-vitro modulatory effects of GS extracts (aqueous, methanol, ethyl acetate, chloroform and n -hexane) and deacylgymnemic acid (DGA) on human CYP1A2, 2C8, 2C9, 2D6 and 3A4 activities in HLM. Probe substrate-based LCMS/MS method was established for all CYPs. The metabolite formations were examined after incubation of probe substrates with HLM in the presence or absence of extracts and DGA. The inhibitory effects of GS extracts and DGA were characterized with kinetic parameters IC50 and Ki values. GS extracts showed differential effect on CYP activities in the following order of inhibitory potency: ethyl acetate > Chloroform > methanol > n -hexane > aqueous > DGA. This differential effect was observed against CYP1A2, 2C9 and less on CYP3A4 and 2C8 but all CYPs were unaffected by aqueous extract and DGA. The ethyl acetate and chloroform extract exhibited moderate inhibition towards CYP1A2 and 3A4. The aqueous extract and DGA however showed negligible inhibition towards all five major human CYPs with very high IC50 values (>90μg/ml). The results of our study revealed that phytoconstituents contained in GS, particularly in ethyl acetate and chloroform extracts, were able to inhibit CYP1A2, 3A4 and 2C9. The presence of relatively small, lipophillic yet slightly polar compounds within the GS extracts may be attributed for inhibition activities. These suggest that the herb or its extracts should be examined for potential pharmacokinetic drug interactions in vivo . Abbreviations used: GS: Gymnema sylvestre , GSE: Gymnema sylvestre extract, DGA: deacyl gymnemic acid, CYP: cytochrome P450, DMSO: dimethylsulphoxide, HLM: human liver microsomes, LC-MS/MS: liquid chromatography tandem mass spectroscopy, NADPH: reduced nicotinamide adeninedinucleotide phosphate, NRS: nicotinamide adeninedinucleotide phosphate regenerating system, CHE: chloroform extract, EAE: ethyl acetate extract, NHE- n -hexane extract, AE: aqueous extract, ME: methanol extract.

Cotinus coggyria: a rich source of antioxidants.

PubMed

Riaz, Tauheeda; Abbasi, Muhammad Athar; Aziz-ur-Rehman; Rubab, Kaniz; Shahzadi, Tayyaba; Ajaib, Muhammad; Khan, Khalid Mohammed

2012-07-01

Methanolic extract of Cotinus coggyria Scop. was mixed in distilled water and partitioned first with the n-hexane, then with chloroform, then ethyl acetate and at the end with n-butanol. The phytochemical screening of plant showed presence of the phenolics, cardiac glycosides and flavonoides in large amount in the chloroform, n-butanol and ethyl acetate soluble fraction. Antioxidant activity of these four fractions and the left behind aqueous fraction was measured by four methods such as: 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, ferric thiocyanate assay, ferric reducing antioxidant power (FRAP) assay and total antioxidant activity. Total phenolics were also measured. Noteworthy antioxidant potential was shown by the chloroform, n-butanol and ethyl acetate soluble fraction showed. Ethyl acetate fraction showed highest % inhibition of the DPPH radical when compared with the other studied fractions i.e. 81.64 ± 1.29% inhibition of the DPPH radical at the concentration of 30 μg/ml. Its IC(50) value was found to be 15.58 ± 0.09 μg/ml, comparative to the butylated hydroxytoluene (BHT), which has IC(50) value 12.6 ± 0.85μg/ml. This fraction also showed the highest lipid peroxidation inhibition (61.41 ± 1.16%), as well as highest values of FRAP (697.76 ± 1.98 μg of trolox equivalents) total antioxidant activity (1.02 ± 0.09) and total phenolic contents (229.34 ± 0.57) comparative to the other studied fractions. The chloroform and n-butanol soluble fraction also showed good results for all the studied antioxidant assays.

Cytotoxic and phytotoxic actions of Heliotropium strigosum.

PubMed

Shah, Syed Majid; Hussain, Sajid; Khan, Arif-Ullah; Shah, Azhar-Ul-Haq Ali; Khan, Haroon; Ullah, Farhat; Barkatullah

2015-05-01

This study describes the cytotoxic and phytotoxic activities of the crude extract of Heliotropium strigosum and its resultant fractions. In brine shrimp toxicology assays, profound cytotoxicity was displayed by ethyl acetate (LD50 8.3 μg/ml) and chloroform (LD50 8.8 μg/ml) fractions, followed by relatively weak crude methanolic extract of H. strigosum (LD50 909 μg/ml) and n-hexane fraction (LD50 1000 μg/ml). In case of phytotoxicity activity against Lemna acquinoctialis, highest phytotoxic effect was showed by ethyl acetate fraction (LD50 91.0 μg/ml), while chloroform fraction, plant crude extract and n-hexane, respectively, caused 50%, 30.76 ± 1.1% and 30.7 ± 1.1% inhibitory action at maximum concentration used, that is, 1000 μg/ml. These data indicates that H. strigosum exhibits cytotoxic and phytotoxic potential, which explore its use as anticancer and herbicidal medicine. The ethyl acetate and chloroform fractions were more potent for the evaluated toxicity effects, thus recommended for isolation and identification of the active compounds. © The Author(s) 2012.

HPLC purification and re-evaluation of chemical identity of two circular bacteriocins, gassericin A and reutericin 6.

PubMed

Arakawa, K; Kawai, Y; Ito, Y; Nakamura, K; Chujo, T; Nishimura, J; Kitazawa, H; Saito, T

2010-04-01

The study aimed for the complete purification and recharacterization of the highly hydrophobic circular bacteriocins, gassericin A and reutericin 6. Gassericin A and reutericin 6 were purified to homogeneity using previously described method and reverse-phase HPLC with an octyl column and eluents of aqueous acetonitrile and 2-propanol. Mass analysis, N-terminal sequencing and bacteriocin assay of the HPLC-purified bacteriocins showed the two bacteriocins had identical seamless circular structures with the same m/z value (5651) of [M + H](+) and both had the same specific activity. D/L-amino acid composition analysis using two distinct methods with the chiral fluorescent derivatization reagents (+)-1-(9-fluorenyl)ethyl chloroformate and O-phthalaldehyde/N-acetyl-L-cystein revealed neither gassericin A nor reutericin 6 contained D-alanine residues contrary to our previous results. Purified gassericin A and reutericin 6 are chemically identical circular molecules containing no D-alanine residues. The HPLC conditions developed in this study will facilitate advanced purification and correct characterization of other highly hydrophobic bacteriocins.

Development of the 2007 Chemical Decontaminant Source Document

DTIC Science & Technology

2009-03-01

Chemical Agent Simulant Specific DEM diethyl malonate MeS methyl salicylate PEG200 Polyethylene glycol 200 TEP triethyl phosphate Group 6...simulants • H-agent simulants o Methyl salicylate (MeS) o Chloroethyl phenyl sulfide (CEPS) o Chloroethyl ethyl sulfide (CEES) • VX simulants... Methyl bromide Ethyl phosphonothioic dichloride Sulfur dioxide Methyl chloroformate Ethyl phosphonic dichloride Sulfuric acid Methyl chlorosilane

In vitro total phenolics, flavonoids contents and antioxidant activity of essential oil, various organic extracts from the leaves of tropical medicinal plant Tetrastigma from Sabah.

PubMed

Hossain, M Amzad; Shah, Muhammad Dawood; Gnanaraj, Charles; Iqbal, Muhammad

2011-09-01

To detect the in vitro total phenolics, flavonoids contents and antioxidant activity of essential oil, various organic extracts from the leaves of tropical medicinal plant Tetrastigma from Sabah. The dry powder leaves of Tetrastigma were extracted with different organic solvent such as hexane, ethyl acetate, chloroform, butanol and aqueous methanol. The total phenolic and total flavonoids contents of the essential oil and various organic extracts such as hexane, ethyl acetate, chloroform, butanol and aqueous ethanol were determined by Folin - Ciocalteu method and the assayed antioxidant activity was determined in vitro models such as antioxidant capacity by radical scavenging activity using α, α-diphenyl- β-picrylhydrazyl (DPPH) method. The total phenolic contents of the essential oil and different extracts as gallic acid equivalents were found to be highest in methanol extract (386.22 mg/g) followed by ethyl acetate (190.89 mg/g), chloroform (175.89 mg/g), hexane (173.44 mg/g), and butanol extract (131.72 mg/g) and the phenolic contents not detected in essential oil. The antioxidant capacity of the essential oil and different extracts as ascorbic acid standard was in the order of methanol extract > ethyl acetate extract >chloroform> butanol > hexane extract also the antioxidant activity was not detected in essential oil. The findings show that the extent of antioxidant activity of the essential oil and all extracts are in accordance with the amount of phenolics present in that extract. Leaves of Tetrastigma being rich in phenolics may provide a good source of antioxidant. Copyright © 2011 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Phytochemical contents and biological evaluation of Ruta chalepennsis L. growing in Saudi Arabia.

PubMed

Alotaibi, Shorok M; Saleem, Monerah S; Al-Humaidi, Jehan G

2018-05-01

Phytochemical screening of Ruta chalepensis L. exhibited the presence of different chemical groups. The dried aerial parts of the plant was total extracted by ethanol and successively using chloroform, ethyl acetate and Butanol, out of the successive extracts four compounds namely, scopletin, kaempferol, quercetin, quercetin 3- O -α-L-rhamno glucopyranosyl (Rutin) were isolated and biological evaluations. Total ethanol and successive extracts; chloroform, ethyl acetate and Butanol were produced excellent antimicrobial activities against gram negative bacteria, gram positive bacteria and fungi. Ethyl acetate extract was the best for inhibition of the microorganism's growth. All extracts (total ethanol, and successive extracts) showed DPPH radical scavenging activity in a concentration-dependent manner. The best antioxidant activity was obtained by ethyl acetate & n -butanol extract (94.28%, IC 50  = 56.6 µg/ml). Also All extracts (total ethanol, and successive extracts) showed anticoagulant activity at higher concentration with prolonged clotting time 6:30 and 4:30 s at 10 mg/ml concentrations, respectively.

Antibacterial activity study of Attacus atlas cocoon against Staphylococcus aureus and Escherichia coli with diffusion and dilution method

NASA Astrophysics Data System (ADS)

Aminah; Nugraheni, E. R.; Yugatama, A.

2018-03-01

The aim of this study was to evaluate the antibacterial activity from Attacus atlas cocoon extract against Gram-negative bacteria (Escherichia coli) and Gram-positive bacteria (Staphylococcus aureus) with 3 diffferent solvents polar, semi-polar and non polar which was ethanol, ethyl acetate and chloroform, also to determine the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of the extract. Cocoon was extracted with maceration method using 3 solvents with ratio of sample and solvent 1:10. Antibacterial activity of the Extracts obtained was evaluated with Agar disk diffusion method. The best result was then continued to determine the MIC and MBC of the extract using broth macro-dilution method. The results show that each of the extracts have antibacterial activity with broad spectrum against two different type of bacteria at concentration of 1 g/mL with different clear zone between these extracts. Clear zone from the biggest to the smallest against Escherichia coli was ethyl acetate (10.5 mm), chloroform (9 mm) and ethanol (8 mm). While against Staphylococcus aureus, was obtained by chloroform (12.5 mm), ethyl acetate (10.5 mm) and ethanol (7 mm). The MIC value of extracts can not be determine. The smallest MBC value against both bacteria was obtained by ethyl acetate with concentration of 3.125% b/v as a bactericidal.

Rapid determination of amino acids in neonatal blood samples based on derivatization with isobutyl chloroformate followed by solid-phase microextraction and gas chromatography/mass spectrometry.

PubMed

Deng, Chunhui; Li, Ning; Zhang, Xiangmin

2004-01-01

The purpose of this study was to develop a simple, rapid and sensitive analytical method for determination of amino acids in neonatal blood samples. The developed method involves the employment of derivatization and a solid-phase microextraction (SPME) technique together with gas chromatography/mass spectrometry (GC/MS). Amino acids in blood samples were derivatized by a mixture of isobutyl chloroformate, methanol and pyridine, and the N(O,S)-alkoxycarbonyl alkyl esters thus formed were headspace extracted by a SPME fiber. Finally, the extracted analytes on the fiber were desorbed and detected by GC/MS in electron impact (EI) mode. L-Valine, L-leucine, L-isoleucine, L-phenylanaline and L-tyrosine in blood samples were quantitatively analyzed by measurement of the corresponding N(O,S)-alkoxycarbonyl alkyl esters using an external standard method. SPME conditions were optimized, and the method was validated. The method was applied to diagnosis of neonatal phenylkenuria (PKU) and maple syrup urine disease (MSUD) by the analyses of five amino acids in blood samples. The results showed that the proposed method is a potentially powerful tool for simultaneous screening for neonatal PKU and MSUD. Copyright (c) 2004 John Wiley & Sons, Ltd.

Human Cytochrome P450 Enzyme Modulation by Gymnema sylvestre: A Predictive Safety Evaluation by LC-MS/MS

PubMed Central

Rammohan, Bera; Samit, Karmakar; Chinmoy, Das; Arup, Saha; Amit, Kundu; Ratul, Sarkar; Sanmoy, Karmakar; Dipan, Adhikari; Tuhinadri, Sen

2016-01-01

Background: Traditionally GS is used to treat diabetes mellitus. Drug-herb interaction of GS via cytochrome P450 enzyme system by substrate cocktail method using HLM has not been reported. Objective: To evaluate the in-vitro modulatory effects of GS extracts (aqueous, methanol, ethyl acetate, chloroform and n-hexane) and deacylgymnemic acid (DGA) on human CYP1A2, 2C8, 2C9, 2D6 and 3A4 activities in HLM. Material and Methods: Probe substrate-based LCMS/MS method was established for all CYPs. The metabolite formations were examined after incubation of probe substrates with HLM in the presence or absence of extracts and DGA. The inhibitory effects of GS extracts and DGA were characterized with kinetic parameters IC50 and Ki values. Results: GS extracts showed differential effect on CYP activities in the following order of inhibitory potency: ethyl acetate > Chloroform > methanol > n-hexane > aqueous > DGA. This differential effect was observed against CYP1A2, 2C9 and less on CYP3A4 and 2C8 but all CYPs were unaffected by aqueous extract and DGA. The ethyl acetate and chloroform extract exhibited moderate inhibition towards CYP1A2 and 3A4. The aqueous extract and DGA however showed negligible inhibition towards all five major human CYPs with very high IC50 values (>90μg/ml). Conclusion: The results of our study revealed that phytoconstituents contained in GS, particularly in ethyl acetate and chloroform extracts, were able to inhibit CYP1A2, 3A4 and 2C9. The presence of relatively small, lipophillic yet slightly polar compounds within the GS extracts may be attributed for inhibition activities. These suggest that the herb or its extracts should be examined for potential pharmacokinetic drug interactions in vivo. Abbreviations used: GS: Gymnema sylvestre, GSE: Gymnema sylvestre extract, DGA: deacyl gymnemic acid, CYP: cytochrome P450, DMSO: dimethylsulphoxide, HLM: human liver microsomes, LC-MS/MS: liquid chromatography tandem mass spectroscopy, NADPH: reduced nicotinamide adeninedinucleotide phosphate, NRS: nicotinamide adeninedinucleotide phosphate regenerating system, CHE: chloroform extract, EAE: ethyl acetate extract, NHE- n-hexane extract, AE: aqueous extract, ME: methanol extract PMID:27761064

Antioxidant activities of extracts and flavonoid compounds from Oxytropis falcate Bunge.

PubMed

Jiang, H; Zhan, W Q; Liu, X; Jiang, S X

2008-12-01

The antioxidant properties of the various extracts and flavonoids prepared from Oxytropis falcate Bunge were investigated by 1,1-diphenyl-2-picryldydrazyl (DPPH) radical-scavenging assay. In the chloroform, ethyl acetate and n-butanol extracts, the ethyl acetate extract exhibited the highest antioxidant activity (IC(50) = 2.05 mg mL(-1)). Furthermore, rhamnocitrin, kaempferol, rhamnetin, 2',4'-dihydroxychalcone and 2',4', beta-trihydroxy-dihydrochalcone were purified from chloroform and ethyl acetate extracts. The radical-scavenging activities of the five compounds were also measured and the results showed that kaempferol (IC(50) = 0.11 mg mL(-1)), rhamnetin (IC(50) = 0.14 mg mL(-1)) and rhamnocitrin (IC(50) = 0.15 mg mL(-1)) exhibited considerable antioxidant activities, but the antioxidant activities of the two dihydrochalcones were very weak. Although these flavonoids are known, this is the first report of antioxidant activity in this plant.

COMPARISON OF PERFLUORINATED CHLOROFORMATES AS DIRECT AQUEOUS SAMPLE DERIVATIZING AGENTS FOR HIGHLY HYDROPHILIC ANALYTES

EPA Science Inventory

Strong oxidants are supposed to produce quite extensive cleavage of the hydrocarbon backbone of natural organic matter, resulting in the release of partly oxidized organic molecules. The identification and detection of these small and highly polar compounds represents a challengi...

Quantitation of fumonisin B1 and B2 in feed using FMOC pre-column derivatization with HPLC and fluorescence detection.

PubMed

Smith, Lori L; Francis, Kyle A; Johnson, Joseph T; Gaskill, Cynthia L

2017-11-01

Pre-column derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) was determined to be effective for quantitation of fumonisins B 1 and B 2 in feed. Liquid-solid extraction, clean-up using immunoaffinity solid phase extraction chromatography, and FMOC-derivatization preceded analysis by reverse phase HPLC with fluorescence. Instrument response was unchanged in the presence of matrix, indicating no need to use matrix-matched calibrants. Furthermore, high method recoveries indicated calibrants do not need to undergo clean-up to account for analyte loss. Established method features include linear instrument response from 0.04-2.5µg/mL and stable derivatized calibrants over 7days. Fortified cornmeal method recoveries from 0.1-30.0μg/g were determined for FB 1 (75.1%-109%) and FB 2 (96.0%-115.2%). Inter-assay precision ranged from 1.0%-16.7%. Method accuracy was further confirmed using certified reference material. Inter-laboratory comparison with naturally-contaminated field corn demonstrated equivalent results with conventional derivatization. These results indicate FMOC derivatization is a suitable alternative for fumonisins B 1 and B 2 quantitation in corn-based feeds. Copyright © 2017 Elsevier Ltd. All rights reserved.

Antimicrobial potential of extracts from Stevia rebaudiana leaves against bacteria of importance in dental caries.

PubMed

Gamboa, Fredy; Chaves, Margarita

2012-01-01

In recent years, the antimicrobial activity of Stevia rebaudiana Bertoni leaf extracts against a large number of microorganisms has been evaluated, but not its activity against microorganisms of importance in dental caries. The aim of this study was to evaluate the antibacterial activity of Stevia rebaudiana Bertoni leaf extracts against cariogenic bacteria. Extracts were obtained from the dried Stevia rebaudiana Bertoni leaves in hexane, methanol, ethanol, ethyl acetate and chloroform. The antimicrobial activity of the 5 extracts against 16 bacterial strains of the genera Streptococcus (n= 12) and Lactobacillus (n= 4) was evaluated by the well diffusion method. Minimal inhibitory concentrations (MIC) of the extracts in hexane, methanol, ethanol, ethyl acetate and chloroform on the 16 bacterial strains were respectively 30 mg/ml, 120 mg/ml, 120 mg/ml, 60 mg/ml and 60 mg/ml. The zones of inhibition present at the MIC were variable, ranging from 9 mm to 17.3 mm. Our results suggest that inhibition zones with a hexane extract are similar to those obtained with ethanol and methanol, but the minimal inhibitory concentration (30 mg/ml) is lower. For the four Lactobacillus species, the inhibition zones obtained between 12.3 and 17.3 mm were somewhat larger with ethyl acetate and chloroform extracts, suggesting they were the most susceptible microorganisms.

In vitro antimicrobial activity of ethanolic fractions of Cryptolepis sanguinolenta

PubMed Central

2012-01-01

Background Following claims that some plants have antimicrobial activities against infectious microbes, the in vitro antimicrobial activities of different solvent fractions of ethanolic extract of Cryptolepis sanguinolenta were evaluated against eight standard bacteria and clinical isolates. Methods The solvent partitioning protocol involving ethanol, petroleum ether, chloroform, ethyl acetate and water, was used to extract various fractions of dried pulverized Cryptolepis sanguinolenta roots. Qualitative phyto-constituents screening was performed on the ethanol extract, chloroform fraction and the water fraction. The Kirby Bauer disk diffusion method was employed to ascertain the antibiogram of the test organisms while the agar diffusion method was used to investigate the antimicrobial properties of the crude plant extracts. The microplate dilution method aided in finding the MICs while the MBCs were obtained by the method of Nester and friends. The SPSS 16.0 version was used to analyze the percentages of inhibitions and bactericidal activities. Results The phytochemical screening revealed the presence of alkaloids, reducing sugars, polyuronides, anthocyanosides and triterpenes. The ethanol extract inhibited 5 out of 8 (62.5%) of the standard organisms and 6 out of 8 (75%) clinical isolates. The petroleum ether fraction inhibited 4 out of 8 (50%) of the standard microbes and 1 out of 8 (12.5%) clinical isolates. It was also observed that the chloroform fraction inhibited the growth of all the organisms (100%). Average inhibition zones of 14.0 ± 1.0 mm to 24.67 ± 0.58 mm was seen in the ethyl acetate fraction which halted the growth of 3 (37.5%) of the standard organisms. Inhibition of 7 (87.5%) of standard strains and 6 (75%) of clinical isolates were observed in the water fraction. The chloroform fraction exhibited bactericidal activity against all the test organisms while the remaining fractions showed varying degrees of bacteriostatic activity. Conclusion The study confirmed that fractions of Cryptolepis sanguinolenta have antimicrobial activity. The chloroform fraction had the highest activity, followed by water, ethanol, petroleum ether and ethyl acetate respectively. Only the chloroform fraction exhibited bactericidal activity and further investigations are needed to ascertain its safety and prospects of drug development. PMID:22709723

Pyrus pashia: A persuasive source of natural antioxidants.

PubMed

Siddiqui, Sabahat Zahra; Ali, Saima; Rehman, Azizur; Rubab, Kaniz; vAbbasi, Muhammad Athar; Ajaib, Muhammad; Z Rasool, Zahid Ghulam

2015-09-01

Pyrus pashia Buch. & Ham. was subjected to extraction with methanol. Methanolic extracts of fruit, bark and leaf were partitioned separately with four organic solvents in order of increasing polarity, asn-hexane, chloroform, ethyl acetate and n-butanol after dissolving in distilled water. Phytochemical screening revealed the presence of phenolics, flavonoides, alkaloids and cardiac glycosides in large amount in chloroform, ethyl acetate and n-butanol soluble fractions. The antioxidant activity of crude methanolic extracts, all the obtained fourorganic fractions and remaining aqueous fractions was evaluated by different methods such as: 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, ferric reducing antioxidant power (FRAP) assay and total antioxidant activity by phosphomolybdenum complex method as well as determination of total phenolics. The results of antioxidant activity exhibited that chloroform soluble fraction of fruit showed the highest value of percent inhibition of DPPH (48.16 ± 0.21 μg/ml) at the concentration of 10 μg/ml. Ethyl acetate soluble fraction displayed the lowest antioxidant activity having IC50 value of bark as (8.64 ± 0.32 μg/ml) relative to butylated hydroxytoluene (BHT), having IC50 of 12.1 ± 0.92 μg/ml. The ethyl acetate soluble fraction of bark revealed the highest FRAPs value (174.618 ± 0.11TE µM/ml) among all the three parts. This fraction also showed the highest value of total antioxidant activity as (1.499 ± 0.90), determined by phosphomolybdenum complex method. Moreover, this fraction also conferred the highest phenolic content (393.19 ± 0.72) as compared to other studied fractions of fruit and leaf.

Liquid chromatography coupled to quadrupole-time of flight tandem mass spectrometry based quantitative structure-retention relationships of amino acid analogues derivatized via n-propyl chloroformate mediated reaction.

PubMed

Kritikos, Nikolaos; Tsantili-Kakoulidou, Anna; Loukas, Yannis L; Dotsikas, Yannis

2015-07-17

In the current study, quantitative structure-retention relationships (QSRR) were constructed based on data obtained by a LC-(ESI)-QTOF-MS/MS method for the determination of amino acid analogues, following their derivatization via chloroformate esters. Molecules were derivatized via n-propyl chloroformate/n-propanol mediated reaction. Derivatives were acquired through a liquid-liquid extraction procedure. Chromatographic separation is based on gradient elution using methanol/water mixtures from a 70/30% composition to an 85/15% final one, maintaining a constant rate of change. The group of examined molecules was diverse, including mainly α-amino acids, yet also β- and γ-amino acids, γ-amino acid analogues, decarboxylated and phosphorylated analogues and dipeptides. Projection to latent structures (PLS) method was selected for the formation of QSRRs, resulting in a total of three PLS models with high cross-validated coefficients of determination Q(2)Y. For this reason, molecular structures were previously described through the use of descriptors. Through stratified random sampling procedures, 57 compounds were split to a training set and a test set. Model creation was based on multiple criteria including principal component significance and eigenvalue, variable importance, form of residuals, etc. Validation was based on statistical metrics Rpred(2),QextF2(2),QextF3(2) for the test set and Roy's metrics rm(Av)(2) and rm(δ)(2), assessing both predictive stability and internal validity. Based on aforementioned models, simplified equivalent were then created using a multi-linear regression (MLR) method. MLR models were also validated with the same metrics. The suggested models are considered useful for the estimation of retention times of amino acid analogues for a series of applications. Copyright © 2015 Elsevier B.V. All rights reserved.

Anti-inflammatory, cyclooxygenase (COX)-2, COX-1 inhibitory, and free radical scavenging effects of Rumex nepalensis.

PubMed

Gautam, Raju; Karkhile, Kailas V; Bhutani, Kamlesh K; Jachak, Sanjay M

2010-10-01

Evaluation of the topical anti-inflammatory activity of chloroform and ethyl acetate extracts of RUMEX NEPALENSIS roots in a TPA-induced acute inflammation mouse model demonstrated a significant reduction in ear edema. The extracts were further tested on purified enzymes for COX-1 and COX-2 inhibition to elucidate their mechanism of action, and a strong inhibition was observed. Six anthraquinones and two naphthalene derivatives were isolated from the ethyl acetate extract. Among the isolated compounds, emodin was found to be a potent inhibitor with slight selectivity towards COX-2, and nepodin exhibited selectivity towards COX-1. Emodin, endocrocin, and nepodin also exhibited significant topical anti-inflammatory activity in mice. Interestingly, nepodin showed better radical scavenging activity than trolox and ascorbic acid against DPPH and ABTS radicals. The strong radical scavenging activity of chloroform and ethyl acetate extracts could be explained by the presence of nepodin as well as by the high phenolic content of the ethyl acetate extract. Thus, the anti-inflammatory effect of R. NEPALENSIS roots was assumed to be mediated through COX inhibition by anthraquinones and naphthalene derivatives and through the radical scavenging activities of naphthalene derivatives. © Georg Thieme Verlag KG Stuttgart · New York.

Screening of plant extracts for anthelmintic activity against Dactylogyrus intermedius (Monogenea) in goldfish (Carassius auratus).

PubMed

Huang, Ai-Guo; Yi, Yang-Lei; Ling, Fei; Lu, Lin; Zhang, Qi-Zhong; Wang, Gao-Xue

2013-12-01

With the aim of finding natural anthelmintic agents against Dactylogyrus intermedius (Monogenea) in goldfish (Carassius auratus), 26 plants were screened for antiparasitic properties using in vivo anthelmintic efficacy assay. The results showed that Caesalpinia sappan, Lysima chiachristinae, Cuscuta chinensis, Artemisia argyi, and Eupatorium fortunei were found to have 100% anthelmintic efficacy at 125, 150, 225, 300, and 500 mg L(-1) after 48 h of exposure. Crude extract of the five plants were further partitioned with petroleum ether, chloroform, ethyl acetate, methanol, and water to obtain anthelmintically active fractions with various polarity. Among these fractions tested, the ethyl acetate extract of L. chiachristinae was found to be the most effective with a 50% effective concentration (EC50) value of 5.1 mg/L after 48 h of exposure. This was followed by ethyl acetate extract of C. chinensis (48 h-EC50 = 8.5 mg L(-1)), chloroform extracts of C. sappan (48 h-EC50 = 15.6 mg L(-1)), methanol extract of C. chinensis (48 h-EC50 = 15.9 mg L(-1)), and chloroform and petroleum ether extract of L. chiachristinae (EC50 values of 17.2 and 21.1 mg/L, respectively), suggesting that these plants, as well as the active fractions, provide potential sources of botanic drugs for the control of D. intermedius in aquaculture.

Indirect enantioseparation of fluoxetine in mouse serum by derivatization with 1R-(-)-menthyl chloroformate followed by ultra high performance liquid chromatography and quadrupole time-of-flight mass spectrometry.

PubMed

Zhao, Jing; Jin, Yan; Shin, Yujin; Jeong, Kyung Min; Lee, Jeongmi

2016-03-01

Here we describe a simple and sensitive analytical method for the enantioselective quantification of fluoxetine in mouse serum using ultra high performance liquid chromatography with quadrupole time-of-flight mass spectrometry. The sample preparation method included a simple deproteinization with acetonitrile in 50 μL of serum, followed by derivatization of the extracts in 50 μL of 2 mM 1R-(-)-menthyl chloroformate at 45ºC for 55 min. These conditions were statistically optimized through response surface methodology using a central composite design. Under the optimized conditions, neither racemization nor kinetic resolution occurred. The derivatized diastereomers were readily resolved on a conventional sub-2 μm C18 column under a simple gradient elution of aqueous methanol containing 0.1% formic acid. The established method was validated and found to be linear, precise, and accurate over the concentration range of 5.0-1000.0 ng/mL for both R and S enantiomers (r(2) > 0.993). Stability tests of the prepared samples at three different concentration levels showed that the R- and S-fluoxetine derivatives were relatively stable for 48 h. No significant matrix effects were observed. Last, the developed method was successfully used for enantiomeric analysis of real serum samples collected at a number of time points from mice administered with racemic fluoxetine. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Correlation of the rates of solvolysis of neopentyl chloroformate-a recommended protecting agent.

PubMed

D'Souza, Malcolm J; Carter, Shannon E; Kevill, Dennis N

2011-02-15

The specific rates of solvolysis of neopentyl chloroformate (1) have been determined in 21 pure and binary solvents at 45.0 °C. In most solvents the values are essentially identical to those for ethyl and n-propyl chloroformates. However, in aqueous-1,1,1,3,3,3-hexafluoro-2-propanol mixtures (HFIP) rich in fluoroalcohol, 1 solvolyses appreciably faster than the other two substrates. Linear free energy relationship (LFER) comparison of the specific rates of solvolysis of 1 with those for phenyl chloroformate and those for n-propyl chloroformate are helpful in the mechanistic considerations, as is also the treatment in terms of the Extended Grunwald-Winstein equation. It is proposed that the faster reaction for 1 in HFIP rich solvents is due to the influence of a 1,2-methyl shift, leading to a tertiary alkyl cation, outweighing the only weak nucleophilic solvation of the cation possible in these low nucleophilicity solvents.

[Growth inhibition of the four species of red tide microalgae by extracts from Enteromorpha prolifera extracted with the five solvents].

PubMed

Sun, Ying-Ying; Liu, Xiao-Xiao; Wang, Chang-Hai

2010-06-01

To study the effects of extracts of Enteromorpha prolifera on the growth of the four species of red tide microalgae (Amphidinium hoefleri, Karenia mikimitoi, Alexandrium tamarense and Skeletonema costatum), the extracts were extracted with five solvents (methanol, acetone, ethyl acetate, chloroform and petroleum ether), respectively. Based on the observation of algal morphology and the measurement of algal density, cell size and the contents of physiological indicators (chlorophyll, protein and polysaccharide), the results showed methanol extracts of E. prolifera had the strongest action. The inhibitory effects of A. hoefleri, K. mikimitoi, A. tamarense and S. costatum by the methanol extracts were 54.0%, 48.1%, 44.0% and 37.5% in day 10, respectively. The extracts of E. prolifera extracted with methanol, acetone and ethyl acetate caused cavities, pieces and pigment reduction in cells, and those with chloroform and petroleum ether caused goffers on cells. The extracts of E. prolifera extracted with all the five solvents decreased athletic ability of the cells, among which those extracted with ethyl acetate, chloroform and petroleum ether decreased cell size of test microalgae. The further investigation found that the methanol extracts significantly decreased contents of chlorophyll, protein and polysaccharide in the cells of those microalgae. The inhibitory effect of chlorophyll, protein and polysaccharide contents of four species of microalgae by the methanol extracts was about 51%. On the basis of the above experiments, dry powder of E. prolifera were extracts with methanol, and extracts were obtained. The methanol extracts were partitioned to petroleum ether phase, ethyl acetate phase, n-butanol phase and distilled water phase by liquid-liquid fractionation, and those with petroleum ether and ethyl acetate significantly inhibited the growth of all test microalgae, and the inhibitory effect of four species of microalgae by those two extracts was above 25% in day 10. Our researches expressed that antialgal substances in E. prolifera extracted with methanol were obtained. And two fractions (petroleum ether phase and ethyl acetate phase) that inhibited the growth of all test microalgae were obtained when the methanol extracts was fractionated by liquid-liquid fractionation.

Human beta-glucuronidase. Measurement of its activity in gallbladder bile devoid of intrinsic interference.

PubMed

Ho, Y C; Ho, K J

1988-04-01

Our purpose is to develop a standard method for preparing the bile for beta-glucuronidase determination by removal of bile acids and conjugated bilirubin which interfere with its activity. The bile acids and conjugated bilirubin in their purified solutions and in the diluted gallbladder biles could be extracted completely with cholestyramine in powder form or tetrahexylammonium chloride (THAC) in chloroform or ethyl acetate. The enzyme was, however, partially precipitated with cholestyramine and denatured by chloroform but not by ethyl acetate. A standard procedure, therefore, includes extraction of the diluted gallbladder bile with THAC in ethyl acetate, followed by determination of the maximal velocity (Vmax) of the enzyme by a kinetic method employing phenolphthalein glucuronide as the substrate. The average Vmax of beta-glucuronidase in the 20 normal gallbladder biles was 165 +/- 86 nmol/min/ml (mean +/- SD), a 23.5-fold increase over the activity before extraction. The measured activity represented the true activity of the enzyme in the bile for recovery of activity of the enzyme added to the bile was practically complete.

Detection of methamphetamine in the presence of nicotine using in situ chemical derivatization and ion mobility spectrometry.

PubMed

Ochoa, Mariela L; Harrington, Peter B

2004-02-15

The detection of methamphetamine in the presence of nicotine has been successfully accomplished using in situ chemical derivatization with propyl chloroformate as the derivatization reagent and ion mobility spectrometry (IMS). The rapid detection of methamphetamine is important for forensic scientists in order to establish a chain of evidence and link criminals to the crime scene. Nicotine is pervasive in clandestine drug laboratories from cigarette smoke residue. It has been demonstrated that nicotine obscures the methamphetamine peaks in ion mobility spectrometers due to their similar charge affinities and ion mobilities, which makes their detection a challenging task. As a consequence, false positive or negative responses may arise. In situ chemical derivatization poses as a sensitive, accurate, and reproducible alternative to remove the nicotine background when detecting nanogram amounts of methamphetamine. The derivatization agent was coated onto the sample disk, and the derivatization product corresponding to propyl methamphetamine carbamate was detected. In the present study, in situ chemical derivatization was demonstrated to be a feasible method to detect methamphetamine hydrochloride as the carbamate derivative, which was baseline-resolved from the nicotine peak. Alternating least squares (ALS) was used to model the datasets. A mixture containing both compounds revealed reduced mobilities of 1.61 cm(2)/V.s and 1.54 cm(2)/V.s for methamphetamine and nicotine, respectively. The reduced mobility of propyl methamphetamine carbamate was found at 1.35 cm(2)/V.s.

A novel fast method for aqueous derivatization of THC, OH-THC and THC-COOH in human whole blood and urine samples for routine forensic analyses.

PubMed

Stefanelli, Fabio; Pesci, Federica Giorgia; Giusiani, Mario; Chericoni, Silvio

2018-04-01

A novel aqueous in situ derivatization procedure with propyl chloroformate (PCF) for the simultaneous, quantitative analysis of Δ 9 -tetrahydrocannabinol (THC), 11-hydroxy-Δ 9 -tetrahydrocannabinol (OH-THC) and 11-nor-Δ 9 -tetrahydrocannabinol-carboxylic acid (THC-COOH) in human blood and urine is proposed. Unlike current methods based on the silylating agent [N,O-bis(trimethylsilyl)trifluoroacetamide] added in an anhydrous environment, this new proposed method allows the addition of the derivatizing agent (propyl chloroformate, PCF) directly to the deproteinized blood and recovery of the derivatives by liquid-liquid extraction. This novel method can be also used for hydrolyzed urine samples. It is faster than the traditional method involving a derivatization with trimethyloxonium tetrafluoroborate. The analytes are separated, detected and quantified by gas chromatography-mass spectrometry in selected ion monitoring mode (SIM). The method was validated in terms of selectivity, capacity of identification, limits of detection (LOD) and quantification (LOQ), carryover, linearity, intra-assay precision, inter-assay precision and accuracy. The LOD and LOQ in hydrolyzed urine were 0.5 and 1.3 ng/mL for THC and 1.2 and 2.6 ng/mL for THC-COOH, respectively. In blood, the LOD and LOQ were 0.2 and 0.5 ng/mL for THC, 0.2 and 0.6 ng/mL for OH-THC, and 0.9 and 2.4 ng/mL for THC-COOH, respectively. This method was applied to 35 urine samples and 50 blood samples resulting to be equivalent to the previously used ones with the advantage of a simpler method and faster sample processing time. We believe that this method will be a more convenient option for the routine analysis of cannabinoids in toxicological and forensic laboratories. Copyright © 2017 John Wiley & Sons, Ltd.

In matrix derivatization of trichloroethylene metabolites in human plasma with methyl chloroformate and their determination by solid-phase microextraction-gas chromatography-electron capture detector.

PubMed

Mudiam, Mohana Krishna Reddy; Jain, Rajeev; Varshney, Meenu; Ch, Ratnasekhar; Chauhan, Abhishek; Goyal, Sudhir Kumar; Khan, Haider A; Murthy, R C

2013-04-15

Trichloroethylene (TCE) is a common industrial chemical that has been widely used as metal degreaser and for many industrial purposes. In humans, TCE is metabolized into dichloroacetic acid (DCA), trichloroacetic acid (TCA) and trichloroethanol (TCOH). A simple and rapid method has been developed for the quantitative determination of TCE metabolites. The procedure involves the in situ derivatization of TCE metabolites with methyl chloroformate (MCF) directly in diluted plasma samples followed by extraction and analysis with solid-phase microextraction (SPME) coupled to gas chromatography-electron capture detector (GC-ECD). Factors which can influence the efficiency of derivatization such as amount of MCF and pyridine (PYR), ratio of water/methanol were optimized. The factors which can affect the extraction efficiencies of SPME were screened using 2(7-4) Placket-Burman Design (PBD). A central composite design (CCD) was then applied to further optimize the most significant factors for optimum SPME extraction. The optimum factors for the SPME extraction were found to be 562.5mg of NaCl, pH at 1 and an extraction time of 22 min. Recoveries and detection limits of all three analytes in plasma were found to be in the range of 92.69-97.55% and 0.036-0.068 μg mL(-1) of plasma, respectively. The correlation coefficients were found to be in the range of 0.990-0.995. The intra- and inter-day precisions for TCE metabolites were found to be in the range of 2.37-4.81% and 5.13-7.61%, respectively. The major advantage of this method is that MCF derivatization allows conversion of TCE metabolites into their methyl esters in very short time (≤30 s) at room temperature directly in the plasma samples, thus makes it a solventless analysis. The method developed was successfully applied to the plasma samples of humans exposed to TCE. Copyright © 2013 Elsevier B.V. All rights reserved.

Free radical scavenging activities measured by electron spin resonance spectroscopy and B16 cell antiproliferative behaviors of seven plants.

PubMed

Calliste, C A; Trouillas, P; Allais, D P; Simon, A; Duroux, J L

2001-07-01

In an effort to discover new antioxidant natural compounds, seven plants that grow in France (most of them in the Limousin countryside) were screened. Among these plants, was the extensively studied Vitis vinifera as reference. For each plant, sequential percolation was realized with five solvents of increasing polarities (hexane, chloroform, ethyl acetate, methanol, and water). Free radical scavenging activities were examined in different systems using electron spin resonance (ESR) spectroscopy. These assays were based on the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH), the hydroxyl radicals generated by a Fenton reaction, and the superoxide radicals generated by the X/XO system. Antiproliferative behavior was studied on B16 melanoma cells. ESR results showed that three plants (Castanea sativa, Filipendula ulmaria, and Betula pendula) possessed, for the most polar fractions (presence of phenolic compounds), high antioxidant activities in comparison with the Vitis vinifera reference. Gentiana lutea was the only one that presented a hydroxyl scavenging activity for the ethyl acetate and chloroform fractions. The antiproliferative test results showed that the same three plants are the most effective, but for the apolar fractions (chloroform and hexane).

Antibacterial, antidiarrhoeal, and cytotoxic activities of methanol extract and its fractions of Caesalpinia bonducella (L.) Roxb leaves

PubMed Central

2013-01-01

Background Caesalpinia bonducella is an important medicinal plant for its traditional uses against different types of diseases. Therefore, the present study investigated the antimicrobial, antidiarrhoeal, and cytotoxic activities of the methanol extract and ethyl acetate, chloroform, and petroleum ether (pet. ether) fractions of C. bonducella leaves. Methods The antibacterial potentialities of methanol extract and its fractions of C. bonducella leaves were investigated by the disc diffusion method against four gram-positive and five gram-negative bacteria at 300, 500 and 800 μg/disc. Kanamycin (30 μg/disc) was used as the standard drug. Antidiarrhoeal activities of leaf extracts were evaluated at two doses (200 and 400 mg/kg) and compared with loperamide in a castor oil-induced diarrhoeal model in rat. The fractions were subjected to a brine shrimp lethality test to evaluate their cytotoxicity. Results The methanol extract and other three fractions exhibited better activities at higher concentrations. Amongst, the chloroform fraction showed maximum activity at all three concentrations (300, 500, and 800 μg/disc) against almost all bacteria. S. aureus and P. aeruginosa showed better sensitivities to all extracts at all three concentrations excluding the pet. ether fraction. Bacillus megaterium and Klebsiella spp. were two bacteria amongst nine that showed lowest sensitivity to the extracts. Maximum zone of inhibition (25-mm) was obtained by the methanol extract at an 800 μg/disc concentration against S. aureus. In the antidiarrhoeal test, all fractions exhibited dose-dependent actions, which were statistically significant (p < 0.05). Ethyl acetate fraction exerted maximum inhibition (51.11%) against defecation, whereas 57.75% inhibition was obtained for loperamide. Moderate cytotoxicity was found for the methanol extract and its three fractions compared with the standard drug vincristine sulfate in the brine shrimp bioassay. In the present study, the LC50 values of the methanol crude extract and ethyl acetate, chloroform, pet. ether fractions and vincristine sulfate were 223.87, 281.84, 112.2, 199.53, and 12.59 μg/mL, respectively. Therefore, the ethyl acetate fraction showed maximum cytotoxicity, whereas minimum cytotoxicity was observed for the chloroform fraction. Conclusion The present study revealed that the ethyl acetate fraction of the C. bonducella leaves has significant antidiarrhoeal properties. The methanol extract and other three fractions of the C. bonducella leaves possess potent antibacterial activities along with moderate cytotoxicities that may lead to new drug development. PMID:23663985

Antibacterial, antidiarrhoeal, and cytotoxic activities of methanol extract and its fractions of Caesalpinia bonducella (L.) Roxb leaves.

PubMed

Billah, Muhammad Mutassim; Islam, Rafikul; Khatun, Hajera; Parvin, Shahnaj; Islam, Ekramul; Islam, Sm Anisul; Mia, Akbar Ali

2013-05-12

Caesalpinia bonducella is an important medicinal plant for its traditional uses against different types of diseases. Therefore, the present study investigated the antimicrobial, antidiarrhoeal, and cytotoxic activities of the methanol extract and ethyl acetate, chloroform, and petroleum ether (pet. ether) fractions of C. bonducella leaves. The antibacterial potentialities of methanol extract and its fractions of C. bonducella leaves were investigated by the disc diffusion method against four gram-positive and five gram-negative bacteria at 300, 500 and 800 μg/disc. Kanamycin (30 μg/disc) was used as the standard drug. Antidiarrhoeal activities of leaf extracts were evaluated at two doses (200 and 400 mg/kg) and compared with loperamide in a castor oil-induced diarrhoeal model in rat. The fractions were subjected to a brine shrimp lethality test to evaluate their cytotoxicity. The methanol extract and other three fractions exhibited better activities at higher concentrations. Amongst, the chloroform fraction showed maximum activity at all three concentrations (300, 500, and 800 μg/disc) against almost all bacteria. S. aureus and P. aeruginosa showed better sensitivities to all extracts at all three concentrations excluding the pet. ether fraction. Bacillus megaterium and Klebsiella spp. were two bacteria amongst nine that showed lowest sensitivity to the extracts. Maximum zone of inhibition (25-mm) was obtained by the methanol extract at an 800 μg/disc concentration against S. aureus. In the antidiarrhoeal test, all fractions exhibited dose-dependent actions, which were statistically significant (p < 0.05). Ethyl acetate fraction exerted maximum inhibition (51.11%) against defecation, whereas 57.75% inhibition was obtained for loperamide. Moderate cytotoxicity was found for the methanol extract and its three fractions compared with the standard drug vincristine sulfate in the brine shrimp bioassay. In the present study, the LC50 values of the methanol crude extract and ethyl acetate, chloroform, pet. ether fractions and vincristine sulfate were 223.87, 281.84, 112.2, 199.53, and 12.59 μg/mL, respectively. Therefore, the ethyl acetate fraction showed maximum cytotoxicity, whereas minimum cytotoxicity was observed for the chloroform fraction. The present study revealed that the ethyl acetate fraction of the C. bonducella leaves has significant antidiarrhoeal properties. The methanol extract and other three fractions of the C. bonducella leaves possess potent antibacterial activities along with moderate cytotoxicities that may lead to new drug development.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Feng; Cooper, S.F.

A novel gas chromatography-mass spectrometric (GC-MS) method was developed to confirm and identify metoprolol and its metabolites by double derivatization with S-(-)menthyl chloroformate [(-)-MCF] and N-methyl(trimethylsilyl-trifluoroacetamide) (MSTFA). This is the first report, which describes the simultaneous identification of metoprolol, its one major acidc and other basic metabolites in human urine based on solid-phase extraction with C{sub 18} reversed-phase cartridges. 12 refs., 4 figs.

Determination of atrazine and its major degradation products in soil pore water by solid-phase extraction, chemical derivatization, and gas chromatography/mass spectrometry

USGS Publications Warehouse

Carter, D.S.

1996-01-01

This report describes a method for the determination of atrazine, desethylatrazine, deisopropylatrazine, didealkylatrazine, and hydroxyatrazine from soil pore waters by use of solid-phase extractionfollowed by chemical derivatization and gas chromatography/mass spectrometry. The analytes are isolated from the pore-water matrix byextraction onto a graphitized carbon-black cartridge. The cartridge is dried under vacuum, and adsorbed analytes are removed by elution with ethyl acetate followed by dichloromethane/methanol (7:3, volume/volume). Water is removed from the ethyl acetate fraction on an anhydrous sodium sulfate column. The combined fractions are solvent exchanged into acetonitrile, evaporated by use of a nitrogen stream, and derivatized by use of N- methyl-N-(tert-butyldimethylsilyl)- trifluoroacetamide. The derivatized extracts are analyzed by capillary-column gaschromatography/electron-impact mass spectrometry in the scan mode. Estimated method detection limits range from 0.03 to 0.07 micrograms per liter. The mean recoveries of all analytes and surrogates determined at 0.74 to 0.82 micrograms per liter in reagent water in soil pore water were 94 percent and 98 percent, respectively. The mean recoveries of all analytes and surrogates determined at 7.4 to 8.2 micrograms per liter in reagent water and in soil pore water were 96 percent and 97 percent,respectively. Recoveries were 90 percent or higher, regardless of analyte concentration or matrix composition, for all compounds excepthydroxyatrazine, whose recoveries were slightly lower (77 percent) at the low concentration.

Anti-angiogenic activity and phytochemical screening of fruit fractions from Vitex agnus castus.

PubMed

Certo, Giovanna; Costa, Rosaria; D'Angelo, Valeria; Russo, Marina; Albergamo, Ambrogina; Dugo, Giacomo; Germanò, Maria Paola

2017-12-01

Although the antitumour activity of Vitex agnus castus fruits has been already addressed, no work has yet assessed their anti-angiogenic potential. To this purpose, several extractive fractions of such fruits were tested on zebrafish embrios by EAP assay, so that only the bioactive fractions could be subsequently tested on the chick chorioallantoic membrane by CAM assay. Bioactive fractions were also phytochemically screened to identify those bioactive compounds responsible for anti-angiogenic activity. A marked inhibition of vessel formation was detected only in zebrafish embryos treated with chloroform or ethyl acetate fractions. Considering CAM assay, chloroform fraction induced a strong reduction of microvasculature and haemoglobin content; while lower anti-angiogenic effects of the ethyl acetate fraction were determined. Phytochemical analyses confirmed the presence of several bioactive anti-angiogenic compounds. Overall, obtained preliminary results highlighted a potential anti-angiogenic activity of V. agnus castus fruits.

Allelopathic potential of Artemisia arborescens: isolation, identification and quantification of phytotoxic compounds through fractionation-guided bioassays.

PubMed

Araniti, Fabrizio; Lupini, Antonio; Sorgonà, Agostino; Conforti, Filomena; Marrelli, Mariangela; Statti, Giancarlo Antonio; Menichini, Francesco; Abenavoli, Maria Rosa

2013-01-01

The aerial part of Artemisia arborescens L. (Asteraceae) was extracted with water and methanol, and both extracts were fractionated using n-hexane, chloroform, ethyl acetate and n-butanol. The potential phytotoxicity of both crude extracts and their fractions were assayed in vitro on seed germination and root growth of lettuce (Lactuca sativa L.), a sensitive species largely employed in the allelopathy studies. The inhibitory activities were analysed by dose-response curves and the ED 50 were estimated. Crude extracts strongly inhibited both germination and root growth processes. The fraction-bioassay indicated the following hierarchy of phytotoxicity for both physiological processes: ethyl acetate ≥ n-hexane > chloroform ≥ n-butanol. On the n-hexane fraction, GC-MS analyses were carried out to characterise and quantify some of the potential allelochemicals. Twenty-one compounds were identified and three of them, camphor, trans-caryophyllene and pulegone were quantified.

Determining the fatty acid composition in plasma and tissues as fatty acid methyl esters using gas chromatography – a comparison of different derivatization and extraction procedures.

PubMed

Ostermann, Annika I; Müller, Maike; Willenberg, Ina; Schebb, Nils Helge

2014-12-01

Analysis of the fatty acid (FA) composition in biological samples is commonly carried out using gas liquid chromatography (GC) after transesterification to volatile FA methyl esters (FAME). We compared the efficacy of six frequently used protocols for derivatization of different lipid classes as well as for plasma and tissue samples. Transesterification with trimethylsulfonium hydroxide (TMSH) led to insufficient derivatization efficacies for polyunsaturated FAs (PUFA, <50%). Derivatization in presence of potassium hydroxide (KOH) failed at derivatizing free FAs (FFAs). Boron trifluoride (BF3) 7% in hexane/MeOH (1:1) was insufficient for the transesterification of cholesterol ester (CE) as well as triacylglycerols (TGs). In contrast, methanolic hydrochloric acid (HCl) as well as a combination of BF3 with methanolic sodium hydroxide (NaOH+BF3) were suitable for the derivatization of FFAs, polar lipids, TGs, and CEs (derivatization rate >80% for all tested lipids). Regarding plasma samples, all methods led to an overall similar relative FA pattern. However, significant differences were observed, for example, for the relative amount of EPA+DHA (n3-index). Absolute FA plasma concentrations differed considerably among the methods, with low yields for KOH and BF3. We also demonstrate that lipid extraction with tert-butyl methyl ether/methanol (MTBE/MeOH) is as efficient as the classical method according to Bligh and Dyer, making it possible to replace (environmentally) toxic chloroform.We conclude that HCl-catalyzed derivatization in combination with MeOH/MTBE extraction is the most appropriate among the methods tested for the analysis of FA concentrations and FA pattern in small biological samples. A detailed protocol for the analysis of plasma and tissues is included in this article.

Study of the Gastroprotective Effect of Extracts and Semipurified Fractions of Chresta martii DC. and Identification of Its Principal Compounds.

PubMed

Franco, E S; Mélo, M E B; Jatobá, B J A; Santana, A L B D; Silva, A A R; Silva, T G; Nascimento, M S; Maia, M B S

2015-01-01

Chresta martii (Asteraceae) is a species widely used by the population of the Xingu region of Sergipe, Brazil, in the form of a decoction (aerial parts) for the treatment of gastrointestinal diseases. The study aims to assess the gastroprotective activity of organic extracts and semipurified fractions and identify the principal compounds present in C. martii responsible for such activity. The organic extracts (cyclohexane: ECCm, ethyl acetate: EACm, and ethanol: EECm) were obtained from the dried aerial parts (500 g) of C. martii. For evaluation of the gastroprotective activity of extracts (50, 100, or 200 mg/kg; p.o.), male Swiss Webster mice (25-30 g) were used which had gastric ulcers induced by indomethacin (40 mg/kg, s.c.) or ethanol (0.2 mL/animal; p.o.). Among the extracts evaluated, EACm exhibited significant (P < 0.05) gastroprotective activity in the models used. The fractionation of EACm was performed in a silica gel column 60 eluted with the following compounds: [chloroform-F1 yield (10%)], [chloroform/ethyl acetate (1/1)-F2 yield (6%)], [ethyl acetate-F3 yield (8%)], and [ethyl/methanol acetate (1/1)-F4 yield (5%)]. Of the fractions described above, the F1 (25 mg/kg; p.o.) had greater gastroprotective activity (P < 0.05) than that displayed by ranitidine (80 mg/kg; p.o.) in the ethanol-induced ulcer model. The refractionation of F1 produced 23 subfractions and from these two yellow amorphous compounds were obtained by recrystallization, Rf: 0.46 and 0.31 (ethyl acetate : chloroform 5 : 5). The compounds isolated were characterized by nuclear magnetic resonance spectroscopy ((1)H-NMR and (13)C-NMR) and identified as flavones: chrysoeriol (yield: 0.43%) and 3',4'-dimethoxyluteolin (yield: 0.58%). Conclusion. Flavone 3',4'-dimethoxyluteolin is the principal compound present in the species C. martii and is probably responsible for gastroprotective activity observed in this species.

"Bligh and Dyer" and Folch Methods for Solid-Liquid-Liquid Extraction of Lipids from Microorganisms. Comprehension of Solvatation Mechanisms and towards Substitution with Alternative Solvents.

PubMed

Breil, Cassandra; Abert Vian, Maryline; Zemb, Thomas; Kunz, Werner; Chemat, Farid

2017-03-27

Bligh and Dyer (B & D) or Folch procedures for the extraction and separation of lipids from microorganisms and biological tissues using chloroform/methanol/water have been used tens of thousands of times and are "gold standards" for the analysis of extracted lipids. Based on the Conductor-like Screening MOdel for realistic Solvatation (COSMO-RS), we select ethanol and ethyl acetate as being potentially suitable for the substitution of methanol and chloroform. We confirm this by performing solid-liquid extraction of yeast ( Yarrowia lipolytica IFP29 ) and subsequent liquid-liquid partition-the two steps of routine extraction. For this purpose, we consider similar points in the ternary phase diagrams of water/methanol/chloroform and water/ethanol/ethyl acetate, both in the monophasic mixtures and in the liquid-liquid miscibility gap. Based on high performance thin-layer chromatography (HPTLC) to obtain the distribution of lipids classes, and gas chromatography coupled with a flame ionisation detector (GC/FID) to obtain fatty acid profiles, this greener solvents pair is found to be almost as effective as the classic methanol-chloroform couple in terms of efficiency and selectivity of lipids and non-lipid material. Moreover, using these bio-sourced solvents as an alternative system is shown to be as effective as the classical system in terms of the yield of lipids extracted from microorganism tissues, independently of their apparent hydrophilicity.

“Bligh and Dyer” and Folch Methods for Solid–Liquid–Liquid Extraction of Lipids from Microorganisms. Comprehension of Solvatation Mechanisms and towards Substitution with Alternative Solvents

PubMed Central

Breil, Cassandra; Abert Vian, Maryline; Zemb, Thomas; Kunz, Werner; Chemat, Farid

2017-01-01

Bligh and Dyer (B & D) or Folch procedures for the extraction and separation of lipids from microorganisms and biological tissues using chloroform/methanol/water have been used tens of thousands of times and are “gold standards” for the analysis of extracted lipids. Based on the Conductor-like Screening MOdel for realistic Solvatation (COSMO-RS), we select ethanol and ethyl acetate as being potentially suitable for the substitution of methanol and chloroform. We confirm this by performing solid–liquid extraction of yeast (Yarrowia lipolytica IFP29) and subsequent liquid–liquid partition—the two steps of routine extraction. For this purpose, we consider similar points in the ternary phase diagrams of water/methanol/chloroform and water/ethanol/ethyl acetate, both in the monophasic mixtures and in the liquid–liquid miscibility gap. Based on high performance thin-layer chromatography (HPTLC) to obtain the distribution of lipids classes, and gas chromatography coupled with a flame ionisation detector (GC/FID) to obtain fatty acid profiles, this greener solvents pair is found to be almost as effective as the classic methanol–chloroform couple in terms of efficiency and selectivity of lipids and non-lipid material. Moreover, using these bio-sourced solvents as an alternative system is shown to be as effective as the classical system in terms of the yield of lipids extracted from microorganism tissues, independently of their apparent hydrophilicity. PMID:28346372

Mosquito larvicidal properties of Orthosiphon thymiflorus (Roth) Sleesen. (Family: Labiatae) against mosquito vectors, Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti (Diptera: Culicidae).

PubMed

Kovendan, K; Murugan, K; Vincent, S; Barnard, Donald R

2012-04-01

To determine the mosquito larvicidal activities of hexane, chloroform, ethyl acetate, acetone and methanol leaf extract of Orthosiphon thymiflorus (O. thymiflorus) against Anopheles stephensi (An. stephensi), Culex quinquefasciatus (Cx. quinquefasciatus) and Aedes aegypti (Ae. aegypti). The larvicidal activity was assayed against three mosquito species at various concentrations ranging from (50-450 ppm) under the laboratory conditions. The LC(50) and LC(90) value of the O. thymiflorus leaf extract was determined by Probit analysis. The LC(50) values of hexane, chloroform, ethyl acetate, acetone and methanol extract of O. thymiflorus third instar larvae of An. stephensi were LC(50)= 201.39, 178.76, 158.06, 139.22 and 118.74 ppm; Cx. quinquefasciatus were LC(50)=228.13, 209.72, 183.35, 163.55 and 149.96 ppm and Ae. aegypti were LC(50)=215.65, 197.91, 175.05, 154.80 and 137.26 ppm, respectively. Maximum larvicidal activity was observed in the methanolic extract followed by acetone, ethyl acetate chloroform and hexane extract. The larval mortality was observed after 24 h exposure. No mortality was observed in control. The present results suggest that the effective plant crude extracts have potential to be used as an ideal eco-friendly approach for the control of mosquito vectors. This study provides the first report on the larvicidal activity of this plant crude solvent extract of against An. stephensi, Cx. quinquefasciatus and Ae. aegypti mosquitoes. Copyright © 2012 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Psychotria viridis: Chemical constituents from leaves and biological properties.

PubMed

Soares, Débora B S; Duarte, Lucienir P; Cavalcanti, André D; Silva, Fernando C; Braga, Ariadne D; Lopes, Miriam T P; Takahashi, Jacqueline A; Vieira-Filho, Sidney A

2017-01-01

The phytochemical study of hexane, chloroform and methanol extracts from leaves of Psychotria viridis resulted in the identification of: the pentacyclic triterpenes, ursolic and oleanolic acid; the steroids, 24-methylene-cycloartanol, stigmasterol and β-sitosterol; the glycosylated steroids 3-O-β-D-glucosyl-β-sitosterol and 3-O-β-D-glucosyl-stigmasterol; a polyunsaturated triterpene, squalene; the esters of glycerol, 1-palmitoylglycerol and triacylglycerol; a mixture of long chain hydrocarbons; the aldehyde nonacosanal; the long chain fat acids hentriacontanoic, hexadecanoic and heptadenoic acid; the ester methyl heptadecanoate; the 4-methyl-epi-quinate and two indole alkaloids, N,N-dimethyltryptamine (DMT) and N-methyltryptamine. The chemical structures were determined by means of spectroscopic (IR, 1H and 13C NMR, HSQC, HMBC and NOESY) and spectrometric (CG-MS and LCMS-ESI-ITTOF) methods. The study of biologic properties of P. viridis consisted in the evaluation of the acetylcholinesterase inhibition and cytotoxic activities. The hexane, chloroform, ethyl acetate and methanol extracts, the substances 24-methylene-cycloartanol, DMT and a mixture of 3-O-β-D-glucosyl-β-sitosterol and 3-O-β-D-glucosyl-stigmasterol showed cholinesterase inhibiting activity. This activity induced by chloroform and ethyl acetate extracts was higher than 90%. The methanol and ethyl acetate extracts inhibit the growth and/or induce the death of the tumor cells strains B16F10 and 4T1, without damaging the integrity of the normal cells BHK and CHO. DMT also demonstrated a marked activity against tumor cell strains B16F10 and 4T1.

Biochemical surface modification of Co-Cr-Mo.

PubMed

Puleo, D A

1996-01-01

Because of the limited mechanical properties of tissue substitutes formed by culturing cells on polymeric scaffolds, other approaches to tissue engineering must be explored for applications that require complete and immediate ability to bear weight, e.g. total joint replacements. Biochemical surface modification offers a way to partially regulate events at the bone-implant interface to obtain preferred tissue responses. Tresyl chloride, gamma-aminopropyltriethoxysilane (APS) and p-nitrophenyl chloroformate (p-NPC) immobilization schemes were used to couple a model enzyme, trypsin, on bulk samples of Co-Cr-Mo. For comparison, samples were simply adsorbed with protein. The three derivatization schemes resulted in different patterns and levels of activity. Tresyl chloride was not effective in immobilizing active enzyme on Co-Cr-Mo. Aqueous silanization with 12.5% APS resulted in optimal immobilized activity. Activity on samples derivatized with 0.65 mg p-NPC cm-2 was four to five times greater than that on samples simple adsorbed with enzyme or optimally derivatized with APS and was about eight times that on tresylated samples. This work demonstrates that, although different methods have different effectiveness, chemical derivatization can be used to alter the amount and/or stability of biomolecules immobilized on the surface of Co-Cr-Mo.

Fast determination of ethylene glycol, 1,2-propylene glycol and glycolic acid in blood serum and urine for emergency and clinical toxicology by GC-FID.

PubMed

Hložek, Tomáš; Bursová, Miroslava; Čabalaa, Radomír

2014-12-01

A simple, cost effective, and fast gas chromatography method with flame ionization detection (GC-FID) for simultaneous measurement of ethylene glycol, 1,2-propylene glycol and glycolic acid was developed and validated for clinical toxicology purposes. This new method employs a relatively less used class of derivatization agents - alkyl chloroformates, allowing the efficient and rapid derivatization of carboxylic acids within seconds while glycols are simultaneously derivatized by phenylboronic acid. The entire sample preparation procedure is completed within 10 min. To avoid possible interference from naturally occurring endogenous acids and quantitation errors 3-(4-chlorophenyl) propionic acid was chosen as an internal standard. The significant parameters of the derivatization have been found using chemometric procedures and these parameters were optimized using the face-centered central composite design. The calibration dependence of the method was proved to be quadratic in the range of 50-5000 mg mL(-1), with adequate accuracy (92.4-108.7%) and precision (9.4%). The method was successfully applied to quantify the selected compounds in serum of patients from emergency units. Copyright © 2014 Elsevier B.V. All rights reserved.

Evaluation of Andrographis paniculata Burm.f. (Family:Acanthaceae) extracts against Culex quinquefasciatus (Say.) and Aedes aegypti (Linn.) (Diptera:Culicidae).

PubMed

Govindarajan, Marimuthu

2011-03-01

To investigate the larvicidal and ovicidal efficacy of different extracts of Andrographis paniculata (A. paniculata) against Culex quinquefasciatus (Cx. quinquefasciatus) Say and Aedes aegypti (Ae. aegypti) L. (Diptera: Culicidae). Larvicidal efficacy of the crude leaf extracts of A. paniculata with five different solvents like benzene, hexane, ethyl acetate, methanol and chloroform was tested against the early third instar larvae of Cx. quinquefasciatus and Ae. aegypti. The ovicidal activity was determined against two mosquito species to various concentrations ranging from 50-300 ppm under the laboratory conditions. The benzene, hexane, ethyl acetate, methanol and chloroform leaf extract of A. paniculata was found to be more effective against Cx. quinquefasciatus than Ae. aegypti. The LC(50) values were 112.19, 137.48, 118.67, 102.05, 91.20 ppm and 119.58, 146.34, 124.24, 110.12, 99.54 ppm respectively. Among five tested solvent, methanol and ethyl acetate crude extract was found to be most effective for ovicidal activity against two mosquito species. The extract of methanol and ethyl acetate exerted 100% mortality at 200 ppm against Cx. quinquefasciatus and at 250 ppm against Ae. aegypti. From the results it can be concluded the crude extract of A. paniculata was a potential for controlling Cx. quinquefasciatus and Ae. aegypti mosquitoes. Copyright © 2011 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Effect of different fractions from hydroalcoholic extract of Black Maca (Lepidium meyenii) on testicular function in adult male rats.

PubMed

Yucra, Sandra; Gasco, Manuel; Rubio, Julio; Nieto, Jessica; Gonzales, Gustavo F

2008-05-01

To evaluate the effect of different fractions of Black Maca (Lepidium meyenii), obtained from the hydroalcoholic extract, on spermatogenesis. Animal study. Animal and laboratory facilities at a university. Forty two adult male rats from the Holtzman strain (3 months old). Hydroalcoholic extract of Black Maca was partitioned with the following solvents: petroleum ether, chloroform, ethyl acetate, n-butanol, and water to obtain each fraction. Forty-two rats were divided in different groups according the fraction administered and vehicle. The hydroalcoholic extract of Black Maca and its fractions and vehicle were given orally by gavage for 7 days. Daily sperm production, epididymal sperm count, and sperm count in the vas deferens. Daily sperm production was higher in the ethyl acetate group compared with all other groups. The epididymal sperm count was higher in rats treated with ethyl acetate fraction compared with rats treated with vehicle (control), petroleum ether, n-butanol, or water fractions. The sperm count in vas deferens was lower in rats treated with ethyl acetate, petroleum ether, or water fractions compared with the control group; thus, the sperm count in vas deferens in rats treated with chloroform and n-butanol fractions was higher than in the petroleum ether group. The greatest effect on spermatogenesis was observed in the ethyl acetate fraction from the hydroalcoholic extract of Black Maca, suggesting that the compounds related to the beneficial effect on sperm production of Black Maca are presented in this fraction. Antioxidant components could play a role in the effect of increased epididymal sperm concentration observed in the model.

Potential of capillary electrophoresis mass spectrometry for the characterization and monitoring of amine-derivatized naphthenic acids from oil sands process-affected water.

PubMed

MacLennan, Matthew S; Tie, Cai; Kovalchik, Kevin; Peru, Kerry M; Zhang, Xinxiang; Headley, John V; Chen, David D Y

2016-11-01

Capillary electrophoresis coupled to mass spectrometry (CE-MS) was used for the analysis of naphthenic acid fraction compounds (NAFCs) of oil sands process-affected water (OSPW). A standard mixture of amine-derivatized naphthenic acids is injected directly onto the CE column and analyzed by CE-MS in less than 15min. Time of flight MS analysis (TOFMS), optimized for high molecular weight ions, showed NAFCs between 250 and 800m/z. With a quadrupole mass analyzer, only low-molecular weight NAFCs (between 100 and 450m/z) are visible under our experimental conditions. Derivatization of NAFCs consisted of two-step amidation reactions mediated by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), or mediated by a mixture of EDC and N-hydroxysuccinimide, in dimethyl sulfoxide, dichloromethane or ethyl acetate. The optimum background electrolyte composition was determined to be 30% (V/V) methanol in water and 2% (V/V) formic acid. NAFCs extracted from OSPW in the Athabasca oil sands region were used to demonstrate the feasibility of CE-MS for the analysis of NAFCs in environmental samples, showing that the labeled naphthenic acids are in the mass range of 350 to 1500m/z. Copyright © 2016. Published by Elsevier B.V.

[Determination of organotin compounds in plastic products by GC/MS after ethyl derivatization with sodium tetraethylborate].

PubMed

Ohno, Hiroyuki; Suzuki, Masako; Nakashima, Shigehito; Aoyama, Taiki; Mitani, Kazunori

2002-08-01

A simultaneous determination method for 9 organotin compounds in polyvinyl chloride (PVC) and silicone products used as kitchen utensils and food packages was developed using ethyl derivatization with sodium tetraethylborate (NaBEt4). Organotin compounds were extracted with acetone-hexane (3:7) from the samples after acidification and the extract was filtered and concentrated at under 40 degrees C. After centrifugal separation, these compounds were derivatized with 2% NaBEt4 solution and determined by GC/MS. This method was applicable for simple routine analysis. Recoveries of spiked compounds were 49.1-118.1% for 3 PVC products and 88.8-102.2% for a siliconized paper. Monooctyltin, dioctyltin and trioctyltin compounds were found in all PVC food containers at the levels of 123-1,380 micrograms/g, 1,770-13,200 micrograms/g and 6.6-139 micrograms/g, respectively. They also were found in 3 gloves, 5 spouts, 1 hose and 5 pipes. Some PVC products contained monomethyltin, dimethyltin, trimethyltin, monobutyltin and dibutyltin compounds at the levels of 97.3-433 micrograms/g, 96.5-5,120 micrograms/g, 8.5-24.9 micrograms/g, 1.2-852 micrograms/g and 1.2-29.4 micrograms/g, respectively.

Urinary Amino Acid Analysis: A Comparison of iTRAQ®-LC-MS/MS, GC-MS, and Amino Acid Analyzer

PubMed Central

Kaspar, Hannelore; Dettmer, Katja; Chan, Queenie; Daniels, Scott; Nimkar, Subodh; Daviglus, Martha L.; Stamler, Jeremiah; Elliott, Paul; Oefner, Peter J.

2009-01-01

Urinary amino acid analysis is typically done by cation-exchange chromatography followed by post-column derivatization with ninhydrin and UV detection. This method lacks throughput and specificity. Two recently introduced stable isotope ratio mass spectrometric methods promise to overcome those shortcomings. Using two blinded sets of urine replicates and a certified amino acid standard, we compared the precision and accuracy of gas chromatography/mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) of propyl chloroformate and iTRAQ® derivatized amino acids, respectively, to conventional amino acid analysis. The GC-MS method builds on the direct derivatization of amino acids in diluted urine with propyl chloroformate, GC separation and mass spectrometric quantification of derivatives using stable isotope labeled standards. The LC-MS/MS method requires prior urinary protein precipitation followed by labeling of urinary and standard amino acids with iTRAQ® tags containing different cleavable reporter ions distinguishable by MS/MS fragmentation. Means and standard deviations of percent technical error (%TE) computed for 20 amino acids determined by amino acid analyzer, GC-MS, and iTRAQ®-LC-MS/MS analyses of 33 duplicate and triplicate urine specimens were 7.27±5.22, 21.18±10.94, and 18.34±14.67, respectively. Corresponding values for 13 amino acids determined in a second batch of 144 urine specimens measured in duplicate or triplicate were 8.39±5.35, 6.23±3.84, and 35.37±29.42. Both GC-MS and iTRAQ®-LC-MS/MS are suited for high-throughput amino acid analysis, with the former offering at present higher reproducibility and completely automated sample pretreatment, while the latter covers more amino acids and related amines. PMID:19481989

Urinary amino acid analysis: a comparison of iTRAQ-LC-MS/MS, GC-MS, and amino acid analyzer.

PubMed

Kaspar, Hannelore; Dettmer, Katja; Chan, Queenie; Daniels, Scott; Nimkar, Subodh; Daviglus, Martha L; Stamler, Jeremiah; Elliott, Paul; Oefner, Peter J

2009-07-01

Urinary amino acid analysis is typically done by cation-exchange chromatography followed by post-column derivatization with ninhydrin and UV detection. This method lacks throughput and specificity. Two recently introduced stable isotope ratio mass spectrometric methods promise to overcome those shortcomings. Using two blinded sets of urine replicates and a certified amino acid standard, we compared the precision and accuracy of gas chromatography/mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) of propyl chloroformate and iTRAQ derivatized amino acids, respectively, to conventional amino acid analysis. The GC-MS method builds on the direct derivatization of amino acids in diluted urine with propyl chloroformate, GC separation and mass spectrometric quantification of derivatives using stable isotope labeled standards. The LC-MS/MS method requires prior urinary protein precipitation followed by labeling of urinary and standard amino acids with iTRAQ tags containing different cleavable reporter ions distinguishable by MS/MS fragmentation. Means and standard deviations of percent technical error (%TE) computed for 20 amino acids determined by amino acid analyzer, GC-MS, and iTRAQ-LC-MS/MS analyses of 33 duplicate and triplicate urine specimens were 7.27+/-5.22, 21.18+/-10.94, and 18.34+/-14.67, respectively. Corresponding values for 13 amino acids determined in a second batch of 144 urine specimens measured in duplicate or triplicate were 8.39+/-5.35, 6.23+/-3.84, and 35.37+/-29.42. Both GC-MS and iTRAQ-LC-MS/MS are suited for high-throughput amino acid analysis, with the former offering at present higher reproducibility and completely automated sample pretreatment, while the latter covers more amino acids and related amines.

Theoretical study on the mechanism of the gas-phase elimination kinetics of alkyl chloroformates

NASA Astrophysics Data System (ADS)

Alcázar, Jackson J.; Marquez, Edgar; Mora, José R.; Cordova-Sintjago, Tania; Chuchani, Gabriel

2016-03-01

The theoretical calculations on the mechanism of the homogeneous and unimolecular gas-phase elimination kinetics of alkyl chloroformates- ethyl chloroformate (ECF), isopropyl chloroformate (ICF), and sec-butyl chloroformate (SCF) - have been carried out by using CBS-QB3 level of theory and density functional theory (DFT) functionals CAM-B3LYP, M06, MPW1PW91, and PBE1PBE with the basis sets 6-311++G(d,p) and 6-311++G(2d,2p). The chlorofomate compounds with alkyl ester Cβ-H bond undergo thermal decomposition producing the corresponding olefin, HCl and CO2. These homogeneous eliminations are proposed to undergo two different types of mechanisms: a concerted process, or via the formation of an unstable intermediate chloroformic acid (ClCOOH), which rapidly decomposes to HCl and CO2 gas. Since both elimination mechanisms may occur through a six-membered cyclic transition state structure, it is difficult to elucidate experimentally which is the most reasonable reaction mechanism. Theoretical calculations show that the stepwise mechanism with the formation of the unstable intermediate chloroformic acid from ECF, ICF, and SCF is favoured over one-step elimination. Reasonable agreements were found between theoretical and experimental values at the CAM-B3LYP/6-311++G(d,p) level.

Antimutagenic and free radical scavenger effects of leaf extracts from Accacia salicina

PubMed Central

2011-01-01

Background Three extracts were prepared from the leaves of Accacia salicina; ethyl acetate (EA), chloroform (Chl) and petroleum ether (PE) extracts and was designed to examine antimutagenic, antioxidant potenty and oxidative DNA damage protecting activity. Methods Antioxidant activity of A. salicina extracts was determined by the ability of each extract to protect against plasmid DNA strand scission induced by hydroxyl radicals. An assay for the ability of these extracts to prevent mutations induced by various oxidants in Salmonella typhimurium TA102 and TA 104 strains was conducted. In addition, nonenzymatic methods were employed to evaluate anti-oxidative effects of tested extracts. Results These extracts from leaf parts of A. salicina showed no mutagenicity either with or without the metabolic enzyme preparation (S9). The highest protections against methylmethanesulfonate induced mutagenicity were observed with all extracts and especially chloroform extract. This extract exhibited the highest inhibitiory level of the Ames response induced by the indirect mutagen 2- aminoanthracene. All extracts exhibited the highest ability to protect plasmid DNA against hydroxyl radicals induced DNA damages. The ethyl acetate (EA) and chloroform (Chl) extracts showed with high TEAC values radical of 0.95 and 0.81 mM respectively, against the ABTS.+. Conclusion The present study revealed the antimutagenic and antioxidant potenty of plant extract from Accacia salicina leaves. PMID:22132863

LFER Studies Evaluating Solvent Effects on an α-Chloro-and two β,β,β-Trichloro-Ethyl Chloroformate Esters

PubMed Central

D’Souza, Malcolm J.; Sandosky, Brandon; Fernandez-Bueno, Gabriel A.; McAneny, Matthew J.; Kevill, Dennis N.

2014-01-01

To provide insight and to identify the occurrence of mechanistic changes in relation to variance in solvent-type, the solvent effects on the rates of solvolysis of three substrates, 2,2,2-trichloro-1,1-dimethylethyl chloroformate, 2,2,2-trichloroethyl chloroformate, and 1-chloroethyl chloroformate, are analyzed using linear free energy relationships (LFERs) such as the extended Grunwald-Winstein equation, and a similarity-based LFER model approach that is based on the solvolysis of phenyl chloroformate. At 25.0 °C, in four common solvents, the α-chloroethyl chloroformate was found to react considerably faster than the two β,β,β-trichloro-substituted analogs. This immense rate enhancement can be directly related to the proximity of the electron-withdrawing α-chlorine atom to the carbonyl carbon reaction center. In the thirteen solvents studied, 1-chloroethyl chloroformate was found to strictly follow a carbonyl addition process, with the addition-step being rate-determining. For the two β,β,β-trichloro-substrates, in aqueous mixtures that are very rich in a fluoroalcohol component, there is compelling evidence for the occurrence of side-by-side addition-elimination and ionization mechanisms, with the ionization pathway being predominant. The presence of the two methyl groups on the α-carbon of 2,2,2-trichloro-1,1-dimethylethyl chloroformate has additive steric and stereoelectronic implications, causing its rate of reaction to be significantly slower than that of 2,2,2-trichloroethyl chloroformate. PMID:24812595

Microwave spectroscopy and curious molecular dynamics of ethyl trifluoroacetate

NASA Astrophysics Data System (ADS)

Bohn, Robert K.; Montgomery, John A.; Harvey Michels, H.; Acharte, Christian

2017-05-01

The first ethyl ester whose structure was determined by microwave spectroscopy is ethyl formate. It exists in two conformations. In the 1970s, that study was used as a model to determine the structures of other ethyl esters, ethyl cyanoformate, chloroformate, and trifluoroacetate. They display the same conformations as ethyl formate. But under the experimental conditions used, Stark modulation with a maximum electric field, static low pressure gas, rapid sweeping, and long detector time constants, each of those esters displays bands of an additional third species. A careful, high resolution study of ethyl cyanoformate only observed two conformers. A model has been proposed that the third species derives from a dense array of torsionally excited states with broadened transitions due to short lifetimes. The present study of ethyl trifluoroacetate in a pulsed jet Fourier Transform spectrometer is intended to clarify the earlier results. Two conformers are observed including all their monosubstituted 13C and 18O isotopologs. In a pulsed jet Fourier Transform spectrometer using argon as the carrier gas, only one conformer is observed. Switching to helium as the carrier gas, another, higher energy conformer is also observed.

Development of a simple and rapid solid phase microextraction-gas chromatography-triple quadrupole mass spectrometry method for the analysis of dopamine, serotonin and norepinephrine in human urine.

PubMed

Naccarato, Attilio; Gionfriddo, Emanuela; Sindona, Giovanni; Tagarelli, Antonio

2014-01-31

The work aims at developing a simple and rapid method for the quantification of dopamine (DA), serotonin (5-HT) and norepinephrine (NE) in human urine. The urinary levels of these biogenic amines can be correlated with several pathological conditions concerning heart disease, stress, neurological disorders and cancerous tumors. The proposed analytical approach is based on the use of solid phase microextraction (SPME) combined with gas chromatography-triple quadrupole mass spectrometry (GC-QqQ-MS) after a fast derivatization of both aliphatic amino and phenolic moieties by propyl chloroformate. The variables influencing the derivatization reaction were reliably optimized by the multivariate approach of "Experimental design". The optimal conditions were obtained by performing derivatization with 100μL of propyl chloroformate and 100μL of pyridine. The extraction ability of five commercially available SPME fibers was evaluated in univariate mode and the best results were obtained using the polyacrylate fiber. The variables affecting the efficiency of SPME analysis were again optimized by the multivariate approach of "Experimental design" and, in particular, a central composite design (CCD) was applied. The optimal values were extraction in 45min at room temperature, desorption temperature at 300°C, no addition of NaCl. Assay of derivatized analytes was performed by using a gas chromatography-triple quadrupole mass spectrometry (GC-QqQ-MS) system in selected reaction monitoring (SRM) acquisition. An evaluation of all analytical parameters demonstrates that the developed method provides satisfactory results. Indeed, very good linearities were achieved in the tested calibration range with correlation coefficient values of 0.9995, 0.9999 and 0.9997 for DA, 5-HT and NE, respectively. Accuracies and RSDs calculated for between-run and tested at concentrations of 30, 200, and 800μg L(-1) were in the range from 92.8% to 103.0%, and from 0.67 to 4.5%, respectively. Finally, the LOD values obtained can be considered very good (0.587, 0.381 and 1.23μg L(-1) for DA, 5-HT and NE, respectively). Copyright © 2013 Elsevier B.V. All rights reserved.

Correlation of the Rates of Solvolysis of Neopentyl Chloroformate—A Recommended Protecting Agent

PubMed Central

D’Souza, Malcolm J.; Carter, Shannon E.; Kevill, Dennis N.

2011-01-01

The specific rates of solvolysis of neopentyl chloroformate (1) have been determined in 21 pure and binary solvents at 45.0 °C. In most solvents the values are essentially identical to those for ethyl and n-propyl chloroformates. However, in aqueous-1,1,1,3,3,3-hexafluoro-2-propanol mixtures (HFIP) rich in fluoroalcohol, 1 solvolyses appreciably faster than the other two substrates. Linear free energy relationship (LFER) comparison of the specific rates of solvolysis of 1 with those for phenyl chloroformate and those for n-propyl chloroformate are helpful in the mechanistic considerations, as is also the treatment in terms of the Extended Grunwald-Winstein equation. It is proposed that the faster reaction for 1 in HFIP rich solvents is due to the influence of a 1,2-methyl shift, leading to a tertiary alkyl cation, outweighing the only weak nucleophilic solvation of the cation possible in these low nucleophilicity solvents. PMID:21541050

Enteromorpha compressa Exhibits Potent Antioxidant Activity

PubMed Central

Shanab, Sanaa M. M.; Shalaby, Emad A.; El-Fayoumy, Eman A.

2011-01-01

The green macroalgae, Enteromorpha compressa (Linnaeus) Nees, Ulva lactuca, and E. linza, were seasonally collected from Abu Qir bay at Alexandria (Mediterranean Sea) This work aimed to investigate the seasonal environmental conditions, controlling the green algal growth, predominance, or disappearance and determining antioxidant activity. The freshly collected selected alga (E. compressa) was subjected to pigment analysis (chlorophyll and carotenoids) essential oil and antioxidant enzyme determination (ascorbate oxidase and catalase). The air-dried ground alga was extracted with ethanol (crude extract) then sequentially fractionated by organic solvents of increasing polarity (petroleum ether, chloroform, ethyl acetate, and water). Antioxidant activity of all extracts was assayed using different methods (total antioxidant, DPPH [2, 2 diphenyl-1-picrylhydrazyl], ABTS [2, 2 azino-bis ethylbenzthiazoline-6-sulfonic acid], and reducing power, and β-carotene linoleic acid bleaching methods). The results indicated that the antioxidant activity was concentration and time dependent. Ethyl acetate fraction demonstrated higher antioxidant activity against DPPH method (82.80%) compared to the synthetic standard butylated hydroxyl toluene (BHT, 88.5%). However, the crude ethanolic extract, pet ether, chloroform fractions recorded lower to moderate antioxidant activities (49.0, 66.0, and 78.0%, resp.). Using chromatographic and spectroscopic analyses, an active compound was separated and identified from the promising ethyl acetate fraction. PMID:21869863

Evaluation of larvicidal activity of Acalypha alnifolia Klein ex Willd. (Euphorbiaceae) leaf extract against the malarial vector, Anopheles stephensi, dengue vector, Aedes aegypti and Bancroftian filariasis vector, Culex quinquefasciatus (Diptera: Culicidae).

PubMed

Kovendan, Kalimuthu; Murugan, Kadarkarai; Vincent, Savariar

2012-02-01

The leaf extract of Acalypha alnifolia with different solvents - hexane, chloroform, ethyl acetate, acetone and methanol - were tested for larvicidal activity against three important mosquitoes such as malarial vector, Anopheles stephensi, dengue vector, Aedes aegypti and Bancroftian filariasis vector, Culex quinquefasciatus. The medicinal plants were collected from the area around Kallar Hills near the Western Ghats, Coimbatore, India. A. alnifolia plant was washed with tap water and shade dried at room temperature. The dried leaves were powdered mechanically using commercial electrical stainless steel blender. The powder 800 g of the leaf material was extract with 2.5 litre of various each organic solvents such as hexane, chloroform, ethyl acetate, acetone, methanol for 8 h using Soxhlet apparatus, and filtered. The crude plant extracts were evaporated to dryness in a rotary vacuum evaporator. The yield of extracts was hexane (8.64 g), chloroform (10.74 g), ethyl acetate (9.14 g), acetone (10.02 g), and methanol (11.43 g). One gram of the each plant residue was dissolved separately in 100 ml of acetone (stock solution) from which different concentrations, i.e., 50, 150, 250, 350 and 450 ppm, was prepared. The hexane, chloroform, ethyl acetate, acetone was moderate considerable mortality; however, the highest larval mortality was methanolic extract observed in three mosquito vectors. The larval mortality was observed after 24 h exposure. No mortality was observed in the control. The early fourth-instar larvae of A. stephensi had values of LC(50) = 197.37, 178.75, 164.34, 149.90 and 125.73 ppm and LC(90) = 477.60, 459.21, 435.07, 416.20 and 395.50 ppm, respectively. The A. aegypti had values of LC(50) = 202.15, 182.58, 160.35, 146.07 and 128.55 ppm and LC(90) = 476.57, 460.83, 440.78, 415.38 and 381.67 ppm, respectively. The C. quinquefasciatus had values of LC(50) = 198.79, 172.48, 151.06, 140.69 and 127.98 ppm and LC(90) = 458.73, 430.66, 418.78, 408.83 and 386.26 ppm, respectively. The results of the leaf extract of A. alnifloia are promising as good larvicidal activity against the mosquito vector, A. stephensi, A. aegypti, C. quinquefasciatus. Therefore, this study provides first report on the larvicidal activities against three species of mosquito vectors of this plant extracts from Southern India.

Determination of atranol and chloroatranol in perfumes using simultaneous derivatization and dispersive liquid-liquid microextraction followed by gas chromatography-mass spectrometry.

PubMed

López-Nogueroles, Marina; Chisvert, Alberto; Salvador, Amparo

2014-05-15

A new analytical method based on simultaneous derivatization and dispersive liquid-liquid microextraction (DLLME) followed by gas chromatography-mass spectrometry (GC-MS), for the determination of the allergenic compounds atranol and chloroatranol in perfumes, is presented. Derivatization of the target analytes by means of acetylation with anhydride acetic in carbonate buffer was carried out. Thereby volatility and detectability were increased for improved GC-MS sensitivity. In addition, extractability by DLLME was also enhanced due to a less polar character of the solutes. A liquid-liquid extraction was performed before DLLME to clean up the sample and to obtain an aqueous sample solution, free of the low polar matrix from the essential oils, as donor phase. Different parameters, such as the nature and volume of both the extraction and disperser solvents, the ionic strength of the aqueous donor phase or the effect of the derivatization reagent volume, were optimized. Under the selected conditions (injection of a mixture of 750μL of acetone as disperser solvent, 100μL of chloroform as extraction solvent and 100μL of anhydride acetic as derivatization reagent) the figures of merit of the proposed method were evaluated. Limits of detection in the low ngmL(-1) range were obtained. Matrix effect was observed in real perfume samples and thus, standard addition calibration is recommended. Copyright © 2014 Elsevier B.V. All rights reserved.

CASTING SLIPS FOR FABRICATION OF REFRACTORY METAL WARE

DOEpatents

Stoddard, S.D.; Nuckolls, D.E.; Cowan, R.E.

1962-09-01

A composition is given for slip casting tungsten metal. The composition consists essentially of tungsten metal with an average particle size of 0.9 micron, an organic vehicle such as methyl chloroform, o-xylene, n-butyl acetate, isobutyl acetate, and 1, 1, 2, 2-tetrachlorethane, and a suspending agent such as ethyl cellulose, with the approximate ratio of said vehicle to the tungsten metal being 12 cc of a solution containing from 5 to about 20 grams of said ethyl cellulose in 400 cc of said organic vehicle per 100 grams of metal. (AEC)

Analytical method for tributyltin and triphenyltin contained in household products-preparing for the revision of authorized analytical method-.

PubMed

Nakashima, Harunobu; Tomiyama, Ken-Ichi; Kawakami, Tsuyoshi; Isama, Kazuo

2010-07-01

In preparing for the revision of the authorized analytical method for tributyltin (TBT) and triphenyltin (TPT), which are banned from using according to the "Act on the Control of Household Products Containing Harmful Substances", an examination was conducted on the detection method of these substances using gas chromatography/mass spectrometry (GC/MS), after derivatizing them (ethyl-derivatizing method and hydrogen-derivatizing method). Ethyl-derivatized compounds had stability, which enabled the detection of TPT with a higher sensitivity. In addition, a preparation suitable for the following analytical objects was established: (1) textile products, (2) water-based products (such as water-based paint), (3) oil-based products (such as wax), and (4) adhesives. Addition-recovery experiments were conducted using the prescribed pretreatment method, when each surrogate substances (TBT-d27, TPT-d15) were added and the data were corrected, good recovery rates (94.5-118.6% in TBT, and 86.6-110.1% in TPT) were obtained. When TBT and TPT in 31 commercially available products were analyzed based on the developed analytical method, an adhesive showed 13.2 microg/g of TBT content, which exceeded the regulatory criterion (1 microg/g as tin). Next, when the same products with different manufacturing date were analyzed, TBT (10.2-10.8 microg/g), which exceeded the regulatory criterion, was detected in 4 products among 8 products, and simultaneously, a high concentration (over 1000 microg/g) of dibutyltin (DBT) was detected. It was suggested that TBT as an impurity of DBT remained, and the manufacturer chose the voluntary recall of the product. The new method is considered sufficiently applicable as a revised method for the conventionally authorized method.

Antimutagenic and free radical scavenger effects of leaf extracts from Accacia salicina.

PubMed

Boubaker, Jihed; Mansour, Hedi Ben; Ghedira, Kamel; Chekir-Ghedira, Leila

2011-12-01

Three extracts were prepared from the leaves of Accacia salicina; ethyl acetate (EA), chloroform (Chl) and petroleum ether (PE) extracts and was designed to examine antimutagenic, antioxidant potenty and oxidative DNA damage protecting activity. Antioxidant activity of A. salicina extracts was determined by the ability of each extract to protect against plasmid DNA strand scission induced by hydroxyl radicals. An assay for the ability of these extracts to prevent mutations induced by various oxidants in Salmonella typhimurium TA102 and TA 104 strains was conducted. In addition, nonenzymatic methods were employed to evaluate anti-oxidative effects of tested extracts. These extracts from leaf parts of A. salicina showed no mutagenicity either with or without the metabolic enzyme preparation (S9). The highest protections against methylmethanesulfonate induced mutagenicity were observed with all extracts and especially chloroform extract. This extract exhibited the highest inhibitiory level of the Ames response induced by the indirect mutagen 2- aminoanthracene. All extracts exhibited the highest ability to protect plasmid DNA against hydroxyl radicals induced DNA damages. The ethyl acetate (EA) and chloroform (Chl) extracts showed with high TEAC values radical of 0.95 and 0.81 mM respectively, against the ABTS(.+). The present study revealed the antimutagenic and antioxidant potenty of plant extract from Accacia salicina leaves. © 2011 Boubaker et al; licensee BioMed Central Ltd.

Isolation of quercetin and mandelic acid from Aesculus indica fruit and their biological activities.

PubMed

Zahoor, Muhammad; Shafiq, Sadaf; Ullah, Habib; Sadiq, Abdul; Ullah, Farhat

2018-06-26

In this study Aesculus indica fruit was subjected to isolation of phytochemicals. Two antioxidants quercetin and Mandelic acid were isolated in pure state. The free radical scavenging and acetyl choline esterase inhibitory potential of the crude extract and sub fractions were also determined. The antioxidant capacity of crude extract, fractions and isolated compounds were determined by DPPH and ABTS methods. Folin-Ciocalteu reagent method was used to estimate the total phenolic contents and were found to be 78.34 ± 0.96, 44.16 ± 1.05, 65.45 ± 1.29, 37.85 ± 1.44 and 50.23 ± 2.431 (mg/g of gallic acid) in crude extract, ethyl acetate, chloroform, n-hexane and aqueous fractions respectively. The flavonoid concentration in crude extract, ethyl acetate, chloroform, n-hexane and aqueous fraction were; 85.30 ± 1.20, 53.80 ± 1.07, 77.50 ± 1.12, 26.30 ± 1.35 and 37.78 ± 1.25 (mg/g of quercetin) respectively. The chloroform fraction was more potent against enzymes, acetyl choline esterase and butyryl choline esterase (IC 50  = 85 and 160 μg/ml respectively). The phenolic compounds in the crude extract and fractions were determined using HPLC standard method. Chlorogenic acid, quercetin, phloroglucinol, rutin, mandelic acid and hydroxy benzoic acid were detected at retention times 6.005, 10.062, 22.623, 30.597, 35.490 and 36.211 in crude extract and different fractions. The ethyl acetate fraction was rich in the targeted compounds and was therefore subjected to column isolation. The HPLC chromatogram of isolated compounds showed single peak at specified retention times which confirms their isolation in pure state. The isolated compounds were then characterized by FTIR and NMR spectrophotometric techniques. The Aesculus indica fruit extracts showed antioxidant and anticholine esterase inhibitory potentials. Two bioactive compounds were isolated in the pure form ethyl acetate fraction. From results it was concluded that the fruit of this plant could be used to minimize oxidative stress caused by reactive oxygen species.

Laboratory and field evaluation of medicinal plant extracts against filarial vector, Culex quinquefasciatus Say (Diptera: Culicidae)

USDA-ARS?s Scientific Manuscript database

In this study, chemical extracts of Jatropha curcas, Hyptis suaveolens, Abutilon indicum, and Leucas aspera were tested for toxicity to larvae of the filariasis vector Culex quinquefasciatus. Respective median lethal concentrations (LC50) for hexane, chloroform, ethyl acetate, and methanol extracts...

Mosquito larvicidal properties of Orthisiphon thymiflorus (Roth) Sleesen. (Family: Labiatae) against mosquito vectors, Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti (Diptera: Culicidae)

USDA-ARS?s Scientific Manuscript database

Objective: To determine the larvicidal activity of hexane, chloroform, ethyl acetate, acetone, and methanol extracts of Orthosiphon thymiflorus leaves against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti. Methods: Larvicidal activity was determined in laboratory bioassays using var...

Ovicidal activity of Ageratina adenophora (Family: Asteraceae) against dengue vector, Aedes aegypti (Diptera: Culicidae)

USDA-ARS?s Scientific Manuscript database

To determine the ovicidal efficacy of different solvent leaf extracts of Ageratina adenophora against dengue vector Aedes aegypti . Methods: The ovicidal efficacy of the crude leaf extracts of A. adenophora with five different solvents (hexane, benzene, chloroform, ethyl acetate, methanol) and was ...

Analysis of psilocybin and psilocin in Psilocybe subcubensis Guzmán by ion mobility spectrometry and gas chromatography-mass spectrometry.

PubMed

Keller, T; Schneider, A; Regenscheit, P; Dirnhofer, R; Rücker, T; Jaspers, J; Kisser, W

1999-01-11

A new method has been developed for the rapid analysis of psilocybin and/or psilocin in fungus material using ion mobility spectrometry. Quantitative analysis was performed by gas chromatography-mass spectrometry after a simple one-step extraction involving homogenization of the dried fruit bodies of fungi in chloroform and derivatization with MSTFA. The proposed methods resulted in rapid procedures useful in analyzing psychotropic fungi for psilocybin and psilocin.

Simple and sensitive analysis of long-chain free fatty acids in milk by fluorogenic derivatization and high-performance liquid chromatography.

PubMed

Lu, Chi-Yu; Wu, Hsin-Lung; Chen, Su-Hwei; Kou, Hwang-Shang; Wu, Shou-Mei

2002-01-02

A highly sensitive high-performance liquid chromatography (HPLC) method is described for the simultaneous determination of some important saturated and unsaturated fatty acids in milk, including lauric (dodecanoic), myristic (tetradecanoic), palmitic (hexadecanoic), stearic (octadecanoic), palmitoleic (hexadecenoic), oleic (octadecenoic), and linoleic acids (octadecadienoic acids). The fatty acids were fluorogenically derivatized with 2-(2-naphthoxy)ethyl 2-(piperidino)ethanesulfonate (NOEPES) as their naphthoxyethyl derivatives. The resulting derivatives were separated by isocratic HPLC and monitored with a fluorometric detector (lambdaex = 235 nm, lambdaem = 350 nm). The fatty acids in milk were extracted with toluene, and the extract with the fatty acids was directly derivatized with NOEPES without solvent replacement. Determination of long-chain free fatty acids in milk is feasible by a standard addition method. A small amount of milk product, 10 microL, is sufficient for the analysis.

Wet in situ transesterification of microalgae using ethyl acetate as a co-solvent and reactant.

PubMed

Park, Jeongseok; Kim, Bora; Chang, Yong Keun; Lee, Jae W

2017-04-01

This study addresses wet in situ transesterification of microalgae for the production of biodiesel by introducing ethyl acetate as both reactant and co-solvent. Ethyl acetate and acid catalyst are mixed with wet microalgae in one pot and the mixture is heated for simultaneous lipid extraction and transesterification. As a single reactant and co-solvent, ethyl acetate can provide higher FAEE yield and more saccharification of carbohydrates than the case of binary ethanol and chloroform as a reactant and a co-solvent. The optimal yield was 97.8wt% at 114°C and 4.06M catalyst with 6.67mlEtOAC/g dried algae based on experimental results and response surface methodology (RSM). This wet in situ transesterification of microalgae using ethyl acetate doesn't require an additional co-solvent and it also promises more economic benefit as combining extraction and transesterification in a single process. Copyright © 2017 Elsevier Ltd. All rights reserved.

Comparison of biodegradation of poly(ethylene glycol)s and poly(propylene glycol)s.

PubMed

Zgoła-Grześkowiak, Agnieszka; Grześkowiak, Tomasz; Zembrzuska, Joanna; Łukaszewski, Zenon

2006-07-01

The biodegradation of poly(ethylene glycol)s (PEGs) and poly(propylene glycol)s (PPGs), both being major by-products of non-ionic surfactants biodegradation, was studied under the conditions of the River Water Die-Away Test. PEGs were isolated from a water matrix using solid-phase extraction with graphitized carbon black sorbent, then derivatized with phenyl isocyanate and determined by HPLC with UV detection. PPGs were isolated from a water matrix by liquid-liquid extraction with chloroform, then derivatized with naphthyl isocyanate and determined by HPLC with fluorescence detection. The primary biodegradation of both PEGs and PPGs reached approximately 99% during the test. The tests show different biodegradation pathways of PEG and PPG. During PEG biodegradation, their chains are shortened leading to the formation of ethylene glycol and diethylene glycol. During PPG biodegradation, no short-chained biodegradation products were found.

A targeted metabolomic protocol for short-chain fatty acids and branched-chain amino acids.

PubMed

Zheng, Xiaojiao; Qiu, Yunping; Zhong, Wei; Baxter, Sarah; Su, Mingming; Li, Qiong; Xie, Guoxiang; Ore, Brandon M; Qiao, Shanlei; Spencer, Melanie D; Zeisel, Steven H; Zhou, Zhanxiang; Zhao, Aihua; Jia, Wei

2013-08-01

Research in obesity and metabolic disorders that involve intestinal microbiota demands reliable methods for the precise measurement of the short-chain fatty acids (SCFAs) and branched-chain amino acids (BCAAs) concentration. Here, we report a rapid method of simultaneously determining SCFAs and BCAAs in biological samples using propyl chloroformate (PCF) derivatization followed by gas chromatography mass spectrometry (GC-MS) analysis. A one-step derivatization using 100 µL of PCF in a reaction system of water, propanol, and pyridine (v/v/v = 8:3:2) at pH 8 provided the optimal derivatization efficiency. The best extraction efficiency of the derivatized products was achieved by a two-step extraction with hexane. The method exhibited good derivatization efficiency and recovery for a wide range of concentrations with a low limit of detection for each compound. The relative standard deviations (RSDs) of all targeted compounds showed good intra- and inter-day (within 7 days) precision (< 10%), and good stability (< 20%) within 4 days at room temperature (23-25 °C), or 7 days when stored at -20 °C. We applied our method to measure SCFA and BCAA levels in fecal samples from rats administrated with different diet. Both univariate and multivariate statistics analysis of the concentrations of these target metabolites could differentiate three groups with ethanol intervention and different oils in diet. This method was also successfully employed to determine SCFA and BCAA in the feces, plasma and urine from normal humans, providing important baseline information of the concentrations of these metabolites. This novel metabolic profile study has great potential for translational research.

Phytohormone profiling of the sweet orange (Citrus sinensis (L.) Osbeck) leaves and roots using GC-MS-based method.

PubMed

Nehela, Yasser; Hijaz, Faraj; Elzaawely, Abdelnaser A; El-Zahaby, Hassan M; Killiny, Nabil

2016-07-20

Phytohormones mainly affect plant development and trigger varied responses to biotic and abiotic stresses. The sensitivity of methods used to profile phytohormones is a vital factor that affects the results. We used an improved GC-MS-based method in the selective ion-monitoring (SIM) mode to study the phytohormone profiling in citrus tissues. One extraction solvent mixture and two derivatization reagents were used, methyl chloroformate (MCF) and N-Methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA). The method showed a low limit of detection and low limit of quantification with high extraction recovery percentage and reproducibility. Overall, we detected 13 phytohormones belonging to six different groups. Auxins, SAs, tJA, and ABA were detected after derivatization with MCF while cytokinins and GAs were detected after derivatization with MSTFA. Cytokinins, SAs, and gibberellins were found in all tissues while auxins and tJA were observed only in the leaves. ABA was found in leaves and roots, but not in root tips. The method we used is efficient, precise, and appropriate to study citrus phytohormonal profiles to understand their crosstalk and responses to environmental and biological stresses. Copyright © 2016 Elsevier GmbH. All rights reserved.

Effect of casting solvent on crystallinity of ondansetron in transdermal films.

PubMed

Pattnaik, Satyanarayan; Swain, Kalpana; Mallick, Subrata; Lin, Zhiqun

2011-03-15

The purpose of the present investigation is to assess the influence of casting solvent on crystallinity of ondansetron hydrochloride in transdermal polymeric matrix films fabricated using povidone and ethyl cellulose as matrix forming polymers. Various casting solvents like chloroform (CHL), dichloromethane (DCM), methanol (MET); and mixture of chloroform and ethanol (C-ETH) were used for fabrication of the transdermal films. Analytical tools like scanning electron microscopy (SEM), X-ray diffraction (XRD) studies, differential scanning calorimetry (DSC), etc. were utilized to characterize the crystalline state of ondansetron in the film. Recrystallisation was observed in all the transdermal films fabricated using the casting solvents other than chloroform. Long thin slab-looking, long wire-like or spherulite-looking crystals with beautiful impinged boundaries were observed in SEM. Moreover, XRD revealed no crystalline peaks of ondansetron hydrochloride in the transdermal films prepared using chloroform as casting solvent. The significantly decreased intensity and sharpness of the DSC endothermic peaks corresponding to the melting point of ondansetron in the formulation (specifically in CHL) indicated partial dissolution of ondansetron crystals in the polymeric films. The employed analytical tools suggested chloroform as a preferred casting solvent with minimum or practically absence of recrystallization indicating a relatively amorphous state of ondansetron in transdermal films. Copyright © 2011 Elsevier B.V. All rights reserved.

Antibacterial and phytochemical studies on Calotropis gigantia (L.) R. Br. latex against selected cariogenic bacteria

PubMed Central

Ishnava, Kalpesh B.; Chauhan, Jenabhai B.; Garg, Akanksha A.; Thakkar, Arpit M.

2011-01-01

In vitro antibacterial potential of the chloroform, ethyl acetate, hexane, methanol and aqueous extracts of Calotropis gigantia (L.) R. Br. was evaluated by using five cariogenic bacteria, Actinomyces viscosus, Lactobacillus acidophilus, Lactobacillus casei, Streptococcus mitis and Streptococcus mutans. Agar well diffusion method and minimum inhibitory concentration (MIC) were used for this purpose. The chloroform extracted fraction of latex showed inhibitory effect against S. mutans and L. acidophilus with MIC value of 0.032 and 0.52 mg/mL, respectively. Qualitative investigation on structure elucidation of bioactive compound using IR, NMR and GC–MS techniques revealed the presence of methyl nonanoate, a saturated fatty acid. PMID:23961166

β-Keto esters from ketones and ethyl chloroformate: a rapid, general, efficient synthesis of pyrazolones and their antimicrobial, in silico and in vitro cytotoxicity studies

PubMed Central

2013-01-01

Background Pyrazolones are traditionally synthesized by the reaction of β-keto esters with hydrazine and its derivatives. There are methods to synthesize β-keto esters from esters and aldehydes, but these methods have main limitation in varying the substituents. Often, there are a number of methods such as acylation of enolates in which a chelating effect has been employed to lock the enolate anion using lithium and magnesium salts; however, these methods suffer from inconsistent yields in the case of aliphatic acylation. There are methods to synthesize β-keto esters from ketones like caboxylation of ketone enolates using carbon dioxide and carbon monoxide sources in the presence of palladium or transition metal catalysts. Currently, the most general and simple method to synthesize β-keto ester is the reaction of dimethyl or ethyl carbonate with ketone in the presence of strong bases which also requires long reaction time, use of excessive amount of reagent and inconsistent yield. These factors lead us to develop a simple method to synthesize β-keto esters by changing the base and reagent. Results A series of β-keto esters were synthesized from ketones and ethyl chloroformate in the presence of base which in turn are converted to pyrazolones and then subjected to cytotoxicity studies towards various cancer cell lines and antimicrobial activity studies towards various bacterial and fungal strains. Conclusion The β-keto esters from ethyl chloroformate was successfully attempted, and the developed method is simple, fast and applicable to the ketones having the alkyl halogens, protecting groups like Boc and Cbz that were tolerated and proved to be useful in the synthesis of fused bicyclic and tricyclic pyrazolones efficiently using cyclic ketones. Since this method is successful for different ketones, it can be useful for the synthesis of pharmaceutically important pyrazolones also. The synthesized pyrazolones were subjected to antimicrobial, docking and cytotoxicity assay against ACHN (human renal cell carcinoma), Panc-1 (human pancreatic adenocarcinoma) and HCT-116 (human colon cancer) cell line, and lead molecules have been identified. Some of the compounds are found to have promising activity against different bacterial and fungal strains tested. PMID:23870758

Hair analysis for delta(9)-THC, delta(9)-THC-COOH, CBN and CBD, by GC/MS-EI. Comparison with GC/MS-NCI for delta(9)-THC-COOH.

PubMed

Baptista, Maria João; Monsanto, Paula Verâncio; Pinho Marques, Estela Gouveia; Bermejo, Ana; Avila, Sofia; Castanheira, Alice Martelo; Margalho, Cláudia; Barroso, Mário; Vieira, Duarte Nuno

2002-08-14

A sensitive analytical method was developed for quantitative analysis of delta(9)-tetrahydrocannabinol (delta(9)-THC), 11-nor-delta(9)-tetrahydrocannabinol-carboxylic acid (delta(9)-THC-COOH), cannabinol (CBN) and cannabidiol (CBD) in human hair. The identification of delta(9)-THC-COOH in hair would document Cannabis use more effectively than the detection of parent drug (delta(9)-THC) which might have come from environmental exposure. Ketamine was added to hair samples as internal standard for CBN and CBD. Ketoprofen was added to hair samples as internal standard for the other compounds. Samples were hydrolyzed with beta-glucuronidase/arylsulfatase for 2h at 40 degrees C. After cooling, samples were extracted with a liquid-liquid extraction procedure (with chloroform/isopropyl alcohol, after alkalinization, and n-hexane/ethyl acetate, after acidification), which was developed in our laboratory. The extracts were analysed before and after derivatization with pentafluoropropionic anhydride (PFPA) and pentafluoropropanol (PFPOH) using a Hewlett Packard gas chromatographer/mass spectrometer detector, in electron impact mode (GC/MS-EI). Derivatized delta(9)-THC-COOH was also analysed using a Hewlett Packard gas chromatographer/mass spectrometer detector, in negative ion chemical ionization mode (GC/MS-NCI) using methane as the reagent gas. Responses were linear ranging from 0.10 to 5.00 ng/mg hair for delta(9)-THC and CBN, 0.10-10.00 ng/mg hair for CBD, 0.01-5.00 ng/mg for delta(9)-THC-COOH (r(2)>0.99). The intra-assay precisions ranged from <0.01 to 12.40%. Extraction recoveries ranged from 80.9 to 104.0% for delta(9)-THC, 85.9-100.0% for delta(9)-THC-COOH, 76.7-95.8% for CBN and 71.0-94.0% for CBD. The analytical method was applied to 87 human hair samples, obtained from individuals who testified in court of having committed drug related crimes. Quantification of delta(9)-THC-COOH using GC/MS-NCI was found to be more convenient than GC/MS-EI. The latter may give rise to false negatives due to the detection limit.

Monitoring the contents of six steroidal and phenolic endocrine disrupting chemicals in chicken, fish and aquaculture pond water samples using pre-column derivatization and dispersive liquid-liquid microextraction with the aid of experimental design methodology.

PubMed

Wu, Hongliang; Li, Guoliang; Liu, Shucheng; Hu, Na; Geng, Dandan; Chen, Guang; Sun, Zhiwei; Zhao, Xianen; Xia, Lian; You, Jinmao

2016-02-01

This research established a sensitive and efficient pre-column derivatization HPLC method based on dispersive liquid-liquid microextraction (DLLME) for the simultaneous determination of six steroidal and phenolic endocrine disrupting chemicals (EDCs). In this study, EDCs were firstly labeled by the derivatization reagent 2-(11H-benzo[a]carbazol-11-yl) ethyl carbonochloridate (BCEC-Cl) and then extracted by DLLME. The response surface methodology was employed to investigate the key parameters of pre-column derivatization and DLLME. Under the optimal conditions, a good linear relationship between the peak area and the concentration of analytes was observed with correlation coefficients of >0.9991. Limits of detection for all EDCs derivatives were achieved within the range of 0.02-0.07 μg L(-1). The proposed method has the advantages of simple operation, low consumption of organic solvent, saving time, low output limit and good selectivity. When applied to several food and water samples analysis, it demonstrated good applicability for the determination of EDCs. Copyright © 2015 Elsevier Ltd. All rights reserved.

A targeted metabolomic protocol for short-chain fatty acids and branched-chain amino acids

PubMed Central

Zheng, Xiaojiao; Qiu, Yunping; Zhong, Wei; Baxter, Sarah; Su, Mingming; Li, Qiong; Xie, Guoxiang; Ore, Brandon M.; Qiao, Shanlei; Spencer, Melanie D.; Zeisel, Steven H.; Zhou, Zhanxiang; Zhao, Aihua; Jia, Wei

2013-01-01

Research in obesity and metabolic disorders that involve intestinal microbiota demands reliable methods for the precise measurement of the short-chain fatty acids (SCFAs) and branched-chain amino acids (BCAAs) concentration. Here, we report a rapid method of simultaneously determining SCFAs and BCAAs in biological samples using propyl chloroformate (PCF) derivatization followed by gas chromatography mass spectrometry (GC-MS) analysis. A one-step derivatization using 100 µL of PCF in a reaction system of water, propanol, and pyridine (v/v/v = 8:3:2) at pH 8 provided the optimal derivatization efficiency. The best extraction efficiency of the derivatized products was achieved by a two-step extraction with hexane. The method exhibited good derivatization efficiency and recovery for a wide range of concentrations with a low limit of detection for each compound. The relative standard deviations (RSDs) of all targeted compounds showed good intra- and inter-day (within 7 days) precision (< 10%), and good stability (< 20%) within 4 days at room temperature (23–25 °C), or 7 days when stored at −20 °C. We applied our method to measure SCFA and BCAA levels in fecal samples from rats administrated with different diet. Both univariate and multivariate statistics analysis of the concentrations of these target metabolites could differentiate three groups with ethanol intervention and different oils in diet. This method was also successfully employed to determine SCFA and BCAA in the feces, plasma and urine from normal humans, providing important baseline information of the concentrations of these metabolites. This novel metabolic profile study has great potential for translational research. PMID:23997757

In vitro assessment of relief to oxidative stress by different fractions of Boerhavia procumbens.

PubMed

Abbasi, Muhammad Athar; Rubab, Kaniz; Rehman, Azizur; Riaz, Tauheeda; Shahzadi, Tayyaba; Khalid, Muniba; Ajaib, Muhammad

2012-04-01

Methanolic extract of Boerhavia procumbens Bank ex Roxb. was partitioned with n-hexane, chloroform, ethyl acetate and n-butanol sequentially after dissolving in distilled water. Phytochemical screening showed presence of phenolics, flavonoides and cardiac glycosides in large amount in chloroform, ethyl acetate and n-butanol soluble fraction. The antioxidant activity of all these fractions and the remaining aqueous fraction was evaluated by four methods such as: 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, ferric reducing antioxidant power (FRAP) assay, total antioxidant activity and ferric thiocyanate assay. Total phenolics were also determined. Some fractions showed noteworthy antioxidant activity. The results of the antioxidant activity revealed that the ethyl acetate soluble fraction showed the highest value of percent inhibition of DPPH (82.54 ± 0.62) at the concentration of 125 μ g/ml. The IC(50) of this fraction was 37.11± 0.23 μg/ml, compared with butylated hydroxytoluene (BHT), which have IC(50) of 12.1 ± 0.92 μ/mL. It also showed the highest FRAP value (251.08 ± 1.46 μg of trolox equivalents) as well as the highest value of lipid peroxidation inhibition (57.21 ± 52%), the highest total antioxidant activity (0.549 ± 0.08) and also the highest total phenolic contents (77.1 ± 0.6) as compared to the studied fractions. Phytochemical screening showed high percentage of phenolics, flavonoides and cardiac glycosides in this fraction.

Monolithic silica spin column extraction and simultaneous derivatization of amphetamines and 3,4-methylenedioxyamphetamines in human urine for gas chromatographic-mass spectrometric detection.

PubMed

Nakamoto, Akihiro; Nishida, Manami; Saito, Takeshi; Kishiyama, Izumi; Miyazaki, Shota; Murakami, Katsunori; Nagao, Masataka; Namura, Akira

2010-02-19

A simple, sensitive, and specific method with gas chromatography-mass spectrometry was developed for simultaneous extraction and derivatization of amphetamines (APs) and 3,4-methylenedioxyamphetamines (MDAs) in human urine by using a monolithic silica spin column. All the procedures, such as sample loading, washing, and elution were performed by centrifugation. APs and MDAs in urine were adsorbed on the monolithic silica and derivatized with propyl chloroformate in the column. Methamphetamine-d(5) was used as an internal standard. The linear ranges were 0.01-5.0 microg mL(-1) for methamphetamine (MA) and 3,4-methylenedioxymethamphetamine (MDMA) and 0.02-5.0 microg mL(-1) for amphetamine (AP) and 3,4-methylenedioxyamphetamine (MDA) (coefficient of correlation > or = 0.995). The recovery of APs and MDAs in urine was 84-94%, and the relative standard deviation of the intra- and interday reproducibility for urine samples containing 0.1, 1.0, and 4.0 microg mL(-1) of APs and MDAs ranged from 1.4% to 13.6%. The lowest detection limit (signal-to-noise ratio > or = 3) in urine was 5 ng mL(-1) for MA and MDMA and 10 ng mL(-1) for AP and MDA. The proposed method can be used to perform simultaneous extraction and derivatization on spin columns that have been loaded with a small quantity of solvent by using centrifugation. Copyright 2009 Elsevier B.V. All rights reserved.

Linker-assisted immunoassay and liquid chromatography/mass spectrometry for the analysis of glyphosate

USGS Publications Warehouse

Lee, E.A.; Zimmerman, L.R.; Bhullar, B.S.; Thurman, E.M.

2002-01-01

A novel, sensitive, linker-assisted enzyme-linked immunosorbent assay (L'ELISA) was compared to on-line solidphase extraction (SPE) with high-performance liquid chromatography/mass spectrometry (HPLC/MS) for the analysis of glyphosate in surface water and groundwater samples. The L'ELISA used succinic anhydride to derivatize glyphosate, which mimics the epitotic attachment of glyphosate to horseradish peroxidase hapten. Thus, L'ELISA recognized the derivatized glyphosate more effectively (detection limit of 0.1 μg/L) and with increased sensitivity (10-100 times) over conventional ELISA and showed the potential for other applications. The precision and accuracy of L'ELISA then was compared with on-line SPE/HPLC/MS, which detected glyphosate and its degradate derivatized with 9-fluorenylmethyl chloroformate using negative-ion electrospray (detection limit 0.1 μg/L, relative standard deviation ±15%). Derivatization efficiency and matrix effects were minimized by adding an isotope-labeled glyphosate (2-13C15N). The accuracy of L'ELISA gave a false positive rate of 18% between 0.1 and 1.0 μg/L and a false positive rate of only 1% above 1.0 μg/L. The relative standard deviation was ±20%. The correlation of L'ELISA and HPLC/MS for 66 surface water and groundwater samples was 0.97 with a slope of 1.28, with many detections of glyphosate and its degradate in surface water but not in groundwater.

Toxic Effects of Couroupita guianensis Against Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae).

PubMed

Baskar, K; Ignacimuthu, S; Jayakumar, M

2015-02-01

Laboratory experiments were conducted to find out the efficacy of different crude extracts and fractions of Couroupita guianensis (Lecythidaceae) against Spodoptera litura (Fabricius). Results revealed that hexane, chloroform and ethyl acetate extracts of C. guianensis showed larvicidal and pupicidal activities against S. litura. Maximum larvicidal activity (68.66%) was observed in hexane extract at 5.0% concentration followed by chloroform and ethyl acetate extracts, and least LC50 value of 2.64% was observed in hexane extract. A 100% pupicidal activity was observed in hexane extract. Based on the efficacy of crude extracts, the effective crude extract (hexane extract) was further fractionated and subjected to screening for biological activities against S. litura. Among the eight fractions isolated from the hexane extract, fraction 8 showed maximum antifeedant activity (81. 8%) and larvicidal activity (76.9%) at 1000-ppm concentration; this fraction showed least LC50 value of 375.92 ppm for larvicidal activity. Cent per cent pupicidal activity was recorded. Reduced midgut and hemolymph protein contents were observed at 1000 ppm of fraction 8. Histopathological studies revealed that fraction 8 severely damaged the midgut cells of S. litura. This fraction could be used to develop botanical formulation to control agricultural pests.

Isolation, purification, toxicity and some physicochemical properties of mycotoxins produced by aspergillus quadrilineatus isolated from acha (Digitaria exilis stapf) in Plateau State of Nigeria.

PubMed

Gbodi, T A

1993-06-01

Some physico-chemical and toxicological studies were carried out on mycotoxins elaborated by Aspergillus quadrilineatus isolated from a grain foodstuff, acha (Digitaria exilis) in the Plateau State of Nigeria. The mycotoxins produced by A quadrilineatus were extractable from rice culture by chloroform. Column chromatographic separations of the crude extract in silica gel using different elution solvents and biological tests showed that the mycotoxins came off in the diethylether, chloroform and mostly in the ethyl acetate fractions. Use of different available mycotoxin standards on silica gel G coated chromatoplates revealed that 1 of the mycotoxins produced by A quadrilineatus was sterigmatocystin. Two other more toxic mycotoxins were isolated and purified from the crude chloroform extract; their column, preparative thin-layer chromatographic, infrared and UV-spectrophotometric characteristics were established. The infrared spectra of the 2 purified mycotoxins suggested that the carbonyl group of their structures were similar to that of aflatoxin.

Corrrelation of the Specific Rates of Solvolysis of Ethyl Fluoroformate Using the Extended Grunwald-Winstein Equation

PubMed Central

Seong, Mi Hye; Kyong, Jin Burm; Lee, Young Hoon; Kevill, Dennis N.

2009-01-01

The specific rates of solvolysis of ethyl fluoroformate have been measured at 24.2 °C in 21 pure and binary solvents. These give a satisfactory correlation over the full range of solvents when the extended Grunwald-Winstein equation is applied. The sensitivities to changes in the NT solvent nucleophilicity scale and the YCl solvent ionizing power scale, and the kF/kCl values are very similar to those for solvolyses of n-octyl fluoroformate, consistent with the addition step of an addition-elimination pathway being rate-determining. For methanolysis, a solvent deuterium isotope effect of 3.10 is compatible with the incorporation of general-base catalysis into the substitution process. For five representative solvents, studies were made at several temperatures and activation parameters determined. The results are also compared with those reported earlier for ethyl chloroformate and mechanistic conclusions are drawn. PMID:19399229

Antimicrobial and anti-inflammatory activities of Pleurostylia capensis Turcz (Loes) (celastraceae).

PubMed

Razwinani, Mapula; Tshikalange, Thilivhali Emmanuel; Motaung, Shirley C K M

2014-01-01

Pleurostylia capensis is a large tree that can reach the maximum height of 20 m long, and it have been traditionally used as cosmetic, for steam bath, ritual body wash, and as a purgative to treat symptoms of witchcraft. Using ethanol, chloroform, dichloromethane (DCM), ethyl acetate (EA), and water extracts, leaves, bark and roots of Pleurostylia capensis were investigated scientifically for their effectiveness in antimicrobial, antioxidant and anti-inflammatory activities using standard methods. The extracts were evaluated for antimicrobial activity against Gram positive (Staphylococcus aureus, Bacillus cereus, and Mycobacterium smegmatis), Gram negative (Escherichia coli, Klebsiella pneumonia, Klebsiella oxytoca, Streptococcus pyogenes, Pseudomonas aeruginosa and Salmonella typhimurium), and Candida albicans. The antioxidant activity was investigated using 2, 2-diphenlyl-1-picrylhadrazyl (DPPH), free radical scavenging assay. The anti-inflammatory activity of P. capensis extracts was evaluated against both cyclooxygenase enzymes (COX 1 and 2). The ethyl acetate extracts of P. capensis showed a strong antimicrobial activity against B. cereus, K. pneumonia, S. pyogenes, and M. smegmatis with MIC value of 0.39 and 0.78 mg/ml. While the ethanol bark extract was most active against M. smegmatis with MIC value of 0.78 mg/ml; the least potent activity was observed with dichloromethane, chloroform and water extracts, with an MIC value ranging from 1.56 mg/ml to 50.0 mg/ml. The plant extracts proved to be good antioxidant agent, whereas extracts of ethanol were the most active, with IC50 ranging from 1.00 to 1.74 µg/ml, which is lower, and in close range to Vitamin C (1.40 µg/ml). Its moderation to potent inhibitory activity was observed in all extracts. Ethanol and dichloromethane extracts were among the most potent when compared to water and petroleum ether extracts. The water extracts showed to be nontoxic on the Hek cell line with an IC50 value of 204.0, and 207.3 µg/ml (roots and bark) respectively. The dichloromethane, ethyl acetate, chloroform and ethanol extracts showed to be toxic on the Hek cell, with IC50 range from 5.94 to 42.91µg/ml. The results obtained indicate the effectiveness of these plants.

Optimisation of resolution in micellar electrokinetic chromatography by multivariate evaluation of electrolytes.

PubMed

Mikaeli, S; Thorsén, G; Karlberg, B

2001-01-12

A novel approach to multivariate evaluation of separation electrolytes for micellar electrokinetic chromatography is presented. An initial screening of the experimental parameters is performed using a Plackett-Burman design. Significant parameters are further evaluated using full factorial designs. The total resolution of the separation is calculated and used as response. The proposed scheme has been applied to the optimisation of the separation of phenols and the chiral separation of (+)-1-(9-anthryl)-2-propyl chloroformate-derivatized amino acids. A total of eight experimental parameters were evaluated and optimal conditions found in less than 48 experiments.

Microwave-assisted synthesis of N-pyrazole ureas and the p38alpha inhibitor BIRB 796 for study into accelerated cell ageing.

PubMed

Bagley, Mark C; Davis, Terence; Dix, Matthew C; Widdowson, Caroline S; Kipling, David

2006-11-21

Microwave irradiation of substituted hydrazines and beta-ketoesters gives 5-aminopyrazoles in excellent yield, which can be transformed to the corresponding N-carbonyl derivatives by treatment with an isocyanate or chloroformate. Derivatization of 4-nitronaphth-1-ol using predominantly microwave heating methods and reaction with an N-pyrazole carbamate provides a rapid route to the N-pyrazole urea BIRB 796 in high purity, as a potent and selective inhibitor of p38alpha mitogen-activated protein kinase for the study of accelerated ageing in Werner syndrome cells.

A practical derivatization LC/MS approach for determination of trace level alkyl sulfonates and dialkyl sulfates genotoxic impurities in drug substances.

PubMed

An, Jianguo; Sun, Mingjiang; Bai, Lin; Chen, Ted; Liu, David Q; Kord, Alireza

2008-11-04

Derivatization LC/MS methodology has been developed for the determination of a group of commonly encountered alkyl esters of sulfonates or sulfates in drug substances at low ppm levels. This general method uses trimethylamine as the derivatizing reagent for ethyl/propyl/isopropyl esters and triethylamine for methyl esters. The resulting quaternary ammonium derivatization products are highly polar (ionic) and can be retained by a hydrophilic interaction liquid chromatography (HILIC) column and readily separated from the main interfering active pharmaceutical ingredient (API) peak that is usually present at very high concentration. The method gives excellent sensitivity for all the alkyl esters at typical target analyte level of 1-2 ppm when the API samples were prepared at 5mg/mL. The recoveries at 1-2 ppm were generally above 85% for all the alkyl esters in the various APIs tested. The injection precisions of the lowest concentration standards were excellent with R.S.D.=0.4-4%. A linear range for concentrations from 0.2 to 20 ppm has been established with R(2)>or=0.99. This general method has been tested in a number of API matrices and used successfully for determination of alkyl sulfonates or dialkyl sulfates in support of API batch releases at GlaxoSmithKline.

Evaluation of larvicidal activity of medicinal plant extracts against three mosquito vectors.

PubMed

Bagavan, A; Rahuman, A Abdul

2011-01-01

To evaluate the mosquito larvicidal activity of plant extracts. The hexane, chloroform, ethyl acetate, acetone, and methanol leaf, flower and seed extracts of Abrus precatorius (A. precatorius), Croton bonplandianum (C. bonplandianum), Cynodon dactylon (C. dactylon), Musa paradisiaca (M. paradisiaca) and Syzygium aromaticum (S. aromaticum) were tested against fourth instar larvae of Anopheles vagus (An. vagus), Armigeres subalbatus (Ar. subalbatus) and Culex vishnui (Cx. vishnui). The highest larval mortality was found in seed ethyl acetate extracts of A. precatorius and leaf extracts of C. bonplandianum, flower chloroform and methanol extracts of M. paradisiaca, and flower bud hexane extract of S. aromaticum against An. vagus with LC(50) values of 19.31, 39.96, 35.18, 79.90 and 85.90 μg/mL; leaf ethyl acetate and methanol extracts of C. dactylon, flower methanol extract of M. paradisiaca, flower bud methanol extract of S. aromaticum against Ar. subalbatus with LC(50) values of 21.67, 32.62, 48.90 and 78.28 μg/mL, and seed methanol of A. precatorius, flower methanol extract of M. paradisiaca, flower bud hexane extract of S. aromaticum against Cx. vishnui with LC(50) values of 136.84, 103.36 and 149.56 μg/mL, respectively. These results suggest that the effective plant crude extracts have the potential to be used as an ideal ecofriendly approach for the control of disease vectors. This study provides the first report on the larvicidal activity of crude solvent extracts of different mosquitoes. Copyright © 2011 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Temperature-Triggered Switchable Helix-Helix Inversion of Poly(phenylacetylene) Bearing l-Valine Ethyl Ester Pendants and Its Chiral Recognition Ability.

PubMed

Zhou, Yanli; Zhang, Chunhong; Qiu, Yuan; Liu, Lijia; Yang, Taotao; Dong, Hongxing; Satoh, Toshifumi; Okamoto, Yoshio

2016-11-21

A phenylacetylene containing the l-valine ethyl ester pendant (PAA-Val) was synthesized and polymerized by an organorhodium catalyst (Rh(nbd)BPh₄) to produce the corresponding one-handed helical cis -poly(phenylacetylene) (PPAA-Val). PPAA-Val showed a unique temperature-triggered switchable helix-sense in chloroform, while it was not observed in highly polar solvents, such as N , N '-dimethylformamide (DMF). By heating the solution of PPAA-Val in chloroform, the sign of the CD absorption became reversed, but recovered after cooling the solution to room temperature. Even after six cycles of the heating-cooling treatment, the helix sense of the PPAA-Val's backbone was still switchable without loss of the CD intensity. The PPAA-Val was then coated on silica gel particles to produce novel chiral stationary phases (CSPs) for high-performance liquid chromatography (HPLC). These novel PPAA-Val based CSPs showed a high chiral recognition ability for racemic mandelonitrile ( α = 2.18) and racemic trans - N , N '-diphenylcyclohexane-1,2-dicarboxamide ( α = 2.60). Additionally, the one-handed helical cis -polyene backbone of PPAA-Val was irreversibly destroyed to afford PPAA-Val-H by heating in dimethyl sulfoxide (DMSO) accompanied by the complete disappearance of the Cotton effect. Although PPAA-Val-H had the same l-valine ethyl ester pendants as its cis -isomer PPAA-Val, it showed no chiral recognition. It was concluded that the one-handed helical cis -polyene backbone of PPAA-Val plays an important role in the chiral recognition ability.

Sensitive determination of thiols in wine samples by a stable isotope-coded derivatization reagent d0/d4-acridone-10-ethyl-N-maleimide coupled with high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry analysis.

PubMed

Lv, Zhengxian; You, Jinmao; Lu, Shuaimin; Sun, Weidi; Ji, Zhongyin; Sun, Zhiwei; Song, Cuihua; Chen, Guang; Li, Guoliang; Hu, Na; Zhou, Wu; Suo, Yourui

2017-03-31

As the key aroma compounds, varietal thiols are the crucial odorants responsible for the flavor of wines. Quantitative analysis of thiols can provide crucial information for the aroma profiles of different wine styles. In this study, a rapid and sensitive method for the simultaneous determination of six thiols in wine using d 0 /d 4 -acridone-10-ethyl-N-maleimide (d 0 /d 4 -AENM) as stable isotope-coded derivatization reagent (SICD) by high performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS) has been developed. Quantification of thiols was performed by using d 4 -AENM labeled thiols as the internal standards (IS), followed by stable isotope dilution HPLC-ESI-MS/MS analysis. The AENM derivatization combined with multiple reactions monitoring (MRM) not only allowed trace analysis of thiols due to the extremely high sensitivity, but also efficiently corrected the matrix effects during HPLC-MS/MS and the fluctuation in MS/MS signal intensity due to instrument. The obtained internal standard calibration curves for six thiols were linear over the range of 25-10,000pmol/L (R 2 ≥0.9961). Detection limits (LODs) for most of analytes were below 6.3pmol/L. The proposed method was successfully applied for the simultaneous determination of six kinds of thiols in wine samples with precisions ≤3.5% and recoveries ≥78.1%. In conclusion, the developed method is expected to be a promising tool for detection of trace thiols in wine and also in other complex matrix. Copyright © 2017 Elsevier B.V. All rights reserved.

Study of the Gastroprotective Effect of Extracts and Semipurified Fractions of Chresta martii DC. and Identification of Its Principal Compounds

PubMed Central

Franco, E. S.; Mélo, M. E. B.; Jatobá, B. J. A.; Santana, A. L. B. D.; Silva, A. A. R.; Silva, T. G.; Nascimento, M. S.; Maia, M. B. S.

2015-01-01

Chresta martii (Asteraceae) is a species widely used by the population of the Xingu region of Sergipe, Brazil, in the form of a decoction (aerial parts) for the treatment of gastrointestinal diseases. The study aims to assess the gastroprotective activity of organic extracts and semipurified fractions and identify the principal compounds present in C. martii responsible for such activity. The organic extracts (cyclohexane: ECCm, ethyl acetate: EACm, and ethanol: EECm) were obtained from the dried aerial parts (500 g) of C. martii. For evaluation of the gastroprotective activity of extracts (50, 100, or 200 mg/kg; p.o.), male Swiss Webster mice (25–30 g) were used which had gastric ulcers induced by indomethacin (40 mg/kg, s.c.) or ethanol (0.2 mL/animal; p.o.). Among the extracts evaluated, EACm exhibited significant (P < 0.05) gastroprotective activity in the models used. The fractionation of EACm was performed in a silica gel column 60 eluted with the following compounds: [chloroform—F1 yield (10%)], [chloroform/ethyl acetate (1/1)—F2 yield (6%)], [ethyl acetate—F3 yield (8%)], and [ethyl/methanol acetate (1/1)—F4 yield (5%)]. Of the fractions described above, the F1 (25 mg/kg; p.o.) had greater gastroprotective activity (P < 0.05) than that displayed by ranitidine (80 mg/kg; p.o.) in the ethanol-induced ulcer model. The refractionation of F1 produced 23 subfractions and from these two yellow amorphous compounds were obtained by recrystallization, Rf: 0.46 and 0.31 (ethyl acetate : chloroform 5 : 5). The compounds isolated were characterized by nuclear magnetic resonance spectroscopy (1H-NMR and 13C-NMR) and identified as flavones: chrysoeriol (yield: 0.43%) and 3′,4′-dimethoxyluteolin (yield: 0.58%). Conclusion. Flavone 3′,4′-dimethoxyluteolin is the principal compound present in the species C. martii and is probably responsible for gastroprotective activity observed in this species. PMID:25866537

In vitro activities of plant extracts from Saudi Arabia against malaria, leishmaniasis, sleeping sickness and Chagas disease.

PubMed

Abdel-Sattar, Essam; Maes, Louis; Salama, Maha Mahmoud

2010-09-01

The in vitro activity of the methanol extracts of 51 plants randomly collected from the Kingdom of Saudi Arabia and some of their fractions (petroleum ether, chloroform, ethyl acetate and aqueous) were evaluated against Plasmodium falciparum, Trypanosoma brucei brucei, T. cruzi and Leishmania infantum, as well as toxicity against MRC-5 fibroblast cells. Ten crude methanolic extracts that demonstrated potent and adequately selective antiprotozoal activity were subjected to solvent fractionation using petroleum ether, ethyl acetate and chloroform. Only three samples showed promising antiprotozoal activity. Argemone ochroleuca (CHCl(3) fraction) showed pronounced activity against P. falciparum(GHA) (IC(50) 0.32 microg/mL) and T. cruzi (IC(50) 0.30 microg/mL) with low cytotoxicity against MRC-5 cells (CC(50) 11.6 microg/mL). Capparis spinosa (EtOAc fraction) showed pronounced activity against P. falciparum(GHA) with an IC(50) 0.50 microg/mL in the absence of toxicity against MRC-5 cell line (CC(50) > 30 microg/mL). Heliotropium curassavicum (CHCl(3) fraction) showed similar activity against P. falciparum (IC(50) 0.65 microg/mL; MRC-5 CC(50) > 30 microg /mL). These three extracts will be subjected for further extensive studies to isolate and identify their active constituents. Copyright 2010 John Wiley & Sons, Ltd.

Comprehensive assessment of phenolics and antiradical potential of Rumex hastatus D. Don. roots

PubMed Central

2014-01-01

Background Roots of Rumex hastatus (Polygonaceae) are traditionally used for the treatment of various ailments including liver and lung diseases. In this study, various solvent extracts of R. hastatus roots, like methanolic, n-hexane, ethyl acetate, chloroform, butanol and aqueous fractions were assessed through their antioxidant properties in vitro and determination of phenolic contents. Methods Several parameters like DPPH˙, ABTS˙+, ˙OH, H2O2, superoxide free radical scavenging, iron chelating power, reducing power, β-carotene bleaching power, antioxidant capacity and total phenolics and flavonoids were evaluated. High Performance liquid Chromatography (HPLC) was also considered. Results Though all the fractions exhibited dose dependant activity. The samples with the highest activity were the butanol and methanol fractions in all the assays except hydrogen peroxide radical scavenging assay where chloroform fraction showed the highest scavenging aptitude. On the other hand, aquous fraction showed significant beta carotene linoleic acid, while n-hexane and ethyl acetate fractions exhibited a lesser antioxidant activity in all the assays. HPLC revealed the presence of rutin, luteolin-7-glucoside, vitexin and luteolin. Conclusion These results have to some extent substantiated the use of R. hastatus roots against different diseases, as an excellent basis of potential antioxidant due to the presence of sufficient amount of phenolics such as rutin and luteolin. PMID:24507200

Comprehensive assessment of phenolics and antiradical potential of Rumex hastatus D. Don. roots.

PubMed

Sahreen, Sumaira; Khan, Muhammad Rashid; Khan, Rahmat Ali

2014-02-08

Roots of Rumex hastatus (Polygonaceae) are traditionally used for the treatment of various ailments including liver and lung diseases. In this study, various solvent extracts of R. hastatus roots, like methanolic, n-hexane, ethyl acetate, chloroform, butanol and aqueous fractions were assessed through their antioxidant properties in vitro and determination of phenolic contents. Several parameters like DPPH˙, ABTS˙(+), ˙OH, H2O2, superoxide free radical scavenging, iron chelating power, reducing power, β-carotene bleaching power, antioxidant capacity and total phenolics and flavonoids were evaluated. High Performance liquid Chromatography (HPLC) was also considered. Though all the fractions exhibited dose dependant activity. The samples with the highest activity were the butanol and methanol fractions in all the assays except hydrogen peroxide radical scavenging assay where chloroform fraction showed the highest scavenging aptitude. On the other hand, aquous fraction showed significant beta carotene linoleic acid, while n-hexane and ethyl acetate fractions exhibited a lesser antioxidant activity in all the assays. HPLC revealed the presence of rutin, luteolin-7-glucoside, vitexin and luteolin. These results have to some extent substantiated the use of R. hastatus roots against different diseases, as an excellent basis of potential antioxidant due to the presence of sufficient amount of phenolics such as rutin and luteolin.

Phytochemical evaluation of Lythrum salicaria extracts and their effects on guinea-pig ileum.

PubMed

Bencsik, Tímea; Barthó, Loránd; Sándor, Viktor; Papp, Nóra; Benkó, Rita; Felinger, Attila; Kilár, Ferenc; Horváth, Györgyi

2013-09-01

n-Hexane, chloroform, ethyl acetate and 50% ethanol in water extracts prepared from the air-dried flowering parts of Lythrum salicaria L. were tested for in vitro pharmacological properties on Guinea-pig ileum, which is suitable for detecting a whole range of neuronal and smooth muscle effects. UHPLC-MS was used to evaluate polyphenol components of the extracts. In the ileum, the most prominent response (46.4% related to 0.5 microM histamine) of the extracts causing smooth muscle contractions were triggered by the 50% ethanol in water extract in a concentration-dependent manner. Atropine, indomethacin and PPADS plus suramin significantly reduced the contractile response caused by this extract. The strongest inhibition was due to atropine. The results suggest that L. salicaria extracts have a moderate muscarinic receptor agonist effect in Guinea-pig ileum and that prostanoids and purinoceptor mechanisms are involved to some extent. Therefore diluted extracts of L. salicaria p.o. could be used as a mild stimulant of gastrointestinal motility. The 50% ethanol in water extract was rich in polyphenols. n-Hexane, chloroform and ethyl acetate extracts failed to contain catechin, caffeic acid, quercetin-3-D-galactoside and rutin, but they all showed spasmogenic effects, and, therefore we do not think that these compounds could be involved in the spasmogenic activity.

Determination of chloropropanols in foods by one-step extraction and derivatization using pressurized liquid extraction and gas chromatography-mass spectrometry.

PubMed

Racamonde, I; González, P; Lorenzo, R A; Carro, A M

2011-09-28

3-Chloropropane-1,2-diol (3-MCPD) and 1,3-dichloro-2-propanol (1,3-DCP) were determined for the first time in bakery foods using pressurized liquid extraction (PLE) combined with in situ derivatization and GC-MS analysis. This one-step protocol uses N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) as silylation reagent. Initially, screening experimental design was applied to evaluate the effects of the variables potentially affecting the extraction process, namely extraction time (min) and temperature (°C), number of cycles, dispersant reagent (diatomaceous earth in powder form and as particulate matter with high pore volume Extrelut NT) and percent of flush ethyl acetate volume (%). To reduce the time of analysis and improve the sensitivity, derivatization of the compounds was performed in the cell extraction. Conditions, such as the volume of BSTFA, temperature and time for the in situ derivatization of analytes using PLE, were optimized by a screening design followed to a Doehlert response surface design. The effect of the in-cell dispersants/adsorbents with diatomaceous earth, Florisil and sodium sulfate anhydrous was investigated using a Box-Behnken design. Using the final best conditions, 1 g of sample dispersed with 0.1 g of sodium sulfate anhydrous and 2.5 g diatomaceous earth was extracted with ethyl acetate. 1 g of Florisil, as clean-up adsorbent, and 70 μL of BSTFA were used for 3 min at 70°C. Under the optimum conditions, the calibration curves showed good linearity (R(2)>0.9994) and precision (relative standard deviation, RSD≤2.4%) within the tested ranges. The limits of quantification for 1,3-DCP and 3-MCDP, 1.6 and 1.7 μg kg(-1), respectively, are far below the established limits in the European and American legislations. The accuracy, precision, linearity, and limits of quantification provided make this analytical method suitable for routine control. The method was applied to the analysis of several toasted bread, snacks, cookies and cereal samples, none of which contained chloropropanols at concentrations above the legislation levels. Copyright © 2011 Elsevier B.V. All rights reserved.

Convenient synthesis of 6-nor-9,10-dihydrolysergic acid methyl ester.

PubMed

Crider, A M; Grubb, R; Bachmann, K A; Rawat, A K

1981-12-01

6-Nor-9,10-dihydrolysergic acid methyl ester (IV) was prepared by demethylation of 9,10-dihydrolysergic acid methyl ester (II) with 2,2,2-trichloroethyl chloroformate, followed by reduction of the intermediate carbamate (III) with zinc in acetic acid. The 6-ethyl-V and 6-n-propyl-VI derivatives were prepared by alkylation of IV with the appropriate halide. All of the ergoline derivatives were evaluated for stereotyped behavior in rats, with 6-nor-6-ethyl-9,10-dihydrolysergic acid methyl ester (V) being active but much less potent than apomorphine. Compound VI was evaluated for its effect on blood pressure; at a dose of 30 mg/kg ip, it significantly lowered, diastolic pressure in normotensive rats.

Dual element ((15)N/(14)N, (13)C/(12)C) isotope analysis of glyphosate and AMPA by derivatization-gas chromatography isotope ratio mass spectrometry (GC/IRMS) combined with LC/IRMS.

PubMed

Mogusu, Emmanuel O; Wolbert, J Benjamin; Kujawinski, Dorothea M; Jochmann, Maik A; Elsner, Martin

2015-07-01

To assess sources and degradation of the herbicide glyphosate [N-(phosphonomethyl) glycine] and its metabolite AMPA (aminomethylphosphonic acid), concentration measurements are often inconclusive and even (13)C/(12)C analysis alone may give limited information. To advance isotope ratio analysis of an additional element, we present compound-specific (15)N/(14)N analysis of glyphosate and AMPA by a two step derivatization in combination with gas chromatography/isotope ratio mass spectrometry (GC/IRMS). The N-H group was derivatized with isopropyl chloroformate (iso-PCF), and remaining acidic groups were subsequently methylated with trimethylsilyldiazomethane (TMSD). Iso-PCF treatment at pH <10 gave too low (15)N/(14)N ratios indicating an incomplete derivatization; in contrast, too high (15)N/(14)N ratios at pH >10 indicated decomposition of the derivative. At pH 10, and with an excess of iso-PCF by 10-24, greatest yields and accurate (15)N/(14)N ratios were obtained (deviation from elemental analyzer-IRMS: -0.2 ± 0.9% for glyphosate; -0.4 ± 0.7% for AMPA). Limits for accurate δ(15)N analysis of glyphosate and AMPA were 150 and 250 ng injected, respectively. A combination of δ(15)N and δ(13)C analysis by liquid chromatography/isotope ratio mass spectrometry (LC/IRMS) (1) enabled an improved distinction of commercial glyphosate products and (2) showed that glyphosate isotope values during degradation by MnO2 clearly fell outside the commercial product range. This highlights the potential of combined carbon and nitrogen isotopes analysis to trace sources and degradation of glyphosate.

[Determination of five representative ultraviolet filters in water by gas chromatography-mass spectrometry].

PubMed

Ding, Yiran; Huang, Yun; Zhao, Tingting; Cai, Qian; Luo, Yu; Huang, Bin; Zhang, Yuxia; Pan, Xuejun

2014-06-01

A method for the determination of five representative organic UV filters: ethylhexyl methoxycinnamate (EHMC), benzophenone-3 (BP-3), 4-methylbenzylidene camphor (4-MBC), octocrylene (OC), homosalate (HMS) in water was investigated. The method was ased on derivatization, solid phase extraction (SPE), followed by determination with gas chromatography-mass spectrometry (GC-MS). The variables involved in the derivatization of BP-3 and HMS were optimized, and SPE conditions were studied. For derivatization, 100 microL N,O-bis(trimethylsilyl) trifluoroacetamide (BSTFA) was used as derivatization reagent and reacted with BP-3 and HMS at 100 degrees C for 100 min. For SPE, the pH value of water sample was adjusted to 3-5. The Oasis HLB cartridges were employed and the solution of ethyl acetate and dichloromethane (1 : 1, v/v) was used as the eluting solvent, and good recoveries of the target compounds were obtained. The limits of detection (LODs) and the limits of quantification (LOQs) for the five target compounds in water samples were 0.5-1.2 ng/L and 1.4-4.0 ng/L, respectively. The recoveries of spiked water samples were 87.85%-102.34% with good repeatability and reproducibility (RSD < 5%, n = 3) for all the target compounds. Finally, the validated method was applied to analysis the representative UV filters in water samples collected from a wastewater treatment plant in Kunming city of Yunnan province.

Characterization of the Chemical Constitution and Profile of Pharmacological Activity of PGB(x).

DTIC Science & Technology

1982-02-26

ischemia. Fed. Proc. 40 , 692 (1980). 3. Burkman, A. M. and Phornchirasilp, S., Prostaglandin Bx enhances the inotropic efficacy (Emax) of...nitroketone 10 was then converted to the sodium salt of the corresponding nitronic acid on treatment with an equimolar amount of sodium metabolite in...bromide in chloroform-ethyl acetate to give unsaturated aldehyde 12 in 40 ’ yield. The reaction of compound 9 with dimethyl (2-oxoheptvl) phosphonate in

[The determination of the herbicide glyphosate and its chief metabolite aminomethylphosphonic acid (AMPA) in drinking water with the aid of HPLC].

PubMed

Gauch, R; Leuenberger, U; Müller, U

1989-01-01

A method for the determination of glyphosate and its major metabolite aminomethylphosphonic acid (AMPA) is described. With a detection limit of 0.02 microgram/l, the method suitably fulfills the requirements of the Swiss legislation (tolerance value of 0.1 micrograms/l water). The compounds are derivatized directly in the original water sample with 9-fluorenylmethyl chloroformate (FMOCC1) in order to obtain extractable and fluorescent derivatives. These are extracted with organic solvents and determined by HPLC using a fluorescence detector. Neither of the compounds could be detected in 151 tap water samples from the Canton of Berne.

A Direct Aqueous Derivatization GSMS Method for Determining Benzoylecgonine Concentrations in Human Urine.

PubMed

Chericoni, Silvio; Stefanelli, Fabio; Da Valle, Ylenia; Giusiani, Mario

2015-09-01

A sensitive and reliable method for extraction and quantification of benzoylecgonine (BZE) and cocaine (COC) in urine is presented. Propyl-chloroformate was used as derivatizing agent, and it was directly added to the urine sample: the propyl derivative and COC were then recovered by liquid-liquid extraction procedure. Gas chromatography-mass spectrometry was used to detect the analytes in selected ion monitoring mode. The method proved to be precise for BZE and COC both in term of intraday and interday analysis, with a coefficient of variation (CV)<6%. Limits of detection (LOD) were 2.7 ng/mL for BZE and 1.4 ng/mL for COC. The calibration curve showed a linear relationship for BZE and COC (r2>0.999 and >0.997, respectively) within the range investigated. The method, applied to thirty authentic samples, showed to be very simple, fast, and reliable, so it can be easily applied in routine analysis for the quantification of BZE and COC in urine samples. © 2015 American Academy of Forensic Sciences.

Automated GC-MS analysis of free amino acids in biological fluids.

PubMed

Kaspar, Hannelore; Dettmer, Katja; Gronwald, Wolfram; Oefner, Peter J

2008-07-15

A gas chromatography-mass spectrometry (GC-MS) method was developed for the quantitative analysis of free amino acids as their propyl chloroformate derivatives in biological fluids. Derivatization with propyl chloroformate is carried out directly in the biological samples without prior protein precipitation or solid-phase extraction of the amino acids, thereby allowing automation of the entire procedure, including addition of reagents, extraction and injection into the GC-MS. The total analysis time was 30 min and 30 amino acids could be reliably quantified using 19 stable isotope-labeled amino acids as internal standards. Limits of detection (LOD) and lower limits of quantification (LLOQ) were in the range of 0.03-12 microM and 0.3-30 microM, respectively. The method was validated using a certified amino acid standard and reference plasma, and its applicability to different biological fluids was shown. Intra-day precision for the analysis of human urine, blood plasma, and cell culture medium was 2.0-8.8%, 0.9-8.3%, and 2.0-14.3%, respectively, while the inter-day precision for human urine was 1.5-14.1%.

Pharmacological effects of the phytochemicals of Anethum sowa L. root extracts.

PubMed

Saleh-E-In, Md Moshfekus; Sultana, Nasim; Hossain, Md Nur; Hasan, Sayeema; Islam, Md Rabiul

2016-11-14

Anethum sowa L. is widely used as an important spice and traditional medicinal plants to treat various ailments. On the basis of scientific ethnobotanical information, this study was undertaken to evaluate the antioxidant, antimicrobial and cytotoxic activity of the crude extracts of Anethum sowa L. roots as well as to identify the classes of phytochemicals by chemical tests. The antioxidant potential of the extracts was ascertained with the stable organic free radical (2, 2-diphenyl-1-picryl-hydrazyl). The agar well diffusion method was used to determine the susceptibility of bacterial and fungal strains of the crude extracts. The minimum inhibitory concentration (MIC) and minimum bactericidal concentrations (MBC) were determined by the microdilution test. Cytotoxic activities were screened using brine shrimps (Artemia salina) lethality assay. Finally, phytochemicals were profiled using standard procedures. A preliminary phytochemical screening of the different crude extracts by methanol, ethyl acetate and chloroform showed the presence of secondary metabolites such as flavonoids, alkaloids, saponin, cardiac glycosides and tannins while cyanogenetic glycosides were not detected. The methanol, ethyl acetate and chloroform extracts displayed high antioxidant activity (IC 50  = 13.08 ± 0.03, 33.48 ± 0.16 and 36.42 ± 0.41 μg/mL, respectively) in the DPPH assay comparable to that of the standard ascorbic acid and BHT (IC 50  = 3.74 ± 0.05 and 11.84 ± 0.29 μg/mL). The cytotoxic activity of the crude ethyl acetate and chloroform extracts possessed excellent activity (LC 50  = 5.03 ± 0.08, 5.23 ± 0.11 and 17.22 ± 0.14 μg/mL, respectively) against brine shrimp larvae after 24 h of treatment and compared with standard vincristine sulphate (LC 50  = 0.46 ± 0.05 μg/mL). The extracts also showed good antimicrobial activity against both Gram-positive and Gram-negative bacteria when compared with two standard antibiotics ciprofloxacin and tetracycline. These results showed that the Anethum sowa root extracts are the important source of the antioxidant, antimicrobial and cytotoxic agent. So, further research is necessary to isolate and characterize of different phytoconstituents for pharmaceutical drug lead molecules and also to verify its traditional uses.

Dengue antiviral activity of polar extract from Melochia umbellata (Houtt) Stapf var. Visenia

NASA Astrophysics Data System (ADS)

Hariani Soekamto, Nunuk; Liong, S.; Fauziah, S.; Wahid, I.; Firdaus; Taba, P.; Ahmad, F.

2018-03-01

A research on the dengue antiviral activity test on the polar bark extract of M. umbelatta (Houtt.) Stapf var. Vicenia have been done to determine the relation to its activity againts brine shrimp Artemia salina. The bark was extracted by maceration with n-hexane, chloroform, and ethylacetate. The activity of the ethyl acetate extract was then tested against A. salina and dengue virus. It was found that the ethyl acetate extract was active to A. salina with the LC50 value of 101.66 μg/mL and also very active to dengue virus with the IC50 value of 1.67μg/mL. It is clear that the toxicity to brine shrimp A. salina has a positive correlation with the dengue anti virus.

[Determination of three phenoxyalkanoic acid herbicides in blood using gas chromatography coupled with solid-phase extraction and derivatization].

PubMed

Xin, Guobin; Tan, Jiayi; Yao, Lijuan; Zhu, Yu; Jiang, Zhaolin; Song, Hui

2008-01-01

A method for the determination of three phenoxyalkanoic acid herbicides, 2,4-dichlorophenoxyacetic acid (2,4-D), 2-(2,4-dichlorophenoxy)-propanoic acid (2,4-DP), and 4-chloro-2-methylphenoxy-acetic acid (MCPA), in blood was developed. The blood sample was diluted with 0.1 mol/L hydrochloric acid, and extracted by solid-phase extraction using porous resin GDX401 as adsorbent and ethyl ether as eluent. The extract was esterified with dichloropropanol in the presence of sulfuric acid as catalyst. The derivatives were analysed by gas chromatography with electron-capture detection. The detection limits of 2,4-D, 2,4-DP and MCPA were 20, 8 and 40 ng/mL, respectively. In quantitative analysis, 2,4-dichlorophenylacetic acid was used as an internal standard. The linear relationships and recoveries were satisfactory. The derivatization of the three herbicides with methanol, ethanol, n-propanol, n-butanol, and trifluoroethanol were also studied, and the analytical methods of these derivatization were compared with that of dichloropropanol as esterifying agent. The method is sensitive enough for the examination of the poison samples in actual.

Hepatoprotective evaluation and isolation of the major secondary metabolites from the ethyl acetate extract of liquid culture filtrate of Chaetomium globosum.

PubMed

Awad, Nagwa E; Kassem, Hanaa A; Hamed, Manal A; El-Feky, Amal M; El-Naggar, Mohamed A A

2018-01-01

The aim of the present study was to evaluate the hepatoprotective activity of ethyl acetate extract of the liquid culture filtrate of Chaetomium globosum fungus (family Chaetomiaceae). Rats were intraperitoneally injected by CCl4 (0.5ml/kg) twice a week for six consecutive weeks. Treatment tacks (250mg/kg) place at the same time of CCl4 induction and with the same duration. The evaluation was done through determination of liver function indices; aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and total serum protein content. In addition, the oxidative stress markers; hepatic glutathione content (GSH), hepatic malondialdehyde (MDA), hepatic superoxide dismutase (SOD), and hepatic total protein were estimated. Moreover, the liver architectures were also examined. Isolation and identification of the main secondary metabolites were identified. Seven volatile compounds were identified from the plain chloroform fraction where, 1-Cyclopentyl-2,2-dimethyl-1-propanol (54.63%) was presented as the major compound. Eleven compounds were also identified from the fraction eluted by chloroform: methanol (85:15). 1,5,5-Trimethyl-6-methylene-1-cyclohexene (25.79%) and Norbornan-2-one (26.84%) are presented as the major compounds of this fraction. In conclusion, the extract recorded hepatoprotective effect by ameliorating the biochemical parameters under investigation. The liver histopathological pictures confirmed our results. Copyright © 2017. Published by Elsevier Masson SAS.

Protective effect of Clerodendrum colebrookianum Walp., on acute and chronic inflammation in rats

PubMed Central

Deb, Lokesh; Dey, Amitabha; Sakthivel, G.; Bhattamishra, Subrat Kumar; Dutta, Amitsankar

2013-01-01

Aim: To evaluate antioxidant, anti-inflammatory potential of the aqueous extracts and its aqueous, n-butanol, ethyl-acetate, and chloroform fractions of Clerodendrum colebrookianum Walp. leaves. Materials and Methods: In this present study, all the test samples were evaluated on in-vivo inflammatory model such as carrageenan and histamine-induced acute-inflammation and cotton pellet induced granuloma formation in albino male rats. Test samples were also employed in in-vitro assays like DPPH* free radical scavenging activity and COX inhibition assay. Results: The test samples at the dose of 200mg/kg/p.o. were found to cause significant inhibition of carrageenan and histamine-induced inflammation and cotton pallet-induced granuloma formation on acute and chronic inflammation in rats. The test samples, except n-butanol fraction, exhibited inhibitory effect for both COX-1 and COX-2, in in-vitro assay but their percentage of inhibition values differs from each other. The test samples (aqueous extracts, aqueous, n-butanol, ethyl-acetate, and chloroform fractions) at 100 μg concentration exhibits 54.37%, 33.88%, 62.85%, 56.28%, and 57.48% DPPH* radical-scavenging effect respectively in in-vitro antioxidant study. Conclusion: These observations established the anti-inflammatory effect of C. colebrookianum leaves in acute and chronic stages of inflammation by free radical scavenging and inhibition of COX-1 and COX-2. PMID:24014914

VIT-CMJ2: Endophyte of Agaricus bisporus in Production of Bioactive Compounds.

PubMed

Gautam, Chandan Kumar; Madhav, Mukund; Sinha, Astha; Jabez Osborne, William

2016-06-01

Agaricus bisporus is an edible basidiomycete fungus. Both the body and the mycelium contain compounds comprising a wide range of antimicrobial molecules, contributing in improvement of immunity and tumor-retardation. The presence of endophytes capable of producing bioactive compounds was investigated in Agaricus bisporus . Endophytes from Agaricus bisporus was isolated on LB agar. The obtained isolates were characterized morphologically and biochemically. Further 16S rRNA sequencing was implemented for molecular analysis of isolates. The isolate was mass produced and the bioactive compounds were extracted using ethyl acetate, chloroform and hexane. Agar well diffusion method was carried out to seek the potential of any antimicrobial activity of the crude bioactive compounds against known pathogens. GC-MS and FT-IR analysis were performed for the identification of bioactive compounds. VIT-CMJ2 was identified as Enterobacter sp. as revealed by 16S rRNA sequencing. Chloroform extract of VIT-CMJ2 showed a maximum zone of inhibition of 19 mm against Salmonella typhi followed by hexane and ethyl acetate extracts. The GC-MS analysis revealed the presence of several bioactive compounds having effective antimicrobial activity like butyl ester, Behenicalcohol, S , S-dioxide derivatives and some others which were later confirmed by FT-IR spectral stretches. The present study shows the insight on the way endophytes interact with Agaricus bisporus ; thereby improving the nutritional profile.

VIT-CMJ2: Endophyte of Agaricus bisporus in Production of Bioactive Compounds

PubMed Central

Gautam, Chandan Kumar; Madhav, Mukund; Sinha, Astha; Jabez Osborne, William

2016-01-01

Background Agaricus bisporus is an edible basidiomycete fungus. Both the body and the mycelium contain compounds comprising a wide range of antimicrobial molecules, contributing in improvement of immunity and tumor-retardation. Objectives The presence of endophytes capable of producing bioactive compounds was investigated in Agaricus bisporus. Materials and Methods Endophytes from Agaricus bisporus was isolated on LB agar. The obtained isolates were characterized morphologically and biochemically. Further 16S rRNA sequencing was implemented for molecular analysis of isolates. The isolate was mass produced and the bioactive compounds were extracted using ethyl acetate, chloroform and hexane. Agar well diffusion method was carried out to seek the potential of any antimicrobial activity of the crude bioactive compounds against known pathogens. GC-MS and FT-IR analysis were performed for the identification of bioactive compounds. Results VIT-CMJ2 was identified as Enterobacter sp. as revealed by 16S rRNA sequencing. Chloroform extract of VIT-CMJ2 showed a maximum zone of inhibition of 19 mm against Salmonella typhi followed by hexane and ethyl acetate extracts. The GC-MS analysis revealed the presence of several bioactive compounds having effective antimicrobial activity like butyl ester, Behenicalcohol, S , S-dioxide derivatives and some others which were later confirmed by FT-IR spectral stretches. Conclusions The present study shows the insight on the way endophytes interact with Agaricus bisporus; thereby improving the nutritional profile. PMID:28959322

Simple high-performance liquid chromatography method for formaldehyde determination in human tissue through derivatization with 2,4-dinitrophenylhydrazine.

PubMed

Yilmaz, Bilal; Asci, Ali; Kucukoglu, Kaan; Albayrak, Mevlut

2016-08-01

A simple high-performance liquid chromatography method has been developed for the determination of formaldehyde in human tissue. FA Formaldehyde was derivatized with 2,4-dinitrophenylhydrazine. It was extracted from human tissue with ethyl acetate by liquid-liquid extraction and analyzed by high-performance liquid chromatography. The calibration curve was linear in the concentration range of 5.0-200 μg/mL. Intra- and interday precision values for formaldehyde in tissue were <6.9%, and accuracy (relative error) was better than 6.5%. The extraction recoveries of formaldehyde from human tissue were between 88 and 98%. The limits of detection and quantification of formaldehyde were 1.5 and 5.0 μg/mL, respectively. Also, this assay was applied to liver samples taken from a biopsy material. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Selective determination of aloin in different matrices by HPTLC densitometry in fluorescence mode.

PubMed

Coran, Silvia A; Bartolucci, Gianluca; Bambagiotti-Alberti, Massimo

2011-01-25

A novel method based on the fluorescence excited solely on aloin by a H₃BO₃ derivatizing procedure, allowed its rapid and selective determination among the co-occurring components in a variety of complex matrices as several Aloes dried extracts and related commercial products. HPTLC LiChrospher silica gel 60 F254S, 20 cm x 10 cm, plates with ethyl formate: CH₃OH:H₂O (100:14.5:10, v/v) as the mobile phase were used. Densitometric determinations were performed in fluorescence mode, exciting wavelength 365 nm, optical filter K540 after derivatization with H₃BO₃. The method was validated giving rise to a dependable and high throughput procedure well suited to routine application. Aloin was quantified in the range of 110-330 ng with RSD of repeatability and intermediate precision not exceeding 2.3% and accuracy inside the acceptance limits. Copyright © 2010 Elsevier B.V. All rights reserved.

Antiulcerogenic activity of crude extract, fractions and populnoic acid isolated from Austroplenckia populnea (Celastraceae).

PubMed

Andrade, Sérgio F; Antoniolli, Daiane; Comunello, Eros; Cardoso, Luis G V; Carvalho, José C T; Bastos, Jairo K

2006-01-01

Many plant crude extracts and their isolated compounds are the most attractive sources of new drugs and show promising results for the treatment of gastric ulcers. Austroplenckia populnea is commonly known as "marmelinho-do campo, mangabeira-brava, mangabarana and vime" and it has been used in folk medicine as anti-dysenteric and anti-rheumatic. Powdered bark wood (3.25 kg) was macerated with aqueous ethanol (96%) and the extract was concentrated under reduced pressure to yield 406 g of crude hydralcoholic extract. The hydralcoholic extract was suspended in aqueous methanol and partitioned with hexane, chloroform and ethyl acetate (EtOAc) in sequence, yielding 8.0 g, 9.5 g and 98.17 g of crude extracts, respectively. Chromatography of the hexane extract over a silica gel column led to the isolation of the triterpene populnoic acid. The oral administration of hydralcoholic, hexane, chloroform and EtOAc extracts (200 mg/kg) decreased the ulcer lesion index (ULI) by 83.15%, 46.87%, 32.2%, 68.12%, respectively. Oral administration of populnoic acid (100 mg/kg) diminished the ULI by 55.29%. All the obtained results were significant in comparison with the negative control, with exception of the chloroform extract.

Determination of hydrazine in drinking water: Development and multivariate optimization of a rapid and simple solid phase microextraction-gas chromatography-triple quadrupole mass spectrometry protocol.

PubMed

Gionfriddo, Emanuela; Naccarato, Attilio; Sindona, Giovanni; Tagarelli, Antonio

2014-07-04

In this work, the capabilities of solid phase microextraction were exploited in a fully optimized SPME-GC-QqQ-MS analytical approach for hydrazine assay. A rapid and easy method was obtained by a simple derivatization reaction with propyl chloroformate and pyridine carried out directly in water samples, followed by automated SPME analysis in the same vial without further sample handling. The affinity of the different derivatized compounds obtained towards five commercially available SPME coatings was evaluated, in order to achieve the best extraction efficiency. GC analyses were carried out using a GC-QqQ-MS instrument in selected reaction monitoring (SRM) acquisition mode which has allowed the achievement of high specificity by selecting appropriate precursor-product ion couples improving the capability in analyte identification. The multivariate approach of experimental design was crucial in order to optimize derivatization reaction, SPME process and tandem mass spectrometry parameters. Accuracy of the proposed protocol, tested at 60, 200 and 800 ng L(-1), provided satisfactory values (114.2%, 83.6% and 98.6%, respectively), whereas precision (RSD%) at the same concentration levels were of 10.9%, 7.9% and 7.7% respectively. Limit of detection and quantification of 4.4 and 8.3 ng L(-1) were obtained. The reliable application of the proposed protocol to real drinking water samples confirmed its capability to be used as analytical tool for routine analyses. Copyright © 2014 Elsevier B.V. All rights reserved.

Vigabatrin in dried plasma spots: validation of a novel LC-MS/MS method and application to clinical practice.

PubMed

Kostić, Nađa; Dotsikas, Yannis; Jović, Nebojša; Stevanović, Galina; Malenović, Anđelija; Medenica, Mirjana

2014-07-01

This paper presents a LC-MS/MS method for the determination of antiepileptic drug vigabatrin in dried plasma spots (DPS). Due to its zwitterionic chemical structure, a pre-column derivatization procedure was performed, aiming to yield enhanced ionization efficiency and improved chromatographic behaviour. Propyl chloroformate, in the presence of propanol, was selected as the best derivatization reagent, providing a strong signal along with reasonable run time. A relatively novel sample collection technique, DPS, was utilized, offering easy sample handling and analysis, using a sample in micro amount (∼5μL). Derivatized vigabatrin and its internal standard, 4-aminocyclohexanecarboxylic acid, were extracted by liquid-liquid extraction (LLE) and determined in positive ion mode by applying two SRM transitions per analyte. A Zorbax Eclipse XDB-C8 column (150×4.6mm, 5μm particle size) maintained at 30°C, was utilized with running mobile phase composed of acetonitrile: 0.15% formic acid (85:15, v/v). Flow rate was 550μL/min and total run time 4.5min. The assay exhibited excellent linearity over the concentration range of 0.500-50.0μg/mL, which is suitable for the determination of vigabatrin level after per os administration in children and youths with epilepsy, who were on vigabatrin therapy, with or without co-medication. Specificity, accuracy, precision, recovery, matrix-effect and stability were also estimated and assessed within acceptance criteria. Copyright © 2014 Elsevier B.V. All rights reserved.

Phytochemical constituents, nutritional values, phenolics, flavonols, flavonoids, antioxidant and cytotoxicity studies on Phaleria macrocarpa (Scheff.) Boerl fruits

PubMed Central

2014-01-01

Background The edible fruits of Phaleria macrocarpa (Scheff.) Boerl are widely used in traditional medicine in Indonesia. It is used to treat a variety of medical conditions such as - cancer, diabetes mellitus, allergies, liver and heart diseases, kidney failure, blood diseases, high blood pressure, stroke, various skin diseases, itching, aches, and flu. Therefore, it is of great interest to determine the biochemical and cytotoxic properties of the fruit extracts. Methods The methanol, hexane, chloroform, ethyl acetate, and water extracts of P. macrocarpa fruits were examined for phytochemicals, physicochemicals, flavonols, flavonoids and phenol content. Its nutritional value (A.O.A.C method), antioxidant properties (DPPH assay) and cytotoxicity (MTT cell proliferation assay) were also determined. Results A preliminary phyotochemical screening of the different crude extracts from the fruits of P. macrocarpa showed the presence secondary metabolites such as of flavonoids, phenols, saponin glycosides and tannins. The ethyl acetate and methanol extracts displayed high antioxidant acitivity (IC50 value of 8.15±0.02 ug/mL) in the DPPH assay comparable to that of the standard gallic acid (IC50 value of 10.8±0.02 ug/mL). Evaluation of cytotoxic activity showed that the crude methanol extract possessed excellent anti-proliferative activity against SKOV-3 (IC50 7.75±2.56 μg/mL) after 72 hours of treatment whilst the hexane and ethyl acetate extracts displayed good cytotoxic effect against both SKOV-3 and MDA-MB231 cell lines. The chloroform extract however, showed selective inhibitory activity in the breast cancer cell line MDA-MB231 (IC50 7.80±1.57 μg/mL) after 48 hours of treatment. There was no cytotoxic effect observed in the Ca Ski cell line and the two normal cell lines (MRC-5 and WRL-68). Conclusion The methanol extract and the ethyl acetate fraction of P. macrocarpa fruits exhibited good nutritional values, good antioxidant and cytotoxic activities, and merits further investigation to identify the specific compound(s) responsible for these activities. PMID:24885709

Optimization in the formaldehyde determination at sub-ppm level from acetals by HPLC-DAD

DOE Office of Scientific and Technical Information (OSTI.GOV)

Medvedovici, A.; David, V.; David, F.

1999-02-01

Carbonylic compounds are mainly monitored as atmospheric pollutants, due to their major contribution to the formation of free radicals and ozone, by means of photolysis. Determination of formaldehyde at sub-ppm level as impurity in acetals using HPLC-DAD is described. Automated on-line precolumn derivatization reaction with 2,4-dinitrophenylhydrazine has been used. Breakdown rates of some industrial scale used acetals (Methylal, Ethylal) to formaldehyde by hydrolysis in aqueous media, according to pH, are described.

Gd@C82 metallofullerenes for neutron capture therapy—fullerene solubilization by poly(ethylene glycol)-block-poly(2-(N, N-diethylamino)ethyl methacrylate) and resultant efficacy in vitro

PubMed Central

Horiguchi, Yukichi; Kudo, Shinpei; Nagasaki, Yukio

2011-01-01

Poly(ethylene glycol)-block-poly(2-(N,N-diethylamino)ethyl methacrylate) (PEG-b-PAMA) was found to solubilize fullerenes such as C60, and this technique was applied to metallofullerenes. Gd@C82 was easily dissolved in water in the presence of PEG-b-PAMA without any covalent derivatization, forming a transparent complex about 20–30 nm in diameter. Low cytotoxicity was confirmed in vitro. Neutron irradiation of cultured cells (colon-26 adenocarcinoma) with Gd@C82-PEG-b-PAMA-complexed nanoparticles showed effective cytotoxicity, indicating the effective emission of gamma rays and internal conversion electrons produced from the neutron capture reaction of Gd. This result suggests a potentially valuable approach to gadolinium-based neutron capture therapy. PMID:27877415

Synthesis of acid addition salt of delta-aminolevulinic acid from 5-bromo levulinic acid esters

DOEpatents

Moens, Luc

2003-06-24

A process of preparing an acid addition salt of delta-aminolevulinc acid comprising: a) dissolving a lower alkyl 5-bromolevulinate and hexamethylenetetramine in a solvent selected from the group consisting of water, ethyl acetate, chloroform, acetone, ethanol, tetrahydrofuran and acetonitrile, to form a quaternary ammonium salt of the lower alkyl 5-bromolevulinate; and b) hydrolyzing the quaternary ammonium salt with an inorganic acid to form an acid addition salt of delta-aminolevulinic acid.

Solubility and dissolution thermodynamics of tetranitroglycoluril in organic solvents at 295-318 K

NASA Astrophysics Data System (ADS)

Zheng, Zhihua; Wang, Jianlong; Hu, Zhiyan; Du, Hongbin

2017-08-01

The solubility data of tetranitroglycoluril in acetone, methanol, ethanol, ethyl acetate, nitromethane and chloroform at temperatures ranging from 295-318 K were measured by gravimetric method. The solubility data of tetranitroglycoluril were fitted with Apelblat semiempirical equation. The dissolution enthalpy, entropy and Gibbs energy of tetranitroglycoluril were calculated using the Van't Hoff and Gibbs equations. The results showed that the Apelblat semiempirical equation was significantly correlated with solubility data. The dissolving process was endothermic, entropy-driven, and nonspontaneous.

40 CFR 439.16 - Pretreatment standards for existing sources (PSES).

Code of Federal Regulations, 2014 CFR

2014-07-01

... Xylenes 3.0 0.7 n-Heptane 3.0 0.7 n-Hexane 3.0 0.7 Methylene chloride 3.0 0.7 Chloroform 0.1 0.03 1,2... 1 Ammonia (as N) 2 84.1 29.4 Acetone 20.7 8.2 4-methyl-2-pentanone 20.7 8.2 Isobutyraldehyde 20.7 8.2 n-Amyl acetate 20.7 8.2 n-Butyl acetate 20.7 8.2 Ethyl acetate 20.7 8.2 Isopropyl acetate 20.7 8.2...

40 CFR 439.16 - Pretreatment standards for existing sources (PSES).

Code of Federal Regulations, 2013 CFR

2013-07-01

... Xylenes 3.0 0.7 n-Heptane 3.0 0.7 n-Hexane 3.0 0.7 Methylene chloride 3.0 0.7 Chloroform 0.1 0.03 1,2... 1 Ammonia (as N) 2 84.1 29.4 Acetone 20.7 8.2 4-methyl-2-pentanone 20.7 8.2 Isobutyraldehyde 20.7 8.2 n-Amyl acetate 20.7 8.2 n-Butyl acetate 20.7 8.2 Ethyl acetate 20.7 8.2 Isopropyl acetate 20.7 8.2...

40 CFR 439.16 - Pretreatment standards for existing sources (PSES).

Code of Federal Regulations, 2012 CFR

2012-07-01

... Xylenes 3.0 0.7 n-Heptane 3.0 0.7 n-Hexane 3.0 0.7 Methylene chloride 3.0 0.7 Chloroform 0.1 0.03 1,2... 1 Ammonia (as N) 2 84.1 29.4 Acetone 20.7 8.2 4-methyl-2-pentanone 20.7 8.2 Isobutyraldehyde 20.7 8.2 n-Amyl acetate 20.7 8.2 n-Butyl acetate 20.7 8.2 Ethyl acetate 20.7 8.2 Isopropyl acetate 20.7 8.2...

Chromatographic methods for determination of S-substituted cysteine derivatives--a comparative study.

PubMed

Kubec, Roman; Dadáková, Eva

2009-10-09

A novel HPLC method for determination of a wide variety of S-substituted cysteine derivatives in Allium species has been developed and validated. This method allows simultaneous separation and quantification of S-alk(en)ylcysteine S-oxides, gamma-glutamyl-S-alk(en)ylcysteines and gamma-glutamyl-S-alk(en)ylcysteine S-oxides in a single run. The procedure is based on extraction of these amino acids and dipeptides by methanol, their derivatization by dansyl chloride and subsequent separation by reversed phase HPLC. The main advantages of the new method are simplicity, excellent stability of derivatives, high sensitivity, specificity and the ability to simultaneously analyze the whole range of S-substituted cysteine derivatives. This method was critically compared with other chromatographic procedures used for quantification of S-substituted cysteine derivatives, namely with two other HPLC methods (derivatization by o-phthaldialdehyde/tert-butylthiol and fluorenylmethyl chloroformate), and with determination by gas chromatography or capillary electrophoresis. Major advantages and drawbacks of these analytical procedures are discussed. Employing these various chromatographic methods, the content and relative proportions of individual S-substituted cysteine derivatives were determined in four most frequently consumed alliaceous vegetables (garlic, onion, shallot, and leek).

Speciation of organotin compounds in urine by GC-MIP-AED and GC-MS after ethylation and liquid-liquid extraction.

PubMed

Zachariadis, G A; Rosenberg, E

2009-04-15

A method for the determination of organotin compounds in urine samples based on liquid-liquid extraction (LLE) in hexane and gas chromatographic separation was developed and optimized. Seven organotin species, namely monobutyltin (MBT), dibutyltin (DBT), tributyltin (TBT), tetrabutyltin (TeBT), monophenyltin (MPhT), diphenyltin (DPhT) and triphenyltin (TPhT), were in situ derivatized by sodium tetraethylborate (NaBEt(4)) to form ethylated less polar derivatives directly in the urine matrix. The critical parameters which have a significant effect on the yield of the successive liquid-liquid extraction procedure were examined, by using standard solutions of tetrabutyltin in hexane. The method was optimized for use in direct analysis of undiluted human urine samples and ways to overcome practical problems such as foam formation during extraction, due to various constituents of urine are discussed. After thorough optimization of the extraction procedure, all examined species could be determined after 3 min of simultaneous derivatization and extraction at room temperature and 5 min phase separation by centrifugation. Gas chromatography with a microwave-induced plasma atomic emission detector (MIP-AED) as element specific detector was employed for quantitative measurements, while a quadrupole mass spectrometric detector (MS) was used as molecular specific detector. The detection limits were between 0.42 and 0.67 microg L(-1) (as Sn) for the quantitative LLE-GC-MIP-AED method and the precision between 4.2% and 11.7%, respectively.

Antibacterial and antifungal activity of Flindersine isolated from the traditional medicinal plant, Toddalia asiatica (L.) Lam.

PubMed

Duraipandiyan, V; Ignacimuthu, S

2009-06-25

The leaves and root of Toddalia asiatica (L.) Lam. (Rutaceae) are widely used as a folk medicine in India. Hexane, chloroform, ethyl acetate, methanol and water extracts of Toddalia asiatica leaves and isolated compound Flindersine were tested against bacteria and fungi. Antibacterial and antifungal activities were tested against bacteria and fungi using disc-diffusion method and minimum inhibitory concentrations (MICs). The compound was confirmed using X-ray crystallography technique. Antibacterial and antifungal activities were observed in ethyl acetate extract. One active principle Flindersine (2,6-dihydro-2,2-dimethyl-5H-pyrano [3,2-c] quinoline-5-one-9cl) was isolated from the ethyl acetate extract. The MIC values of the compound against bacteria Bacillus subtilis (31.25 microg/ml), Staphylococcus aureus (62.5 microg/ml), Staphylococcus epidermidis (62.5 microg/ml), Enterococcus faecalis (31.25 microg/ml), Pseudomonas aeruginosa (250 microg/ml), Acinetobacter baumannii (125 microg/ml) and fungi Trichophyton rubrum 57 (62.5 microg/ml), Trichophyton mentagrophytes (62.5 microg/ml), Trichophyton simii (62.5 microg/ml), Epidermophyton floccosum (62.5 microg/ml), Magnaporthe grisea (250 microg/ml) and Candida albicans (250 microg/ml) were determined. Ethyl acetate extract showed promising antibacterial and antifungal activity and isolated compound Flindersine showed moderate activity against bacteria and fungi.

Rapid and cost-effective determination of acrylamide in coffee by planar chromatography and fluorescence detection after derivatization with dansulfinic acid.

PubMed

Alpmann, Alexander; Morlock, Gertrud

2009-01-01

A new method has been developed for the determination of acrylamide in ground coffee by planar chromatography using prechromatographic in situ derivatization with dansulfinic acid. After pressurized fluid extraction of acrylamide from the coffee samples, the extracts were passed through activated carbon and concentrated. These extracts were applied onto a silica gel 60 HPTLC plate and oversprayed with dansulfinic acid. By heating the plate, acrylamide was derivatized into the fluorescent product dansylpropanamide. The chromatographic separation with ethyl acetate-tert.-butyl methyl ether (8 + 2, v/v) mobile phase was followed by densitometric quantification at 254/>400 nm using a 4 point calibration via the standard addition method over the whole system for which acrylamide was added at different concentrations at the beginning of the extraction process. The method was validated for commercial coffee. The linearity over the whole procedure showed determination coefficients between 0.9995 and 0.9825 (n = 6). Limit of quantitation at a signal-to-noise ratio of 10 was determined to be 48 microg/kg. The within-run precision (relative standard deviation, n = 6) of the chromatographic method was 3%. Commercial coffee samples analyzed showed acrylamide contents between 52 and 191 microg/kg, which was in correlation with amounts reported in previous publications.

Preparation of corncob grits as a carrier for immobilizing yeast cells for ethanol production.

PubMed

Lee, Sang-Eun; Lee, Choon Geun; Kang, Do Hyung; Lee, Hyeon-Yong; Jung, Kyung-Hwan

2012-12-01

In this study, DEAE-corncobs [delignified corncob grits derivatized with 2-(diethylamino)ethyl chloride hydrochloride (DEAE·HCl)] were prepared as a carrier to immobilize yeast (Saccharomyces cerevisiae) for ethanol production. The immobilized yeast cell reactor produced ethanol under optimized DEAE·HCl derivatization and adsorption conditions between yeast cells and the DEAE-corncobs. When delignified corncob grit (3.0 g) was derivatized with 0.5M DEAE·HCl, the yeast cell suspension (OD600 = 3.0) was adsorbed at >90% of the initial cell OD600. This amount of adsorbed yeast cells was estimated to be 5.36 mg-dry cells/g-DEAE corncobs. The Qmax (the maximum cell adsorption by the carrier) of the DEAE-corncobs was estimated to be 25.1 (mg/g), based on a Languir model biosorption isotherm experiment. When we conducted a batch culture with medium recycling using the immobilized yeast cells, the yeast cells on DEAE-corncobs produced ethanol gradually, according to glucose consumption, without cells detaching from the DEAE-corncobs. We observed under electron microscopy that the yeast cells grew on the surface and in the holes of the DEAEcorncobs. In a future study, DEAE-corncobs and the immobilized yeast cell reactor system will contribute to bioethanol production from biomass hydrolysates.

A study of the relationship between the chemical structures and the fluorescence quantum yields of coumarins, quinoxalinones and benzoxazinones for the development of sensitive fluorescent derivatization reagents.

PubMed

Azuma, Kentaro; Suzuki, Sachiko; Uchiyama, Seiichi; Kajiro, Toshi; Santa, Tomofumi; Imai, Kazuhiro

2003-04-01

To develop new fluorescent derivatization reagents, we investigated the relationship between the chemical structures and the fluorescence quantum yields (phi(f)) of coumarins, quinoxalinones and benzoxadinones. Forty-six compounds were synthesized and their fluorescence spectra were measured in n-hexane, ethyl acetate, methanol and water. The energy levels of these compounds were calculated by combination of the semi-empirical AM1 and INDO/S (CI = all) methods. The deltaE(Tn(n,pi*), S1(pi,pi*)) (the energy gap between the Tn(n,pi*) and S1(pi,pi*) states) values were well correlated with the phi(f) values, which enables us to predict the phi(f) values from their chemical structures. Based on this relationship, 3-phenyl-7-N-piperazinoquinoxalin-2(1H)-one (PQ-Pz) and 7-(3-(S)-aminopyrrolidin-1-yl)-3-phenylquinoxalin-2-(1H)-one (PQ-APy) were developed as fluorescent derivatization reagents for carboxylic acids. The derivatives of the carboxylic acids with PQ-Pz and PQ-APy showed large phi(f) values even in polar solvents, suggesting that these reagents are suitable for the microanalysis of biologically important carboxylic acids by reversed phase HPLC.

Determination of fatty acids in bio-samples based on the pre-column fluorescence derivatization with 1,3,5,7-tetramethyl-8-butyrethylenediamine-difluoroboradiaza-s-indacene by high performance liquid chromatography.

PubMed

Wang, Fei-Hua; Xiong, Xu-Jie; Guo, Xiao-Feng; Wang, Hong; Zhang, Hua-Shan

2013-05-24

1,3,5,7-Tetramethyl-8-butyrethylenediamine-difluoroboradiaza-s-indacene (TMBB-EDAN) has been designed and synthesized as a highly fluorescent labeling reagent for carboxylic acids. By using TMBB-EDAN, a sensitive and rapid method based on high performance liquid chromatography-fluorescence detection for the determination of twelve fatty acids (FAs) in bio-samples has been developed. Under optimized conditions, these FAs were tagged with TMBB-EDAN in the presence of 1-ethyl-3-(3-dimethyla-minopropyl) carbodiamide at 20°C for 30min and then the baseline separation was achieved on a C18 column with a linear gradient elution in 26min. With fluorescence detection at λex/λem=490nm/510nm, the linear ranges of FAs were from 3.0 to 300nM and the detection limits with a signal-to-noise ratio of 3 were in the 0.2-0.4nM range. The proposed method offers advantages of milder derivatization condition and much better sensitivity for the determination of FAs, when compared to the reported fluorescence derivatization-based methods. Copyright © 2013 Elsevier B.V. All rights reserved.

Chemical constituents and antibacterial activity of Melastoma malabathricum L.

PubMed

Wong, Keng-Chong; Hag Ali, Dafaalla Mohamed; Boey, Peng-Lim

2012-01-01

The aqueous methanolic extracts of Melastoma malabathricum L. exhibited antibacterial activity when assayed against seven microorganisms by the agar diffusion method. Solvent fractionation afforded active chloroform and ethyl acetate fractions from the leaves and the flowers, respectively. A phytochemical study resulted in the identification of ursolic acid (1), 2α-hydroxyursolic acid (2), asiatic acid (3), β-sitosterol 3-O-β-D-glucopyranoside (4) and the glycolipid glycerol 1,2-dilinolenyl-3-O-β-D-galactopyanoside (5) from the chloroform fraction. Kaempferol (6), kaempferol 3-O-α-L-rhamnopyranoside (7), kaempferol 3-O-β-D-glucopyranoside (8), kaempferol 3-O-β-D-galactopyranoside (9), kaempferol 3-O-(2″,6″-di-O-E-p-coumaryl)-β-D-galactopyranoside (10), quercetin (11) and ellagic acid (12) were found in the ethyl acetate fraction. The structures of these compounds were determined by chemical and spectral analyses. Compounds 1-4, the flavonols (6 and 11) and ellagic acid (12) were found to be active against some of the tested microorganisms, while the kaempferol 3-O-glycosides (7-9) did not show any activity, indicating the role of the free 3-OH for antibacterial activity. Addition of p-coumaryl groups results in mild activity for 10 against Staphylococcus aureus and Bacillus cereus. Compounds 2-5, 7 and 9-12 are reported for the first time from M. malabathricum. Compound 10 is rare, being reported only once before from a plant, without assignment of the double bond geometry in the p-coumaryl moiety.

Evaluation of antibacterial, antioxidant and nootropic activities of Tiliacora racemosa Colebr. leaves: In vitro and in vivo approach.

PubMed

T, Vivek Kumar; M, Vishalakshi; M, Gangaraju; Das, Parijat; Roy, Pratiti; Banerjee, Anindita; Dutta Gupta, Sayan

2017-02-01

The antibacterial and antioxidant potential of Tiliacora racemosa leaf extracts in various solvents (methanolic, hexane, chloroform and ethyl acetate) was determined. Additionally, the presence of bisbenzylisoquinoline alkaloids in the plant prompted us to evaluate the nootropic activity of the methanolic extract in mice. Further, we seek to verify the nootropic effect by examining the anticholinesterase inhibition potential of the methanolic extract. The leaf extracts in various solvents were evaluated for their antibacterial and antioxidant activity by agar diffusion technique and α, α-diphenyl-β-picrylhydrazyl (DPPH) free radical scavenging method, respectively. The ex vivo acetylcholine esterase inhibitory activity of the methanolic extract was carried out by Ellman's method in male Wistar rats. The nootropic capacity of the methanolic extract was examined in Swiss albino mice by utilizing the diazepam induced acute amnesic model. The chloroform/n-hexane and ethyl acetate fraction showed promising antioxidant and antibacterial (Gram positive and Gram negative bacteria) property, respectively. The methanolic extract was able to diminish the amnesic effect induced by diazepam (1mg/kg i.p.) in mice. The extract also showed significant acetyl cholinesterase inhibition in rats. The findings prove that the memory enhancing capability is due to increased acetyl choline level at the nerve endings. The strong antioxidant nature and potential nootropic activity shown by the extract suggests its future usage in the treatment of neurodegenerative disorders such as dementia and Alzheimer. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Conformational and spectroscopic study of xanthogen ethyl formates, ROC(S)SC(O)OCH2CH3. Isolation of CH3CH2OC(O)SH

NASA Astrophysics Data System (ADS)

Juncal, Luciana C.; Cozzarín, Melina V.; Romano, Rosana M.

2015-03-01

ROC(S)SC(O)OCH2CH3, with R = CH3sbnd , (CH3)2CHsbnd and CH3(CH2)2sbnd , were obtained through the reaction between potassium xanthate salts, ROC(S)SK, and ethyl chloroformate, ClC(O)OCH2CH3. The liquid compounds were identified and characterized by 1H and 13C NMR and mass spectrometry. The conformations adopted by the molecules were studied by DFT methods. 6 conformers were theoretically predicted for R = CH3sbnd and (CH3)2CHsbnd , while the conformational flexibility of the n-propyl substituent increases the total number of feasible rotamers to 21. For the three molecules, the conformers can be associated in 3 groups, being the most stable the AS forms - the Cdbnd S double bond anti (A) with respect to the Csbnd S single bond and the Ssbnd C single bond syn (S) with respect to the Cdbnd O double bond - followed by AA and SS conformers. The vibrational spectra were interpreted in terms of the predicted conformational equilibrium, presenting the ν(Cdbnd O) spectral region signals corresponding to the three groups of conformers. A moderated pre-resonance Raman enhancement of the ν(Cdbnd S) vibrational mode of CH3(CH2)2OC(S)SC(O)OCH2CH3 was detected, when the excitation radiation approaches the energy of a n → π∗ electronic transition associated with the Cdbnd S chromophore. UV-visible spectra in different solvents were measured and interpreted in terms of TD-DFT calculations. The unknown molecule CH3CH2OC(O)SH was isolated by the UV-visible photolysis of CH3OC(S)SC(O)OCH2CH3 isolated in Ar matrix, and also obtained as a side-product of the reaction between potassium xanthate salts, ROC(S)SK, and ethyl chloroformate, ClC(O)OCH2CH3.

Synthesis, photophysical, structural and electronic properties of novel regioisomerically pure 1,7-disubstituted perylene-3,4,9,10-tetracarboxylic monoimide dibutylester derivatives

NASA Astrophysics Data System (ADS)

Ozser, Mustafa E.; Mohiuddin, Obaidullah

2018-04-01

Three new isomerically pure 1,7-disubstituted perylene-3,4,9,10-tetracarboxylic monoimide dibutylester derivatives namely; N-[2-(diethylamino)ethyl]-1,7-dibromoperylene-3,4,9,10-tetracarboxy monoimide dibutylester (PMD-1), N-[2-(diethylamino)ethyl]-1,7-di(4-tert-butylphenoxy)perylene-3,4,9,10-tetracarboxy monoimide dibutylester (PMD-2) and N-[2-(diethylamino)ethyl]-1,7-di(pyrrolidinyl)perylene-3,4,9,10-tetracarboxy monoimide dibutylester (PMD-3), have been synthesized and their electronic absorption spectra, and steady-state fluorescence were investigated experimentally as well as by using density functional theory (DFT) calculations. All three compounds show good solubility in toluene and chloroform. Attachment of two tert-butylphenoxy groups at the 1,7-positions in PMD-2 resulted in a red shifted absorption band with an absorption maximum at 518 nm. UV/Vis absorption spectrum of PMD-3 bearing electron donating pyrrolidinyl groups at the 1,7-positions shows a broad absorption band within the visible region, extending to red region. Absorption maximum of lowest energy transition now shifts to 653 nm. In addition to the S0 → S1 absorption bands, bands belonging to S0 → S2 electronic transitions were observed at 378, 386, and 411 nm for PMD-1, PMD-2, and PMD-3 respectively. Compounds PMD-1, PMD-2 and PMD-3 display low fluorescence quantum yields of 0.027, 0.040 and 0.001, respectively in chloroform. DFT calculations revealed that the attachment of electron donating groups at 1,7-positions of perylene core, results in an increase in frontier orbitals energy levels. Observed energy increase in HOMO level is larger in each case, compared to the energy increase in LUMO levels, due to the higher contribution of side groups to HOMO. DFT calculated band gaps for PMD-1, PMD-2 and PMD-3 are 2.68, 2.63 and 2.29 eV respectively.

Conformational and spectroscopic study of xanthogen ethyl formates, ROC(S)SC(O)OCH₂CH₃. Isolation of CH₃CH₂OC(O)SH.

PubMed

Juncal, Luciana C; Cozzarín, Melina V; Romano, Rosana M

2015-03-15

ROC(S)SC(O)OCH2CH3, with R=CH3-, (CH3)2CH- and CH3(CH2)2-, were obtained through the reaction between potassium xanthate salts, ROC(S)SK, and ethyl chloroformate, ClC(O)OCH2CH3. The liquid compounds were identified and characterized by (1)H and (13)C NMR and mass spectrometry. The conformations adopted by the molecules were studied by DFT methods. 6 conformers were theoretically predicted for R=CH3- and (CH3)2CH-, while the conformational flexibility of the n-propyl substituent increases the total number of feasible rotamers to 21. For the three molecules, the conformers can be associated in 3 groups, being the most stable the AS forms - the C=S double bond anti (A) with respect to the C-S single bond and the S-C single bond syn (S) with respect to the C=O double bond - followed by AA and SS conformers. The vibrational spectra were interpreted in terms of the predicted conformational equilibrium, presenting the ν(C=O) spectral region signals corresponding to the three groups of conformers. A moderated pre-resonance Raman enhancement of the ν(C=S) vibrational mode of CH3(CH2)2OC(S)SC(O)OCH2CH3 was detected, when the excitation radiation approaches the energy of a n→π∗ electronic transition associated with the C=S chromophore. UV-visible spectra in different solvents were measured and interpreted in terms of TD-DFT calculations. The unknown molecule CH3CH2OC(O)SH was isolated by the UV-visible photolysis of CH3OC(S)SC(O)OCH2CH3 isolated in Ar matrix, and also obtained as a side-product of the reaction between potassium xanthate salts, ROC(S)SK, and ethyl chloroformate, ClC(O)OCH2CH3. Copyright © 2014 Elsevier B.V. All rights reserved.

Antibacterial, antifungal and antioxidant activity of Olea africana against pathogenic yeast and nosocomial pathogens.

PubMed

Masoko, Peter; Makgapeetja, David M

2015-11-17

Olea africana leaves are used by Bapedi people to treat different ailments. The use of these leaves is not validated, therefore the aim of this study is to validate antimicrobial properties of this plant. The ground leaves were extracted using solvents of varying polarity (hexane, chloroform, dichloromethane (DCM), ethyl acetate, acetone, ethanol, methanol, butanol and water). Thin layer chromatography (TLC) was used to analyse the chemical constituents of the extracts. The TLC plates were developed in three different solvent systems, namely, benzene/ethanol/ammonium solution (BEA), chloroform/ethyl acetate/formic acid (CEF) and ethyl acetate/methanol/water (EMW). The micro-dilution assay and bioautography method were used to evaluate the antibacterial activity of the extracts against Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis and Staphylococcus aureus and the antifungal activity against Candida albicans and Cryptococcus neoformans. Methanol was the best extractant, yielding a larger amount of plant material whereas hexane yielded the least amount. In phytochemical analyses, more compounds were observed in BEA, followed by EMW and CEF. Qualitative 2, 2- diphenylpacryl-1-hydrazyl (DPPH) assay displayed that all the extracts had antioxidant activity. Antioxidant compounds could not be separated using BEA solvent system while with CEF and EMW enabled antioxidant compounds separation. The minimum inhibitory concentrations (MIC) values against test bacteria ranged between 0.16 and 2.50 mg/mL whereas against fungi, MIC ranged from 0.16 to 0.63 mg/mL. Bioautography results demonstrated that more than one compound was responsible for antimicrobial activity in the microdilution assay as the compounds were located at different Rf values. The results indicate that leaf extracts of Olea africana contain compounds with antioxidant, antibacterial and antifungal activities. Therefore, further studies are required to isolate the active compounds and perform other tests such as cytotoxicity. Olea africana may be a potential source of antimicrobial compounds.

Determination of free amino compounds in betalainic fruits and vegetables by gas chromatography with flame ionization and mass spectrometric detection.

PubMed

Kugler, Florian; Graneis, Stephan; Schreiter, Pat P-Y; Stintzing, Florian C; Carle, Reinhold

2006-06-14

Amino acids and amines are the precursors of betalains. Therefore, the profiles of free amino compounds in juices obtained from cactus pears [Opuntia ficus-indica (L.) Mill. cv. Bianca, cv. Gialla, and cv. Rossa], pitaya fruits [Selenicereus megalanthus (K. Schumann ex Vaupel) Moran, Hylocereus polyrhizus (Weber) Britton & Rose, and Hylocereus undatus (Haworth) Britton & Rose], and in extracts from differently colored Swiss chard [Beta vulgaris L. ssp. cicla (L.) Alef. cv. Bright Lights] petioles and red and yellow beets (B. vulgaris L. ssp. vulgaris var. conditiva Alef. cv. Burpee's Golden) were investigated for the first time. Amino compounds were derivatized with propyl chloroformate. While gas chromatography (GC) with mass spectrometry was used for peak assignment, GC flame ionization detection was applied for quantification of individual compounds. Whereas proline was the major free amino compound of cactus pear and pitaya fruit juices, glutamine dominated in Swiss chard stems and beets, respectively. Interestingly, extremely high concentrations of dopamine were detected in Swiss chard stems and beets. Furthermore, the cleavage of betaxanthins caused by derivatization in alkaline reaction solutions is demonstrated for the first time. Amino acids and amines thus released might increase the actual free amino compound contents of the respective sample. To evaluate the contribution of betaxanthin cleavage to total amino acid and amine concentration, isolated betaxanthins were derivatized according to the "EZ:faast" method prior to quantification of the respective amino compounds released. On a molar basis, betaxanthin contribution to overall amino compound contents was always below 6.4%.

Synthesis and optoelectronic properties of dodecyl substituted diphenylamine and pyridine based conjugated molecule

NASA Astrophysics Data System (ADS)

Priyanka, V.; Vijai Anand, A. S.; Mahesh, K.; Karpagam, S.

2017-11-01

The new donor-acceptor type conjugated moiety, namely 3-([4-(2-Cyano-2pyridine-2yl-vinyl)-phenyl]-dodecyl-amino)-phenyl)-2-pyridine-2-yl-acrylonitrile (DPA-PA) has been synthesized according to the Knoevenagel condensation. Here dodecyloxy diphenylamine moiety acts as an electron donor and cyano-pyridyl moiety acts as an electron acceptor. These moieties are recently showing great interest in optoelectronic applications. The structure of the DPA-PA was confirmed by FT-IR, 1H NMR. The final product showed great solubility in common organic solvents such as toluene, tetrahydrofuran, ethyl acetate, dichloromethane, chloroform etc due to the dodecyl chain. The absorption maximum of DPA-PA appeared at 433 nm in chloroform solution. The optical band gap is 2.2 eV calculated from thin film absorption edge (550 nm). The photoluminescence spectra exhibited a maximum peak at 513 nm with greenish fluorescence in chloroform solution and at 541 nm as the thin film state. The emission spectra of thin film state are 28 nm red shifted with broadening peak. The lower electrochemical band gap 1.55 eV was observed by cyclic voltammetry. This type of low band gap materials has much attention for their various potential applications in optoelectronic devices.

Communication: Potentials of mean force study of ionic liquid ion pair aggregation in polar covalent molecule solvents

NASA Astrophysics Data System (ADS)

Bandlamudi, Santosh Rathan Paul; Benjamin, Kenneth M.

2018-05-01

Molecular dynamics (MD) simulations were conducted for 1-ethyl-3-methylimidazolium methylsulfate [EMIM][MeSO4] dissolved in six polar covalent molecules [acetic acid, acetone, chloroform, dimethyl sulfoxide (DMSO), isopropyl alcohol, and methanol] to understand the free energies of ionic liquid (IL) ion pairing/aggregation in the limit of infinite dilution. Free energy landscapes or potentials of mean force (PMF) were computed using umbrella sampling and the weighted histogram analysis method. The PMF studies showed the strongest IL ion pairing in chloroform, and the strength of IL ion pairing decreases in the order of chloroform, acetone, propanol, acetic acid, DMSO, and methanol. In the limit of infinite dilution, the free energy curves for IL ion aggregation in co-solvents were characterized by two distinct minima [global (˜3.6 Å) and local (˜5.7 Å)], while free energy values at these minima differed significantly for IL in each co-solvent. The PMF studies were extended for determining the free energy of IL ion aggregation as a function of concentration of methanol. Studies showed that as the concentration of methanol increased, the free energy of ion aggregation decreased, suggesting greater ion pair stability, in agreement with previously reported MD clustering and radial distribution function data.

Skin penetration and kinetics of pristine fullerenes (C{sub 60}) topically exposed in industrial organic solvents

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xia, Xin R., E-mail: xia@ncsu.ed; Monteiro-Riviere, Nancy A.; Riviere, Jim E.

2010-01-01

Pristine fullerenes (C{sub 60}) in different solvents will be used in many industrial and pharmaceutical manufacturing and derivatizing processes. This report explores the impact of solvents on skin penetration of C{sub 60} from different types of industrial solvents (toluene, cyclohexane, chloroform and mineral oil). Yorkshire weanling pigs (n = 3) were topically dosed with 500 muL of 200 mug/mL C{sub 60} in a given solvent for 24 h and re-dosed daily for 4 days to simulate the worst scenario in occupational exposures. The dose sites were tape-stripped and skin biopsies were taken after 26 tape-strips for quantitative analysis. When dosedmore » in toluene, cyclohexane or chloroform, pristine fullerenes penetrated deeply into the stratum corneum, the primary barrier of skin. More C{sub 60} was detected in the stratum corneum when dosed in chloroform compared to toluene or cyclohexane. Fullerenes were not detected in the skin when dosed in mineral oil. This is the first direct evidence of solvent effects on the skin penetration of pristine fullerenes. The penetration of C{sub 60} into the stratum corneum was verified using isolated stratum corneum in vitro; the solvent effects on the stratum corneum absorption of C{sub 60} were consistent with those observed in vivo. In vitro flow-through diffusion cell experiments were conducted in pig skin and fullerenes were not detected in the receptor solutions by 24 h. The limit of detection was 0.001 mug/mL of fullerenes in 2 mL of the receptor solutions.« less

AAS and spectrophotometric methods for the determination metoprolol tartrate in tablets

NASA Astrophysics Data System (ADS)

Alpdoğan, Güzin; Sungur, Sidika

1999-11-01

Sensitive and specific atomic adsorption spectroscopy (AAS) and spectrophotometric methods have been developed for the determination of beta adrenergic blocking drug, metoprolol tartrate.The method is based on the formation of Cu(II) dithiocarbamate complex by derivatization of the secondary amino group of metoprolol with CS 2 and CuCl 2 in the presence of ammonia.The copper-bis(dithiocarbamate) complex was extracted into chloroform and the concentration of metoprolol tartrate was determined directly by spectrophotometric and indirectly by AAS measurement of copper.The two methods developed were applied to the assay of metoprolol tartrate in commercial tablet formulations.The methods were compared statistically with each other and with the high performance liquid chromatography (HPLC) method of USPXXII using t- and F-tests.

Stereoselective determination of amino acids in beta-amyloid peptides and senile plaques.

PubMed

Thorsén, G; Bergquist, J; Westlind-Danielsson, A; Josefsson, B

2001-06-01

A novel method for the determination of the enantiomeric composition of peptides is presented. In this paper, the focus has been on beta-amyloid peptides from deceased Alzheimer's disease patients. The peptides are hydrolyzed using mineral acid. The free amino acids are derivatized with the chiral reagent (+)- or (-)-1-(9-anthryl)-2-propyl chloroformate and subsequently separated using micellar electrokinetic chromatography (MEKC) and detected using laser-induced fluorescence (LIF) detection. The high separation efficiency of the MEKC-LIF system, yielding approximately 1 million theoretical plates/m for most amino acids, facilitates the simultaneous chiral determination of nine amino acids. The samples that have been analyzed were standard 1-40 beta-amyloid peptides, in vitro precipitated beta-amyloid fibrils, and human senile plaque samples.

In vitro antioxidant activity of Retama monosperma (L.) Boiss.

PubMed

Belmokhtar, Zoubir; Harche, Meriem Kaid

2014-01-01

The relationship between the antioxidant activity and the phenolic contents (total polyphenol, flavonoid and condensed tannin) of Retama monosperma (Fabaceae), used commonly in the traditional medicine of Mediterranean regions, was investigated. The antioxidant activities of the various fractions (toluene, chloroform, ethyl acetate and butanol) of the hydromethanolic extract of the seeds, stems and flowers have been evaluated using in vitro 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) radical scavenging activities and Phosphomolybdic acid assays and were compared to ascorbic acid. A significant high Pearson's correlations between flavonoid content and antioxidant activities (r = 0.91) with Phosphomolybdic acid assays and (r = - 0.79) with IC50 DPPH radical scavenging activities. However, there was no correlation between condensed tannin and antioxidant activities. The results obtained in the present study indicate that the ethyl acetate fraction of seeds is a potential source of natural antioxidant for R. monosperma.

[Determination of emamectin benzoate residue in vegetables by high performance liquid chromatography with fluorescence detection].

PubMed

Zhang, Yan; Wu, Yinliang; Hu, Jiye; Wang, Hongwei; Pan, Canping; Liu, Fengmao

2008-01-01

A method was developed for the determination of emamectin benzoate residue in cabbage and mushroom using solid-phase extraction (SPE) and high performance liquid chromatography (HPLC) with fluorescence detection. The sample was extracted with ethyl acetate. Further cleanup was performed on a propylsulfonic acid solid phase extraction cartridge, followed by the derivatization with trifluoroacetic anhydride in the presence of N-methylimidazole. The amount of derivatized emamectin benzoate was determined by fluorescence detector after separation by HPLC. The detection limit was 0.10 microg/kg for cabbage and mushroom samples. The recoveries of emamectin benzoate in cabbage and mushroom samples were 78.6%-84.9%. The inter-day relative standard deviation (RSD) and intra-day RSD were 2.7%-6.0% and 3.1%-8.9%, respectively, at the fortified levels of 1.0-20.0 microg/kg. The calibration curve of emamectin benzoate in vegetables at the concentration range of 0.002 mg/L to 0.10 mg/L was linear (r = 0.9999).

Chloroformate derivatization for tracing the fate of Amino acids in cells and tissues by multiple stable isotope resolved metabolomics (mSIRM).

PubMed

Yang, Ye; Fan, Teresa W-M; Lane, Andrew N; Higashi, Richard M

2017-07-11

Amino acids have crucial roles in central metabolism, both anabolic and catabolic. To elucidate these roles, steady-state concentrations of amino acids alone are insufficient, as each amino acid participates in multiple pathways and functions in a complex network, which can also be compartmentalized. Stable Isotope-Resolved Metabolomics (SIRM) is an approach that uses atom-resolved tracking of metabolites through biochemical transformations in cells, tissues, or whole organisms. Using different elemental stable isotopes to label multiple metabolite precursors makes it possible to resolve simultaneously the utilization of these precursors in a single experiment. Conversely, a single precursor labeled with two (or more) different elemental isotopes can trace the allocation of e.g. C and N atoms through the network. Such dual-label experiments however challenge the resolution of conventional mass spectrometers, which must distinguish the neutron mass differences among different elemental isotopes. This requires ultrahigh resolution Fourier transform mass spectrometry (UHR-FTMS). When combined with direct infusion nano-electrospray ion source (nano-ESI), UHR-FTMS can provide rapid, global, and quantitative analysis of all possible mass isotopologues of metabolites. Unfortunately, very low mass polar metabolites such as amino acids can be difficult to analyze by current models of UHR-FTMS, plus the high salt content present in typical cell or tissue polar extracts may cause unacceptable ion suppression for sources such as nano-ESI. Here we describe a modified method of ethyl chloroformate (ECF) derivatization of amino acids to enable rapid quantitative analysis of stable isotope labeled amino acids using nano-ESI UHR-FTMS. This method showed excellent linearity with quantifiable limits in the low nanomolar range represented in microgram quantities of biological specimens, which results in extracts with total analyte abundances in the low to sub-femtomole range. We have applied this method to profile amino acids and their labeling patterns in 13 C and 2 H doubly labeled PC9 cell extracts, cancerous and non-cancerous tissue extracts from a lung cancer patient and their protein hydrolysates as well as plasma extracts from mice fed with a liquid diet containing 13 C 6 -glucose (Glc). The multi-element isotopologue distributions provided key insights into amino acid metabolism and intracellular pools in human lung cancer tissues in high detail. The 13 C labeling of Asp and Glu revealed de novo synthesis of these amino acids from 13 C 6 -Glc via the Krebs cycle, specifically the elevated level of 13 C 3 -labeled Asp and Glu in cancerous versus non-cancerous lung tissues was consistent with enhanced pyruvate carboxylation. In addition, tracking the fate of double tracers, ( 13 C 6 -Glc +  2 H 2 -Gly or 13 C 6 -Glc +  2 H 3 -Ser) in PC9 cells clearly resolved pools of Ser and Gly synthesized de novo from 13 C 6 -Glc ( 13 C 3 -Ser and 13 C 2 -Gly) versus Ser and Gly derived from external sources ( 2 H 3 -Ser, 2 H 2 -Gly). Moreover the complex 2 H labeling patterns of the latter were results of Ser and Gly exchange through active Ser-Gly one-carbon metabolic pathway in PC9 cells. Copyright © 2017 Elsevier B.V. All rights reserved.

Weed growth inhibitors from Aspergillus fischeri TISTR 3272.

PubMed

Phattanawasin, P; Pojchanakom, K; Sotanaphun, U; Piyapolrungroj, N; Zungsontiporn, S

2007-12-01

Chloroform and ethyl acetate extracts of Aspergillus fischeri TISTR 3272 showed good growth inhibitory activity on Mimosa pigra and Echinochloa crus-galli. Bioassay-directed fractionation of the active extracts led to the isolation of five known compounds, (+)-terrein (1), (-)-6-hydroxymellein (2), two diketopiperazines (cyclo-(S-Pro-S-Leu) (3) and cyclo-(S-Pro-S-Val) (4)) and butyrolactone I (5). Compounds 2-5 were reported for the first time in this fungus. Their structural determinations were based on analyses of spectroscopic data and their weed growth inhibitory effects were assessed.

Installation Restoration Program. Stage 3. McClellan Air Force Base. Quality Assurance Project Plan

DTIC Science & Technology

1990-03-01

Dibromomethane 5.0 trans- 1,2-Dichloropropene 5.0 1,2-Dibromoethane 4.8 total- 1,4-Dichlorobutene 2.6 Methyl methacrylate 28 1,1, 1,2-Trichloropropane 3.9...trans- 1,2-DCE); • Methyl ethyl ketone (MEK); • Carbon disulfide; • Dichlorobenzene; * Chloroform; and * Methylene chloride. Contaminants reported in soil...2-Hexanone 10 4- Methyl -2-pentanone 10 Tetrachloroethene 4.1 1, 1,2,2-Tetrachloroethane 5.0 Toluene 5.0 Chlorobenzene 5.0 Ethylbenzene 5.0 Styrene 3.0

Synthesis of some N-substituted indole derivatives and their biological activities.

PubMed

el-Diwani, H; Nakkady, S S; Hishmat, O H; el-Shabrawy, O A; Mahmoud, S S

1992-03-01

Acylation of 2,3-diphenyl-5-methoxy-indole using ethyl chloroformate or chloroacetyl chloride in dimethylformamide and sodium hydride yielded the N-substituted derivatives 1 and 2, respectively. While Friedel-Crafts acylation using chloroacetyl chloride afforded di-4,6-chloroacetyl derivative 3, the reaction of the N-chloroacetyl derivative 2 with amines, hydrazines, urea, semicarbazide hydrochloride, thiophenol, benzimidazole-2-thiol, thiosemicarbazide, 2-mercaptoethanol and thioglycolic acid was studied. Several of the compounds were tested for their effect on arterial blood pressure, antiinflammatory and ulcerogenic activities.

Pesticidal and pest repellency activities of rhizomes of Drynaria quercifolia (J. Smith) against Tribolium castaneum (Herbst).

PubMed

Khan, Alam; Islam, Md Hedayetul; Islam, Md Ekramul; Al-Bari, Md Abdul Alim; Parvin, Mst Shahnaj; Sayeed, Mohammed Abu; Islam, Md Nurul; Haque, Md Ekramul

2014-10-01

Tribolium castaneum (Herbst) is a harmful pest of stored grain and flour-based products in tropical and subtropical region. In the present study, rhizome of Drynaria quercifolia (J. Smith) was evaluated for pesticidal and pest repellency activities against T. castaneum, using surface film method and filter paper disc method, respectively. In addition, activity of the isolated compound 3,4-dihydroxybenzoic acid was evaluated against the pest. Chloroform soluble fraction of ethanol extract of rhizome of D. quercifolia showed significant pesticidal activity at doses 0.88 to 1.77 mg/cm(2) and significant pest repellency activity at doses 0.94 to 0.23 mg/cm(2). No pesticidal and pest repellency activity was found for petroleum ether, ethyl acetate and methanol soluble fractions of ethanol extract as well as for 3,4-dihydroxybenzoic acid. Considering our findings it can be concluded that chloroform soluble fraction of rhizome of D. quercifolia is useful in controlling T. castaneum of stored grain and flour-based products.

Larvicidal activity of medicinal plant extracts against Anopheles subpictus & Culex tritaeniorhynchus.

PubMed

Kamaraj, C; Bagavan, A; Elango, G; Zahir, A Abduz; Rajakumar, G; Marimuthu, S; Santhoshkumar, T; Rahuman, A Abdul

2011-07-01

Mosquitoes transmit serious human diseases, causing millions of deaths every year and the development of resistance to chemical insecticides resulting in rebounding vectorial capacity. Plants may be alternative sources of mosquito control agents. The present study assessed the role of larvicidal activities of hexane, chloroform, ethyl acetate, acetone, and methanol dried leaf and bark extracts of Annona squamosa L., Chrysanthemum indicum L., and Tridax procumbens L. against the fourth instar larvae of malaria vector, Anopheles subpictus Grassi and Japanese encephalitis vector, Culex tritaeniorhynchus Giles (Diptera: Culicidae). Larvicidal activities of three medicinal plant extracts were studied in the range of 4.69 to 1000 mg/l in the laboratory bioassays against early 4 th instar larvae of An. subpictus and Cx. tritaeniorhynchus. The mortality data were subjected to probit analysis to determine the lethal concentrations (LC50 and LC90) to kill 50 and 90 per cent of the treated larvae of the respective species. All plant extracts showed moderate effects after 24 h of exposure; however, the highest toxic effect of bark methanol extract of A. squamosa, leaf ethyl acetate extract of C. indicum and leaf acetone extract of T. procumbens against the larvae of An. subpictus (LC 50 = 93.80, 39.98 and 51.57 mg/l) and bark methanol extract of A. squamosa, leaf methanol extract of C. indicum and leaf ethyl acetate extract of T. procumbens against the larvae of Cx. tritaeniorhynchus (LC50 =104.94, 42.29 and 69.16 mg/l) respectively. Our data suggest that the bark ethyl acetate and methanol extract of A. squamosa, leaf ethyl acetate and methanol extract of C. indicum, acetone and ethyl acetate extract of T. procumbens have the potential to be used as an ecofriendly approach for the control of the An. subpictus, and Cx. tritaeniorhynchus.

Lipid peroxidation inhibition and antiradical activities of some leaf fractions of Mangifera indica.

PubMed

Badmus, Jelili A; Adedosu, Temitope O; Fatoki, John O; Adegbite, Victor A; Adaramoye, Oluwatosin A; Odunola, Oyeronke A

2011-01-01

This study was undertaken to assess in vitro lipid peroxidation inhibitions and anti-radical activities of methanolic, chloroform, ethyl acetate and water fractions of Mangifera indica leaf. Inhibition of Fe(2+)-induced lipid peroxidation (LPO) in egg, brain, and liver homogenates, 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl (OH-) radical scavenging activities were evaluated. Total phenol was assessed in all fractions, and the reducing power of methanolic fraction was compared to gallic acid and ascorbic acid. The results showed that Fe2+ induced significant lipid peroxidation (LPO) in all the homogenates. Ethyl acetate fraction showed the highest percentage inhibition of LPO in both egg yolk (68.3%) and brain (66.3%), while the aqueous fraction exerted the highest inhibition in liver homogenate (89.1%) at a concentration of 10 microg/mL. These observed inhibitions of LPO by these fractions were higher than that of ascorbic acid used as a standard. The DPPH radical scavenging ability exhibited by ethyl acetate fraction was found to be the highest with IC50 value of 1.5 microg/mL. The ethyl acetate and methanolic fractions had the highest OH- radical scavenging ability with the same IC50 value of 5 microg/mL. The total phenol content of ethyl acetate fraction was the highest with 0.127 microg/mg gallic acid equivalent (GAE). The reductive potential of methanolic fraction showed a concentration-dependent increase. This study showed that inhibition of LPO and the DPPH and OH- radicals scavenging abilities of Mangifera indica leaf could be related to the presence of phenolic compounds. Therefore, the ethyl acetate fraction of the leaf may be a good source of natural antioxidative agent.

Extracts of Actinidia arguta stems inhibited LPS-induced inflammatory responses through nuclear factor-κB pathway in Raw 264.7 cells.

PubMed

Kim, Hae-Young; Hwang, Kwang Woo; Park, So-Young

2014-11-01

The inflammatory response protects our body from bacteria and tumors, but chronic inflammation driven by the persistent activation of macrophages can lead to serious adverse effects including gastrointestinal problems, cardiac disorders, and a sore throat. Part of the ongoing research is focused on searching for antiinflammatory compounds from natural sources, so we investigated the effects of hardy kiwis (Actinidia arguta, Lauraceae) stems on inflammation induced by lipopolysaccharide (LPS) in Raw 264.7 cells to test the hypothesis that antiinflammatory effects of A. arguta stems were exerted through the inhibition of the nuclear factor (NF)-κB pathway. The methanol extract of A. arguta (20 μg/mL) stems lowered nitric oxide production in LPS-stimulated Raw 264.7 cells by 40%. It was then partitioned with hexane, chloroform, ethyl acetate, butanol, and water based on the polarity of each compound. Among the 5 layers, the chloroform layer had the greatest inhibitory effect on LPS-stimulated nitric oxide production and inducible nitric oxide synthase mRNA expression in Raw 264.7 cells. However, the levels of prostaglandin E2 and cyclooxygease 2 were not altered. On the other hand, treatment of cells with the chloroform layer of A. arguta before LPS stimulation also reduced them RNA expression of proinflammatory cytokines including tumor necrosis factor α and interleukin 1β. Nuclear translocation of NF-κB p50 and p65 subunits induced by LPS was also inhibited by treatment with the chloroform layer of A. arguta. This was accompanied with the reduced phosphorylation of mitogen-activated protein kinases including extracellular signal-regulated protein kinase 1/2, c-Jun N-terminal protein kinase, and p38. Taken together, these results suggest that chloroform layer of A. arguta exerted antiinflammatory effects by the inhibition of mitogen-activated protein kinase phosphorylation and nuclear translocation of NF-κB.

Determination of Parabens by Injection-Port Derivatization Coupled With Gas-Chromatography-Mass Spectrometry and Matrix Solid Phase Dispersion

NASA Astrophysics Data System (ADS)

Djatmika, Rosalina; Ding, Wang-Hsien; Sulistyarti, Hermin

2018-01-01

A rapid determination of four parabens preservatives (methyl paraben, ethyl paraben, propyl paraben, and butyl paraben) in marketed seafood is presented. Analytes were extracted and purified using matrix solid-phase dispersion (MSPD) method, followed by Injection port acylation gas chromatography-mass spectrometry (GC-MS) with acetic anhydride reagent. In this method, acylation of parabens was performed by acetic anhydride at GC injection-port generating reduction of the time-consuming sample-processing steps, and the amount of toxic reagents and solvents. The parameters affecting this method such as injection port temperature, purge-off time and acylation (acetic anhydride) volume were studied. In addition, the MSPD influence factors (including the amount of dispersant and clean-up co-sorbent, as well as the volume of elution solvent) were also investigated. After MSPD method and Injection port acylation applied, good linearity of analytes was achieved. The limits of quantitation (LOQs) were 0.2 to 1.0 ng/g (dry weight). Compared with offline derivatization commonly performed, injection port acylation employs a rapid, simple, low-cost and environmental-friendly derivatization process. The optimized method has been successfully applied for the analysis of parabens in four kind of marketed seafood. Preliminary results showed that the total concentrations of four selected parabens ranged from 16.7 to 44.7 ng/g (dry weight).

Phytochemical Screening, Alpha-Glucosidase Inhibition, Antibacterial and Antioxidant Potential of Ajuga bracteosa Extracts.

PubMed

Hafeez, Kokab; Andleeb, Saiqa; Ghousa, Tahseen; Mustafa, Rozina G; Naseer, Anum; Shafique, Irsa; Akhter, Kalsoom

2017-01-01

Ajuga bracteosa, a medicinal herb, is used by local community to cure a number of diseases such as inflammation, jaundice bronchial asthma, cancer and diabetes. The aim of present work was to evaluate the antioxidant potential, in vitro antidiabetic and antimicrobial effects of A. bracteosa. n-hexane, ethyl acetate, chloroform, acetone, methanol and aqueous extracts of Ajuga bracteosa roots, were prepared via maceration. Antibacterial activity was carried out by agar well diffusion method. Quantitative and qualitative phytochemical screening was done. The antioxidant activity was determined by iron (II) chelating activity, iron reducing power, DPPH, and ABTS free radical scavenging methods, Antidiabetic activity was evaluated through inhibition of α-glucosidase assay. Phytochemical analysis showed the presence of phenols, flavonoids, tannins, saponins, quinines, terpenoids, xanthoproteins, glycosides, carbohydrates, steroids, phytosterols and amino acids. DPPH and ABTS potential values were recorded as 61.92% to 88.84% and 0.11% to 38.82%, respectively. Total phenolic and total flavonoid contents were expressed as gallic acid and rutin equivalents. Total iron content was expressed as FeSO4 equivalents. Chloroform and n-hexane extracts showed significant enzyme inhibition potential with IC50 values of 29.92 μg/ml and 131.7 μg/ml respectively. Aqueous extract showed maximum inhibition of E. coli, S. typhimurium, E. amnigenus, S. pyogenes, and S. aureus, (18.0±1.0 mm, 12.5±0.7 mm, 17.0±0.0 mm, 11.0±0.0 mm and 15.3±2.0 mm mm), respectively. Similarly, n-hexane extract showed maximum inhibition of E. coli, E. amnigenus, S. aureus (11.6±1.5 mm; 11.3±1.5 mm; 13.3±0.5 mm). This study also shows that n-hexane, chloroform, ethyl acetate and aqueous extracts of A. bracteosa root possess α-glucosidase inhibitory activities and therefore it may be used as hypoglycemic agents in the management of postprandial hyperglycemia. Ajuga bracteosa root extracts may provide a basis for development of antioxidant, antimicrobial and antidiabetic drugs. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Quantitation of total homocysteine in human plasma by derivatization to its N(O,S)-propoxycarbonyl propyl ester and gas chromatography-mass spectrometry analysis.

PubMed

Sass, J O; Endres, W

1997-08-01

Much evidence supports the hypothesis that mild or moderate hyperhomocysteinaemia represents an important and independent risk factor for occlusive vascular diseases. Therefore, the accurate and reliable determination of total plasma homocysteine has gained major importance for risk assessment. Furthermore, it can help in the detection of folate and vitamin B12 deficiency. This has prompted us to develop a sensitive gas chromatography-mass spectrometry (GC-MS) method in order to quantify total homocysteine in human plasma. Prior to chromatography, reduced homocysteine was released from disulfide bonds by incubation with excess dithiothreitol and converted into its N(O,S)-propoxycarbonyl propyl ester by derivatization with n-propyl chloroformate. Aminoethylcysteine served as internal standard. The method proved to be highly linear over the entire concentration range examined (corresponding to 0-266 microM homocysteine) and showed intra-assay and inter-assay variation (relative standard deviations) of approximately 5 and 5-10%, respectively. External quality control by comparison with duplicate analysis performed on a HPLC-based system revealed satisfactory correlation. The newly developed GC-MS based method provides simple, reliable and fast quantification of total homocysteine and requires only inexpensive chemicals, which are easy to obtain.

Potential Health Benefits and Metabolomics of Camel Milk by GC-MS and ICP-MS.

PubMed

Ahamad, Syed Rizwan; Raish, Mohammad; Ahmad, Ajaz; Shakeel, Faiyaz

2017-02-01

None of the research reports reveals the metabolomics and elemental studies on camel milk. Recent studies showed that camel milk possesses anticancer and anti-inflammatory activity. Metabolomics and elemental studies were carried out in camel milk which showed us the pathways and composition that are responsible for the key biological role of camel milk. Camel milk was dissolved in methanol and chloroform fraction and then vortexed and centrifuged. Both the fractions were derivatized by N,O-bis-(trimethylsilyl)trifluoroacetamide (BSTFA) and TMCS after nitrogen purging and analyzed by GC-MS. Camel milk was also analyzed by ICP-MS after microwave digestion. We found that higher alkanes and fatty acids are present in the chloroform fraction and amino acids, sugars and fatty acid derivatives are present in aqueous fractions. All the heavy metals like As, Pb, Cd, Co, Cu, and Ni were in the safe limits in terms of maximum daily intake of these elements. Na, K, Mg, and Ca were also present in the safe limits in terms of maximum daily intake of these elements. These results suggested that the camel milk drinking is safe and there is no health hazard. The present data of GC-MS and ICP-MS correlate the activities related to camel milk.

Simultaneous derivatisation and preconcentration of parabens in food and other matrices by isobutyl chloroformate and dispersive liquid-liquid microextraction followed by gas chromatographic analysis.

PubMed

Jain, Rajeev; Mudiam, Mohana Krishna Reddy; Chauhan, Abhishek; Ch, Ratnasekhar; Murthy, R C; Khan, Haider A

2013-11-01

A simple, rapid and economical method has been proposed for the quantitative determination of parabens (methyl, ethyl, propyl and butyl paraben) in different samples (food, cosmetics and water) based on isobutyl chloroformate (IBCF) derivatisation and preconcentration using dispersive liquid-liquid microextraction in single step. Under optimum conditions, solid samples were extracted with ethanol (disperser solvent) and 200 μL of this extract along with 50 μL of chloroform (extraction solvent) and 10 μL of IBCF was rapidly injected into 2 mL of ultra-pure water containing 150 μL of pyridine to induce formation of a cloudy state. After centrifugation, 1 μL of the sedimented phase was analysed using gas chromatograph-flame ionisation detector (GC-FID) and the peaks were confirmed using gas chromatograph-positive chemical ionisation-mass spectrometer (GC-PCI-MS). Method was found to be linear over the range of 0.1-10 μg mL(-1) with square of correlation coefficient (R(2)) in the range of 0.9913-0.9992. Limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.029-0.102 μg mL(-1) and 0.095-0.336 μg mL(-1) with a signal to noise ratio of 3:1 and 10:1, respectively. Copyright © 2013 Elsevier Ltd. All rights reserved.

Antifungal potential of Avicennia schaueriana Stapf & Leech. (Acanthaceae) against Cladosporium and Colletotrichum species.

PubMed

Fardin, K M; Young, M C M

2015-07-01

There is significant interest in research to develop plant extracts with fungicidal activities that are less harmful to the environment and human health than synthetic fungicides. This study aimed to evaluate the antifungal activity of the extracts of Avicennia schaueriana against Colletotrichum and Cladosporium species and to identify the compounds responsible for the activity. Leaves and stems of A. schaueriana were extracted with ethanol and partitioned with petroleum ether, chloroform and ethyl acetate. The antifungal activity of such extracts was tested by bioautography against Cladosporium sphaerospermum, Cladosporium cladosporioides and Colletotrichum lagenarium. Ethanolic extracts, petroleum ether and chloroform fractions of stems had the highest antifungal activity with several active bands (Rf = 0·72 and Rf = 0·55). In the agar dilution assay, ethanolic extract, petroleum ether and chloroform fractions of stems were the most efficacious, presenting 85, 62 and 63% growth inhibition of Colletotrichum gloeopsporioides and minimum inhibitory concentration values between 1 and 1·5 mg ml(-1) , respectively. Analysis carried out using gas chromatography coupled to a mass spectrometry of petroleum ether and chloroform fractions allowed the identification of fatty acids methyl esters, lupeol and naphthoquinones such as lapachol, α-lapachone, naphtho[2,3-b]furan-4,9-dione, 2-isopropyl- and avicenol-C. We may infer that the antifungal activity of A. schaueriana is due to the abundance of these compounds. This study shows that Avicennia schaueriana extracts have a high potential for the growth inhibition of Colletotrichum and Cladosporium ssp. and will provide a starting point for discovering new natural products with antifungal activity. Their development is of particular interest to organic production systems where synthetic fungicides cannot be used. © 2015 The Society for Applied Microbiology.

Characterization of three agave species by gas chromatography and solid-phase microextraction-gas chromatography-mass spectrometry.

PubMed

Peña-Alvarez, Araceli; Díaz, Laura; Medina, Alejandra; Labastida, Carmen; Capella, Santiago; Vera, Luz Elena

2004-02-20

Steam distillation (SD) extraction-solid-phase microextraction coupled to GC-MS was developed for the determination of terpenes and Bligh-Dyer extraction-derivatization coupled with GC for the determination of fatty acids such as ethyl esters were used. It was found that the three different Agave species have the same profile of fatty acids; the quantity of these compounds is different in each Agave variety. On the other hand, different terpenes were identified in the three Agave plants studied: nine in A. salmiana, eight in A. angustifolia and 32 in A. tequilana Weber var. azul.

Solid sorbent air sampling and analytical procedure for methyl-, dimethyl-, ethyl-, and diethylamine

DOE Office of Scientific and Technical Information (OSTI.GOV)

Elskamp, C.J.; Schultz, G.R.

1986-01-01

A sampling and analytical procedure for methyl-, dimethyl-, ethyl-, and diethylamine was developed in order to avoid problems typically encountered in the sampling and analysis of low molecular weight aliphatic amines. Samples are collected with adsorbent tubes containing Amberlite XAD-7 resin coated with the derivatizing reagent, NBD chloride (7-chloro-4-nitrobenzo-2-oxa-1,3-diazole). Analysis is performed by high performance liquid chromatography with the use of a fluorescence and/or UV/visible detector. All four amines can be monitored simultaneously, and neither collection nor storage is affected by humidity. Samples are stable at room temperature for at least two weeks. The methodology has been tested for eachmore » of the four amines at sample loadings equivalent to air concentration ranges of 0.5 to 30 ppm for a sample volume of 10 liters. The method shows promise for determining other airborne primary and secondary low molecular weight aliphatic amines.« less

Determination of fatty acid ethyl esters (FAEE) and ethyl glucuronide (EtG) in hair: a promising way for retrospective detection of alcohol abuse during pregnancy?

PubMed

Pragst, Fritz; Yegles, Michel

2008-04-01

The retrospective detection of alcohol consumption during pregnancy is an important part of the diagnosis of the fetal alcohol syndrome. A promising way to solve this problem can be the determination of fatty acid ethyl esters (FAEE) or/and ethyl glucuronide (EtG) in hair of the mothers. In this article, the present state in analytical determination and interpretation of FAEE and EtG concentrations in hair are reviewed. Both FAEE and EtG are minor metabolites of ethanol and as direct alcohol markers very specific for alcohol. They are durably deposited in hair, which enables taking advantage of the long diagnostic time window of this sample material. In the last years, specific and sensitive methods for determination of both alcohol markers in hair were developed. Headspace solid phase microextraction in combination with gas chromatography-mass spectroscopy after hair extraction with an n-heptane/dimethylsulfoxide mixture proved to be a favorable technique for determination of four characteristic FAEE (ethyl myristate, ethyl palmitate, ethyl oleate, and ethyl stearate). EtG is extracted from hair by water and analyzed either by gas chromatography-mass spectroscopy with negative chemical ionization after cleanup with solid phase extraction and derivatization with pentafluoropropionic anhydride or by liquid chromatography-mass spectroscopy-mass spectroscopy. The detection limits of the single FAEE as well as of EtG are in the range of 1 to 10 pg/mg. FAEE as well as EtG were determined in a larger number of hair samples of teetotalers, social drinkers, patients in alcohol withdrawal treatment, and death cases with previous known heavy drinking. From the results, the following criteria were derived: strict abstinence is excluded or improbable at C FAEE >0.2 ng/mg or C EtG >7 pg/mg. Moderate social drinkers should have C FAEE <0.5 ng/mg and C EtG <25 pg/mg; above these values, alcohol abuse is probable. Until now, there has been no evaluation in context of FAS diagnosis; however, a successful application for this purpose can be expected from the good experience in driving ability examination.

Hydroquinone; A Novel Bioactive Compound from Plant-Derived Smoke Can Cue Seed Germination of Lettuce.

PubMed

Kamran, Muhammad; Khan, Abdul L; Ali, Liaqat; Hussain, Javid; Waqas, Muhammad; Al-Harrasi, Ahmed; Imran, Qari M; Kim, Yoon-Ha; Kang, Sang-Mo; Yun, Byung-Wook; Lee, In-Jung

2017-01-01

Plant-derived smoke has been known to play an important role in distribution and growth of vegetation. Using a proficiently designed furnace, we extracted smoke from the leaves of four plant viz. Helianthus annuus , Aloe vera , Ginkgo biloba , and Cymbopogon jwarancusa . Smoke dilutions obtained from these plants were obtained in different concentrations to identify potential lettuce growth promoting smoke solution. Results revealed that smoke obtained from G. biloba significantly enhanced the lettuce seed germination. This solution was then partitioned into ethyl acetate, dichloromethane, n -hexane, chloroform and ether fractions. Ethyl acetate fraction was found to be potent to enhance seed germination. This fraction was subjected to column chromatography and spectroscopic techniques to obtain compound 1 . This compound was identified as hydroquinone using 1D and 2D NMR techniques. At low concentrations (5, 10, and 20 ppm), compound 1 enhanced the lettuce seed germination; however, higher concentrations inhibited its growth as compared to control.

Hydroquinone; A novel bioactive compound from plant-derived smoke can cue seed germination of lettuce

NASA Astrophysics Data System (ADS)

Kamran, Muhammad; Khan, Abdul L.; Ali, Liaqat; Hussain, Javid; Waqas, Muhammad; Al-Harrasi, Ahmed; Imran, Qari M.; Kim, Yoon-Ha; Kang, Sang-Mo; Yun, Byung-Wook; Lee, In-Jung

2017-05-01

Plant-derived smoke has been known to play an important role in distribution and growth of vegetation. Using a proficiently designed furnace, we extracted smoke from the leaves of four plant viz. Helianthus annuus, Aloe vera, Ginkgo biloba, and Cymbopogon jwarancusa. Smoke dilutions obtained from these plants were obtained in different concentrations to identify potential lettuce growth promoting smoke solution. Results revealed that smoke obtained from Ginkgo biloba significantly enhanced the lettuce seed germination. This solution was then partitioned into ethyl acetate, dichloromethane, n-hexane, chloroform and ether fractions. Ethyl acetate fraction was found to be potent to enhance seed germination. This fraction was subjected to column chromatography and spectroscopic techniques to obtain compound 1. This compound was identified as hydroquinone using 1D and 2D NMR techniques. At low concentrations (5, 10 and 20 ppm), compound 1 enhanced the lettuce seed germination; however, higher concentrations inhibited its growth as compared to control.

Hydroquinone; A Novel Bioactive Compound from Plant-Derived Smoke Can Cue Seed Germination of Lettuce

PubMed Central

Kamran, Muhammad; Khan, Abdul L.; Ali, Liaqat; Hussain, Javid; Waqas, Muhammad; Al-Harrasi, Ahmed; Imran, Qari M.; Kim, Yoon-Ha; Kang, Sang-Mo; Yun, Byung-Wook; Lee, In-Jung

2017-01-01

Plant-derived smoke has been known to play an important role in distribution and growth of vegetation. Using a proficiently designed furnace, we extracted smoke from the leaves of four plant viz. Helianthus annuus,Aloe vera,Ginkgo biloba, and Cymbopogon jwarancusa. Smoke dilutions obtained from these plants were obtained in different concentrations to identify potential lettuce growth promoting smoke solution. Results revealed that smoke obtained from G. biloba significantly enhanced the lettuce seed germination. This solution was then partitioned into ethyl acetate, dichloromethane, n-hexane, chloroform and ether fractions. Ethyl acetate fraction was found to be potent to enhance seed germination. This fraction was subjected to column chromatography and spectroscopic techniques to obtain compound 1. This compound was identified as hydroquinone using 1D and 2D NMR techniques. At low concentrations (5, 10, and 20 ppm), compound 1 enhanced the lettuce seed germination; however, higher concentrations inhibited its growth as compared to control. PMID:28553632

Application of off-line two-dimensional high-performance countercurrent chromatography on the chloroform-soluble extract of Cuscuta auralis seeds.

PubMed

Rho, Taewoong; Yoon, Kee Dong

2018-05-01

In this study, the chloroform-soluble extract of Cuscuta auralis was separated successfully using off-line two-dimensional high-performance countercurrent chromatography, yielding a γ-pyrone, two alkaloids, a flavonoid, and four lignans. The first-dimensional countercurrent separation using a methylene chloride/methanol/water (11:6:5, v/v/v) system yielded three subfractions (fractions I-III). The second-dimensional countercurrent separations, conducted on fractions I-III using n-hexane/ethyl acetate/methanol/water/acetic acid (5:5:5:5:0, 3:7:3:7:0, and 1:9:1:9:0.01, v/v/v/v/v) systems, gave maltol (1), (-)-(13S)-cuscutamine (2), (+)-(13R)-cuscutamine (3), (+)-pinoresinol (4), (+)-epipinoresinol (5), kaempferol (6), piperitol (7), and (9R)-hydroxy-d-sesamin (8). To the best of our knowledge, maltol was identified for the first time in Cuscuta species. Furthermore, this report details the first full assignment of spectroscopic data of two cuscutamine epimers, (-)-(13S)-cuscutamine and (+)-(13R)-cuscutamine. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Comparative studies on extracts from Hericium erinaceus by different polarity reagents to gain higher antioxidant activities.

PubMed

Jiang, Shengjuan; Wang, Yuliang; Zhang, Xiaolong

2016-07-01

Hericium erinaceus (H. erinaceus) is a source of exogenous antioxidants that has been traditionally used in China for the prevention and treatment of oxidative stress-associated disease. In the present study, the bioactive compounds of H. erinaceus were extracted with the following eight representative reagents: n-Hexane, xylene, chloroform, anhydrous ether, ethyl acetate, acetone, anhydrous ethanol and distilled water. The in vitro antioxidant activities were also evaluated. All of the extracted compounds exhibited reducing power and scavenging activity against 1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide anion free radicals. In addition, the antioxidant capacities varied with the used chemical reagents and exhibited dose-dependent effects. Extracts from anhydrous ethanol, chloroform and acetone were capable of inhibiting lipid peroxidation. The anhydrous ethanol extracts were observed to have significant levels of antioxidant compounds since they had a strong reducing power, high scavenging rates against DPPH and superoxide anion-free radicals (>90%), and high inhibition rates on lipid peroxidation (>60%). The present study will provide reference data for the antioxidant applications of H. erinaceus in pharmaceutical use and disease prevention.

Comparative studies on extracts from Hericium erinaceus by different polarity reagents to gain higher antioxidant activities

PubMed Central

JIANG, SHENGJUAN; WANG, YULIANG; ZHANG, XIAOLONG

2016-01-01

Hericium erinaceus (H. erinaceus) is a source of exogenous antioxidants that has been traditionally used in China for the prevention and treatment of oxidative stress-associated disease. In the present study, the bioactive compounds of H. erinaceus were extracted with the following eight representative reagents: n-Hexane, xylene, chloroform, anhydrous ether, ethyl acetate, acetone, anhydrous ethanol and distilled water. The in vitro antioxidant activities were also evaluated. All of the extracted compounds exhibited reducing power and scavenging activity against 1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide anion free radicals. In addition, the antioxidant capacities varied with the used chemical reagents and exhibited dose-dependent effects. Extracts from anhydrous ethanol, chloroform and acetone were capable of inhibiting lipid peroxidation. The anhydrous ethanol extracts were observed to have significant levels of antioxidant compounds since they had a strong reducing power, high scavenging rates against DPPH and superoxide anion-free radicals (>90%), and high inhibition rates on lipid peroxidation (>60%). The present study will provide reference data for the antioxidant applications of H. erinaceus in pharmaceutical use and disease prevention. PMID:27347087

Adulticidal and repellent properties of indigenous plant extracts against Culex quinquefasciatus and Aedes aegypti (Diptera: Culicidae).

PubMed

Govindarajan, Marimuthu; Sivakumar, Rajamohan

2012-05-01

Several diseases are associated to the mosquito-human interaction. Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikunguniya fever, West Nile virus, and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. The adulticidal and repellent activities of crude hexane, ethyl acetate, benzene, chloroform, and methanol extracts of leaf of Eclipta alba and Andrographis paniculata were assayed for their toxicity against two important vector mosquitoes, viz., Culex quinquefasciatus and Aedes aegypti (Diptera: Culicidae). The adult mortality was observed after 24 h of exposure. All extracts showed moderate adulticide effects; however, the highest adult mortality was found in methanol extract of A. paniculata against the adults of C. quinquefasciatus and A. aegypti with the LC(50) and LC(90) values were 149.81, 172.37 ppm and 288.12, 321.01 ppm, respectively. The results of the repellent activity of hexane, ethyl acetate, benzene, chloroform, and methanol extract of E. alba and A. paniculata plants at three different concentrations of 1.0, 2.5, and 5.0 mg/cm(2) were applied on skin of forearm in man and exposed against adult female mosquitoes. In this observation, these two plant crude extracts gave protection against mosquito bites without any allergic reaction to the test person, and also, the repellent activity is dependent on the strength of the plant extracts. These results suggest that the leaf solvent plant extracts have the potential to be used as an ideal ecofriendly approach for the control of mosquitoes. This is the first report on the mosquito adulticidal and repellent activities of the reported E. alba and A. paniculata plants.

Mosquitocidal Effect of Glycosmis pentaphylla Leaf Extracts against Three Mosquito Species (Diptera: Culicidae)

PubMed Central

Ramkumar, Govindaraju; Karthi, Sengodan; Muthusamy, Ranganathan; Suganya, Ponnusamy; Natarajan, Devarajan; Kweka, Eliningaya J.; Shivakumar, Muthugounder S.

2016-01-01

Background The resistance status of malaria vectors to different classes of insecticides used for public health has raised concern for vector control programmes. Alternative compounds to supplement the existing tools are important to be searched to overcome the existing resistance and persistence of pesticides in vectors and the environment respectively. The mosquitocidal effects of Glycosmis pentaphylla using different solvents of acetone, methanol, chloroform and ethyl acetate extracts against three medically important mosquito vectors was conducted. Methods Glycosmis pentaphylla plant leaves were collected from Kolli Hills, India. The WHO test procedures for larval and adult bioassays were used to evaluate extracts against mosquito vectors, and the chemical composition of extracts identified using GC-MS analysis. Results The larvicidal and adulticidal activity of G. pentaphylla plant extracts clearly impacted the three species of major mosquitoes vectors. Acetone extracts had the highest larvicidal effect against An. stephensi, Cx. quinquefasciatus and Ae. aegypti with the LC50 and LC90 values of 0.0004, 138.54; 0.2669, 73.7413 and 0.0585, 303.746 mg/ml, respectively. The LC50 and LC90 adulticide values of G. pentaphylla leaf extracts in acetone, methanol, chloroform and ethyl acetate, solvents were as follows for Cx. quinquefasciatus, An. stephensi and Ae. Aegypti: 2.957, 5.458, 2.708, and 4.777, 3.449, 6.676 mg/ml respectively. The chemical composition of G. pentaphylla leaf extract has been found in 20 active compounds. Conclusions The plant leaf extracts of G. pentaphylla bioactive molecules which are effective and can be developed as an eco-friendly approach for larvicides and adulticidal mosquitoes vector control. Detailed identification and characterization of mosquitocidal effect of individual bioactive molecules ingredient may result into biodegradable effective tools for the control of mosquito vectors. PMID:27391146

Antiplasmodial activity of extracts of Tridax procumbens and Phyllanthus amarus in in vitro Plasmodium falciparum culture systems.

PubMed

Appiah-Opong, R; Nyarko, A K; Dodoo, D; Gyang, F N; Koram, K A; Ayisi, N K

2011-12-01

Aqueous extracts of Tridax procumbens (TP) (Compositae) and Phyllanthus amarus (PA) (Euphorbiaceae) are used in traditional medicine in Ghana to treat malaria. Previous studies have demonstrated the anti-trypanosoma, anti-bacterial and anti-HIV effects of TP and PA. To assess the antiplasmodial activity of extracts of TP and PA. Aqueous extracts of TP and PA were prepared. A portion of each was freeze-dried and the remaining extracted sequentially with ethyl acetate and chloroform. Ethanolic extracts were also prepared. The antiplasmodial activity of the extracts was assessed with the 3H-hypoxanthine assay using chloroquine-resistant (Dd2) Plasmodium falciparum parasites. Chloroquine was used as the reference drug. The modified tetrazolium-based colorimetric assay was also used to evaluate the red blood cell (RBC)-protective/antiplasmodial activities and cytotoxicities of the extracts. Results showed that TP and PA have antiplasmodial activities. The aqueous and ethanolic extracts of PA were the most active, yielding EC50 values of 34.9 µg/ml and 31.2 µg/ml, respectively in the tetrazolium-based assay. The TP and PA produced and IC50 values of 24.8 µg/ml and 11.7 µg/ml, respectively in the hypoxanthine assay. Protection of human RBCs against P. falciparum damage by the extracts highly correlated with their antiplasmodial activities. None of the extracts, within the concentration range (1.9-500 µg/ml) studied produced any overt toxicity to human RBCs. The results indicate that both PA and TP have activities against chloroquine-resistant P. falciparum (Dd2) parasites. The antiplasmodial principles extracted into water and ethanol but not chloroform or ethyl acetate.

Mosquitocidal Effect of Glycosmis pentaphylla Leaf Extracts against Three Mosquito Species (Diptera: Culicidae).

PubMed

Ramkumar, Govindaraju; Karthi, Sengodan; Muthusamy, Ranganathan; Suganya, Ponnusamy; Natarajan, Devarajan; Kweka, Eliningaya J; Shivakumar, Muthugounder S

2016-01-01

The resistance status of malaria vectors to different classes of insecticides used for public health has raised concern for vector control programmes. Alternative compounds to supplement the existing tools are important to be searched to overcome the existing resistance and persistence of pesticides in vectors and the environment respectively. The mosquitocidal effects of Glycosmis pentaphylla using different solvents of acetone, methanol, chloroform and ethyl acetate extracts against three medically important mosquito vectors was conducted. Glycosmis pentaphylla plant leaves were collected from Kolli Hills, India. The WHO test procedures for larval and adult bioassays were used to evaluate extracts against mosquito vectors, and the chemical composition of extracts identified using GC-MS analysis. The larvicidal and adulticidal activity of G. pentaphylla plant extracts clearly impacted the three species of major mosquitoes vectors. Acetone extracts had the highest larvicidal effect against An. stephensi, Cx. quinquefasciatus and Ae. aegypti with the LC50 and LC90 values of 0.0004, 138.54; 0.2669, 73.7413 and 0.0585, 303.746 mg/ml, respectively. The LC50 and LC90 adulticide values of G. pentaphylla leaf extracts in acetone, methanol, chloroform and ethyl acetate, solvents were as follows for Cx. quinquefasciatus, An. stephensi and Ae. Aegypti: 2.957, 5.458, 2.708, and 4.777, 3.449, 6.676 mg/ml respectively. The chemical composition of G. pentaphylla leaf extract has been found in 20 active compounds. The plant leaf extracts of G. pentaphylla bioactive molecules which are effective and can be developed as an eco-friendly approach for larvicides and adulticidal mosquitoes vector control. Detailed identification and characterization of mosquitocidal effect of individual bioactive molecules ingredient may result into biodegradable effective tools for the control of mosquito vectors.

Changes in total phenol, flavonoid contents and anti-Lactobacillus activity of Callisia fragrans due to extraction solvent

NASA Astrophysics Data System (ADS)

Le, Thom; Cao, Diem Kieu; Pham, Thanh Vy; Huynh, Tan Dat; Ta, Nhat Thuy Anh; Nguyen, Ngoc Thao Linh; Nguyen, Huu Thanh; Le, Hue Huong; Bui, Anh Vo; Truong, Dieu-Hien

2018-04-01

Callisia fragrans is a wonder herb with many medicinal properties such as burn, dental diseases, cancer diseases and arthritis in folk medicine. It is noted that the phytochemical constituents and antimicrobial activity of traditional plants depend on not only the extracting method but also the solvent used for extraction. In this study, the effect of five extraction solvents (i.e., distilled water, 80% methanol, 80% ethanol, 80% ethyl acetate, and 80% chloroform) on yield, total phenolic content (TPC) and total flavonoid content (TFC) of Callisia leaves was determined. Besides, changes in anti-Lactobacillus fermentum activity of C. fragrans freeze-dried extract was also evaluated using disk-diffusion method. The recovery percentage of extractable yield of fresh leaves are ranged from 11.93% w/w for distilled water extract to 16.60% w/w for aqueous ethanol extracts. The yield of 80% aqueous methanol extract (16.27% w/w) is only slightly less than that of the ethanol extract. Significant differences were observed among TPC and TFC obtaining by 80% methanol (0.0522% and 0.0335% w/w, respectively) compared to other solvents (p < 0.05). TPC and TFC of C. fragrans extracts increase in the following order: distilled water < 80% chloroform < 80% ethyl acetate < 80% ethanol < 80% methanol. The results revealed that 80% aqueous methanol Calissia extracts has moderate inhibition (9.0 mm of inhibition zone for 1.5 mg/mL of extracts) of L. fermentum compared to standard antibacterial agent. Based on the study results, it can be concluded that the yield, TPC and TFC of C. frgrans extract varied with the extracting solvent. It also showed that Callisia extracts can prevent dental caries by inhibiting the growth of L. fermentum, towards new insights for treatment of dental caries.

Anti-anxiety activity of successive extracts of Angelica archangelica Linn. on the elevated T-maze and forced swimming tests in rats.

PubMed

Kumar, Dinesh; Bhat, Zulfiqar Ali; Shah, M Y

2012-09-01

Angelica archangelica Linn. is widely used in food and liquor preparations and also in Kashmiri folk medicine to reduce anxiety. We evaluated the anxiolytic effect of successive extracts of A. archangelica linn. (SAE) on rats tested in the elevated T-maze test (an animal model of generalized anxiety) at doses that exhibit antidepressant-like activity in humans. A. archangelica (1 kg) was subjected to successive extraction in a soxhlet apparatus with solvents [petroleum ether (40-60 degrees C), chloroform, ethyl acetate, methanol and decoction with water] in order of increasing polarity (yield: 6.9%, 7.3%, 5.1%, 11.88% and 8.2% w/w, respectively). SAE were evaluated for anxiolytic effects using the elevated T-maze and forced swimming tests in rats. Oral dosing of diazepam (1 mg/kg) and extracts (50, 100 and 200 mg/kg) clearly showed an anxiolytic-like profile in the elevated T-maze test: it increased one-way escape and decreased inhibitory avoidance on the first, third and seventh day. In the forced swimming test, imipramine and SAE showed antidepressant- and anxiolytic-like effects as reflected by increased climbing time, swimming time and decreased immobility time on the first, third and seventh day. Aqueous and methanol extracts showed the most, petroleum ether (40-60 degrees C) and chloroform intermediate, and ethyl acetate the least anxiolytic activity (*P<0.05, **P<0.01, ***P< 0.001) in both models. These results suggest the anti-anxiety activity of various extracts of A. archangelica and strongly justify its use in traditional Indian medicine for the treatment of anxiety.

The synthesis, structure and properties of N-acetylated derivatives of ethyl 3-amino-1H-pyrazole-4-carboxylate.

PubMed

Kusakiewicz-Dawid, Anna; Masiukiewicz, Elzbieta; Rzeszotarska, Barbara; Dybała, Izabela; Kozioł, Anna Eugenia; Broda, Małgorzata Anna

2007-05-01

Ethyl 3-amino-1H-pyrazole-4-carboxylate (1) was yielded through total synthesis and reacted with acetic anhydride to give the acetylated products 2-6. Compounds 1-6 were studied with HPLC, X-ray, FT-IR, (1)H-NMR, (13)C-NMR and MS. Acetylation was carried out in solvents of various polarity, namely; chloroform; dioxane; DMF; acetic anhydride, at room temperature and at boiling points; and in the presence and absence of DMAP. The acetylated products are mainly nitrogen atoms in the ring. The position of the ring proton in the solution was based on NOESY; multinuclear HMBC, HSQC spectra and calculations. For equivalent amounts (1-1.5 mol) of acetic anhydride at room temperature two products of monoacetylation are produced in the ring: 2 and 3, ca. 2 : 1 and at the same time only small amount of the third product of monoacetylated, 5 in DMF, as well the product diacetylated, 4. The greatest amount of the product 4 is produced during the reaction with chloroform. However, in this solvent and in dioxane no product 5 is produced. Compound 2 is, largely, formed in dimethylformamide, in the presence DMAP, 0.2 eq. In the presence of this catalytic base, for the first hour, there is a mixture 2 and 3 to the ratio ca. 95 : 5. With 8 eq of Ac(2)O at reflux, after another hour, the compounds 3, 4 and 6 appear about equal amounts. After a longer time, the compound, which appears most in this mixture is triacetylated derivative 6. The structural and spectroscopic characteristics of compounds 1-6 have been given and the methods for their preparation have been provided.

In vitro antimicrobial and antimycobacterial activity and HPLC-DAD screening of phenolics from Chenopodium ambrosioides L.

PubMed

Jesus, Roberta S; Piana, Mariana; Freitas, Robson B; Brum, Thiele F; Alves, Camilla F S; Belke, Bianca V; Mossmann, Natália Jank; Cruz, Ritiel C; Santos, Roberto C V; Dalmolin, Tanise V; Bianchini, Bianca V; Campos, Marli M A; Bauermann, Liliane de Freitas

The main objective of this study was to demonstrate the antimicrobial potential of the crude extract and fractions of Chenopodium ambrosioides L., popularly known as Santa-Maria herb, against microorganisms of clinical interest by the microdilution technique, and also to show the chromatographic profile of the phenolic compounds in the species. The Phytochemical screening revealed the presence of cardiotonic, anthraquinone, alkaloids, tannins and flavonoids. The analysis by HPLC-DAD revealed the presence of rutin in the crude extract (12.5±0.20mg/g), ethyl acetate (16.5±0.37mg/g) and n-butanol (8.85±0.11mg/g), whereas quercetin and chrysin were quantified in chloroform fraction (1.95±0.04 and 1.04±0.01mg/g), respectively. The most promising results were obtained with the ethyl acetate fraction, which inhibited a greater number of microorganisms and presented the lowest values of MIC against Staphylococcus aureus and Enterococcus faecalis (MIC=0.42mg/mL), Pseudomonas aeruginosa (MIC=34.37mg/mL), Paenibacillus apiarus (MIC=4.29mg/mL) and Paenibacillus thiaminolyticus (MIC=4.29mg/mL). Considering mycobacterial inhibition, the best results were obtained by chloroform fraction against M. tuberculosis, M. smegmatis, and M. avium (MIC ranging from 156.25 to 625μg/mL). This study proves, in part, that the popular use of C. ambrosioides L. can be an effective and sustainable alternative for the prevention and treatment of diseases caused by various infectious agents. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Piscicidal activity of leaf and bark extract of Thevetia peruviana plant and their biochemical stress response on fish metabolism.

PubMed

Singh, S K; Yadav, R P; Singh, A

2010-11-01

The leaf and bark of Thevetia peruviana (Family: Apocynaceae) plant was administered for 24 h to the freshwater fish Catla catla (Hamilton) to evaluate their piscicidal activity in laboratory and cemented pond condition. The LC0 values of lef and bark extracts of different solvents (i.e., acetone, diethyl ether, ethyl alcohol, chloroform and carbon tetrachloride) of this plant to fish Catla catla were determined. The LC50 values of acetone leaf extract of Thevetia peruviana plant is 88.80 mg/L (24h) in laboratory condition and 529.38 mg/L (24h) in cemented pond condition; acetone bark extract of this plant is 99.43 mg/L (24h) in laboratory condition and 591.78 mg/L (24h) in cemented pond condition against freshwater fish Catla catla. Similar trend was also observed in case of other solvent (i.e., diethyl ether, ethyl alcohol, chloroform and carbon tetrachloride) of leaf and bark extracts of Thevetia peruviana plant against freshwater fish Catla catla in laboratory and cemented pond conditions. The acetone leaf and bark extract of this plant was very effective in comparison to other solvent extract in both the conditions. So, the biochemical analysis is taken only acetone leaf and bark extract of Thevetia peruviana plant in laboratory condition. Exposure of sub-lethal doses (40% and 80% of LC,) of acetone leaf and bark extract of this plant over 24 h caused significant (P < 0.05) alterations in total protein, free amino acids, DNA & RNA, protease and acid and alkaline phosphatase activity in muscle, liver and gonadal tissues of fish Catla catla in laboratory condition.

Antioxidant and antidermatophytic activities of essential oil and extracts of Metasequoia glyptostroboides Miki ex Hu.

PubMed

Bajpai, Vivek K; Yoon, Jung In; Chul Kang, Sun

2009-06-01

This study was undertaken to assess the antioxidant and antidermatophytic potential of the essential oil and extracts (hexane, chloroform, ethyl acetate and methanol) of Metasequoia glyptostroboides Miki ex Hu. Antioxidant activity was evaluated by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. The free radical scavenging activities of the oil and ethyl acetate extract were found to be superior (IC(50)=9.1 and 14.24 microg/ml, respectively) as compared to butylatedhydroxyanisole (BHA), (IC(50)=18.27 microg/ml). Also the ethyl acetate extract revealed the highest phenolic contents (93.26 mg/g of dry wt) as compared to the other extracts. Further, oil (1250 microg/disc) and extracts (1750 microg/disc) revealed 35.33-67.66 and 18.0-53.3% antidermatophytic effect, respectively, along with their respective MIC values (62.5-500 and 250-4000 microg/ml) against Trichophyton rubrum KCTC 6345, T. rubrum KCTC 6375, T. rubrum KCTC 6352, T. mentagrophytes KCTC 6085, T. mentagrophytes KCTC 6077, T. mentagrophytes KCTC 6316, Microsporum canis KCTC 6591, M. canis KCTC 6348 and M. canis KCTC 6349. The oil also had a strong detrimental effect on spore germination as well as concentration and time-dependent kinetic inhibition of M. canis KCTC 6591.

Effect of different solvents extracts and mode of action of Loktanella spp. Gb03 on toxic dinoflagellate

NASA Astrophysics Data System (ADS)

Hameed, Anmar; Usup, Gires; Ahmad, Asmat

2016-11-01

This study was aimed to evaluate the algicidal activity of Loktanella sp. Gb-03 bacterial extracts against toxic dinoflagellate, using various polar and non-polar solvents. For this purpose, six different solvent extracts were prepared (i.e. methanol, ethyl acetate, hexane, chloroform, acetonitrile and water). Ratio of 1:100 (v:v) (extract to dinoflagellate culture) of each extract was used for preliminary algicidal activity screening against toxic dinoflagellate Coolia malaynesis. Dinoflagellate cells at the stationary phase (1.0 × 103 cells/ mL) were treated with 1% (v/v) of each extract by using 24-well microplate. The plates were then incubated for 24 hours at dinoflagellate culture condition (under a light intensity of 140 µmol m-2s-1 and 12:12 hours light:dark photoperiod). The result of algicidal activity screening showed that all 6 extracts from Loktanella sp. Gb-03 had different ranges of algicidal activity against the toxic dinoflagellates. Ethyl acetate extract showed the highest activity against C. malaynesis and also other harmful dinoflagellate (Alexandrium sp. Alexandrium leei, Alexandrium affine, Alexandrium tamiyavanichi, Alexandrium tamarense, Gambierdiscus belizeanus, and Ostreopsis). This study was the first to explore the algicidal activity of Loktanella sp. Gb-03 extracts against toxic dinoflagellate with ethyl acetate as the best solvent to extract algicidal active compounds.

New seminal variety of Stevia rebaudiana: Obtaining fractions with high antioxidant potential of leaves.

PubMed

Milani, Paula G; Formigoni, Maysa; Dacome, Antonio S; Benossi, Livia; Costa, Cecília E M DA; Costa, Silvio C DA

2017-01-01

The aim of this study was to determine the composition and antioxidant potential of leaves of a new variety of Stevia rebaudiana (Stevia UEM-13). Stevia leaves of UEM-13 contain rebaudioside A as the main glycoside, while most wild Stevia plants contain stevioside. Furthermore can be multiplied by seed, which reduces the cost of plant culture techniques as other clonal varieties are multiplied by buds, requiring sophisticated and expensive seedling production systems. Ethanol and methanol were used in the extraction to determine the bioactive compounds. The methanolic extract was fractionated sequentially with hexane, chloroform, ethyl acetate and isobutanol, and the highest concentration of phenolic compounds and flavonoids was obtained in the ethyl acetate fraction (524.20 mg galic acid equivalent/g; 380.62 µg quercetin equivalent/g). The glycoside content varied greatly among the fractions (0.5% - 65.3%). Higher antioxidant potential was found in the methanol extract and the ethyl acetate fraction with 93.5% and 97.32%, respectively. In addition to being an excellent source for obtaining of extracts rich in glycoside, this new variety can also be used as raw material for the production of extracts or fractions with a significant amount of antioxidant activity and potential to be used as additives in food.

Cosmeceutical effect of ethyl acetate fraction of Kombucha tea by intradermal administration in the skin of aged mice.

PubMed

Pakravan, Nafiseh; Mahmoudi, Elaheh; Hashemi, Seyed-Ali; Kamali, Jamal; Hajiaghayi, Reza; Rahimzadeh, Mitra; Mahmoodi, Vajiheh

2017-11-19

Natural ingredients have been always an interesting approach to prolong youthful appearance of skin. One of the natural compounds is Kombucha tea (KT), which has been mainly used as an energy drink in Asian countries for a long time. Previous reports indicated that it has pharmaceutical and favorable wound repairing effects. The beneficial properties of KT are thought to be mainly due to the presence of fermentation products such as flavonoids and other polyphenols with inhibition of hydrolytic and oxidative enzymes and anti-inflammatory effects. These properties prompted us to study the anti-aging potential of KT and investigate its effective fraction in aged mice, METHODS: Kombucha tea was fractionated into chloroform, butanol, and ethyl acetate, and flavonoid content was determined. Young and old mice were used as control. KT ethyl acetate fraction (KEAf), which had the highest flavonoid content, was intradermally administered to old mice. Administration of KEAf significantly increased the collagen content, NAD + /NADH level, and concomitantly improved skin connective tissue abnormalities in the aged skin. No sensitivity or irritation was observed. This finding suggested that KEAf can be a suitable candidate as a cosmetic product to improve aging-related skin abnormalities and regeneration of aged skin. © 2017 Wiley Periodicals, Inc.

Comparative evaluation of different extracts of leaves of Psidium guajava Linn. for hepatoprotective activity.

PubMed

Roy, Chanchal K; Das, Amit Kumar

2010-01-01

The study was designed to evaluate the hepatoprotective activity of different extracts (petroleum ether, chloroform, ethyl acetate, methanol and aqueous) of P. guajava in acute experimental liver injury induced by carbon tetrachloride and paracetamol. The effects observed were compared with a known hepatoprotective agent, silymarin (100 mg/kg p.o.). In the acute liver damage induced by different hepatotoxins, P. guajava methanolic leaf extract (200 mg/kg, p.o.) significantly reduced the elevated serum levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and bilirubin in carbon tetrachloride and paracetamol induced hepatotoxicity. P. guajava ethyl acetate leaf extract (200 mg/kg, p.o.) significantly reduced the elevated serum levels of aspartate aminotransferase, alanine aminotransferase and bilirubin in carbon tetrachloride induced hepatotoxicity whereas P. guajava aqueous leaf extract (200 mg/kg, p.o.) significantly reduced the elevated serum levels of alkaline phosphatase, alanine aminotransferase and bilirubin in carbon tetrachloride induced hepatotoxicity. P. guajava ethyl acetate and aqueous leaf extracts (200 mg/kg, p.o.) significantly reduced the elevated serum levels of aspartate aminotransferase in paracetamol induced hepatotoxicity. Histological examination of the liver tissues supported the hepatoprotection. It is concluded that the methanolic extract of leaves of Psidium guajava plant possesses better hepatoprotective activity compared to other extracts.

High performance thin layer chromatography fingerprint analysis of guava (Psidium guajava) leaves

NASA Astrophysics Data System (ADS)

Astuti, M.; Darusman, L. K.; Rafi, M.

2017-05-01

High-performance thin layer chromatography (HPTLC) fingerprint analysis is commonly used for quality control of medicinal plants in term of identification and authentication. In this study, we have been developed HPTLC fingerprint analysis for identification of guava (Psidium guajava) leaves raw material. A mixture of chloroform, acetone, and formic acid in the ratio 10:2:1 was used as the optimum mobile phase in HPTLC silica plate and with 13 bands were detected. As reference marker we chose gallic acid (Rf = 0.21) and catechin (Rf = 0.11). The two compound were detected as pale black bands at 366 nm after derivatization with sulfuric acid 10% v/v (in methanol) reagent. Validation of the method was met within validation criteria, so the developed method could be used for quality control of guava leaves.

In vitro antimicrobial and antiprotozoal activities, phytochemical screening and heavy metals toxicity of different parts of Ballota nigra.

PubMed

Ullah, Najeeb; Ahmad, Ijaz; Ayaz, Sultan

2014-01-01

The study was done to assess the phytochemicals (flavonoids, terpenoids, saponins, tannin, alkaloids, and phenol) in different parts (root, stem, and leaves) of Ballota nigra and correlated it to inhibition of microbes (bacteria and fungi), protozoan (Leishmania), and heavy metals toxicity evaluation. In root and stem flavonoids, terpenes and phenols were present in ethanol, chloroform, and ethyl acetate soluble fraction; these were found to be the most active inhibiting fractions against all the tested strains of bacteria, fungi, and leishmania. While in leaves flavonoids, terpenes, and phenols were present in ethanol, chloroform, and n-butanol fractions which were the most active fractions against both types of microbes and protozoan (leishmania) in in vitro study. Ethanol and chloroform fractions show maximum inhibition against Escherichia coli (17 mm). The phytochemical and biological screenings were correlated with the presence of heavy metals in selected plant Ballota nigra. Cr was found above permissible value (above 1.5 mg/kg) in all parts of the plant. Ni was above WHO limit in B. nigra root and leaves (3.35 ± 1.20 mg/kg and 5.09 ± 0.47 mg/kg, respectively). Fe was above permissible value in all parts of B. nigra (above 20 mg/kg). Cd was above permissible value in all parts of the plant (above 0.3 mg/kg). Pb was above WHO limit (above 2 mg/kg) in all parts of Ballota nigra.

In Vitro Antimicrobial and Antiprotozoal Activities, Phytochemical Screening and Heavy Metals Toxicity of Different Parts of Ballota nigra

PubMed Central

Ullah, Najeeb; Ahmad, Ijaz; Ayaz, Sultan

2014-01-01

The study was done to assess the phytochemicals (flavonoids, terpenoids, saponins, tannin, alkaloids, and phenol) in different parts (root, stem, and leaves) of Ballota nigra and correlated it to inhibition of microbes (bacteria and fungi), protozoan (Leishmania), and heavy metals toxicity evaluation. In root and stem flavonoids, terpenes and phenols were present in ethanol, chloroform, and ethyl acetate soluble fraction; these were found to be the most active inhibiting fractions against all the tested strains of bacteria, fungi, and leishmania. While in leaves flavonoids, terpenes, and phenols were present in ethanol, chloroform, and n-butanol fractions which were the most active fractions against both types of microbes and protozoan (leishmania) in in vitro study. Ethanol and chloroform fractions show maximum inhibition against Escherichia coli (17 mm). The phytochemical and biological screenings were correlated with the presence of heavy metals in selected plant Ballota nigra. Cr was found above permissible value (above 1.5 mg/kg) in all parts of the plant. Ni was above WHO limit in B. nigra root and leaves (3.35 ± 1.20 mg/kg and 5.09 ± 0.47 mg/kg, respectively). Fe was above permissible value in all parts of B. nigra (above 20 mg/kg). Cd was above permissible value in all parts of the plant (above 0.3 mg/kg). Pb was above WHO limit (above 2 mg/kg) in all parts of Ballota nigra. PMID:25054139

Determination of efficacy of fingermark enhancement reagents; the use of propyl chloroformate for the derivatization of fingerprint amino acids extracted from paper.

PubMed

Mink, Tineke; Voorhaar, Annelies; Stoel, Reinoud; de Puit, Marcel

2013-09-01

The analysis of the constituents of fingerprints has been described numerous times, mainly with the purpose of determining the aging effect on fingerprints or showing the differences between donors or groups of donors. In this paper we describe the use of derivatized amino acids to determine the efficacy of the visualization reagents 1,8-diazafluoren-9-one (DFO) and ninhydrin. At present certain conditions are used for the application of these reagents, as determined by trial-and-error investigations, to the effect on fingerprints. The recovery of amino acids from a porous surface can be used as a measure for the efficacy of a visualization agent. In this paper we describe a method for the determination of the amount of amino acid left after reaction with well known fingerprint visualization reagents. This will allow a more scientific approach to method development for fingermark enhancement techniques. Furthermore, investigations on the influence of the concentration of fingermark amino acids, the order of application of and exposure time to reagents and the influence of age of the amino acids were carried out. These studies have resulted in a broader understanding of the mechanism involved in visualization of fingermarks using DFO and ninhydrin. Copyright © 2013 Forensic Science Society. Published by Elsevier Ireland Ltd. All rights reserved.

Comprehensive analysis of the lipophilic reactive carbonyls present in biological specimens by LC/ESI-MS/MS.

PubMed

Tomono, Susumu; Miyoshi, Noriyuki; Ohshima, Hiroshi

2015-04-15

A new analytical method has been developed for profiling lipophilic reactive carbonyls (RCs) such as aldehydes and ketones in biological samples using liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) with selected reaction monitoring (SRM). The method consists of several phases, including (1) extraction of lipophilic RCs with a chloroform/methanol mixture; (2) derivatization of the extracted RCs with dansyl hydrazine (DH); and (3) SRM detection of the characteristic product ion of the 5-dimethylaminonaphthalene-1-sulfonyl moiety (m/z 236.1). The analytical results were expressed as RC maps, which allowed for the occurrence and levels of different lipophilic RCs to be visualized. We also developed a highly reproducible and accurate method to extract, purify and derivatize RCs in small volumes of biological specimens. This method was applied to the detection of free RCs in mice plasma samples, and resulted in the detection of more than 400 RCs in samples obtained from C57BL/6J mice. Thirty-four of these RCs were identified by comparison with authentic RCs. This method could be used to investigate the levels of RCs in biological and environmental samples, as well as studying the role of lipid peroxidation in oxidative stress related-disorders and discovering new biomarkers for the early diagnosis of these diseases. Copyright © 2015 Elsevier B.V. All rights reserved.

Larvicidal activity of medicinal plant extracts against Anopheles subpictus & Culex tritaeniorhynchus

PubMed Central

Kamaraj, C.; Bagavan, A.; Elango, G.; Zahir, A. Abduz; Rajakumar, G.; Marimuthu, S.; Santhoshkumar, T.; Rahuman, A. Abdul

2011-01-01

Background & objectives: Mosquitoes transmit serious human diseases, causing millions of deaths every year and the development of resistance to chemical insecticides resulting in rebounding vectorial capacity. Plants may be alternative sources of mosquito control agents. The present study assessed the role of larvicidal activities of hexane, chloroform, ethyl acetate, acetone, and methanol dried leaf and bark extracts of Annona squamosa L., Chrysanthemum indicum L., and Tridax procumbens L. against the fourth instar larvae of malaria vector, Anopheles subpictus Grassi and Japanese encephalitis vector, Culex tritaeniorhynchus Giles (Diptera: Culicidae). Methods: Larvicidal activities of three medicinal plant extracts were studied in the range of 4.69 to 1000 mg/l in the laboratory bioassays against early 4th instar larvae of An. subpictus and Cx. tritaeniorhynchus. The mortality data were subjected to probit analysis to determine the lethal concentrations (LC50 and LC90) to kill 50 and 90 per cent of the treated larvae of the respective species. Results: All plant extracts showed moderate effects after 24 h of exposure; however, the highest toxic effect of bark methanol extract of A. squamosa, leaf ethyl acetate extract of C. indicum and leaf acetone extract of T. procumbens against the larvae of An. subpictus (LC50 = 93.80, 39.98 and 51.57 mg/l) and bark methanol extract of A. squamosa, leaf methanol extract of C. indicum and leaf ethyl acetate extract of T. procumbens against the larvae of Cx. tritaeniorhynchus (LC50 =104.94, 42.29 and 69.16 mg/l) respectively. Interpretation & Conclusions: Our data suggest that the bark ethyl acetate and methanol extract of A. squamosa, leaf ethyl acetate and methanol extract of C. indicum, acetone and ethyl acetate extract of T. procumbens have the potential to be used as an ecofriendly approach for the control of the An. subpictus, and Cx. tritaeniorhynchus. PMID:21808141

Capillary electrophoresis coupled with chloroform-acetonitrile extraction for rapid and highly selective determination of cysteine and homocysteine levels in human blood plasma and urine.

PubMed

Ivanov, Alexander Vladimirovich; Bulgakova, Polina Olegovna; Virus, Edward Danielevich; Kruglova, Maria Petrovna; Alexandrin, Valery Vasil'evich; Gadieva, Viktoriya Aleksandrovna; Luzyanin, Boris Petrovich; Kushlinskii, Nikolai Evgen'evich; Fedoseev, Anatolij Nikolaevich; Kubatiev, Aslan Amirkhanovich

2017-10-01

A rapid and selective method has been developed for highly sensitive determination of total cysteine and homocysteine levels in human blood plasma and urine by capillary electrophoresis (CE) coupled with liquid-liquid extraction. Analytes were first derivatized with 1,1'-thiocarbonyldiimidazole and then samples were purified by chloroform-ACN extraction. Electrophoretic separation was performed using 0.1 M phosphate with 30 mM triethanolamine, pH 2, containing 25 μM CTAB, 2.5 μM SDS, and 2.5% polyethylene glycol 600. Samples were injected into the capillary (with total length 32 cm and 50 μm id) at 2250 mbar*s and subsequent injection was performed for 30 s with 0.5 M KОН. The total analysis time was less than 9 min, accuracy was 98%, and precision was <2.6%. The LOD was 0.2 μM for homocysteine and 0.5 μM for cysteine. The use of liquid-liquid extraction allowed the precision and sensitivity of the CE method to be significantly increased. The validated method was applied to determine total cysteine and homocysteine content in human blood plasma and urine samples obtained from healthy volunteers and patients with kidney disorders. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Determination of tylosin residues in different animal tissues by high performance liquid chromatography.

PubMed

Prats, C; Francesch, R; Arboix, M; Pérez, B

2002-01-05

A HPLC method to determine and quantify tylosin residues from calves, pigs and poultry is reported. This procedure permitted tylosin to be separated from muscle, liver, kidney and fat after a simple extraction with chloroform or ethyl acetate under basic conditions. The analytical methodology showed a high specificity and sensitivity and an adequate precision and accuracy with a limit of quantification of 50 microg/kg. Eight calves were administered 20 mg/kg/day of tylosin for 5 days and slaughtered at 7 and 14 days post-administration. Results showed that at the 14th day tylosin levels were lower than the MRL in all target tissues.

Thermal Degradation of Small Molecules: A Global Metabolomic Investigation.

PubMed

Fang, Mingliang; Ivanisevic, Julijana; Benton, H Paul; Johnson, Caroline H; Patti, Gary J; Hoang, Linh T; Uritboonthai, Winnie; Kurczy, Michael E; Siuzdak, Gary

2015-11-03

Thermal processes are widely used in small molecule chemical analysis and metabolomics for derivatization, vaporization, chromatography, and ionization, especially in gas chromatography mass spectrometry (GC/MS). In this study the effect of heating was examined on a set of 64 small molecule standards and, separately, on human plasma metabolite extracts. The samples, either derivatized or underivatized, were heated at three different temperatures (60, 100, and 250 °C) at different exposure times (30 s, 60 s, and 300 s). All the samples were analyzed by liquid chromatography coupled to electrospray ionization mass spectrometry (LC/MS) and the data processed by XCMS Online ( xcmsonline.scripps.edu ). The results showed that heating at an elevated temperature of 100 °C had an appreciable effect on both the underivatized and derivatized molecules, and heating at 250 °C created substantial changes in the profile. For example, over 40% of the molecular peaks were altered in the plasma metabolite analysis after heating (250 °C, 300s) with a significant formation of degradation and transformation products. The analysis of 64 small molecule standards validated the temperature-induced changes observed on the plasma metabolites, where most of the small molecules degraded at elevated temperatures even after minimal exposure times (30 s). For example, tri- and diorganophosphates (e.g., adenosine triphosphate and adenosine diphosphate) were readily degraded into a mono-organophosphate (e.g., adenosine monophosphate) during heating. Nucleosides and nucleotides (e.g., inosine and inosine monophosphate) were also found to be transformed into purine derivatives (e.g., hypoxanthine). A newly formed transformation product, oleoyl ethyl amide, was identified in both the underivatized and derivatized forms of the plasma extracts and small molecule standard mixture, and was likely generated from oleic acid. Overall these analyses show that small molecules and metabolites undergo significant time-sensitive alterations when exposed to elevated temperatures, especially those conditions that mimic sample preparation and analysis in GC/MS experiments.

Miscibility and in vitro osteocompatibility of biodegradable blends of poly[(ethyl alanato) (p-phenyl phenoxy) phosphazene] and poly(lactic acid-glycolic acid)

PubMed Central

Deng, Meng; Nair, Lakshmi S.; Nukavarapu, Syam P.; Kumbar, Sangamesh G.; Jiang, Tao; Krogman, Nicholas R.; Singh, Anurima; Allcock, Harry R.; Laurencin, Cato T.

2007-01-01

Previously we demonstrated the ability of ethyl glycinato substituted polyphosphazenes to neutralize the acidic degradation products and control the degradation rate of poly(lactic acid-glycolic acid) by blending. In this study, blends of high strength poly[(50% ethyl alanato) (50% p-phenyl phenoxy) phosphazene] (PNEA50PhPh50) and 85:15 poly(lactic acid-glycolic acid) (PLAGA) were prepared using a mutual solvent approach. Three different solvents, methylene chloride (MC), chloroform (CF) and tetrahydrofuran (THF) were studied to investigate solvent effects on blend miscibility. Three different blends were then fabricated at various weight ratios namely 25:75 (BLEND25), 50:50 (BLEND50), and 75:25 (BLEND75) using THF as the mutual solvent. The miscibility of the blends was evaluated by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and Fourier transform infrared spectroscopy (FTIR). Among these, BLEND25 was miscible while BLEND50 and BLEND75 were partially miscible. Furthermore, BLEND25 formed apatite layers on its surface as evidenced in a biomimetic study performed. These novel blends showed cell adhesion and proliferation comparable to PLAGA. However, the PNEA50PhPh50 component in the blends was able to increase the phenotypic expression and mineralized matrix synthesis of the primary rat osteoblasts (PRO) in vitro. Blends of high strength poly[(50% ethyl alanato) (50% p-phenyl phenoxy) phosphazene] (PNEA50PhPh50) and 85:15 poly(lactic acid-glycolic acid) (PLAGA) are promising biomaterials for a variety of musculoskeletal applications. PMID:17942150

Gas chromatography-mass spectrometry of carbonyl compounds in cigarette mainstream smoke after derivatization with 2,4-dinitrophenylhydrazine.

PubMed

Dong, Ji-Zhou; Moldoveanu, Serban C

2004-02-20

An improved gas chromatography-mass spectrometry (GC-MS) method was described for the analysis of carbonyl compounds in cigarette mainstream smoke (CMS) after 2,4-dinitrophenylhydrazine (DNPH) derivatization. Besides formaldehyde, acetaldehyde, acetone, acrolein, propionaldehyde, methyl ethyl ketone, butyraldehyde, and crotonaldehyde that are routinely analyzed in cigarette smoke, this technique separates and allows the analysis of several C4, C5 and C6 isomeric carbonyl compounds. Differentiation could be made between the linear and branched carbon chain components. In cigarette smoke, the branched chain carbonyls are found at higher level than the linear chain carbonyls. Also, several trace carbonyl compounds such as methoxyacetaldehyde were found for the first time in cigarette smoke. For the analysis, cigarette smoke was collected using DNPH-treated pads, which is a simpler procedure compared to conventional impinger collection. Thermal decomposition of DNPH-carbonyl compounds was minimized by the optimization of the GC conditions. The linear range of the method was significantly improved by using a standard mixture of DNPH-carbonyl compounds instead of individual compounds for calibration. The minimum detectable quantity for the carbonyls ranged from 1.4 to 5.6 microg/cigarette.

Simultaneous in-cell derivatization pressurized liquid extraction for the determination of multiclass preservatives in leave-on cosmetics.

PubMed

Sanchez-Prado, Lucia; Lamas, J Pablo; Lores, Marta; Garcia-Jares, Carmen; Llompart, Maria

2010-11-15

An effective one-step sample preparation methodology for the determination of multiclass preservatives in cosmetics has been developed, applying, for the first time to this kind of matrix, pressurized liquid extraction (PLE) and a very simple, cheap, and fast derivatization procedure: acetylation with acetic anhydride and pyridine. A multifactorial experimental design has been used to evaluate and optimize the main experimental parameters potentially affecting the extraction process. In the final conditions the sample was mixed with Florisil as the dispersing sorbent and extracted with ethyl acetate for 15 min at 120 °C. One of the main goals of this work was to demonstrate the possibility of carrying out direct cosmetic preservative acetylation by simply adding the derivatization reagents into the PLE cell. The extract was then analyzed by GC/MS without any further cleanup or concentration step. The accuracy, precision, linearity, and detection limits (LODs) were evaluated to assess the performance of the proposed method. Quantitative recoveries were obtained, and relative standard deviation values were lower than 10% in all cases. The obtained LODs ranged from 0.000004% to 0.0001% (w/w), values far below the established restrictions in the European Cosmetics Regulation, making this multicomponent analytical method suitable for routine control. Finally, several cosmetic products such as moisturizing and antiwrinkle creams and lotions, hand creams, sunscreen and after-sun creams, baby lotions, and hair care products were analyzed. All the samples contained several of the target cosmetic ingredients, in some cases at quite high concentrations, although the actual European Cosmetics Regulation was fulfilled in all cases.

Simultaneous analyses of cocaine, cocaethylene, and their possible metabolic and pyrolytic products.

PubMed

Cardona, Patrick S; Chaturvedi, Arvind K; Soper, John W; Canfield, Dennis V

2006-02-10

A method was developed for simultaneously analyzing cocaine (COC), benzoylecgonine (BZE), norbenzoylecgonine (BNE), norcocaine (NCOC), ecgonine (ECG), ecgonine methyl ester (EME), m-hydroxybenzoylecgonine (HBZE), anhydroecgonine methyl ester (AEME), cocaethylene (CE), norcocaethylene (NCE), and ecgonine ethyl ester (EEE) in blood, urine, and muscle. Available deuterated analogs of these analytes were used as internal standards. Proteins from blood and muscle homogenate were precipitated with cold acetonitrile. After the removal of acetonitrile by evaporation, the supernatants and urine were subjected to solid-phase extraction. The eluted analytes were converted to their hydrochloride salts and derivatized with pentafluoropropionic anhydride and 2,2,3,3,3-pentafluoro-1-propanol. The derivatized products were analyzed by a gas chromatograph (GC)/mass spectrometer by selected ion monitoring. The limit of detection (LOD) for COC, BZE, NCOC, EME, CE, NCE, and EEE was 2ng/ml, while the LODs for BNE, ECG, HBZE, and AEME were 25, 640, 50, and 13 ng/ml, respectively. This method was successfully applied in analyzing 13 case samples from aviation accident pilot fatalities and motor vehicle operators. AEME concentrations found in the 13 samples were consistent with those produced solely by the GC inlet pyrolysis of COC controls in blood. Anhydroecgonine cannot be used as a marker for the abuse of COC by smoking because it is also pyrolytically produced from COC metabolites on the GC inlet. The developed method can be effectively adopted for analyzing COC and related compounds in urine, blood, and muscle by a single extraction with increased sensitivity through formation of hydrochloride salts and using a one-step derivatization.

Simultaneous derivatization and lighter-than-water air-assisted liquid-liquid microextraction using a homemade device for the extraction and preconcentration of some parabens in different samples.

PubMed

Farajzadeh, Mir Ali; Aghdam, Mehri Bakhshizadeh; Mogaddam, Mohammad Reza Afshar; Nabil, Ali Akbar Alizadeh

2018-06-06

Simultaneous derivatization and air-assisted liquid-liquid microextraction using an organic solvent lighter than water has been developed for the extraction of some parabens in different samples with the aid of a newly designed device for collecting the extractant. For this purpose, the sample solution is transferred into a glass test tube and a few microliters of acetic anhydride (as a derivatization agent) and p-xylene (as an extraction solvent) are added to the solution. After performing the procedure, the homemade device consists of an inverse funnel with a capillary tube placed into the tube. In this step, the collected extraction solvent and a part of the aqueous solution are transferred into the device and the organic phase indwells in the capillary tube of the device. Under the optimal conditions, limits of detection and quantification for the analytes were obtained in the ranges of 0.90-2.7 and 3.0-6.1 ng mL -1 , respectively. The enrichment and enhancement factors were in the ranges of 370-430 and 489-660, respectively. The method precision, expressed as the relative standard deviation, was within the ranges of 4-6% (n = 6) and 4-9% (n = 4) for intra- and inter-day precisions, respectively. The proposed method was successfully used for the determination of methyl-, ethyl-, and propyl parabens in cosmetic, hygiene, and food samples, and personal care products. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Development of an indirect ELISA for the determination of ethyl carbamate in Chinese rice wine.

PubMed

Luo, Lin; Lei, Hong-Tao; Yang, Jin-Yi; Liu, Gong-Liang; Sun, Yuan-Ming; Bai, Wei-Dong; Wang, Hong; Shen, Yu-Dong; Chen, Sui; Xu, Zhen-Lin

2017-01-15

The widespread occurrence of ethyl carbamate (EC, 89.09 Da), a group 2A carcinogen, in fermented foods and alcoholic beverages has raised worldwide public health concern. Immunoassay for EC is unavailable due to the simple and small structure of EC. In this work, an initial attempt to produce antibody specific for EC, by using 4-((ethoxycarbonyl)amino)butanoic acid as hapten, was made but failed. However, since EC can easily react with 9-xanthydrol to form xanthyl ethyl carbamate (XEC), two haptens based on XEC structure were designed and synthesized. Polyclonal antibody against XEC, instead of EC was obtained and then used to develop a competitive indirect ELISA for EC via a pre-analysis derivatization. After optimization, the ciELISA was applied in analyzing Chinese rice wine with detection limit of 166 μg/L, and negligible cross-reactivity with EC analogs. Recoveries of EC in fortified samples were from 84.4% to 100.9%, with coefficients of variation below 10%. Results for analysis of real samples by the ci-ELISA correlated well with that by reference method GC-MS, suggesting the good accuracy and reproducibility of the proposed method. This is the first report of an immunoassay capable of detecting EC, which is suitable for monitoring EC in a large amount of samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Strategies for the capillary electrophoretic separation of indole alkaloids in Psilocybe semilanceata.

PubMed

Pedersen-Bjergaard, S; Rasmussen, K E; Sannes, E

1998-01-01

While the hallucinogenic mushrooms Psilocybe semilanceata have previously been analyzed for the indole alkaloids psilocybin and baeocystin by capillary zone electrophoresis (CZE) at pH 11.5, the present work focused on the development of an alternative and complementary capillary electrophoretic method for their identification. Owing to their structural similarity and zwitterionic nature, the compounds were difficult to resolve based on different interactions with cationic or anionic micelles. However, while the attempts with micellar electrokinetic chromatography (MEKC) were unsuccessful, rapid derivatization with propyl chloroformate and reanalysis by CZE at pH 11.5 was effective to support identification of the two indole alkaloids. Psilocin was difficult to analyze by CZE at pH 11.5 owing to comigration with the electroosmotic flow. For this compound, the pH of the running buffer was reduced to 7.2 to effectively enhance the electrophoretic mobility.

Effect of particle size in preparative reversed-phase high-performance liquid chromatography on the isolation of epigallocatechin gallate from Korean green tea.

PubMed

Kim, Jung Il; Hong, Seung Bum; Row, Kyung Ho

2002-03-08

To isolate epigallocatechin gallate (EGCG) of catechin compounds from Korean green tea (Bosung, Chonnam), a C18 reversed-phase preparative column (250x22 mm) packed with packings of three different sizes (15, 40-63, and 150 microm) was used. The sample extracted with water was partitioned with chloroform and ethyl acetate to remove the impurities including caffeine. The mobile phases in this experiment were composed of 0.1% acetic acid in water, acetonitrile, methanol and ethyl acetate. The injection volume was fixed at 400 microl and the flow rate was increased as the particle size becomes larger. The isolation of EGCG with particle size was compared at a preparative scale and the feasibility of separation of EGCG at larger particle sizes was confirmed. The optimum mobile phase composition for separating EGCG was experimentally obtained at the particle sizes of 15 and 40-63 microm in the isocratic mode, but EGCG was not purely separated at the particle size of 150 microm.

Larvicidal and repellent properties of Streptomyces sp. VITJS4 crude extract against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae).

PubMed

Naine, S Jemimah; Devi, C Subathra

2014-01-01

The aim of the present study was to assess the larvicidal and repellent properties of marine Streptomyces sp. VITJS4 crude extracts. The marine soil samples were collected from the Puducherry coast, Tamil Nadu, India. The isolate Streptomyces sp. VITJS4 was taxonomically characterized and identified. The ethyl acetate crude extract tested for larvicidal property showed 100% mortality for all the 3 species after 24 h exposure against the early fourth instar larvae of malarial vector--Anopheles stephensi at 50% and 90% lethal concentration (LC50 = 132.86, LC90 396.14 ppm); dengue vector--Aedes aegypti (LC50 = 112.78, LC90 336.42 ppm) and filariasis vector--Culex quinquefasciatus (LC50 = 156.53, LC90 468.37 ppm). The Streptomyces sp. VITJS4 solvent extracts of hexane, ethyl acetate, benzene, chloroform and methanol were tested for repellent activity against A. stephensi, A. aegypti and C. quinquefasciatus. The ethyl acetate extract showed complete protection for 210 min at 6 mg/cm2 against these mosquito bites. The crude extract was analyzed further for Fourier Transform-infrared spectroscopy (FT-IR) analysis. In addition to the importance of bioactive compounds, the utilization of Streptomyces sp. VITJS4 crude extracts revealed effective larvicidal and repellent activity against the vectors, which perhaps represents a promising tool in the management of mosquito control.

Chemical composition, antibacterial and antioxidant activities of leaf essential oil and extracts of Metasequioa glyptostroboides Miki ex Hu.

PubMed

Bajpai, Vivek K; Al-Reza, Sharif M; Choi, Ung Kyu; Lee, Jong Hwi; Kang, Sun Chul

2009-08-01

The aims of this study were to analyze the chemical composition of leaf essential oil of Metasequioa glyptostroboides Miki, and to test the efficacy of oil and extracts (hexane, chloroform, ethyl acetate and methanol) against food spoilage and food-borne pathogenic bacteria and their antioxidant activity. The GC-MS analysis revealed 49 compounds representing 94.62% of the total oil containing 2-butaneone (30.6%), cyclopentane (15.1%), beta-myrcene (13.29%), cyclobutane (7.67%), furan (3%), valeramide (2.81%), borneol (1.2%), beta-farnesene (1.67%), thymol (1.44%) and alpha-pinene (1.46%) as major components. The oil (1000 microg/disc), and extracts (1500 microg/disc) exhibited promising antibacterial effect as a diameter of zones of inhibition (10-18 and 7-13 mm), respectively. MIC values of oil and the extracts were ranged 125-2000 and 250 to <2000 microg/ml, respectively. Also the oil had strong antibacterial effect on the viable counts. Scanning electron microscopic study demonstrated potential detrimental effect of the oil on the morphology of S. aureus KCTC1916. The free radical scavenging activities of the oil and ethyl acetate extract were found to be 11.32 and 19.12 microg/ml, respectively. Also the ethyl acetate extract revealed the highest phenolic contents (85.17 mg/g of dry wt) as compared to the other extracts.

Effects of different extracts of Eucalyptus camaldulensis on Trichomonas vaginalis parasite in culture medium

PubMed Central

Hassani, Solmaz; Asghari, Gholamreza; Yousefi, Hossseinali; Kazemian, Afsaneh; Rafieiean, Mahmood; Darani, Hossein Yousofi

2013-01-01

Background: Trichomonas vaginalis is considered one of the main causes of vulvovaginitis in women. Metronidazole with vast side effects is now the drug of choice for treatment of this infection. In an attempt to find an alternative drug, the effect of Eucalyptus camaldulensis on this parasite was shown in previous studies. In this investigation, the effect of different extracts of this plant on T. vaginalis in culture medium has been investigated. Materials and Methods: Five different extracts including total extract, diethyl ether, chloroform, ethyl acetate, and water fractions were prepared. The extracts were dried using vacuum rotary evaporator and then they were used for in vitro anti-trichomonas experiments. Results: Crude extract of E. camaldulensis showed 80% growth inhibition (GI) in a concentration of 12.5 mg/ml during 24 h. Diethyl ether extract in a concentration of 25 mg/ml showed 100% GI during 24 h. With ethyl acetate extract, 100% GI was detected with the minimum concentration of 12.5 mg/ml in the first 24 h. Finally, water extract in a concentration of 50 mg/ml showed 80% and 100% GI after 48 and 72 h, respectively. Conclusion: Ethyl acetate fraction is the extract which showed the highest percentage of GI (100%) with the least concentration (12.5 mg/ml) after 24 and 48 h. PMID:24516847

Anti-tuberculosis and cytotoxic evaluation of the seaweed Sargassum boveanum.

PubMed

Akbari, Vajihe; Zafari, Saeed; Yegdaneh, Afsaneh

2018-02-01

Marine seaweeds produce a variety of compounds with different biological activities, including antituberculosis and anticancer effects. The aim of this study was to investigate anti-tuberculosis activity of Sargassum boveanum ( S. boveanum ) and cytotoxicity of different fractions of this seaweed. S. boveanum was collected from Persian Gulf. The plant was extracted by maceration with methanol-ethyl acetate solvent. The extract was evaporated and partitioned by Kupchan method to yield hexane, tricholoroethane, chloroform, and butanol partitions. The anti-tuberculosis activity of the crude extract and toxicity of the fractions were investigated using green fluorescent protein reporter microplate assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay methods, respectively. The cell survivals of HeLa cell were decreased by increasing the concentration of the extracts. The IC 50 values of hexane, tricholoroethane, chloroform, and butanol partitions were 150.3 ± 23.10, 437.0 ± 147.3, 110.4 ± 33.67, and 1025.0 ± 15.20 μg/mL, respectively. The crude extract was not active against tuberculosis. This study reveals that different partitions of S. boveanum have cytotoxic activity against the cancer cell lines.

Simultaneous determination of azathioprine and 6-mercaptopurine in serum by reversed-phase high-performance liquid chromatography.

PubMed

Tsutsumi, K; Otsuki, Y; Kinoshita, T

1982-09-10

The simultaneous determination of azathioprine and its metabolite 6-mercaptopurine in serum by reversed-phase high-performance liquid chromatography is described. 6-Mercaptopurine was converted to a derivative, 6-mercaptopurine-N-ethylmaleimide, which is stable against autoxidation, on reaction with N-ethylmaleimide. Since the N-ethylmaleimide derivative was more hydrophobic than the parent compound, it could be extracted into ethyl acetate together with azathioprine and the derivative was retained on the reversed-phase column better than 6-mercaptopurine. In addition, 6-mercaptopurine-N-ethylmaleimide absorbed at the same wavelength (280 nm) as azathioprine. Consequently, this derivatization procedure enabled the simultaneous extraction, separation, and detection of these compounds.

Enantioselective separation of amino acids as biomarkers indicating life in extraterrestrial environments.

PubMed

Pietrogrande, Maria Chiara

2013-10-01

Traces of prebiotic amino acids, i.e., the building blocks of proteins, are excellent biomarkers that could provide evidence of extinct or extant life in extra-terrestrial environments. In particular, characterization of the enantiomeric excess of amino acids gives relevant information about the biotic or abiotic origin of molecules, because it is generally assumed that life elsewhere could be based on either L or D amino acids, but not both. The analytical procedures used in in-situ space missions for chiral discrimination of amino acids must meet severe requirements imposed by flight conditions: short analysis time, low energy consumption, robustness, storage for long periods under extreme conditions, high efficiency and sensitivity, automation, and remote-control operation. Such methods are based on gas chromatography, high-pressure liquid chromatography, and capillary electrophoresis, usually coupled with mass spectrometry; of these, gas chromatography-mass spectrometry (GC-MS) is the only such combination yet used in space missions. Preliminary in-situ sample derivatization is required before GC-MS analysis to convert amino acids into volatile and thermally stable compounds. The silylation reagent most commonly used, N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide, is unsuitable for detection of homochirality, and alternative derivatization techniques have been developed that preserve the stereochemical configuration of the original compounds and are compatible with spaceflight conditions. These include the reagent N,N-dimethylformamide dimethylacetal, which has already been used in the Rosetta mission, a mixture of alkyl chloroformate, ethanol, and pyridine, a mixture of perfluorinated anhydrides and perfluoro alcohols, and hexafluoroacetone, the first gaseous derivatizing agent. In all the space instruments, solvent extraction of organic matter and chemical derivatization have been combined in a single automatic and remote-controlled procedure in a chemical reactor. Liquid-based separation systems have been used in space missions. In particular, microchip capillary electrophoresis, based on microfluidic lab-on-a-chip systems, enables high-performance chemical analysis of amino acids with low mass and volume equipment and low power and reagent consumption. Coupling with laser-induced fluorescence detectors results in ultra-low limits of detection. This critical review describes applications of the on-board instruments used in the Rosetta mission to comets and in the more recent Mars exploration program, i.e., the Mars Science Laboratory and ExoMars missions.

Sensitivity of infectious bovine rhinotracheitis virus to ether.

PubMed

Crandell, R A; Melloh, A J; Sorlie, P

1975-12-01

The sensitivity of 12 field isolates of infectious boviine rhinotracheitis (IBR) virus and four commercial modified-live infectious bovine rhinotracheitis virus vaccine strains was determined after exposure to 20% ethyl ether (anesthetic) for 16 h at 4 c. The infectivity of five field strains was reduced by varying degrees, whereas seven were found to be resistant. Three vaccine strains were moderately sensitive, and one strain was resistant. Four of the sensitive field strains were isolated from the conjunctiva and the other was isolated from the surface of the epiglottis of natural infective cattle. Strains of virus isolated from fetal tissue and nasal cavity were resistant. All viruses were readily inactivated by chloroform treatment.

Quantitative profiling of O-glycans by electrospray ionization- and matrix-assisted laser desorption ionization-time-of-flight-mass spectrometry after in-gel derivatization with isotope-coded 1-phenyl-3-methyl-5-pyrazolone.

PubMed

Sić, Siniša; Maier, Norbert M; Rizzi, Andreas M

2016-09-07

The potential and benefits of isotope-coded labeling in the context of MS-based glycan profiling are evaluated focusing on the analysis of O-glycans. For this purpose, a derivatization strategy using d0/d5-1-phenyl-3-methyl-5-pyrazolone (PMP) is employed, allowing O-glycan release and derivatization to be achieved in one single step. The paper demonstrates that this release and derivatization reaction can be carried out also in-gel with only marginal loss in sensitivity compared to in-solution derivatization. Such an effective in-gel reaction allows one to extend this release/labeling method also to glycoprotein/glycoform samples pre-separated by gel-electrophoresis without the need of extracting the proteins/digested peptides from the gel. With highly O-glycosylated proteins (e.g. mucins) LODs in the range of 0.4 μg glycoprotein (100 fmol) loaded onto the electrophoresis gel can be attained, with minor glycosylated proteins (like IgAs, FVII, FIX) the LODs were in the range of 80-100 μg (250 pmol-1.5 nmol) glycoprotein loaded onto the gel. As second aspect, the potential of isotope coded labeling as internal standardization strategy for the reliable determination of quantitative glycan profiles via MALDI-MS is investigated. Towards this goal, a number of established and emerging MALDI matrices were tested for PMP-glycan quantitation, and their performance is compared with that of ESI-based measurements. The crystalline matrix 2,6-dihydroxyacetophenone (DHAP) and the ionic liquid matrix N,N-diisopropyl-ethyl-ammonium 2,4,6-trihydroxyacetophenone (DIEA-THAP) showed potential for MALDI-based quantitation of PMP-labeled O-glycans. We also provide a comprehensive overview on the performance of MS-based glycan quantitation approaches by comparing sensitivity, LOD, accuracy and repeatability data obtained with RP-HPLC-ESI-MS, stand-alone nano-ESI-MS with a spray-nozzle chip, and MALDI-MS. Finally, the suitability of the isotope-coded PMP labeling strategy for O-glycan profiling of biological important proteins is demonstrated by comparative analysis of IgA immunoglobulins and two coagulation factors. Copyright © 2016 Elsevier B.V. All rights reserved.

Metabolite extraction from adherently growing mammalian cells for metabolomics studies: optimization of harvesting and extraction protocols.

PubMed

Dettmer, Katja; Nürnberger, Nadine; Kaspar, Hannelore; Gruber, Michael A; Almstetter, Martin F; Oefner, Peter J

2011-01-01

Trypsin/ethylenediaminetetraacetic acid (EDTA) treatment and cell scraping in a buffer solution were compared for harvesting adherently growing mammalian SW480 cells for metabolomics studies. In addition, direct scraping with a solvent was tested. Trypsinated and scraped cell pellets were extracted using seven different extraction protocols including pure methanol, methanol/water, pure acetone, acetone/water, methanol/chloroform/water, methanol/isopropanol/water, and acid-base methanol. The extracts were analyzed by GC-MS after methoximation/silylation and derivatization with propyl chloroformate, respectively. The metabolic fingerprints were compared and 25 selected metabolites including amino acids and intermediates of energy metabolism were quantitatively determined. Moreover, the influence of freeze/thaw cycles, ultrasonication and homogenization using ceramic beads on extraction yield was tested. Pure acetone yielded the lowest extraction efficiency while methanol, methanol/water, methanol/isopropanol/water, and acid-base methanol recovered similar metabolite amounts with good reproducibility. Based on overall performance, methanol/water was chosen as a suitable extraction solvent. Repeated freeze/thaw cycles, ultrasonication and homogenization did not improve overall metabolite yield of the methanol/water extraction. Trypsin/EDTA treatment caused substantial metabolite leakage proving it inadequate for metabolomics studies. Gentle scraping of the cells in a buffer solution and subsequent extraction with methanol/water resulted on average in a sevenfold lower recovery of quantified metabolites compared with direct scraping using methanol/water, making the latter one the method of choice to harvest and extract metabolites from adherently growing mammalian SW480 cells.

Ovicidal and adulticidal potential of leaf and seed extract of Albizia lebbeck (L.) Benth. (Family: Fabaceae) against Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi (Diptera: Culicidae).

PubMed

Govindarajan, Marimuthu; Rajeswary, Mohan

2015-05-01

Several diseases are associated to the mosquito-human interaction. Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikungunya fever, West Nile virus, and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. In the present study, hexane, benzene, chloroform, ethyl acetate, and methanol extracts of leaf and seed of Albizia lebbeck were assayed for their toxicity against three important vector mosquitoes, viz., Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi. The mean percent hatchability of the eggs was observed after 48 h post-treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. All the five solvent extracts showed moderate ovicidal activity; however, the methanol extract showed the highest ovicidal activity. One hundred percent mortality was observed at 250, 200, and 150 ppm for leaf methanol extract and 375, 300, and 225 ppm for seed methanol extract of A. lebbeck against C. quinquefasciatus, Ae. aegypti, and An. stephensi, respectively. The adulticidal activity of plant leaf and seed extracts showed moderate toxic effect on the adult mosquitoes after 24 h of exposure period. However, the highest adulticidal activity was observed in the leaf methanol extract of A. lebbeck against An. stephensi where the LC₅₀ and LC₉₀ values were 65.12 and 117.70 ppm, respectively. Compared to leaf extracts, seeds have low potency against three mosquito species. No mortality was recorded in the control. Our data suggest that the crude hexane, benzene, chloroform, ethyl acetate, and methanol solvent extracts of A. lebbeck have the potential to be used as an eco-friendly approach for the control of the An. stephensi, Ae. aegypti, and C. quinquefasciatus. These results suggest that the leaf and seed extracts have the potential to be used as an ideal eco-friendly approach for the control of mosquitoes.

Chemical composition, iron bioavailability, and antioxidant activity of Kappaphycus alvarezzi (Doty).

PubMed

Fayaz, Mohamed; Namitha, K K; Murthy, K N Chidambara; Swamy, M Mahadeva; Sarada, R; Khanam, Salma; Subbarao, P V; Ravishankar, G A

2005-02-09

Kappaphycus alvarezzi, an edible seaweed from the west coast of India, was analyzed for its chemical composition. It was found that K. alvarezzi is rich in protein (16.24% w/w) and contains a high amount of fiber (29.40% w/w) and carbohydrates (27.4% w/w). K. alvarezzi showed vitamin A activity of 865 mug retinal equivalents/100 g of sample. It contained a higher quantity of unsaturated fatty acids (44.50% of the total), in which relative percentage of oleic acid was 11%, cis-heptadecanoic acid 13.50%, and linoleic acid 2.3% and 37.0% of saturated fatty acids (mainly heptadecanoic acid). K. alvarezziwas also found to be good source of minerals, viz 0.16% of calcium, 0.033% of iron, and 0.016% of zinc, which are essential for various vital biological activities. Bioavailability of iron by in vitro methods showed a higher efficiency in intestinal conditions than in stomach conditions. Ascorbic acid influenced higher bioavailability of iron. Successive extracts of n-hexane, acetone, ethyl acetate, ethanol, and direct extractables of chloroform/methanol (1:1 and 2:1) were screened for antioxidant activity using a beta-carotene linoleic acid model system (B-CLAMS), DPPH (alpha,alpha-diphenyl-beta-picrylhydrazyl) model system and hydroxyl radical scavenging activity. The chloroform/methanol (2:1) extract has shown 82.5% scavenging activity at 1000 ppm. Acetone fraction extracts at the 1000 ppm level showed 63.31% antioxidant activity in beta-carotene linoleic acid system. The acetone extract showed 46.04% scavenging activity at 1000 ppm concentration. In the case of hydroxyl radical scavenging activity, all the extracts showed better activity at the concentrations of 25 and 50 ppm, where at the 50 ppm level ethyl acetate extract showed 76.0%, acetone 75.12%, and hexane 71.15% activity, respectively. Results of this study suggest the utility of K. alvarezzi (Eucheuma) for various nutritional products, including antioxidant for use as health food or nutraceutical supplement.

A gas chromatography-mass spectrometry method for the determination of delta-aminolevulinic acid in plant leaves.

PubMed

Hijaz, Faraj; Killiny, Nabil

2016-05-20

Delta-aminolevulinic (δ-ALA) acid is an important intermediate for tetrapyrroles biosynthesis and it has recently received great attention in plant physiology and human toxicology. However, the colorimetric method which is the most common method for determination of δ-ALA is time consuming and is not specific. In this study, a method for determination of δ-ALA in plant tissues was developed based on the trimethylsilyl (TMS) derivative of the pyrrole formed from the reaction of δ-ALA with ethyl acetoacetate via Knorr condensation. The δ-ALA in the HCl extract was reacted with ethyl acetoacetate to form a pyrrole. Then, the pyrrole compound was extracted using ethyl acetate and the solvent was evaporated to dryness. The dried sample was derivatized to its TMS ester and analyzed using GC-MS. The concentration of δ-ALA in citrus leaves incubated with levulinic acid was also determined by the conventional colorimetric method. The linear range was 10-200ppm in the full scan mode and 0.1-20ppm in the selected ion monitoring (SIM). The limit of detection was 6ppm in the full scan and 0.05ppm in SIM mode, representing a four-fold increase in sensitivity compared to the colorimetric method. The GC-MS method developed in this study is simple, accurate, sensitive, and could also be used to measure δ-ALA in other biological samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Field validation of the dnph method for aldehydes and ketones. Final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Workman, G.S.; Steger, J.L.

1996-04-01

A stationary source emission test method for selected aldehydes and ketones has been validated. The method employs a sampling train with impingers containing 2,4-dinitrophenylhydrazine (DNPH) to derivatize the analytes. The resulting hydrazones are recovered and analyzed by high performance liquid chromatography. Nine analytes were studied; the method was validated for formaldehyde, acetaldehyde, propionaldehyde, acetophenone and isophorone. Acrolein, menthyl ethyl ketone, menthyl isobutyl ketone, and quinone did not meet the validation criteria. The study employed the validation techniques described in EPA method 301, which uses train spiking to determine bias, and collocated sampling trains to determine precision. The studies were carriedmore » out at a plywood veneer dryer and a polyester manufacturing plant.« less

Proximate composition, phytochemical analysis, and in vitro antioxidant potentials of extracts of Annona muricata (Soursop).

PubMed

Agu, Kingsley C; Okolie, Paulinus N

2017-09-01

Numerous bioactive compounds and phytochemicals have been reported to be present Annona muricata (Soursop). Some of these chemical compounds have been linked to the ethnomedicinal properties of the plant and its antioxidant properties. The aim of this study was to assess the proximate composition, phytochemical constituents and in vitro antioxidant properties of A. muricata using standard biochemical procedures. The defatted Annona muricata crude methanolic extracts of the different parts of the plant were used for the estimation of proximate composition and phytochemical screening. The crude methanolic extracts of the different parts of the plant were also fractionated using solvent-solvent partitioning. Petroleum ether, chloroform, ethyl acetate, methanol, and methanol-water (90:10) were the solvents used for the fractionation. The different fractions obtained were then used to perform in vitro antioxidant analyses including, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability, ferric reducing properties, and hydroxyl radical scavenging ability. The leaf methanolic extract had a higher lipid content, whereas its chloroform fraction demonstrated a better ability to quench DPPH free radical. The root-bark methanol-water, leaf methanol, fruit pulp chloroform, and leaf petroleum ether fractions demonstrated potent ferric reducing properties. The leaf and stem-bark petroleum ether fractions demonstrated better hydroxyl-free radical scavenging abilities. The leaf and fruit pulp of Annona muricata have a very potent antioxidant ability compared to the other parts of the plant. This can be associated with the rich phytochemicals and other phytoconstituents like phenols, flavonoids, alkaloids, and essential lipids, etc. Significant correlations were observed between the antioxidant status and phytochemicals present. These results thus suggest that some of the reported ethnomedicinal properties of this plant could be due to its antioxidant potentials.

Photophysical and electrochemical investigation of highly conjugated pyridine based diphenylamine materials

NASA Astrophysics Data System (ADS)

Mahesh, K.; Priyanka, V.; Vijai Anand, A. S.; Karpagam, S.

2018-02-01

Three simple and small donor-acceptor type conjugated moieties, namely (2Z, 2‧Z)-3,3'-((hexylazanediyl)bis (4,1-phenylene))bis (2-(pyridin-2-yl)acrylonitrile) (DPA-PA-1), (2Z, 2‧Z)-3,3'-((dodecylazanediyl)bis (4,1-phenylene))bis (2-(pyridin-2-yl)acrylonitrile) (DPA-PA-2), (2Z,2‧Z)-3,3'-((allylazanediyl)bis (4,1-phenylene))bis (2-(pyridin-2-yl)acrylonitrile) DPA-PA-3 have been synthesized according to the Knoevenagel condensation. Here alkyl (hexyl, dodecyl, allyl) diphenylamine (DPA) moieties acts as an electron donor and pyridine-acetonitrile (PA) moiety acts as an electron acceptor. These moieties are recently showing great interest in optoelectronic applications. The structures of the DPA-PA-1-3 were confirmed by FT-IR, 1H NMR and 13C NMR. The final products showed great solubility in common organic solvents such as toluene, tetrahydrofuran, ethyl acetate, dichloromethane, chloroform etc due to the alkyl chains. The absorption maximum of DPA-PA-1-3 appeared at 440, 433, 447 nm in chloroform solution. The optical band gaps are 2.33, 2.29, and 2.25 eV calculated from thin film absorption edges. The photoluminescence spectra of three molecules were exhibited a maximum peak at 511, 513, 529 nm with greenish fluorescence in chloroform solution and 553, 541, 554 nm as in thin film state. DPA-PA-1-3 showed a delay fluorescence decay time (τ1) of 35, 16 and 14 μs respectively. The lower electrochemical band gaps 1.90 and 1.80 eV was observed by cyclic voltammetry. The morphological images were indicated that spherical shaped particles were observed with lower surface roughness. These types of low bandgap materials have much attention for their various potential applications in optoelectronic devices.

Neuroprotective effect of Alpinia galanga (L.) fractions on Aβ(25-35) induced amnesia in mice.

PubMed

Hanish Singh, J C; Alagarsamy, V; Diwan, Prakash V; Sathesh Kumar, S; Nisha, J C; Narsimha Reddy, Y

2011-10-31

The rhizomes of Alpinia galanga (L.) Willd (Zingiberaceae), a ginger substitute for flavouring food was traditionally used as nervine tonic and stimulant. This investigation is designed to screen cognitive improvement of Alpinia galanga (AG) fractions in Alzheimer's type of amnesia in mice induced by Aβ((25-35)). Alzheimer's disease induced mice treated with fractions (n-hexane, chloroform and ethyl acetate) of AG in 200 and 400mg/kg. Neurotoxicity was induced by intracerebroventricular injection of Aβ((25-35)) on the 14th day of 21 days drug treatment. Open field and water maze were carried to determine habituation memory and hippocampal memory. Na(+)/K(+)-ATPase, acetylcholinesterase (AChE) and antioxidant enzymes (SOD, GPx, catalase and vitamin C) were determined in brain tissue homogenate to estimate the brain biochemical changes and its anti-amnesic potential with intensity of oxidative stress signaling. Further bioactive (chloroform) fraction was eluted through column chromatography to identify the lead molecules. Increased habituation memory and decreased escape latency in behavioral parameter are the indicative of the cognitive enhancement after treatment with Alpinia galanga fractions. Increment in Na(+)/K(+)-ATPase and antioxidant activity depicts brain membrane integrity improvement and free radical scavenging property. AChE level was decreased to improve the cognition by enhancing cholinergic transmission. Anti-amnesic effect was exerted by various fractions of Alpinia galanga. Among all fractions, preeminent neuroprotection was exerted by chloroform fraction, which has compound, 1'δ-1'-acetoxyeugenol acetate and it may be a potential therapeutic agent for Alzheimer's type of amnesia. These results further motivate us to explore the activity of lead compound's anti-amnesic effect on transgenic mice model of AD. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

Bactericidal effect of extracts and metabolites of Robinia pseudoacacia L. on Streptococcus mutans and Porphyromonas gingivalis causing dental plaque and periodontal inflammatory diseases.

PubMed

Patra, Jayanta Kumar; Kim, Eun Sil; Oh, Kyounghee; Kim, Hyeon-Jeong; Dhakal, Radhika; Kim, Yangseon; Baek, Kwang-Hyun

2015-04-08

The mouth cavity hosts many types of anaerobic bacteria, including Streptococcus mutans and Porphyromonas gingivalis, which cause periodontal inflammatory diseases and dental caries. The present study was conducted to evaluate the antibacterial potential of extracts of Robinia pseudoacacia and its different fractions, as well as some of its natural compounds against oral pathogens and a nonpathogenic reference bacteria, Escherichia coli. The antibacterial activity of the crude extract and the solvent fractions (hexane, chloroform, ethyl acetate and butanol) of R. pseudoacacia were evaluated against S. mutans, P. gingivalis and E. coli DH5α by standard micro-assay procedure using conventional sterile polystyrene microplates. The results showed that the crude extract was more active against P. gingivalis (100% growth inhibition) than against S. mutans (73% growth inhibition) at 1.8 mg/mL. The chloroform and hexane fractions were active against P. gingivalis, with 91 and 97% growth inhibition, respectively, at 0.2 mg/mL. None of seven natural compounds found in R. pseudoacacia exerted an antibacterial effect on P. gingivalis; however, fisetin and myricetin at 8 µg/mL inhibited the growth of S. mutans by 81% and 86%, respectively. The crude extract of R. pseudoacacia possesses bioactive compounds that could completely control the growth of P. gingivalis. The antibiotic activities of the hexane and chloroform fractions suggest that the active compounds are hydrophobic in nature. The results indicate the effectiveness of the plant in clinical applications for the treatment of dental plaque and periodontal inflammatory diseases and its potential use as disinfectant for various surgical and orthodontic appliances.

Chemical composition, antioxidant and antigenotoxic activities of different fractions of Gentiana asclepiadea L. roots extract

PubMed Central

Mihailovic, Vladimir; Matic, Sanja; Mišic, Danijela; Solujic, Slavica; Stanic, Snežana; Katanic, Jelena; Mladenovic, Milan; Stankovic, Nevena

2013-01-01

The aim of this study was to evaluate the antioxidant and antigenotoxic activities of chloroform, ethyl acetate and n-butanol fractions obtained from Gentiana asclepiadea L. roots methanolic extract. The main secondary metabolites sweroside, swertiamarin and gentiopicrine were quantified in G. asclepiadea root extracts using HPLC-DAD analysis. Amount of total phenols, flavonoids, flavonols and gallotannins was also determined. The antigenotoxic potential of extracts from roots of G. asclepiadea was assessed using the standard in vivo procedure for the detection of sex linked recessive lethal mutations in Drosophila melanogaster males treated with ethyl methanesulfonate (EMS). The results showed that the most abundant secoiridoid in G. asclepiadea roots was gentiopicrine and its content in the n-butanol fraction (442.89 mg/g) was the highest. Among all extracts, ethyl acetate fraction showed the highest antioxidant activity, as well as total phenolics (146.64 GAE/g), flavonoids (44.62 RUE/g), flavonols (22.71 RUE/g) and gallotannins (0.99 mg GAE/g) content. All the fractions showed antioxidant activity using in vitro model systems and the results have been correlated with total phenolics, flavonoids, flavonols and gallotannins content. In addition to antioxidant activity, G. asclepiadea root extract fractions possess an antigenotoxic effect against DNA damage induced by alkylation with EMS. The antioxidant activity exhibited by G. asclepiadea depended on the phenolic compounds content of the tested extracts, while there was no significant difference in the antigenotoxic potential between fractions. PMID:26622219

Miscibility and in vitro osteocompatibility of biodegradable blends of poly[(ethyl alanato) (p-phenyl phenoxy) phosphazene] and poly(lactic acid-glycolic acid).

PubMed

Deng, Meng; Nair, Lakshmi S; Nukavarapu, Syam P; Kumbar, Sangamesh G; Jiang, Tao; Krogman, Nicholas R; Singh, Anurima; Allcock, Harry R; Laurencin, Cato T

2008-01-01

Previously we demonstrated the ability of ethyl glycinato substituted polyphosphazenes to neutralize the acidic degradation products and control the degradation rate of poly(lactic acid-glycolic acid) (PLAGA) by blending. In this study, blends of high strength poly[(50% ethyl alanato) (50% p-phenyl phenoxy) phosphazene] (PNEA(50)PhPh(50)) and 85:15 PLAGA were prepared using a mutual solvent approach. Three different solvents, methylene chloride (MC), chloroform (CF) and tetrahydrofuran (THF) were studied to investigate solvent effects on blend miscibility. Three different blends were then fabricated at various weight ratios namely 25:75 (BLEND25), 50:50 (BLEND50), and 75:25 (BLEND75) using THF as the mutual solvent. The miscibility of the blends was evaluated by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and Fourier transform infrared spectroscopy (FTIR). Among these, BLEND25 was miscible while BLEND50 and BLEND75 were partially miscible. Furthermore, BLEND25 formed apatite layers on its surface as evidenced in a biomimetic study performed. These novel blends showed cell adhesion and proliferation comparable to PLAGA. However, the PNEA(50)PhPh(50) component in the blends was able to increase the phenotypic expression and mineralized matrix synthesis of the primary rat osteoblasts (PRO) in vitro. Blends of high strength PNEA(50)PhPh(50) and 85:15 PLAGA are promising biomaterials for a variety of musculoskeletal applications.

Antimicrobial activity of Carpolobia lutea extracts and fractions.

PubMed

Nwidu, Lucky L; Nwafor, Paul A; Vilegas, Wagner

2012-01-01

Carpolobia lutea (G. Don) (Polygalaceae) is a tropical medicinal plant putative in traditional medicines against gonorrhea, gingivitis, infertility, antiulcer and malaria. The present study evaluated the antimicrobial, antifungal and antihelicobacter effects of extracts C. lutea leaf, stem and root. The extracts were examined using the disc-diffusion and Microplates of 96 wells containing Muller-Hinton methods against some bacterial strains: Eschericia coli (ATCC 25922), E. coli (ATCC10418), Pseudomonas aeruginosa (ATCC 27853), Staphylococcus aureus (ATCC 25923), Staphyllococus aureus (ATCC 6571), Enterococcus faecalis (ATCC 29212) and Bacillus subtilis (NCTC 8853) and four clinical isolates: one fungi (Candida albican) and three bacteria (Salmonella, Sheigella and staphylococcus aureus). The Gram-positive bacteria: Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 29212), Bacillus subtilis (ATCC 19659) and the Gram-negative bacteria: Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), Cândida albicans (ATCC 18804) and Helicobacter pylori (ATCC 43504). Some of these extracts were found to be active against some tested strains but activity against H. pylori was >1000mg/ml and good fungistatic activity against C. albican. The MIC against C. albican is in the order n-HF > CHF > ETF= EAF.The order of potency of fraction was the ethanol root > n-HF leaf > ethanol fraction stem > chloroform fraction leaf = ethyl acetate fraction leaf. Polyphenols were demonstrated in ethanol fraction, ethyl acetate fraction, crude ethyl acetate extract and ethanol extract, respectively. These polyphenols isolated may partly explain and support the use of C. lutea for the treatment of infectious diseases in traditional Ibibio medicine of Nigeria.

A comparative study of monosaccharide composition analysis as a carbohydrate test for biopharmaceuticals.

PubMed

Harazono, Akira; Kobayashi, Tetsu; Kawasaki, Nana; Itoh, Satsuki; Tada, Minoru; Hashii, Noritaka; Ishii, Akiko; Arato, Teruyo; Yanagihara, Shigehiro; Yagi, Yuki; Koga, Akiko; Tsuda, Yuriko; Kimura, Mikiko; Sakita, Masashi; Kitamura, Satoshi; Yamaguchi, Hideto; Mimura, Hisashi; Murata, Yoshimi; Hamazume, Yasuki; Sato, Takayuki; Natsuka, Shunji; Kakehi, Kazuaki; Kinoshita, Mitsuhiro; Watanabe, Sakie; Yamaguchi, Teruhide

2011-05-01

The various monosaccharide composition analysis methods were evaluated as monosaccharide test for glycoprotein-based pharmaceuticals. Neutral and amino sugars were released by hydrolysis with 4-7N trifluoroacetic acid. The monosaccharides were N-acetylated if necessary, and analyzed by high-performance liquid chromatography (HPLC) with fluorometric or UV detection after derivatization with 2-aminopyridine, ethyl 4-aminobenzoate, 2-aminobenzoic acid or 1-phenyl-3-methyl-5-pyrazolone, or high pH anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Sialic acids were released by mild acid hydrolysis or sialidase digestion, and analyzed by HPLC with fluorometric detection after derivatization with 1,2-diamino-4,5-methylenedioxybenzene, or HPAEC-PAD. These methods were verified for resolution, linearity, repeatability, and accuracy using a monosaccharide standard solution, a mixture of epoetin alfa and beta, and alteplase as models. It was confirmed that those methods were useful for ensuring the consistency of glycosylation. It is considered essential that the analytical conditions including desalting, selection of internal standards, release of monosaccharides, and gradient time course should be determined carefully to eliminate interference of sample matrix. Various HPLC-based monosaccharide analysis methods were evaluated as a carbohydrate test for glycoprotein pharmaceuticals by an inter-laboratory study. Copyright © 2011 The International Association for Biologicals. Published by Elsevier Ltd. All rights reserved.

Determination of ethyl glucuronide in hair: a rapid sample pretreatment involving simultaneous milling and extraction.

PubMed

Mönch, Bettina; Becker, Roland; Nehls, Irene

2014-01-01

A combination of simultaneous milling and extraction known as micropulverized extraction was developed for the quantification of the alcohol marker ethyl glucuronide (EtG) in hair samples using a homogeneous reference material and a mixer mill. Best extraction results from 50 mg of hair were obtained with 2-mL plastic tubes containing two steel balls (∅ = 5 mm), 0.5 mL of water and with an oscillating frequency of 30 s(-1) over a period of 30 min. EtG was quantified employing a validated GC-MS procedure involving derivatization with pentafluoropropionic acid anhydride. This micropulverization procedure was compared with dry milling followed by separate aqueous extraction and with aqueous extraction after manual cutting to millimeter-size snippets. Micropulverization yielded 28.0 ± 1.70 pg/mg and was seen to be superior to manually cutting (23.0 ± 0.83 pg/mg) and equivalent to dry grinding (27.7 ± 1.71 pg/mg) with regard to completeness of EtG extraction. The option to process up to 20 samples simultaneously makes micropulverization especially valuable for the high throughput of urgent samples.

Manipulation of Rumen Microbial Fermentation by Polyphenol Rich Solvent Fractions from Papaya Leaf to Reduce Green-House Gas Methane and Biohydrogenation of C18 PUFA.

PubMed

Jafari, Saeid; Meng, Goh Yong; Rajion, Mohamed Ali; Jahromi, Mohammad Faseleh; Ebrahimi, Mahdi

2016-06-08

Different solvents (hexane, chloroform, ethyl acetate, butanol, and water) were used to identify the effect of papaya leaf (PL) fractions (PLFs) on ruminal biohydrogenation (BH) and ruminal methanogenesis in an in vitro study. PLFs at a concentration of 0 (control, CON) and 15 mg/250 mg dry matter (DM) were mixed with 30 mL of buffered rumen fluid and were incubated for 24 h. Methane (CH4) production (mL/250 mg DM) was the highest (P < 0.05) for CON (7.65) and lowest for the chloroform fraction (5.41) compared to those of other PLFs at 24 h of incubation. Acetate to propionate ratio was the lowest for PLFs compared to that of CON. Supplementation of the diet with PLFs significantly (P < 0.05) decreased the rate of BH of C18:1n-9 (oleic acid; OA), C18:2n-6 (linoleic acid; LA), and C18:3n-3 (α-linolenic acid; LNA) compared to that of CON after 24 h of incubation. Real time PCR indicated that total protozoa and total methanogen population in PLFs decreased (P < 0.05) compared to those of CON.

Performance tests of three types of air-sampling bags on organic solvent vapor retention.

PubMed

Fukui, Yoshinari; Kanemaru, Ai; Nagasawa, Yasuhiro; Kawakami, Takuya; Iwata, Toyoto; Murata, Katsuyuki; Ohashi, Fumiko; Ikeda, Masayuki

2013-01-01

Performance of two new air sampling bags [the transparent bag (TP bag) and the semi-transparent bag (ST bag)] was examined as possible surrogates for the traditional PVF bag (the Ref bag). Solvent vapor mixture of butyl acetate, chloroform, ethyl acetate, isopropyl alcohol and toluene at administrative control levels were introduced to each bag (n=5 for each of the three types), and the decay in the concentrations (by%) was followed by use of a gas auto-sampler - FID-GC system. A trend of time-dependent decay was noted for all types including the Ref bag. When the performance was compared, the TP bag was equal to or even better than the Ref bag. In contrast, the performance of the ST bag was comparable to that of the other two types of bags with regard to toluene and chloroform when the storage time was short, but poorer than others for the other three solvents throughout the test period. The TP bag may be a bag of choice when the storage time is extended (e.g., up to 48 h) although this bag is physically less robust and requires careful handling. The ST bag may be used when analysis will be completed within 24 h.

Antiulcerogenic activity of fractions and 3,15-dioxo-21alpha-hydroxy friedelane isolated from Maytenus robusta (Celastraceae).

PubMed

de Andrade, Sérgio Faloni; Comunello, Eros; Noldin, Vânia Floriani; Monache, Franco Delle; Cechinel Filho, Valdir; Niero, Rivaldo

2008-01-01

The hexane, chloroform, ethyl acetate and aqueous-soluble fractions from leaves of Maytenus robusta (Celastraceae) were evaluated for their protective actions against ethanol-induced gastric lesions in rats. The treatment with all fractions (150 mg/kg) and omeprazol (30 mg/kg) significantly reduced the lesion index, the total lesion area, and the percentage of lesion, in comparison with the control group (p<0.05). Since the ethyl acetate-soluble fraction was found to be most active in the pylorus ligated model, this fraction was further investigated and resulted in the isolation of triterpene 3,15-dioxo-21alpha-hydroxy friedelane. The triterpene was evaluated in the HCl/ethanol-induced ulcer model in mice. In this assay, both the groups treated with 3,15-dioxo-21alpha-hydroxy friedelane and omeprazol, at a dose of 30 mg/kg, presented a significant reduction in lesion index, total lesion area, and in the percentage of the lesion, when compared with the control group (p<0.05). The result suggests that the antiulcer effect observed in the extract and fractions may be attributed, at least in part, to this compound. Further experiments are underway to determine which antiulcer mechanisms involved in gastroprotection.

Marangoni-Benard Convection in a Evaporating Liquid Thin Layer

NASA Technical Reports Server (NTRS)

Chai, An-Ti; Zhang, Nengli

1996-01-01

Marangoni-Benard convection in evaporating liquid thin layers has been investigated through flow visualization and temperature profile measurement. Twelve liquids, namely ethyl alcohol, methanol, chloroform, acetone, cyclohexane, benzine, methylene chloride, carbon tetrachloride, ethyl acetate, n-pentane, silicone oil (0.65 cSt.), and freon-113, were tested and convection patterns in thin layers of these samples were observed. Comparison among these tested samples shows that some liquids are sensitive to surface contamination from aluminum powder but some are not. The latter is excellent to be used for the investigation of surface-tension driven convection through visualization using the tracer. Two sample liquids, alcohol and freon-113 were particularly selected for systematic study. It was found that the wavelength of Benard cells would not change with thickness of the layer when it evaporates at room temperature. Special attention was focused on cases in which a liquid layer was cooled from below, and some interesting results were obtained. Convection patterns were recorded during the evaporation process and the patterns at certain time frame were compared. Benard cells were observed in thin layers with a nonlinear temperature profile and even with a zero or positive temperature gradient. Wavelength of the cells was found to increase as the evaporation progressed.

Effect of solvents on the extraction of natural pigments and adsorption onto TiO2 for dye-sensitized solar cell applications

NASA Astrophysics Data System (ADS)

Al-Alwani, Mahmoud A. M.; Mohamad, Abu Bakar; Kadhum, Abd. Amir H.; Ludin, Norasikin A.

2015-03-01

Nine solvents, namely, n-hexane, ethanol, acetonitrile, chloroform, ethyl-ether, ethyl-acetate, petroleum ether, n-butyl alcohol, and methanol were used to extract natural dyes from Cordyline fruticosa, Pandannus amaryllifolius and Hylocereus polyrhizus. To improve the adsorption of dyes onto the TiO2 particles, betalain and chlorophyll dyes were mixed with methanol or ethanol and water at various ratios. The adsorption of the dyes mixed with titanium dioxide (TiO2) was also observed. The highest adsorption of the C.fruticosa dye mixed with TiO2 was achieved at ratio 3:1 of methanol: water. The highest adsorption of P.amaryllifolius dye mixed with TiO2 was observed at 2:1 of ethanol: water. H.polyrhizus dye extracted by water and mixed with TiO2 demonstrated the highest adsorption among the solvents. All extracted dye was adsorbed onto the surface of TiO2 based on Fourier Transform Infrared Spectroscopy (FTIR) analysis. The inhibition of crystallinity of TiO2 was likewise investigated by X-ray analysis. The morphological properties and composition of dyes were analyzed via SEM and EDX.

Characterization and structure elucidation of antibacterial compound of Streptomyces sp. ECR77 isolated from east coast of India.

PubMed

Thirumurugan, D; Vijayakumar, R

2015-05-01

Forty marine actinobacteria were isolated from the sediments of east coast (Bay of Bengal) region of Tamilnadu, India. Morphologically distinct colonies were primarily tested against fish pathogenic bacteria such as Vibrio cholerae, V. parahaemolyticus, V. alginolyticus, Pseudomonas fluorescens and Aeromonas hydrophila by cross-streak plate method. The secondary metabolites produced by the highly potential strain cultured on starch casein broth were extracted separately with various solvents such as alcohol, ethyl acetate, methanol, petroleum ether and chloroform. The antibacterial assay of the bioactive compounds was tested against the fish pathogenic bacteria by well diffusion method. Of the various solvents used, the ethyl acetate extract of the isolate had good antibacterial activity. The potential strain was identified as Streptomyces labedae by phenotypic, 16S rRNA gene sequence and phylogenetic analysis. Purification of the biologically active compounds by column chromatography led to isolation of 27 fractions. The biologically active fraction was re-chromatographed on a silica gel column to obtain a single active compound, namely N-isopentyltridecanamide. The structure of the compounds was elucidated on the basis of ultra violet, Fourier transform infrared and nuclear magnetic resonance spectra.

Quantitation of pregabalin in dried blood spots and dried plasma spots by validated LC-MS/MS methods.

PubMed

Kostić, Nađa; Dotsikas, Yannis; Jović, Nebojša; Stevanović, Galina; Malenović, Anđelija; Medenica, Mirjana

2015-05-10

In this paper, novel LC-MS/MS methods for the determination of antiepileptic drug pregabalin in dried matrix spots (DMS) are presented. This attractive technique of sample collection in micro amount was utilized in the form of dried blood spots (DBS) and dried plasma spots (DPS). Following a pre-column derivatization procedure, using n-propyl chloroformate in the presence of n-propanol, and consecutive liquid-liquid extraction, derivatized pregabalin and its internal standard, 4-aminocyclohexanecarboxylic acid, were detected in positive ion mode by applying two SRM transitions per analyte. A YMC-Pack Octyl column (50mm×4.0mm, 3μm particle size) maintained at 30°C, was utilized with running mobile phase composed of acetonitrile: 0.15% formic acid (85:15, v/v). Flow rate was 550μL/min and total run time 2min. Established methods were fully validated over the concentration range of 0.200-20.0μg/mL for DBS and 0.400-40.0μg/mL for DPS, respectively, while specificity, accuracy, precision, recovery, matrix-effect, stability, dilution integrity and spot homogeneity were found within acceptance criteria. Validated methods were applied for the determination of pregabalin levels in dried blood and plasma samples obtained from patients with epilepsy, after per os administration of commercial capsules. Comparison of drug level in blood and plasma, as well as correction steps undertaken in order to overcome hematocrit issue, when analyzing DBS, are also given. Copyright © 2015 Elsevier B.V. All rights reserved.

Chiral separation of amino acids in biological fluids by micellar electrokinetic chromatography with laser-induced fluorescence detection.

PubMed

Thorsén, G; Bergquist, J

2000-08-18

A method is presented for the chiral analysis of amino acids in biological fluids using micellar electrokinetic chromatography (MEKC) and laser-induced fluorescence (LIF). The amino acids are derivatized with the chiral reagent (+/-)-1-(9-anthryl)-2-propyl chloroformate (APOC) and separated using a mixed micellar separation system. No tedious pre-purification of samples is required. The excellent separation efficiency and good detection capabilities of the MEKC-LIF system are exemplified in the analysis of urine and cerebrospinal fluid. This is the first time MEKC has been reported for chiral analysis of amino acids in biological fluids. The amino acids D-alanine, D-glutamine, and D-aspartic acid have been observed in cerebrospinal fluid, and D-alanine and D-glutamic acid in urine. To the best of our knowledge no measurements of either D-alanine in cerebrospinal fluid or D-glutamic acid in urine have been presented in the literature before.

Identification of fentanyl metabolites in rat urine by gas chromatography-mass spectrometry with stable-isotope tracers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Goromaru, T.; Matsuura, H.; Furuta, T.

The metabolites of fentanyl (l), which has been widely used as a neuroleptic analgesic agent, were identified in urine of rats by gas chromatography-mass spectrometry combined with a stable-isotope tracer technique. After the oral administration of an equimolar mixture of l and deuterium-labeled l (l/l-d5), the urinary metabolites were extracted with chloroform at pH 9.0. Extracts were derivatized and analyzed by GC/MS. Metabolites were identified by the presence of doublet ion peaks separated by 5 amu, and chemical structures were established from analyses of fragmentation pathways. The metabolites were identified as 4-N-(N-propionylanilino)-piperidine, 4-N-(N-hydroxypropionylanilino)piperidine, 4-N-(N-propionylanilino) hydroxypiperidine, 1-(2-phenethyl)-4-N-(N-hydroxypropionylanilino)piperidine and 1-(2-phenethyl)-4-N-(N-propionylanilino)hydroxypiperidine. These metabolites,more » together with unchanged l, were also detected in urine of rats receiving l/l-d5 intravenously, by selected-ion monitoring of the specific cluster ions.« less

An AAA-DDD triply hydrogen-bonded complex easily accessible for supramolecular polymers.

PubMed

Han, Yi-Fei; Chen, Wen-Qiang; Wang, Hong-Bo; Yuan, Ying-Xue; Wu, Na-Na; Song, Xiang-Zhi; Yang, Lan

2014-12-15

For a complementary hydrogen-bonded complex, when every hydrogen-bond acceptor is on one side and every hydrogen-bond donor is on the other, all secondary interactions are attractive and the complex is highly stable. AAA-DDD (A=acceptor, D=donor) is considered to be the most stable among triply hydrogen-bonded sequences. The easily synthesized and further derivatized AAA-DDD system is very desirable for hydrogen-bonded functional materials. In this case, AAA and DDD, starting from 4-methoxybenzaldehyde, were synthesized with the Hantzsch pyridine synthesis and Friedländer annulation reaction. The association constant determined by fluorescence titration in chloroform at room temperature is 2.09×10(7)  M(-1) . The AAA and DDD components are not coplanar, but form a V shape in the solid state. Supramolecular polymers based on AAA-DDD triply hydrogen bonded have also been developed. This work may make AAA-DDD triply hydrogen-bonded sequences easily accessible for stimuli-responsive materials. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Brazilian red propolis: phytochemical screening, antioxidant activity and effect against cancer cells.

PubMed

de Mendonça, Izabel Cristina Gomes; Porto, Isabel Cristina Celerino de Moraes; do Nascimento, Ticiano Gomes; de Souza, Naiana Soares; Oliveira, José Marcos dos Santos; Arruda, Rodolfo Elleson dos Santos; Mousinho, Kristiana Cerqueira; dos Santos, Aldenir Feitosa; Basílio-Júnior, Irinaldo Diniz; Parolia, Abhishek; Barreto, Francisco Stefânio

2015-10-14

The implementation of new public healthcare models that stimulate the use of natural products from traditional medicine, as a so-called integrated medicine, refers to an approach that use best of both conventional medicine and traditional medicine. Propolis is a widely used natural product by different ancient cultures and known to exhibit biological activities beneficial for health. The large number of studies conducted with propolis had shown that its chemical composition differs as a function of the climate, plant diversity and bee species and plays an important role on its therapeutic properties. The aim of this study was to analyse the phytochemical profile of the ethanolic extract of red propolis (EEP) and its fractionation, antioxidant action of EEP and its fractions hexane, cloroform and ethyl acetate and cytotoxic activity of EEP on human tumour cell lines SF-295 (glioblastoma), OVCAR-8 (ovary) and HCT-116 (colon). EEP was obtained by maceration with absolute ethanol, then it was concentrated in rotaevaporator up to complete evaporation of the solvent. The crude extract was fractionated with hexane, ethyl acetate, chloroform and methanol and they were subjected to phytochemical screening and total phenolic compounds. Antioxidant activity of EEP and fractions was done by means of the 2,2-diphenyl-1-picryhydrazyl (DPPH) method. Biomarkers of red propolis were identified by LC-Orbitrap-FTMS. To assess cytotoxic activity of the extract, cells were exposed to EEP over 72 h. Cell viability was assessed by means of MTT assay. The percentage of cell growth inhibition (IC50) was analysed by means of non-linear regression, and the absorbance values of the various investigated concentrations were subjected to one-factor analysis of variance (ANOVA) followed by Tukey's or Tamhane's tests (α = 0.05). The results obtained using phytochemical screening and LC-Orbitrap-FTMS indicated the presence of phlobaphene tannins, catechins, chalcones, aurones, flavonones, flavonols, xanthones, pentacyclic triterpenoids and guttiferones in Brazilian red propolis. EEP and its hexane, chloroform and ethyl acetate fractions obtained by liquid-liquid partitioning exhibited satisfactory antioxidant percentages. EEP (IC50 < 34.27 μg/mL) exhibited high levels of cytotoxicity on all human tumour cell lines tested when compared to negative control. C-Orbitrap-FTMS was useful to establish the chemical profile of the red propolis. Brazilian red propolis has antioxidant properties and decreases substantially the percentage of cell survival of human tumour cells; thus, it has potential to serve as an anticancer drug.

Microwave-assisted decomplexation and in-situ headspace in-syringe dynamic derivatization of dimethylamine borane with high performance liquid chromatography-fluorescence detection.

PubMed

Muniraj, Sarangapani; Lee, Hua-Kwang; Hsiech, Chunming; Jen, Jen-Fon

2018-02-16

A rapid, sensitive, selective, and simple method for monitoring dimethylamine borane (DMAB) in aqueous sample is proposed by combining microwave-assisted de-complexation, headspace liquid phase in-situ derivatization extraction, and high-performance liquid chromatography-fluorescence detection for the determination of DMAB in samples. The present procedure involves de-complexation of DMAB using microwave irradiation, evolution of dimethylamine (DMA) to the headspace from an alkalized sample solution, and dynamic headspace liquid-phase derivatization extraction (Dy-HS-LPDE) of DMA with 9-fluorenylmethyl chloroformate in a syringe barrel. In addition to the optimal Dy-HS-LPDE and chromatographic parameters described in our previous study, the de-complexation of DMAB by thermal and microwave-assisted procedures and evolution of DMA into the headspace from an alkalized solution and modification of the Dy-HS-LPDE method are thoroughly investigated. The results indicate that complete de-complexation was obtained at 70 °C for 5 min, 30 °C for 10 min, or using microwave irradiation for 30 s at any applied power. It indicates that the DMAB complex easily undergoes de-complexation under microwave irradiation. The linearity range was 0.01-0.5 mg L -1 for DMAB and 0.0077-0.38 mg L -1 for DMA, with a coefficient of determination of 0.9995, and limit of detection of 3 μg L -1 (limit of quantitation of 10 μg L -1 ) for DMAB. The recoveries of DMAB are 95.3% (3.0% RSD) for waste water when spiked 0.05 mg L -1 and 93.5% (5.4% RSD) for the samples spiked with copper and nickel salts (5 mM each in the spiked waste sample). The whole analytical procedure can be completed within 25 min. The results confirm that the present method is a rapid, sensitive, selective, automated, low-cost and eco-friendly procedure to identify DMAB in samples. Copyright © 2018 Elsevier B.V. All rights reserved.

Occurrence of free fatty acids in the phloem sap of different citrus varieties.

PubMed

Valim, Maria Filomena; Killiny, Nabil

2017-06-03

Candidatus Liberibacter asiaticus is a phloem restricted bacterium that causes citrus greening disease or huanglongbing (HLB), a major treat to commercial citrus production in Florida. It is transmitted by the Asian citrus psyllid, a phloem sap-feeding insect. Studies conducted on the composition of citrus phloem sap revealed the presence amino acids, organic acids and sugars and of low amounts of free fatty acids. In the present study, the phloem sap of 12 citrus varieties with different degrees of tolerance to HLB were extracted with ethyl acetate and analyzed by GC-MS after derivatization with boron trifluoride, a fatty acid-specific reagent. Nine free fatty acids were detected in all varieties. Of the 9 fatty acids detected, only capric acid was significantly different among varieties.

Uncovering the structure of human red hair pheomelanin: benzothiazolylthiazinodihydroisoquinolines as key building blocks.

PubMed

Greco, Giorgia; Panzella, Lucia; Verotta, Luisella; d'Ischia, Marco; Napolitano, Alessandra

2011-04-25

Biomimetic oxidation of the pheomelanin precursor 5-S-cysteinyldopa in the presence of Zn(2+) ions led to the isolation of two isomeric products, one of which could be identified as the benzothiazolylthiazinodihydroisoquinoline 5, while the other proved too unstable for a complete characterization. Both these products were converted into more stable oxidized forms, which after ethylchloroformate derivatization were characterized as the ethyl ester/ethoxycarbonyl isoquinolines 8 and 9. Compound 5 exhibited absorption characteristics similar to those of red hair pheomelanin, including a main band around 400 nm in acids. Similarly to red hair pheomelanin and synthetic pigments, 5 afforded on chemical degradation a thiazolylpyridinecarboxylic acid fragment. Model chemical studies allowed the proposal of a formation mechanism for the benzothiazole and dihydroisoquinoline systems in compound 5.

Characterization of Compounds with Tumor-Cell Proliferation Inhibition Activity from Mushroom (Phellinus baumii) Mycelia Produced by Solid-State Fermentation.

PubMed

Zhang, Henan; Shao, Qian; Wang, Wenhan; Zhang, Jingsong; Zhang, Zhong; Liu, Yanfang; Yang, Yan

2017-04-27

The inhibition of tumor-cell proliferationbyan organicsolvent extract from the solid-state fermentation of Phellinus baumii mycelia inoculated in rice medium was investigated in vitro. The active compounds inhibiting tumor-cell proliferation were characterized. Results revealed that all (petroleum ether, chloroform, ethyl acetate, and butanol) fractions inhibited tumor-cell proliferation in a dose-dependent fashion. The ethyl acetate extract had the highest inhibitory effecton tumor-cell proliferation, and the butanol fraction had the lowest. Six compounds were isolated and purified from the ethyl acetate extract of P. baumii mycelia by the tandem application of silica-gel column chromatography (SGCC), high-speed countercurrent chromatography (HSCCC), and preparative HPLC. These compounds were identified by NMR and electrospray ionization-mass spectrometry (ESI-MS) spectroscopic methods as ergosterol (RF1), ergosta-7,22-dien-3β-yl pentadecanoate (RF3), 3,4-dihydroxy benzaldehyde(RF6), inoscavinA (RF7), baicalein(RF10), and 24-ethylcholesta-5,22-dien-3β-ol (RF13). To further clarify the activity of these compounds, the cell-proliferation-inhibition tests of these compounds on various tumor cells were carried out and evaluatedin vitro. Results suggested that compounds RF6, RF7, and RF10 had potent inhibition effects on the proliferation of a series of tumor cell lines, including K562, L1210, SW620, HepG2, LNCaP, and MCF-7cells. These findings indicated that P. baumii mycelia produced by solid-state fermentation in rice canbe used to obtain active compounds with the ability to inhibittumor-cell proliferation.

Experimental consideration of capillary chromatography based on tube radial distribution of ternary mixture carrier solvents under laminar flow conditions.

PubMed

Jinno, Naoya; Hashimoto, Masahiko; Tsukagoshi, Kazuhiko

2011-01-01

A capillary chromatography system has been developed based on the tube radial distribution of the carrier solvents using an open capillary tube and a water-acetonitrile-ethyl acetate mixture carrier solution. This tube radial distribution chromatography (TRDC) system works under laminar flow conditions. In this study, a phase diagram for the ternary mixture carrier solvents of water, acetonitrile, and ethyl acetate was constructed. The phase diagram that included a boundary curve between homogeneous and heterogeneous solutions was considered together with the component ratios of the solvents in the homogeneous carrier solutions required for the TRDC system. It was found that the TRDC system performed well with homogeneous solutions having component ratios of the solvents that were positioned near the homogeneous-heterogeneous solution boundary of the phase diagram. For preparing the carrier solutions of water-hydrophilic/hydrophobic organic solvents for the TRDC system, we used for the first time methanol, ethanol, 1,4-dioxane, and 1-propanol, instead of acetonitrile (hydrophilic organic solvent), as well as chloroform and 1-butanol, instead of ethyl acetate (hydrophobic organic solvent). The homogeneous ternary mixture carrier solutions were prepared near the homogeneous-heterogeneous solution boundary. Analyte mixtures of 2,6-naphthalenedisulfonic acid and 1-naphthol were separated with the TRDC system using these homogeneous ternary mixture carrier solutions. The pressure change in the capillary tube under laminar flow conditions might alter the carrier solution from homogeneous in the batch vessel to heterogeneous, thus affecting the tube radial distribution of the solvents in the capillary tube.

Synthesis, spectroscopic, and molecular structure characterizations of some azo derivatives of 2-hydroxyacetophenone

NASA Astrophysics Data System (ADS)

Albayrak, Çiğdem; Gümrükçüoğlu, İsmail E.; Odabaşoğlu, Mustafa; İskeleli, Nazan Ocak; Ağar, Erbil

2009-08-01

Some novel azo compounds were prepared by the reaction of 2-hydroxyacetophenone with aniline and its substituted derivatives. The structures of synthesized azo compounds were determined by IR, UV-Vis, 1H NMR and 13C NMR spectroscopic techniques and the structures of some of these compounds were also determined by X-ray diffraction studies. Structural analysis using IR in solid state shows that the azo form is favoured in the azo compounds whereas UV-Vis analysis of the azo compounds in solution has shown that there is a azo and ionic form. The azo compounds in the basic solvents dimethylformamide (DMF) and dimethylsulfoxide (DMSO) are both azo and ionic form while these compounds in ethyl alcohol (EtOH) and chloroform (CHCl 3) are only azo form.

Biologically active traditional medicinal herbs from Balochistan, Pakistan.

PubMed

Zaidi, Mudassir A; Crow, Sidney A

2005-01-04

The biological activities of the following four important medicinal plants of Balochistan, Pakistan were checked; Grewia erythraea Schwein f. (Tiliaceae), Hymenocrater sessilifolius Fisch. and C.A. Mey (Lamiaceae), Vincetoxicum stocksii Ali and Khatoon (Asclepiadaceae) and Zygophyllum fabago L. (Zygophyllaceae). The methanolic extracts were fractionated into hexane, ethyl acetate, chloroform, butanol and water. The antifungal and antibacterial activities of these plants were determined against 12 fungal and 12 bacterial strains by agar well diffusion and disk diffusion assays. The extract of Zygophyllum fabago was found to be highly effective against Candida albicans and Escherichia coli. The extract of Vincetoxicum stocksii was also found to be significantly active against Candida albicans, Bacillus subtilis and Bacillus cereus. Extracts of Hymenocrater sessilifolius and Grewia erythraea showed good activity only against Pseudomonas aeruginosa.

Recovery of catechin compounds from Korean tea by solvent extraction.

PubMed

Row, Kyung Ho; Jin, Yinzhe

2006-03-01

Catechin compounds from Korean green tea as potential sources of anticancer and antioxidant components were target materials in this work. The methodologies of solvent extraction and partition were utilized to recover catechin compounds from green tea. The optimum experimental condition was obtained by optimizing operating factors, such as, the extraction solvent, extraction time and operating temperature. After extracting the green tea with water at 80 degrees C for 40 min, the extract was partitioned with water/chloroform, which was best suited to remove caffeine impurity from the extract. Further, the resulting extract was partitioned water/ethyl acetate to deeply purify the catechin compounds of EGC, EC, EGCG and ECG. The experimental result in this work could be extended to preparative HPLC to obtain EGCG on commercial scale.

Modification of agonist binding moiety in hybrid derivative 5/7-{[2-(4-aryl-piperazin-1-yl)-ethyl]-propyl-amino}-5,6,7,8-tetrahydro-naphthalen-1-ol/-2-amino versions: Impact on functional activity and selectivity for dopamine D2/D3 receptors

PubMed Central

Gopishetty, Bhaskar; Zhang, Suhong; Kharkar, Prashant S.; Antonio, Tamara; Reith, Maarten; Dutta, Aloke K.

2013-01-01

The goal of the present study was to explore, in our previously developed hybrid template, the effect of introduction of additional heterocyclic rings (mimicking catechol hydroxyl groups as bioisosteric replacement) on selectivity and affinity for the D3 versus D2 receptor. In addition, we wanted to explore the effect of derivatization of functional groups of the agonist binding moiety in compounds developed by us earlier from the hybrid template. Binding affinity (Ki) of the new compounds was measured with tritiated spiperone as the radioligand and HEK-293 cells expressing either D2 or D3 receptors. Functional activity of selected compounds was assessed in the GTPγS binding assay. In the imidazole series, compound 10a exhibited the highest D3 affinity whereas the indole derivative 13 exhibited similar high D3 affinity. Functionalization of the amino group in agonist (+)-9d with different sulfonamides derivatives improved the D3 affinity significantly with (+)-14f exhibiting the highest affinity. However, functionalization of the hydroxyl and amino groups of 15 and (+)-9d, known agonist and partial agonist, to sulfonate ester and amide in general modulated the affinity. In both cases loss of agonist potency resulted from such derivatization. PMID:23623679

Gas chromatographic–mass spectrometric fragmentation study of phytoestrogens as their trimethylsilyl derivatives: Identification in soy milk and wastewater samples

USGS Publications Warehouse

Ferrar, Imma; Barber, Larry B.; Thurman, E. Michael

2009-01-01

An analytical method for the identification of eight plant phytoestrogens (biochanin A, coumestrol, daidzein, equol, formononetin, glycitein, genistein and prunetin) in soy products and wastewater samples was developed using gas chromatography coupled with ion trap mass spectrometry (GC/MS–MS). The phytoestrogens were derivatized as their trimethylsilyl ethers with trimethylchlorosilane (TMCS) and N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA). The phytoestrogens were isolated from all samples with liquid–liquid extraction using ethyl acetate. Daidzein-d4 and genistein-d4 labeled standards were used as internal standards before extraction and derivatization. The fragmentation patterns of the phytoestrogens were investigated by isolating and fragmenting the precursor ions in the ion-trap and a typical fragmentation involved the loss of a methyl and a carbonyl group. Two characteristic fragment ions for each analyte were chosen for identification and confirmation. The developed methodology was applied to the identification and confirmation of phytoestrogens in soy milk, in wastewater effluent from a soy-milk processing plant, and in wastewater (influent and effluent) from a treatment plant. Detected concentrations of genistein ranged from 50,000 μg/L and 2000 μg/L in soy milk and in wastewater from a soy-plant, respectively, to 20 μg/L and <1 μg/L for influent and effluent from a wastewater treatment plant, respectively.

High Performance Thin layer Chromatography: Densitometry Method for Determination of Rubraxanthone in the Stem Bark Extract of Garcinia cowa Roxb.

PubMed

Hamidi, Dachriyanus; Aulia, Hilyatul; Susanti, Meri

2017-01-01

Garcinia cowa is a medicinal plant widely grown in Southeast Asia and tropical countries. Various parts of this plant have been used in traditional folk medicine. The bark, latex, and root have been used as an antipyretic agent, while fruit and leaves have been used as an expectorant, for indigestion and improvement of blood circulation. This study aims to determine the concentration of rubraxanthone found in ethyl acetate extract of the stem bark of G. cowa by the high-performance thin-layer chromatography (HPTLC). HPTLC method was performed on precoated silica gel G 60 F254 plates using an HPTLC system with a developed mobile-phase system of chloroform: ethyl acetate: methanol: formic acid (86:6:3:5). A volume of 5 μL of standard and sample solutions was applied to the chromatographic plates. The plates were developed in saturated mode of twin trough chamber at room temperature. The method was validated based on linearity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), and specificity. The spots were observed at ultraviolet 243 nm. The linearity of rubraxanthone was obtained between 52.5 and 157.5 ppm/spot. The LOD and LOQ were found to be 4.03 and 13.42 ppm/spot, respectively. The proposed method showed good linearity, precision, accuracy, and high sensitivity. Therefore, it may be applied for the quantification of rubraxanthone in ethyl acetate extract of the stem bark of G. cowa . High performance thin layer chromatography (HPTLC) method provides rapid qualitative and quantitative estimation of rubraxanthone as a marker com¬pound in G. cowa extract used for commercial productRubraxanthone found in ethyl acetate extracts of G. cowa was successfully quantified using HPTLC method. Abbreviations Used : TLC: Thin-layer chromatography, HPTLC: High-performance thin-layer chromatography, LOD: Limit of detection, LOQ: Limit of quantification, ICH: International Conference on Harmonization.

Derivatization of organophosphorus nerve agent degradation products for gas chromatography with ICPMS and TOF-MS detection.

PubMed

Richardson, Douglas D; Caruso, Joseph A

2007-06-01

Separation and detection of seven V-type (venomous) and G-type (German) organophosphorus nerve agent degradation products by gas chromatography with inductively coupled plasma mass spectrometry (GC-ICPMS) is described. The nonvolatile alkyl phosphonic acid degradation products of interest included ethyl methylphosphonic acid (EMPA, VX acid), isopropyl methylphosphonic acid (IMPA, GB acid), ethyl hydrogen dimethylamidophosphate sodium salt (EDPA, GA acid), isobutyl hydrogen methylphosphonate (IBMPA, RVX acid), as well as pinacolyl methylphosphonic acid (PMPA), methylphosphonic acid (MPA), and cyclohexyl methylphosphonic acid (CMPA, GF acid). N-(tert-Butyldimethylsilyl)-N-methyltrifluroacetamide with 1% TBDMSCl was utilized to form the volatile TBDMS derivatives of the nerve agent degradation products for separation by GC. Exact mass confirmation of the formation of six of the TBDMS derivatives was obtained by GC-time of flight mass spectrometry (TOF-MS). The method developed here allowed for the separation and detection of all seven TBDMS derivatives as well as phosphate in less than ten minutes. Detection limits for the developed method were less than 5 pg with retention times and peak area precisions of less than 0.01 and 6%, respectively. This method was successfully applied to river water and soil matrices. To date this is the first work describing the analysis of chemical warfare agent (CWA) degradation products by GC-ICPMS.

Antioxidant activity and total phenolic and flavonoids content variations of leaves extracts of white Horehound (Marrubium vulgare Linné) from three geographical origins.

PubMed

Bouterfas, K; Mehdadi, Z; Elaoufi, M M; Latreche, A; Benchiha, W

2016-11-01

To elucidate the effect of the sampling location of Marrubium vulgare L. leaves on phenolic contents and antioxidant proprieties of flavonoids extracts. M. vulgare L. leaves were collected from three different geographical locations belonging to northwest Algeria: Tessala (mountain region), M'sila forest (coastal region), and Ain Skhouna (steppe region). The flavonoid extraction was achieved using organic solvents with different polarities (methanol, chloroform, ethyl acetate, and hexane). Folin-Ciocalteu colorimetric method was used for quantification of total phenolic contents, and aluminum chloride assay for quantification of total flavonoid contents. The antioxidant properties of flavonoids extract were studied by free l,l-diphenyl-2-picrylhydrazyl radical-scavenging technique. Total phenolic and flavonoids concentrations varied respectively between 40.7 and 160mg gallic acid equivalents/g and 27.4 and 66.3mg catechin equivalents/g. The DPPH free radical-scavenging activity shows that the antioxidant activity of the flavonoid extracts varied significantly (P<0.001) depending on the type of the organic solvent used, and the sampling location. The methanol, chloroform and ethyl acetate extracts exhibited the highest percentages of inhibition unlike to the aqueous and hexane extracts. These percentages are ranged from 54.8 to 98.8% at 1000μg/mL. In general, M'sila forest flavonoids extracts showed the highest free radical inhibition capacity; followed by those of Ain Skhouna and Tessala Mountain. The inhibitory concentration 50 (IC 50 ) ranged from 33.7 to 774μg/mL and often exceeded those recorded by phenolic standards (ascorbic acid, gallic acid, caffeic acid, tannic acid and catechin). The phytochemical screening revealed the presence of some flavonoid classes, such as flavans and flavanols. The results suggested a potent antioxidant activity of M. vulgare flavonoids extracts, which may find its application in feature research for the food and the pharmacological industries. Copyright © 2016 Académie Nationale de Pharmacie. Published by Elsevier Masson SAS. All rights reserved.

Quantitative treatment of the solvent effects on the electronic absorption and fluorescence spectra of acridines and phenazines. The ground and first excited singlet-state dipole moments

NASA Astrophysics Data System (ADS)

Aaron, Jean Jacques; Maafi, Mounir; Párkányi, Cyril; Boniface, Christian

1995-04-01

Electronic absorption and fluorescence excitation and emission spectra of four acridines (acridine, Acridine Yellow, 9-aminoacridine and proflavine) and three phenazines (phenazine, neutral Red and safranine) are determined at room temperature (298 K) in several solvents of various polarities (dioxane, chloroform, ethyl ether, ethyl acetate, 1-butanol, 2-propanol, ethanol, methanol, dimethylformamide, acetonitrile and dimethyl sulfoxide). The effect of the solvent upon the spectral characteristics of the above compounds, is studied. In combination with the ground-state dipole moments of these compounds, the spectral data are used to evaluate their first excited singlet-state dipole moments by means of the solvatochromic shift method (Bakhshiev's and Kawski-Chamma-Viallet's correlations). The theoretical ground and excited singlet-state dipole moments for acridines and phenazines are also calculated as a vector sum of the π-component (obtained by the PPP method) and the σ-component (obtained from σ-bond moments). For most acridines and phenazines under study, the experimental excited singlet-state dipole moments are found to be higher than their ground state counterpart. The application of the Kamlet-Abboud-Taft solvatochromic parameters to the solvent effect on spectral properties of acridine and phenazine derivatives is discussed.

Phenolic-rich extract from the Costa Rican guava (Psidium friedrichsthalianum) pulp with antioxidant and anti-inflammatory activity. Potential for COPD therapy

PubMed Central

Flores, Gema; Dastmalchi, Keyvan; Wu, Shi-Biao; Whalen, Kathleen; Dabo, Abdoulaye J.; Reynertson, Kurt A.; Foronjy, Robert F.; D’Armiento, Jeanine M.; Kennelly, Edward J.

2016-01-01

The potential therapeutic effects of Costa Rican guava (Psidium friedrichsthalianum) extracts for chronic obstructive pulmonary disease were examined. The ethyl acetate fraction displayed the highest antioxidant activity, as compared to the hexane, chloroform, and n-butanol fractions, as well as the crude extract. This fraction was evaluated for its anti-inflammatory activity response relationship against interleukin-8 (IL-8) and inhibition of matrix metalloproteinase-1 (MMP-1) expression before and after treatment with cigarette smoke. The ethyl acetate fraction exhibited inhibitory activity against IL-8 production and MMP-1 expression, showing the most potent inhibitory activities in both assays at 100 μg/mL, and nine compounds (1–9) were found. Phenolic compounds 1-O-trans-cinnamoyl-β-D-glucopyranose (2), ellagic acid (3), myricetin (4), quercitrin (7), and quercetin (9) were identified using standard compounds or literature reports from related species. Compounds 1, 5, 6, and 8 were tentatively identified as 1,5-dimethyl citrate (1), sinapic aldehyde 4-O-β-D-glucopyranose (5), 3,3′,4-tri-O-methylellagic acid-4′-O-D-glucopyranoside (6), and 1,3-O-diferuloylglycerol (8), All nine compounds are reported for the first time in Costa Rican guava. PMID:23790863

Validated determination of primulasaponins in primula root by a high-performance-thin-layer-chromatography densitometric approach.

PubMed

Coran, Silvia A; Mulas, Stefano

2012-11-01

A novel HPTLC-densitometric method was developed for separation and quantitation of primulasaponin I and II in different matrices. HPTLC silica gel 60 F254(S), 20 cm × 10 cm, plates with ethyl acetate:water:formic acid (5:1:1 v/v) as the mobile phase were used. Densitometric determinations were performed in reflectance mode at 540 nm after derivatization with vanillin reagent. The method was validated giving rise to a dependable and high throughput procedure well suited to routine applications. Primulasaponins were quantified in the range of 150-450 ng with RSD of repeatability and intermediate precision between 0.8 and 1.4% and accuracy within the acceptance limits. The method was tested on commercial herbal medicinal preparations claiming to contain primula root extract. Copyright © 2012 Elsevier B.V. All rights reserved.

Some Strychnos spinosa (Loganiaceae) leaf extracts and fractions have good antimicrobial activities and low cytotoxicities.

PubMed

Isa, Adamu Imam; Awouafack, Maurice Ducret; Dzoyem, Jean Paul; Aliyu, Mohammed; Magaji, Rabiu AbduSsalam; Ayo, Joseph Olusegun; Eloff, Jacobus Nicolaas

2014-11-27

Strychnos spinosa Lam. is a deciduous tree used in traditional medicine to treat infectious diseases. This study is designed to determine the antimicrobial, antioxidant and cytotoxic activities of extracts and fractions from leaves of S. spinosa. Extracts were obtained by maceration with acetone, methanol and dichloromethane/methanol (1/1) while fractions were prepared by liquid-liquid fractionation of the acetone extract. A broth serial microdilution method with tetrazolium violet as growth indicator was used to determine the minimum inhibitory concentration (MIC) against fungi, Gram-positive and Gram-negative bacteria. The antioxidant activity was determined using free-radical-scavenging assays, and the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide reduction assay was used to determine cytotoxicity. Four extracts and five fractions had good to weak antimicrobial activity with MICs ranging from 0.04 to >1.25 mg/ml against both fungi and bacteria. The chloroform and ethyl acetate fractions had an MIC of 0.08 mg/ml against Aspergillus fumigatus. The n-butanol fraction had an MIC of 0.04 mg/ml against Cryptococcus neoformans. The hexane and chloroform fractions had an MIC of 0.08 mg/ml against Staphylococcus aureus. The antioxidant activities were much lower than that of the positive controls. Except for the alkaloid extract, all the extracts and fractions had free-radical-scavenging activity (IC50 ranging from 33.66 to 314.30 μg/ml). The cytotoxicity on Vero cells was reasonable to low with LC50 values ranging between 30.56 and 689.39 μg/ml. The acetone extract and the chloroform fraction had the highest antibacterial activity. By solvent-solvent fractionation it was possible to increase the activity against A. fumigatus and to decrease the cytotoxicity leading to a potentially useful product to protect animals against aspergillosis. Our results therefore support the use of S. spinosa leaves in traditional medicine to treat infectious diseases.

Antimycobacterial activity of selected medicinal plants traditionally used in Sudan to treat infectious diseases.

PubMed

Abuzeid, Nadir; Kalsum, Sadaf; Koshy, Richin John; Larsson, Marie; Glader, Mikaela; Andersson, Henrik; Raffetseder, Johanna; Pienaar, Elsje; Eklund, Daniel; Alhassan, Muddathir S; AlGadir, Haidar A; Koko, Waleed S; Schön, Thomas; Ahmed Mesaik, M; Abdalla, Omer M; Khalid, Asaad; Lerm, Maria

2014-11-18

The emergence of multidrug-resistant strains of Mycobacterium tuberculosis underscores the need for continuous development of new and efficient methods to determine the susceptibility of isolates of Mycobacterium tuberculosis in the search for novel antimycobacterial agents. Natural products constitute an important source of new drugs, and design and implementation of antimycobacterial susceptibility testing methods are necessary to evaluate the different extracts and compounds. In this study we have explored the antimycobacterial properties of 50 ethanolic extracts from different parts of 46 selected medicinal plants traditionally used in Sudan to treat infectious diseases. Plants were harvested and ethanolic extracts were prepared. For selected extracts, fractionation with hydrophilic and hydrophobic solvents was undertaken. A luminometry-based assay was used for determination of mycobacterial growth in broth cultures and inside primary human macrophages in the presence or absence of plant extracts and fractions of extracts. Cytotoxicity was also assessed for active fractions of plant extracts. Of the tested extracts, three exhibited a significant inhibitory effect on an avirulent strain of Mycobacterium tubercluosis (H37Ra) at the initial screening doses (125 and 6.25µg/ml). These were bark and leaf extracts of Khaya senegalensis and the leaf extract of Rosmarinus officinalis L. Further fractions of these plant extracts were prepared with n-hexane, chloroform, ethyl acetate, n-butanol, ethanol and water, and the activity of these extracts was retained in hydrophobic fractions. Cytotoxicity assays revealed that the chloroform fraction of Khaya senegalensis bark was non-toxic to human monocyte-derived macrophages and other cell types at the concentrations used and hence, further analysis, including assessment of IC50 and intracellular activity was done with this fraction. These results encourage further investigations to identify the active compound(s) within the chloroform fraction of Khaya senegalensis bark. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

Chemical composition and biological activities of extracts and essential oil of Boswellia dalzielii leaves.

PubMed

Kohoude, Midéko Justin; Gbaguidi, Fernand; Agbani, Pierre; Ayedoun, Marc-Abel; Cazaux, Sylvie; Bouajila, Jalloul

2017-12-01

Boswellia dalzielii Hutch. (Burseraceae) is an aromatic plant. The leaves are used for beverage flavouring. This study investigates the chemical composition and biological activities of various extracts. The essential oil was prepared via hydrodistillation. Identification and quantification were realized via GC-MS and GC-FID. Consecutive extractions (cyclohexane, dichloromethane, ethyl acetate and methanol) were carried out and various chemical groups (phenolics, flavonoids, tannins, antocyanins and sugar) were quantified. The volatile compounds of organic extracts were identified before and after derivatization. Antioxidant, antihyperuricemia, anti-Alzheimer, anti-inflammatory and anticancer activities were evaluated. In the essential oil, 50 compounds were identified, including 3-carene (27.72%) and α-pinene (15.18%). 2,5-Dihydroxy acetophenone and β-d-xylopyranose were identified in the methanol extract. Higher phenolic (315.97 g GAE/kg dry mass) and flavonoid (37.19 g QE/kg dry mass) contents were observed in the methanol extract. The methanol extract has presented remarkable IC 50  =   6.10 mg/L for antiDPPH, 35.10 mg/L for antixanthine oxidase and 28.01 mg/L for anti-5-lipoxygenase. For acetylcholinesterase inhibition, the best IC 50 (76.20 and 67.10 mg/L) were observed, respectively, with an ethyl acetate extract and the essential oil. At 50 mg/L, the dichloromethane extract inhibited OVCAR-3 cell lines by 65.10%, while cyclohexane extract inhibited IGROV-1 cell lines by 92.60%. Biological activities were fully correlated with the chemical groups of the extracts. The ethyl acetate and methanol extracts could be considered as potential alternatives for use in dietary supplements for the prevention or treatment of diseases because of these extracts natural antioxidant, antihyperuricemic and anti-inflammatory activities.

[Study on solubility of Chinese herbal compound by solubility parameter].

PubMed

Wu, Dezhi; Chen, Lihua; Wang, Sen; Zhu, Weifeng; Guan, Yongmei

2010-02-01

To demonstrate the solubility of Chinese herbal compound with solubility parameters. The solubility parameters of Liangfu effective components and Liangfu compound were determined by inverse gas chromatograph (IGC) and group contribution. Hansen ball was plotting by HSPiP, which could be used to investigate the solubility of Liangfu effective components and Liangfu compound in different solvents. And the results were verified by approximate solubility. Liangfu effective components and Liangfu compound could be dissolved in chloroform, ethyl acetate, acetone, octanol and ether, and were slightly soluble in glycerol, methanol, ethanol and propanediol, but could not be dissolved in water. They were all liposoluble, and the results were the same as the test results of the approximate solubility. The solubility of Chinese herbal compound can be expressed by solubility parameters, and it is accurate, convenient and visual.

Antimicrobial activity of Ulopterol isolated from Toddalia asiatica (L.) Lam.: a traditional medicinal plant.

PubMed

Karunai Raj, M; Balachandran, C; Duraipandiyan, V; Agastian, P; Ignacimuthu, S

2012-03-06

The leaves of Toddalia asiatica (L.) Lam. (Rutaceae) are widely used in folk medicine in India to treat various ailments like cough, malaria, indigestion, influenza lung diseases and rheumatism, fever, stomach ailments, cholera and diarrhea. In our earlier communication we have reported the antimicrobial study on the various extracts of the leaves and the isolation and identification of Flindersine, a quinolone alkaloid as the major active principle. In the present study, we report the antibacterial and antifungal activities of Ulopterol, a coumarin isolated as another major active antimicrobial principle. The leaves were successively extracted with hexane, chloroform, ethyl acetate, methanol and water. The extracts were studied for their antimicrobial activity against selected bacteria and fungi by using disc-diffusion method. The ethyl acetate extract which was found to possess highest antimicrobial activity was subjected to activity guided fractionation by column chromatography over silica gel. This resulted in the isolation of the coumarin, Ulopetrol, an active principle besides Flindersine which was reported by us earlier. The structure of the compound was elucidated using physical and spectroscopic data. Flindersine and Ulopterol were quantified by HPLC. Ulopterol showed activity against the bacteria viz. Staphylococcus epidermidis, Enterobacter aerogenes, Shigella flexneri, Klebsiella pneumoniae (ESBL-3967), Escherichia coli (ESBL-3984) and fungi viz. Aspergillus flavus, Candida krusei and Botrytis cinerea. Quantification by HPLC showed the content of Flindersine and Ulopterol to be 0.361% and 0.266% respectively on dry weight basis of the leaves. Ethyl acetate extract (successive extraction) contained Ulopterol, a coumarin, besides Flindersine, a quinolone alkaloid, as a major active principle in the antimicrobial studies. This is the first report of the antimicrobial activity of Ulopterol and also its first report from the plant. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

Blechnum Orientale Linn - a fern with potential as antioxidant, anticancer and antibacterial agent

PubMed Central

2010-01-01

Background Blechnum orientale Linn. (Blechnaceae) is used ethnomedicinally for the treatment of various skin diseases, stomach pain, urinary bladder complaints and sterilization of women. The aim of the study was to evaluate antioxidant, anticancer and antibacterial activity of five solvent fractions obtained from the methanol extract of the leaves of Blechnum orientale Linn. Methods Five solvent fractions were obtained from the methanol extract of B. orientale through successive partitioning with petroleum ether, chloroform, ethyl acetate, butanol and water. Total phenolic content was assessed using Folin-Ciocalteu's method. The antioxidant activity was determined by measuring the scavenging activity of DPPH radicals. Cytotoxic activity was tested against four cancer cell lines and a non-malignant cell using MTT assay. Antibacterial activity was assessed using the disc diffusion and broth microdilution assays. Standard phytochemical screening tests for saponins, tannins, terpenoids, flavonoids and alkaloids were also conducted. Results The ethyl acetate, butanol and water fractions possessed strong radical scavenging activity (IC50 8.6-13.0 μg/ml) and cytotoxic activity towards human colon cancer cell HT-29 (IC50 27.5-42.8 μg/ml). The three extracts were also effective against all Gram-positive bacteria tested: Bacillus cereus, Micrococcus luteus, methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA) and Stapylococcus epidermidis(minimum inhibitory concentration MIC 15.6-250 μg/ml; minimum bactericidal concentration MBC 15.6-250 μg/ml). Phytochemical analysis revealed the presence of flavonoids, terpenoids and tannins. Ethyl acetate and butanol fractions showed highest total phenolic content (675-804 mg gallic acid equivalent/g). Conclusions The results indicate that this fern is a potential candidate to be used as an antioxidant agent, for colon cancer therapy and for treatment of MRSA infections and other MSSA/Gram-positive bacterial infectious diseases. PMID:20429956

Proteomics analysis of altered proteins in kidney of mice with aristolochic acid nephropathy using the fluorogenic derivatization-liquid chromatography-tandem mass spectrometry method.

PubMed

Lin, Chia-En; Chang, Wen-Shin; Lee, Jen-Ai; Chang, Ting-Ya; Huang, Yu-Shen; Hirasaki, Yoshiro; Chen, Hung-Shing; Imai, Kazuhiro; Chen, Shih-Ming

2018-03-01

Aristolochic acid (AA) causes interstitial renal fibrosis, called aristolochic acid nephropathy (AAN). There is no specific indicator for diagnosing AAN, so this study aimed to investigate the biomarkers for AAN using a proteomics method. The C3H/He female mice were given ad libitum AA-distilled water (0.5 mg/kg/day) and distilled water for 56 days in the AA and normal groups, respectively. The AA-induced proteins in the kidney were investigated using a proteomics study, including fluorogenic derivatization with 7-chloro-N-[2-(dimethylamino)ethyl]-2,1,3-benzoxadiazole-4-sulfonamide, followed by high-performance liquid chromatography analysis and liquid chromatography tandem mass spectrometry with a MASCOT database searching system. There were two altered proteins, thrombospondin type 1 (TSP1) and G protein-coupled receptor 87 (GPR87), in the kidney of AA-group mice on day 56. GPR87, a tumorigenesis-related protein, is reported for the first time in the current study. The renal interstitial fibrosis was certainly induced in the AA-group mice under histological examination. Based on the results of histological examination and the proteomics study, this model might be applied to AAN studies in the future. TSP1 might be a novel biomarker for AAN, and the further role of GPR87 leading to AA-induced tumorigenesis should be researched in future studies. Copyright © 2017 John Wiley & Sons, Ltd.

Tin speciation in the femtogram range in open ocean seawater by gas chromatography/inductively coupled plasma mass spectrometry using a shield torch at normal plasma conditions.

PubMed

Tao, H; Rajendran, R B; Quetel, C R; Nakazato, T; Tominaga, M; Miyazaki, A

1999-10-01

A sensitive method for the determination of ultratrace organotin species in seawater is described. The merits and demerits of derivatization methods using Grignard reagent or sodium tetraethylborate (NaBEt4) were evaluated in terms of derivatization efficiency, applicability to the programmed temperature vaporization (PTV) method, and procedural blanks. The sensitivity of the gas chromatography/inductively coupled plasma mass spectrometry (GC/ICPMS) was improved by more than 100-fold by operating the shield torch at normal plasma conditions, compared with that obtained without using it. The absolute detection limit as tin reached subfemtogram (fg) levels. Furthermore, the detection limit in terms of relative concentration was improved 100-fold by using the PTV method, which enabled the injection of a large sample volume of as much as 100 microL without loss of analyte. When the organotin species in seawater were extracted into hexane with a preconcentration factor of 1000 after ethylation with NaBEt4 and a 100 microL aliquot of the extract was injected into the GC, the instrumental detection limit in relative concentration reached 0.01 pg/L in original seawater. Sources of contamination of organotin species during the sample preparation were examined, and a purification method of NaBEt4 was developed. Finally, the method was successfully applied to open ocean seawater samples containing organotin species at the level of 1-100 pg/L.

Tricaproin Isolated From Simarouba glauca Inhibits the Growth of Human Colorectal Carcinoma Cell Lines by Targeting Class-1 Histone Deacetylases

PubMed Central

Jose, Asha; Chaitanya, Motamari V. N. L.; Kannan, Elango; Madhunapantula, SubbaRao V.

2018-01-01

While anticancer properties of Simarouba glauca (SG, commonly known as Paradise tree) are well documented in ancient literature, the underlying mechanisms leading to cancer cell death begin to emerge very recently. The leaves of SG have been used as potential source of anticancer agents in traditional medicine. Recently attempts have been made to isolate anticancer agents from the leaves of SG using solvent extraction, which identified quassinoids as the molecules with tumoricidal activity. However, it is not known whether the anti-cancer potential of SG leaves is just because of quassinoids alone or any other phytochemicals also contribute for the potency of SG leaf extracts. Therefore, SG leaves were first extracted with hexane, chloroform, ethyl acetate, 70% ethanol, water and anti-cancer potential (for inhibiting colorectal cancer (CRC) cells HCT-116 and HCT-15 proliferation) determined using Sulforhodamine-B (SRB) assay. The chloroform fraction with maximal anticancer activity was further fractionated by activity-guided isolation procedure and structure of the most potent compound determined using spectral analysis. Analysis of the structural characterization data showed the presence of tricaproin (TCN). TCN inhibited CRC cells growth in a time- and dose dependent manner but not the normal cell line BEAS-2B. Mechanistically, TCN reduced oncogenic Class-I Histone deacetylases (HDACs) activity, followed by inducing apoptosis in cells. In conclusion, the anti-cancer potential of SG is in part due to the presence of TCN in the leaves. PMID:29593526

An eco-friendly, quick and cost-effective method for the quantification of acrylamide in cereal-based baby foods.

PubMed

Cengiz, Mehmet Fatih; Gündüz, Cennet Pelin Boyacı

2014-09-01

The presence of acrylamide in cereal-based baby foods is a matter of great concern owing to its possible health effects. Derivatization followed by gas chromatography/mass spectrometry (GC/MS) is one of the most common methods to quantify acrylamide. However, it requires the use of toxic chemicals and is time-consuming. The aim of this study was to develop an eco-friendly, rapid and inexpensive method for the determination of acrylamide in cereal-based baby foods. The method involves defatting with n-hexane, extraction into water, precipitation of proteins, bromination, extraction into ethyl acetate and injection into a GC/MS system. The effects of defatting, precipitation, treatment with triethylamine, addition of internal standard and column selection were reviewed. A flow chart for acrylamide analysis was prepared. To evaluate the applicability of the method, 62 different cereal-based baby foods were analyzed. The levels of acrylamide ranged from not detected (below the limit of detection) to 660 µg kg(-1). The method is more eco-friendly and less expensive because it consumes very little solvent relative to other methods using bromine solutions and ethyl acetate. In addition, sample pre-treatment requires no solid phase extraction or concentration steps. The method is recommended for the determination of trace acrylamide in complex cereal-based baby food products. © 2014 Society of Chemical Industry.

Combining mass spectrometry diagnostic and density functional theory calculations for a better understanding of the plasma polymerization of ethyl lactate.

PubMed

Ligot, S; Guillaume, M; Gerbaux, P; Thiry, D; Renaux, F; Cornil, J; Dubois, P; Snyders, R

2014-04-17

The focus of this work is on the growth mechanism of ethyl lactate-based plasma polymer film (ELPPF) that could be used as barrier coatings. In such an application, the ester density of the plasma polymer has to be controlled to tune the degradation rate of the material. Our strategy consists of correlating the plasma chemistry evaluated by RGA mass spectrometry and understanding, via DFT calculations, the chemistry of the synthesized thin films. The theoretical calculations helped us to understand the plasma chemistry in plasma ON and OFF conditions. From these data it is unambiguously shown that the signal m/z 75 can directly be correlated with the precursor density in the plasma phase. The combination of XPS and chemical derivatization experiments reveal that the ester content in the ELPFF can be tailored from 2 to 18 at. % by decreasing the RF power, which is perfectly correlated with the evolution of the plasma chemistry. Our results also highlight that the ELPPF chemistry, especially the ester content, is affected by the plasma mode of operation (continuous or pulsed discharge, at similar injected mean power) for similar ester content in the plasma. This could be related to different energy conditions at the interface of the growing films that could affect the sticking coefficient of the ester-bearing fragments.

Determination of nitrosourea compounds in brain tissue by gas chromatography and electron capture detection.

PubMed

Hassenbusch, S J; Colvin, O M; Anderson, J H

1995-07-01

A relatively simple, high-sensitivity gas chromatographic assay is described for nitrosourea compounds, such as BCNU [1,3-bis(2-chloroethyl)-1-nitrosourea] and MeCCNU [1-(2-chloroethyl)-3-(trans-4-methylcyclohexyl)-1-nitrosourea], in small biopsy samples of brain and other tissues. After extraction with ethyl acetate, secondary amines in BCNU and MeCCNU are derivatized with trifluoroacetic anhydride. Compounds are separated and quantitated by gas chromatography using a capillary column with temperature programming and an electron capture detector. Standard curves of BCNU indicate a coefficient of variance of 0.066 +/- 0.018, a correlation coefficient of 0.929, and an extraction efficiency from whole brain of 68% with a minimum detectable amount of 20 ng in 5-10 mg samples. The assay has been facile and sensitive in over 1000 brain biopsy specimens after intravenous and intraarterial infusions of BCNU.

Procedure for detecting and confirming pentobarbital residues in dog food by gas chromatography/mass spectrometry.

PubMed

Adam, L A; Reeves, V B

1998-01-01

The method described detects and confirms presence of pentobarbital residues in dry, extruded feeds at concentrations of 5-20 ppb. Dried feed is ground to a uniform powder and shaken overnight in methanol. A portion of the methanolic extract is evaporated, and the residue is reconstituted in phosphate-buffered saline. The aqueous extract is cleaned with a solid-phase extraction cartridge designed to extract barbiturate residues from biological matrixes. Dimethyl sulfoxide, tetramethylammonium hydroxide, and iodomethane are added to derivatize pentobarbital, 1,3-Dimethyl-pentobarbital is then acidified with dilute hydrochloric acid and extracted with isooctane. The organic layer is transferred and evaporated under a stream of nitrogen. The residue is reconstituted in a small volume of ethyl acetate for analysis by gas chromatography/mass spectrometry. The limit of detection is approximately 0.7 ppb. The method was validated with pentobarbital-fortified feed samples containing high concentrations of meat and bone meal.

Toxicology of 2,4-dichlorophenoxyacetic acid (2,4-D) and its determination in serum and brain tissue using gas chromatography-electron-capture detection.

PubMed

Oliveira, G H; Palermo-Neto, J

1995-01-01

A gas-liquid chromatographic method with an electron-capture detector was applied for 2,4-dichlorophenoxyacetic acid (2,4-D) determination in the serum and brain tissue of rats acutely intoxicated with the dimethylamine salt of 2,4-D. After extraction with ethyl ether, 2,4-D derivatization was performed using 2-chloroethanol and BCI3. The average recovery values found for serum and brain tissue were 98.5 +/- 4.8 and 93.3 +/- 7.5, respectively. The sensitivity limit of the method was 250 ng/mL for serum and 300 ng/g for brain tissue. The toxic effects of 2,4-D in rats were observed within one-half hour after its oral administration. Results suggest that the toxic mechanism of 2,4-D is related to an action on the central nervous system.

Anti-Inflammatory Effect of Malva sylvestris, Sida cordifolia, and Pelargonium graveolens Is Related to Inhibition of Prostanoid Production.

PubMed

Martins, Cleverson Antonio Ferreira; Campos, Michel Leandro; Irioda, Ana Carolina; Stremel, Dile Pontarolo; Trindade, Angela Cristina Leal Badaró; Pontarolo, Roberto

2017-11-03

The ability of plant extracts and preparations to reduce inflammation has been proven by different means in experimental models. Since inflammation enhances the release of specific mediators, inhibition of their production can be used to investigate the anti-inflammatory effect of plants widely used in folk medicine for this purpose. The study was performed for leaves and flowers of Malva sylvestris , and leaves of Sida cordifolia and Pelargonium graveolens . These are three plant species known in Brazil as Malva. The anti-inflammatory activity of extracts and fractions (hexane, chloroform, ethyl acetate, and residual) was evaluated by quantitation of prostaglandins (PG) PGE₂, PGD₂, PGF 2α , and thromboxane B₂ (the stable nonenzymatic product of TXA₂) concentration in the supernatant of lipopolysaccharide (LPS)- induced RAW 264.7 cells. Inhibition of anti-inflammatory mediator release was observed for plants mainly in the crude extract, ethyl acetate fraction, and residual fraction. The results suggest superior activity of S. cordifolia , leading to significantly lower values of all mediators after treatment with its residual fraction, even at the lower concentration tested (10 μg/mL). M. sylvestris and P. graveolens showed similar results, such as the reduction of all mediators after treatment, with leaf crude extracts (50 μg/mL). These results suggest that the three species known as Malva have anti-inflammatory properties, S. cordifolia being the most potent.

Antioxidant, genotoxic and antigenotoxic activities of daphne gnidium leaf extracts

PubMed Central

2012-01-01

Background Plants play a significant role in maintaining human health and improving the quality of human life. They serve humans well as valuable components of food, as well as in cosmetics, dyes, and medicines. In fact, many plant extracts prepared from plants have been shown to exert biological activity in vitro and in vivo. The present study explored antioxidant and antigenotoxic effects of Daphne gnidium leaf extracts. Methods The genotoxic potential of petroleum ether, chloroform, ethyl acetate, methanol and total oligomer flavonoid (TOF) enriched extracts from leaves of Daphne gnidium, was assessed using Escherichia coli PQ37. Likewise, the antigenotoxicity of the same extracts was tested using the “SOS chromotest test”. Antioxidant activities were studied using non enzymatic and enzymatic method: NBT/Riboflavine and xantine oxidase. Results None of the different extracts produced a genotoxic effect, except TOF extract at the lowest tested dose. Our results showed that D. gnidium leaf extracts possess an antigenotoxic effect against the nitrofurantoin a mutagen of reference. Ethyl acetate and TOF extracts were the most effective in inhibiting xanthine oxidase activity. While, methanol extract was the most potent superoxide scavenger when tested with the NBT/Riboflavine assay. Conclusions The present study has demonstrated that D. gnidium leaf extract possess antioxidant and antigenotoxic effects. These activities could be ascribed to compounds like polyphenols and flavonoid. Further studies are required to isolate the active molecules. PMID:22974481

The BUME method: a new rapid and simple chloroform-free method for total lipid extraction of animal tissue

NASA Astrophysics Data System (ADS)

Löfgren, Lars; Forsberg, Gun-Britt; Ståhlman, Marcus

2016-06-01

In this study we present a simple and rapid method for tissue lipid extraction. Snap-frozen tissue (15-150 mg) is collected in 2 ml homogenization tubes. 500 μl BUME mixture (butanol:methanol [3:1]) is added and automated homogenization of up to 24 frozen samples at a time in less than 60 seconds is performed, followed by a 5-minute single-phase extraction. After the addition of 500 μl heptane:ethyl acetate (3:1) and 500 μl 1% acetic acid a 5-minute two-phase extraction is performed. Lipids are recovered from the upper phase by automated liquid handling using a standard 96-tip robot. A second two-phase extraction is performed using 500 μl heptane:ethyl acetate (3:1). Validation of the method showed that the extraction recoveries for the investigated lipids, which included sterols, glycerolipids, glycerophospholipids and sphingolipids were similar or better than for the Folch method. We also applied the method for lipid extraction of liver and heart and compared the lipid species profiles with profiles generated after Folch and MTBE extraction. We conclude that the BUME method is superior to the Folch method in terms of simplicity, through-put, automation, solvent consumption, economy, health and environment yet delivering lipid recoveries fully comparable to or better than the Folch method.

Preparative separation of phenolic compounds from Halimodendron halodendron by high-speed counter-current chromatography.

PubMed

Wang, Jihua; Gao, Haifeng; Zhao, Jianglin; Wang, Qi; Zhou, Ligang; Han, Jianguo; Yu, Zhu; Yang, Fuyu

2010-08-31

Three phenolic compounds, p-hydroxybenzoic acid (1), isorhamnetin-3-O-β-D-rutinoside (2), and 3,3'-di-O-methylquercetin (5), along with a phenolic mixture were successfully separated from the ethyl acetate crude extract of Halimodendron halodendron by high-speed counter-current chromatography (HSCCC) with chloroform-methanol-water-acetic acid (4:3:2:0.05, v/v) as the two-phase solvent system. The phenolic mixture from HSCCC was further separated by preparative HPLC and purified by Sephadex LH-20 to afford quercetin (3) and 3-O-methylquercetin (4). Seven hundred mg of ethyl acetate crude extract was separated by HSCCC to obtain six fractions which were then analyzed by high performance liquid chromatography (HPLC). The HSCCC separation obtained total of 80 mg of the mixture of quercetin (3) and 3-O-methylquercetin (4) (26.43% and 71.89%, respectively) in fraction 2, 14 mg of 3,3'-di-O-methylquercetin (5) at 95.14% of purity in fraction 3, 15 mg of p-hydroxybenzoic acid (1) at 92.83% of purity in fraction 5, 12 mg of isorhamnetin-3-O-β-D-rutinoside (2) at 97.99% of purity in fraction 6. This is the first time these phenolic compounds have been obtained from H. halodendron, and their chemical structures identified by means of physicochemical and spectrometric analysis.

Assessment of antioxidant potential, total phenolics and flavonoids of different solvent fractions of monotheca buxifolia fruit.

PubMed

Jan, Shumaila; Khan, Muhammad Rashid; Rashid, Umbreen; Bokhari, Jasia

2013-10-01

This study was conducted to investigate the antioxidant potential of methanol extract and its derived fractions (hexane, ethyl acetate, butanol, and aqueous) of fruits of Monotheca buxifolia (Falc.) Dc., a locally used fruit in Pakistan. Dried powder of the fruit of M. buxifolia was extracted with methanol and the resultant was fractionated with solvents having escalating polarity; n-hexane, chloroform, ethyl acetate, n-butanol and the residual soluble aqueous fraction. Total phenolic and total flavonoid contents were estimated for the methanol and various fractions. These fractions were also subjected to various in vitro assays to estimate the scavenging activity for 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), superoxide, hydroxyl, hydrogen peroxide and reductive ability for ferric ions and phosphomolybdate assay. The n-butanol, aqueous and methanol fractions possessed high amount of phenolics and flavonoids compared with other fractions, and subsequently showed a pronounced scavenging activity on DPPH, ABTS, superoxide, hydroxyl and hydrogen peroxide radicals and had a potent reductive ability on ferric ion and phosphomolybdate assay. There was a found significant correlation between total phenolic and flavonoid contents and EC50 of DPPH, superoxide, hydrogen peroxide radical and phosphomolybdate assays, whereas a nonsignificant correlation was found with the hydroxyl radical and ABTS radical assay. M. buxifolia fruit can be used as natural antioxidant source to prevent damage associated with free radicals.

HPLC-DAD identification of polyphenols from ethyl acetate extract of Amaranthus spinosus leaves and determination of their antioxidant and antinociceptive effects.

PubMed

Rjeibi, Ilhem; Ben Saad, Anouar; Sdayria, Jazia; Feriani, Anouar; Ncib, Sana; Allagui, Mohamed Salah; Hfaiedh, Najla; Souid, Sami

2018-04-20

Amaranthus spinosus has been consumed traditionally to prevent various diseases including abdominal pain. In this study, the phytochemical composition, antioxidant and analgesic activities of an ethyl acetate extract of A. spinosus leaves (ASEA) were evaluated. The ASEA had the highest concentrations of total phenols (462.2 mg GAE/g DW), condensed tannin (5.01 mg CE/g DW) and total flavonoid contents (30.07 mg CE/g DW) compared to the chloroform, n-hexane, n-butanol and water extracts. Similarly, ASEA showed the most effective total antioxidant activity (45.45 µg/mL), DPPH scavenging activity (27.32 µg/mL) and hydrogen peroxide scavenging activity (30.60 µg/mL). ASEA with the doses of 200-600 mg/kg (p.o.) clearly demonstrated antinociceptive effects by reducing acetic acid-induced abdominal contortions with a maximal inhibition of 79.57% at 600 mg/kg and increasing latencies of the hot-plate paw-licking response. The tested doses also significantly (p < 0.001) decreased the reaction time in the formalin test at the neurogenic and inflammatory phases. ASEA contained ten polyphenols with caffeic acid being the predominant polyphenol. Overall, this study gave evidence that A. spinosus is a new antioxidant and analgesic agent, and justified its traditional use for the treatment of pain.

Terminalia ferdinandiana extracts as inhibitors of Giardia duodenalis proliferation: a new treatment for giardiasis.

PubMed

Rayan, P; Matthews, B; McDonnell, P A; Cock, I E

2015-07-01

Giardisis is a debilitating disease caused by gastrointestinal parasites of the genus Giardia. High-antioxidant T. ferdinandiana fruit extracts were investigated for the ability to block Giardia duodenalis growth. Methanolic and aqueous extracts had the most potent growth inhibitory activity (IC50 values of approximately 700 and 140 μg/ml, respectively). Ethyl acetate and chloroform extracts also inhibited G. duodenalis growth, albeit with lower potency. The hexane extract was completely devoid of G. duodenalis growth inhibitory activity. All extracts were nontoxic in the Artemia fransiscana bioassay. Nontargeted HPLC-quadrupole time-of-flight (QTOF) mass spectroscopy (with screening against three compound databases) putatively identified 17 compounds in all of the inhibitory extracts but not in the inactive hexane extract. The low toxicity of the Terminalia ferdinandiana fruit extracts and their potent G. duodenalis growth inhibitory bioactivity indicate their potential as medicinal agents in the treatment and prevention of this disease.

Screening for acetylcholinesterase inhibition and antioxidant activity of selected plants from Croatia.

PubMed

Jukic, Mila; Burcul, Franko; Carev, Ivana; Politeo, Olivera; Milos, Mladen

2012-01-01

The methanol, ethyl acetate and chloroform extracts of selected Croatian plants were tested for their acetylcholinesterase (AChE) inhibition and antioxidant activity. Assessment of AChE inhibition was carried out using microplate reader at 1 mg mL⁻¹. Antioxidant capacities were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging test and ferric reducing/antioxidant power assay (FRAP). Total phenol content (TPC) of extracts were determined using Folin-Ciocalteu colorimetric method. Out of 48 extracts, only methanolic extract of the Salix alba L. cortex exerted modest activity towards AChE, reaching 50.80% inhibition at concentration of 1 mg mL⁻¹. All the other samples tested had activity below 20%. The same extract performed the best antioxidative activity using DPPH and FRAP method, too. In essence, among all extracts used in the screening, methanolic extracts showed the best antioxidative activity as well as highest TPC.

Antibacterial Constituents of Hainan Morinda citrifolia (Noni) Leaves.

PubMed

Zhang, Wei-Min; Wang, Wei; Zhang, Jing-Jing; Wang, Zhi-Rong; Wang, Yu; Hao, Wang-Jun; Huang, Wu-Yang

2016-05-01

Noni (Morinda citrifolia L.) is an edible and medicinal plant distributed in Hainan, China. The antibacterial activities of the extracts of water (WE), petroleum ether (PEE), ethyl acetate (EAE), chloroform (CE), and n-butanol (BE) were assayed by the disk diffusion method. The results showed that the extracts from Noni leaves possessed antibacterial effects against Bacillus subtilis, Escherichia coli, Proteus vulgaris, and Staphylococcus aureus. Among 5 different extracts, the BE produced the best antibacterial activity. The samples were first extracted by ethanol, and the primary compounds in the BE fraction of ethanol extract was further isolated and identified. Six phenolic compounds, including 5, 15-dimethylmorindol, ferulic acid, p-hydroxycinamic acid, methyl 4-hydroxybenzoate, methyl ferulate, and methyl 4-hydroxycinnamate, were identifiedby NMR. The results indicated that the phenolic compounds might significantly contribute to antibacterial activities of Noni leaves. © 2016 Institute of Food Technologists®

Screening for fractions of Oxytropis falcata Bunge with antibacterial activity.

PubMed

Jiang, H; Hu, J R; Zhan, W Q; Liu, X

2009-01-01

Preliminary studies with the four extracts of Oxytropis falcate Bunge exhibited that the chloroform and ethyl acetate extracts showed stronger antibacterial activities against the nine tested Gram-positive and Gram-negative bacteria. The HPLC-scanned and bioassay-guided fractionation led to the isolation and identification of the main flavonoid compounds, i.e. rhamnocitrin, kaempferol, rhamnetin, 2',4'-dihydroxychalcone and 2',4',beta-trihydroxy-dihydrochalcon. Except 2',4',beta-trihydroxy-dihydrochalcon, four other compounds had good antibacterial activities. The minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs) of the four compounds ranged between 125 and 515 microg mL(-1). Staphylococcus aureus was the most susceptible to these compounds, with MIC and MBC values from 125 to 130 microg mL(-1). This is the first report of antibacterial activity in O. falcate Bunge. In this study, evidence to evaluate the biological functions of O. falcate Bunge is provided, which promote the rational use of this herb.

An interesting two-phase solvent system and its use in preparative isolation of aconitines from aconite roots by counter-current chromatography.

PubMed

Han, Quan-Bin; Tang, Wai-Lun; Dong, Cai-Xia; Xu, Hong-Xi; Jiang, Zhi-Hong

2013-04-01

Two-phase solvent system plays crucial role in successful separation of organic compounds using counter-current chromatography (CCC). An interesting two-phase solvent system, composed of chloroform/ethyl acetate/methanol/water, is reported here, in which both phases contain sufficient organic solvents to balance their dissolving capacities. Adjusting the solvent system to get satisfactory partition coefficients (K values) for target compounds becomes relatively simple. This solvent system succeeded in sample preparation of aconitine (8.07 mg, 93.69%), hypaconitine (7.74 mg, 93.17%), mesaconitine (1.95 mg, 94.52%) from raw aconite roots (102.24 mg, crude extract), benzoylmesaconine (34.79 mg, 98.67%) from processed aconite roots (400.01 mg, crude extract), and yunaconitine (253.59 mg, 98.65%) from a crude extract of Aconitum forrestii (326.69 mg, crude extract). © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Hydrogen-bonded structures from adamantane-based catechols

NASA Astrophysics Data System (ADS)

Kawahata, Masatoshi; Matsuura, Miku; Tominaga, Masahide; Katagiri, Kosuke; Yamaguchi, Kentaro

2018-07-01

Adamantane-based bis- and tris-catechols were synthesized to examine the effect of hydrogen bonds on the arrangement and packing of the components in the crystalline state. Single-crystal X-ray crystallographic analysis revealed that hydrogen bonds formed by the hydroxyl groups of catechol groups play essential roles in the production of various types of unique structures. 1,3-Bis(3,4-dihydroxyphenyl)adamantane (1) provided hydrogen-bonded network structures composed of helical chains in crystal from chloroform/methanol, and layer structures in crystal from ethyl acetate/hexane. The complexation of 1 with 1,3,5-trinitrobenzene or 1,2,4,5-tetracyanobenzene resulted in the formation of co-crystals, respectively. One-dimensional hydrogen-bonded structures were constructed from the adamantane-based molecules, which participated in charge-transfer interactions with guests. 1,3,5-Tris(3,4-dihydroxyphenyl)adamantane also afforded crystal, and the components were assembled into infinite polymers.

Antibacterial, antifungal, antispasmodic and Ca++ antagonist effects of Caesalpinia bonducella.

PubMed

Khan, Hidayat-Ullah; Ali, Irshad; Khan, Arif-Ullah; Naz, Rubina; Gilani, Anwarul Hassan

2011-02-01

Caesalpinia bonducella F. (Leguminosae) has been used as a folk medicine for a variety of ailments. The crude extract of C. bonducella and its fractions were studied for antibacterial, antifungal, antispasmodic and Ca++ antagonistic properties. The strongest antibacterial effect was displayed by the n-butanol (72%) and ethyl acetate (80%) fractions, followed by the crude extract (46% and 42%), against Escherichia coli and Bacillus subtilis, respectively. The plant extract and its fractions showed mild to excellent activity in antifungal bioassays, with maximum antifungal activity against Candida glaberata (80%) and Aspergillus flavus (70%) by the n-butanol and chloroform fractions, followed by the crude extract (70% and 65%). Caesalpinia bonducella extract caused concentration-dependent inhibition of spontaneous and high K+ (80 mM)-induced contractions of isolated rabbit jejunum preparations, similar to that caused by Verapamil. These results indicate that C. bonducella exhibits antibacterial, antifungal, spasmolytic and Ca++ channel blocking actions.

Antioxidant Activity in the Extracts of Two Edible Aroids

PubMed Central

Mandal, P.; Misra, T. K.; Singh, I. D.

2010-01-01

Two neglected species of Araceae, Alocasia macrorhiza (Linn.) G. Don and Alocasia fornicata (Roxb.) Schott are important as food and ethno medicine in Asia and Africa. Their bioefficacy is documented in the Ayurveda. The solvent extracts of different edible parts of these two species like rhizomes, leaves, roots and stolons were screened for in vitro antioxidant properties using standard procedures. The successive extracts in hexane, benzene, toluene, chloroform, diethyl ether, ethyl acetate and water fraction exhibited IC50 values in the following order, roots>rhizome>leaves for Alocasia macrorhiza and leaves>stolon for Alocasia fornicate, respectively in 2,2-diphenyl-1-picryl hydrazyl antioxidant inhibition assay. Maximum antioxidant activity was observed in diethyl ether extracts for both species. The IC50 values were comparable with those of quercetine and ascorbic acid as standards. These results suggest that the two aroid species have antioxidant activity in their edible parts and should be extracted using diethyl ether solvent. PMID:20582198

Inhibition of norsolorinic acid accumulation to Aspergillus parasiticus by marine actinomycetes

NASA Astrophysics Data System (ADS)

Yan, Peisheng; Shi, Cuijuan; Shen, Jihong; Wang, Kai; Gao, Xiujun; Li, Ping

2014-11-01

Thirty-six strains of marine actinomycetes were isolated from a sample of marine sediment collected from the Yellow Sea and evaluated in terms of their inhibitory activity on the growth of Aspergillus parasiticus and the production of norsolorinic acid using dual culture plate assay and agar diffusion methods. Among them, three strains showed strong antifungal activity and were subsequently identified as Streptomyces sp. by 16S rRNA gene sequencing analysis. The supernatant from the fermentation of the MA01 strain was extracted sequentially with chloroform and ethyl acetate, and the activities of the extracts were determined by tip culture assay. The assay results show that both extracts inhibited mycelium growth and toxin production, and the inhibitory activities of the extracts increased as their concentrations increased. The results of this study suggest that marine actinomycetes are biologically important for the control of mycotoxins, and that these bacteria could be used as novel biopesticides against mycotoxins.

Antipyretic and antinociceptive activity of Diospyros lotus L. in animals

PubMed Central

Rauf, Abdur; Uddin, Ghias; Siddiqui, Bina S.; Muhammad, Naveed; Khan, Haroon

2014-01-01

Objective To evaluate pharmacologically the traditional use of Diospyros lotus as antipyretic and antinociceptive in various animal models. Methods In vivo experimental models were used in this study. Antipyretic activity of extract/fractions was evaluated in brewer's yeast induced hyperthermic mice while antinociceptive activity was studied in acetic acid induced writhing test at 50 and 100 mg/kg i.p. Results The crude extract strongly ameliorated the induced pyrexia during various assessment times. Upon fractionation, the antipyretic effects were strongly augmented by the chloroform and ethyl acetate fractions of the plant. However, hexane and butanol fractions were insignificant in their effect as antipyretic. The extract showed marked inhibition on the noxious simulation induced by post acetic acid injection. The effect was strongly supported by other fraction expect hexane. Conclusions In short, our study scientifically validated the traditional use of the plant as antipyretic. PMID:25183115

Simultaneous quantification of cannabinoids and metabolites in oral fluid by two-dimensional gas chromatography mass spectrometry

PubMed Central

Milman, Garry; Barnes, Allan J.; Lowe, Ross H.; Huestis, Marilyn A.

2010-01-01

Development and validation of a method for simultaneous identification and quantification of Δ9-tetrahydrocannabinol (THC), cannabidiol (CBD), cannabinol (CBN), and metabolites 11-hydroxy-THC (11-OH-THC) and 11-nor-9-carboxy-THC (THCCOOH) in oral fluid. Simultaneous analysis was problematic due to different physicochemical characteristics and concentration ranges. Neutral analytes, such as THC and CBD, are present in ng/mL, rather than pg/mL concentrations, as observed for the acidic THCCOOH biomarker in oral fluid. THCCOOH is not present in cannabis smoke, definitively differentiating cannabis use from passive smoke exposure. THC, 11-OH-THC, THCCOOH, CBD, and CBN quantification was achieved in a single oral fluid specimen collected with the Quantisal™ device. One mL oral fluid/buffer solution (0.25mL oral fluid and 0.75mL buffer) was applied to conditioned CEREX® Polycrom™ THC solid phase extraction (SPE) columns. After washing, THC, 11-OH-THC, CBD, and CBN were eluted with hexane/acetone/ethyl acetate (60:30:20, v/v/v), derivatized with N, O-bis-(trimethylsilyl) trifluoroacetamide and quantified by two-dimensional gas chromatography electron ionization mass spectrometry (2D-GCMS) with cold trapping. Acidic THCCOOH was separately eluted with hexane/ethyl acetate/acetic acid (75:25:2.5, v/v/v), derivatized with trifluoroacetic anhydride and hexafluoroisopropanol, and quantified by the more sensitive 2D-GCMS–electron capture negative chemical ionization (NCI-MS). Linearity was 0.5-50ng/mL for THC, 11-OH-THC, CBD and 1-50ng/mL for CBN. The linear dynamic range for THCCOOH was 7.5–500pg/mL. Intra-and inter-assay imprecision as percent RSD at three concentrations across the linear dynamic range were 0.3%-6.6%. Analytical recovery was within 13.8% of target. This new SPE 2D-GCMS assay achieved efficient quantification of five cannabinoids in oral fluid, including pg/mL concentrations of THCCOOH by combining differential elution, 2D-GCMS with electron ionization and negative chemical ionization. This method will be applied to quantification of cannabinoids in oral fluid specimens from individuals participating in controlled cannabis and Sativex® (50% THC and 50% CBD) administration studies, and during cannabis withdrawal. PMID:20083251

Selenium Derivatization of Nucleic Acids for Crystallography

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jiang,J.; Sheng, J.; Carrasco, N.

2007-01-01

The high-resolution structure of the DNA (5'-GTGTACA-C-3') with the selenium derivatization at the 2'-position of T2 was determined via MAD and SAD phasing. The selenium-derivatized structure (1.28 {angstrom} resolution) with the 2'-Se modification in the minor groove is isomorphorous to the native structure (2.0 {angstrom}). To directly compare with the conventional bromine derivatization, we incorporated bromine into the 5-postion of T4, determined the bromine-derivatized DNA structure at 1.5 {angstrom} resolution, and found that the local backbone torsion angles and solvent hydration patterns were altered in the structure with the Br incorporation in the major groove. Furthermore, while the native andmore » Br-derivatized DNAs needed over a week to form reasonable-size crystals, we observed that the Se-derivatized DNAs grew crystals overnight with high-diffraction quality, suggesting that the Se derivatization facilitated the crystal formation. In addition, the Se-derivatized DNA sequences crystallized under a broader range of buffer conditions, and generally had a faster crystal growth rate. Our experimental results indicate that the selenium derivatization of DNAs may facilitate the determination of nucleic acid X-ray crystal structures in phasing and high-quality crystal growth. In addition, our results suggest that the Se derivatization can be an alternative to the conventional Br derivatization.« less

Investigation on Antibacterial and Antioxidant Activities, Phenolic and Flavonoid Contents of Some Thai Edible Plants as an Alternative for Antibiotics

PubMed Central

Lee, J. H.; Cho, S.; Paik, H. D.; Choi, C. W.; Nam, K. T.; Hwang, S. G.; Kim, S. K.

2014-01-01

This study was aimed to examine the antibacterial and antioxidative properties of seven edible plants from Thailand to develop alternative antibiotics as feed additives. The plants include Citrus aurantifolia Swingle (Lime) fruits and its leaves, Sesbania grandiflora L. (Agati sesbania) leaves, Piper sarmentosum Roxb (Wild betal) leaves, Curcuma domestica Valeton (Turmeric) roots, Morinda citrifolia L. (Beach mulberry) leaves, Cassia siamea britt (Siamea cassia) leaves, and Cocos nucifera L. (Coconut) peels. The plants were extracted by methanol, n-hexane, chloroform, ethyl acetate, butanol and water. Antibacterial activities with minimum inhibitory concentration (MIC) were determined by agar diffusion assay against Escherichia coli, Burkholderia sp., Haemopilus somnus, Haemopilus parasuis, and Clostridium perfringens that were considered pathogenic strains in livestock infection. Methanol extracts of C. aurantifolia Swingle fruits and leaves showed the broadest spectrum of antibacterial activities except for C. perfringens. Butanol extract of S. grandiflora L. leaves showed the strongest activity against Burkholderia sp. with MIC, 135 μg/mL. P. sarmentosum Roxb leaves showed antibacterial activities against E. coli, Burkholderia sp. and H. parasuis. Ethyl acetate and water extracts from C. domesitca Valeton roots showed MIC of 306 μg/mL and 183 μg/mL, respectively against only C. perfringens. Antioxidative activity was determined by 2-diphenyl-2-picryl hydrazyl photometric assay. The methanol extracts of C. aurantifolia Swingle fruits and P. sarmentosum Roxb leaves showed the highest antioxidant activity among all the extracts with 3.46 mg/mL and 2.70 mg/mL effective concentration 50% (EC50) values, respectively. Total contents of phenolics and flavonoids were measured from the plant extracts. Methanol extracts of S. grandiflora L. and chloroform extracts of C. domestica Valeton were found to have the highest amount of total phenolics, 41.7 and 47.8 μg/mL, respectively. Flavonoid content of methanol extracts in S. grandiflora L. T was 22.5 μg/mL and the highest among plant extracts tested. These results indicated that C. aurantifolia Swingle, S. grandiflora L., P. sarmentosum Roxb, and C. domestica Valeton have antibacterial and antioxidant activities and can be used as alternative antibiotics or potential feed additives for the control of animal pathogenic bacteria. PMID:25178298

Free Radical Scavenging Properties of Annona squamosa

PubMed

Vikas, Biba; Akhil B, S; P, Remani; Sujathan, K

2017-10-26

Annona squamosa has extensively been used in the traditional and folkloric medicine and found to possess many biological activities. Different solvents, petroleum ether, chloroform, ethyl acetate and methanol extracts of Annona squamosa seeds (ASPE, ASCH, ASEA, ASME) have been used to prepare plant extracts. The present investigations dealt with the free radical scavenging activity of four extracts using various techniques such as total reducing power estimation, total phenolic count, 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging effect, evaluation of ABTS cation decolorisation capacity, FRAP assay, hdroxyl radical scavenging assay, super oxide assay and Nitric oxide radical scavenging assay of the extracts. The results showed that the four extracts of Annona squamosa showed significant reducing power in four extracts. The total phenolic contents in petroleum ether, chloroform, ethyl acetate, methanol extracts and positive control were 0.64±0.17, 0.54±0.27, 0.49±0.24, 0.57±0.22 and 0.66±0.33. The antioxidant capacity by ABTS assay of ASPE, ASCH, ASEA, ASME and positive control, trolox showed 77.75±0.5,73.25±1.7,78.5± 1.2 , 80 ± 0.8 μg/ml and 94.2 ± 0.9 respectively. The (50 % scavenging activity) SA50 of ASPE and ASCH, ASEA and ASME was found to be 34.4 μg/ml, 43.8 μg/ml 34.7 μg/m and 28.8 μg/ml respectively by DPPH assay. The percentage of hydroxyl radical scavenging increased with the increasing concentration of the extracts. ASPE, ASCH, ASEA and ASME showed superoxide radical scavenging activity, as indicated by their values 66 ± 0.5, 68 ± 1 ,63 ± 1 and 70 ± 0.5 μg/ml respectively compared to gallic acid which was 97 ± 0.5 μg/ml. The values for scavenging of nitric oxide for ASPE, ASCH, ASEA and ASME were 91.0 ± 1.0, 66.75 ± 0.5, 71.75 ± 1.1 and 75.75 ± 1.15 μg/ml while value for standard ascorbic acid was 91.0 ± 1.0 μg/ml. The results revealed strong antioxidants in four extracts may lead to the development of potent antioxidant agents from Annona squamosa seeds. Creative Commons Attribution License

Characterization of oxidized coal surfaces: Quarterly report, May 1988--September 1988

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hercules, D. M.

1988-01-01

Laser mass spectra have been obtained of 2-fluoro-1-methylpyridinium (FMP) derivatized coal consistent with a prime objective of this research. This reagent, specific for the hydroxy functionality, produced major peaks at m/z 184 and 199 in the spectra of Pocahontas coal following derivatization. A gas phase reactor was built in order to enhance coal derivatization. Gas phase derivatization was accomplished on model compounds. Derivatization was carried out on Illinois No. 6 coal and analyzed with DRIFTS to determine the extent of derivatization. Oxidation indices of the oxidized coal decreased upon reacting with 2,4-dinitrophenylhydrazine, a derivatizing agent specific for carbonyl groups, confirmingmore » that the derivatization reaction had taken place. 5 refs., 5 figs.« less

Characterization of dicarboxylic naphthenic acid fraction compounds utilizing amide derivatization: Proof of concept.

PubMed

Kovalchik, Kevin A; MacLennan, Matthew S; Peru, Kerry M; Ajaero, Chukwuemeka; McMartin, Dena W; Headley, John V; Chen, David D Y

2017-12-30

The characterization of naphthenic acid fraction compounds (NAFCs) in oil sands process affected water (OSPW) is of interest for both toxicology studies and regulatory reasons. Previous studies utilizing authentic standards have identified dicarboxylic naphthenic acids using two-dimensional gas chromatography hyphenated to time-of-flight mass spectrometry (GC × GC/TOFMS). The selective derivatization of hydroxyl groups has also recently aided in the characterization of oxy-NAFCs, and indirectly the characterization of dicarboxylic NAFCs. However, there has been no previous report of derivatization being used to directly aid in the standard-free characterization of NAFCs with multiple carboxylic acid functional groups. Herein we present proof-of-concept for the characterization of dicarboxylic NAFCs utilizing amide derivatization. Carboxylic acid groups in OSPW extract and in a dicarboxylic acidstandard were derivatized to amides using a previously described method. The derivatized extract and derivatized standard were analyzed by direct-injection positive-mode electrospray ionization ((+)ESI) high-resolution mass spectrometry (HRMS), and the underivatized extract was analyzed by (-)ESI MS. Tandem mass spectrometry (MS/MS) was carried out on selected ions of the derivatized standard and derivatized OSPW. Data analysis was carried out using the Python programming language. The distribution of monocarboxylic NAFCs observed in the amide-derivatized OSPW sample by (+)ESI-MS was generally similar to that seen in underivatized OSPW by (-)ESI-MS. The dicarboxylic acid standard shows evidence of being doubly derivatized, although the second derivatization appears to be inefficient. Furthermore, a spectrum of potential diacid NAFCs is presented, identified by both charge state and derivatization mass. Interference due to the presence of multiple derivatization products is noted, but can be eliminated using on-line separation or an isotopically labelled derivatization reagent. Proof of concept for the characterization of dicarboxylic NAFCs utilizing amide derivatization is demonstrated. Furthermore, (+)ESI-HRMS of the derivatized monocarboxylic NAFCS yields similar information to (-)ESI-MS analysis of underivatized NAFCs, with the benefit of added selectivity for carboxylic acid species and the characterization of diacids. Copyright © 2017 John Wiley & Sons, Ltd.

Acute toxicity, brine shrimp cytotoxicity and relaxant activity of fruits of callistemon citrinus curtis.

PubMed

Ali, Niaz; Ahmed, Ghayour; Shah, Syed Wadood Ali; Shah, Ismail; Ghias, Mehreen; Khan, Imran

2011-10-24

Callistemon citrinus Curtis belongs to family Myrtaceae that has a great medicinal importance. In our previous work, fruits of Callistemon citrinus were reported to have relaxant (antispasmodic) activity. The current work describes the screening of fractions of the crude methanol extract for tracing spasmolytic constituents so that it shall help us for isolation of bioactive compounds. Acute toxicity and brine shrimp cytotoxicity of crude methanol extract are also performed to standardize it. The crude methanol extract was obtained by maceration with distilled water (500 ml) three times and fractionated successively with n-hexane, chloroform, ethyl acetate and n-butanol (300 ml of each solvent). Phytochemical analysis for crude methanol extract was performed. Acute toxicity studies were performed in mice. Brine shrimp cytotoxicity studies were performed to determine its cytotoxicity and standardize it. In other series of experiments, rabbits' jejunum preparations were used in screening for possible relaxant activities of various fractions. They were applied in concentrations of 0.01, 0.03, 0.1, 0.3, 1.0, 3.0, 5.0 and 10.0 mg/ml on spontaneous rabbits' jejunum preparations. In similar fashion, fractions were also tested on KCl (80 mM) -induced contractions. Calcium chloride curves were constructed in K-rich Tyrode's solution. The effects of various fractions were tested on calcium chloride curves at concentrations 1.0, 3.0, 5.0 and 10.0 mg/ml. Curves of verapamil used as reference drug at concentration 0.1 μM and 0.3 μM were also constructed. The curves were compared with their respective controls for possible right shift. Methanol extract tested strongly positive for saponins and tannins. However, it tested mild positive for presence of proteins, amino acids, carbohydrates and phenolic compounds. LD(50) value for crude methanol extract is 476.25 ± 10.3 (470-481, n = 4) mg/ml. Similarly, EC(50) value for brine shrimp cytotoxicity is 65.5 ± 7.28 (60.8- 69.4, n = 4) mg/ml. All the fractions relaxed the spontaneous and KCl-induced contractions. EC(50) values (mg/ml) for effects of ethyl acetate fraction on spontaneous and KCl induced contractions are 2.62 ± 0.78 (2.15-3.0, n = 4) and 3.72 ± 0.86 (3.38-4.28, n = 4) respectively. Respective EC(50) values (mg/ml) for n-butanol fraction are 3.59 ± 0.2(3.07-3.9, n = 4) for spontaneous, and 5.57 ± 0.2 (5.07-6.11, n = 4) for KCl- induced contractions. EC(50) value for control calcium chloride curve (without extract) is -2.73 ± 0.19 (-2.6 - -2.81, n = 4) while EC(50) for curves treated with 5.0 mg/ml of chloroform is -2.22 ± 0.02 (-2.16 - -2.3, n = 4). EC(50) value for ethyl acetate treated (1.0 mg/ml) tissues is -1.95 ± 0.10 (-1.88 - -2.0, n = 4) vs. control EC(50) = -2.71 ± 0.08 (-2.66 - -2.76, n = 4). All the fractions, except n-hexane, showed a right shift like that of verapamil (EC(50) = -1.72 ± 0.15 (-1.62 - -1.8, n = 4) vs. Control EC(50) = -2.41 ± 0.06 (-2.38 - - 2.44, n = 4), a standard drug that blocks voltage operated calcium channels. Relaxant constituents were more concentrated in ethylacetate fraction followed by chloroform, n -butanol and aqueous fractions that warrant for its isolation. The crude methanol extract is safe at concentration 250 mg/ml or below and results of brine shrimp cytotoxicity assay imply the plant specie may be a source of cytotoxic agents.

Linkage-specific sialic acid derivatization for MALDI-TOF-MS profiling of IgG glycopeptides.

PubMed

de Haan, Noortje; Reiding, Karli R; Haberger, Markus; Reusch, Dietmar; Falck, David; Wuhrer, Manfred

2015-08-18

Glycosylation is a common co- and post-translational protein modification, having a large influence on protein properties like conformation and solubility. Furthermore, glycosylation is an important determinant of efficacy and clearance of biopharmaceuticals such as immunoglobulin G (IgG). Matrix-assisted laser desorption/ionization (MALDI)-time-of-flight (TOF)-mass spectrometry (MS) shows potential for the site-specific glycosylation analysis of IgG at the glycopeptide level. With this approach, however, important information about glycopeptide sialylation is not duly covered because of in-source and metastable decay of the sialylated species. Here, we present a highly repeatable sialic acid derivatization method to allow subclass-specific MALDI-TOF-MS analysis of tryptic IgG glycopeptides. The method, employing dimethylamidation with the carboxylic acid activator 1-ethyl-3-(3-dimethylamino)propyl)carbodiimide (EDC) and the catalyst 1-hydroxybenzotriazole (HOBt), results in different masses for the functionally divergent α2,3- and α2,6-linked sialic acids. Respective lactonization and dimethylamidation leads to their direct discrimination in MS and importantly, both glycan and peptide moieties reacted in a controlled manner. In addition, stabilization allowed the acquisition of fragmentation spectra informative with respect to glycosylation and peptide sequence. This was in contrast to fragmentation spectra of underivatized samples, which were dominated by sialic acid loss. The method allowed the facile discrimination and relative quantitation of IgG Fc sialylation in therapeutic IgG samples. The method has considerable potential for future site- and sialic acid linkage-specific glycosylation profiling of therapeutic antibodies, as well as for subclass-specific biomarker discovery in clinical IgG samples derived from plasma.

Therapeutic drug monitoring of topiramate with a new HPLC method, SPE extraction and high sensitivity pre-column fluorescent derivatization.

PubMed

Bolner, Andreas; De Riva, Valentina; Galloni, Elisabetta; Perini, Francesco

2014-01-01

Topiramate is a 2nd generation antiepileptic drug (AED) recently approved by the FDA for migraine prophylaxis. Its pharmacological activity already appears significant at low doses. Unfortunately, the difficulty in determining the drug in serum at low concentrations hampers the completion of accurate pharmacokinetic studies in humans. Only chromatographic methods allow reaching the necessary sensitivities. Almost all of the HPLC methods proposed were based on the preliminary extraction of topiramate from the sample using organic solvents. In our study, the conditions for purifying topiramate through solid-liquid technique in disposable cartridges (SPE) packed with C18 reversed phase were examinated and optimised. After a pre-column derivatization step with 9-fluorenylmethyl chloroformate (FMOC-Cl) and internal standard addition, topiramate was analysed on a CN column with sodium phosphate buffer 50 mmol/L (pH 2.5) containing acetonitrile (60:40, v/v) as the mobile phase. The column effluent was monitored with a fluorescence detector (excitation and emission 1 260 and 315 nm, respectively). 122 samples from our routine laboratory work were analysed in order to confirm the existence of a relationship between topiramate dose and serum concentration and to evaluate the effect of concomitant therapies with enzyme-inducing AEDs. Sensitivity (2 ng/mL), precision (CV within assay of 3.8% and between assays of 6.6%), linearity and accuracy of the method were better than other analytical procedures previously reported. Serum topiramate levels in the group with enzyme-inducing AEDs showed a reduction with respect to the group with non-enzyme-inducing AEDs and the correlation between doses and mean serum concentration gives a linear trend (r2 = 0.916). The efficacy of SPE extraction together with the method's reliability proved very advantageous for pharmacokinetics studies and, in principle, for therapeutic drug monitoring and toxicological investigations.

Calibrating compound-specific stable nitrogen isotope analysis in avian eggs as bioarchives for paleoreconstruction

NASA Astrophysics Data System (ADS)

Preciado, C., Jr.

2016-12-01

Compound-specific stable isotope analysis (CSIA) has become a powerful tool for reconstructing consumer-resource relationships in modern and ancient systems. Stable nitrogen isotope analysis (δ15N) of "trophic" and "source" amino acids provides independent proxies of trophic position and the δ15N value at the base of the food web, respectively. When applied to avian egg tissues (e.g., shell protein, membrane), which can be preserved in the archaeological record, this approach can be used to address complex questions in food web architecture and biogeochemical cycling. In this study, we examined how sample-processing protocols affected the chromatography and reliability of δ15N values of individual amino acids in avian (chicken and penguin) egg components (shell protein, membrane, yolk, and albumen) via gas chromatography-combustion-isotope ratio mass spectrometry. "Unprocessed" egg tissues underwent standard acid hydrolysis protocols prior to derivatization, and resulted in poor chromatography with highly variable δ15N values across replicates. "Processed" samples were eluted through cation exchange columns (Dowex 50WX*400) prior to derivatization, followed by a P-buffer (KH2PO4 + Na2HPO4)-chloroform centrifugation extraction to remove confounding peaks and matrix impurities. These additional procedures greatly improved the chromatography of the "processed" samples, revealing better peak separation and baseline integration as well as lower δ15N variability. Additionally, a standard lipid extraction was necessary for yolk but not membrane, albumen, and shell. While the additional procedures applied to the "columned" samples did result in a significant reduction in sample yield ( 20%), it was non-fractionating and thus only affected the total sample sizes necessary for δ15N CSIA (shell protein 50mg and membrane, yolk, and albumen 0.5mg). The protocols developed here will streamline CSIA of egg tissues for future work in avian ecology.

Condensation of the isoprenoid and amino precursors in the biosynthesis of domoic acid.

PubMed

Savage, Thomas J; Smith, G Jason; Clark, Amy T; Saucedo, Portia N

2012-01-01

Understanding how environmental signals regulate production of domoic acid in blooms of Pseudo-nitzschia spp. at a molecular level requires description of the biochemical pathway to this kainoid neurotoxin. Precursor feeding studies have suggested domoic acid arises from the condensation of the C(10) isoprenoid geranyl diphosphate with glutamate, but the specific reactions leading to domoic acid from these precursors remain undescribed. Here, we develop a method to derivatize domoic acid with propyl chloroformate that enables gas chromatography-mass spectrometry (GC-MS) analysis to measure incorporation of stable isotopes into domoic acid generated in cultures incubated with isotopically-labeled substrates. We apply this method to demonstrate that both (2)H from [1-(2)H(2)]geraniol are incorporated into domoic acid, suggesting that the condensation of geranyl diphosphate with an amino group occurs by nucleophilic substitution of the diphosphate rather than by oxidation of geraniol to the aldehyde before reaction with an amino group to form an imine. Ultimately, these and similar studies will facilitate the identification of DA biosynthetic enzymes and genes which will enable the study of how environmental factors regulate DA biosynthesis at the molecular level. Copyright © 2011 Elsevier Ltd. All rights reserved.

A fast and simple solid phase microextraction coupled with gas chromatography-triple quadrupole mass spectrometry method for the assay of urinary markers of glutaric acidemias.

PubMed

Naccarato, Attilio; Gionfriddo, Emanuela; Elliani, Rosangela; Sindona, Giovanni; Tagarelli, Antonio

2014-10-30

The analysis of characteristic urinary acidic markers such as glutaric, 3-hydroxyglutaric, 2-hydroxyglutaric, adipic, suberic, sebacic, ethylmalonic, 3-hydroxyisovaleric and isobutyric acid constitutes the recommended follow-up testing procedure for glutaric acidemia type 1 (GA-1) and type 2 (GA-2). The goal of the work herein presented is the development of a fast and simple method for the quantification of these biomarkers in human urine. The proposed analytical approach is based on the use of solid phase microextraction (SPME) combined with gas chromatography-triple quadrupole mass spectrometry (GC-QqQ-MS) afterward a rapid derivatization of acidic moieties by propyl chloroformate, propanol and pyridine. Trueness and precision of the proposed protocol, tested at 5, 30 and 80mgl -1 , provided satisfactory values: recoveries were in the range between 72% and 116% and the relative standard deviations (RSD%) were between 0.9% and 18% (except for isobutyric acid at 5mgl -1 ). The LOD values achieved by the proposed method ranged between 1.0 and 473μgl -1 . Copyright © 2014 Elsevier B.V. All rights reserved.

[Simultaneous determination of cocaine and its metabolite ecgonine methyl ester in human blood using microwave extraction-gas chromatography].

PubMed

Wang, Xiaobo; Ye, Nengsheng; Wang, Jifen; Gu, Xuexin

2010-07-01

A method was developed for the simultaneous determination of cocaine (COC) and its metabolite ecgonine methyl ester (EME) in human blood using gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC-FID). The blood sample was prepared by microwave extraction (MWE). The optimal parameters of MWE were as follows: 6 mL of chloroform-isopropanol (9: 1, v/v) mixture as extraction solvent, the pH value of the sample was adjusted at 10.0 with 0.05 mol/L Na2CO3-NaHCO3 buffer, the extraction was performed at 40 degrees C for 6 min. The COC and EME in the extract were qualified using GC-MS and quantitated using GC-FID. The average recoveries of COC and EME were from 79.91% to 99.85%, the relative standard deviations were less than 3.10%, and the limits of detection (LOD) were 60 and 40 mg/L, respectively. In the method COC and EME were detected without derivatization. The method is rapid, accurate and sensitive, and can be used for the simultaneous determination of COC and EME in blood samples.

Generation of a natural glycan microarray using 9-fluorenylmethyl chloroformate (FmocCl) as a cleavable fluorescent tag.

PubMed

Song, Xuezheng; Lasanajak, Yi; Rivera-Marrero, Carlos; Luyai, Anthony; Willard, Margaret; Smith, David F; Cummings, Richard D

2009-12-15

Glycan microarray technology has become a successful tool for studying protein-carbohydrate interactions, but a limitation has been the laborious synthesis of glycan structures by enzymatic and chemical methods. Here we describe a new method to generate quantifiable glycan libraries from natural sources by combining widely used protease digestion of glycoproteins and Fmoc chemistry. Glycoproteins including chicken ovalbumin, bovine fetuin, and horseradish peroxidase (HRP) were digested by Pronase, protected by FmocCl, and efficiently separated by 2D-HPLC. We show that glycans from HRP glycopeptides separated by HPLC and fluorescence monitoring retained their natural reducing end structures, mostly core alpha1,3-fucose and core alpha1,2-xylose. After simple Fmoc deprotection, the glycans were printed on NHS-activated glass slides. The glycans were interrogated using plant lectins and antibodies in sera from mice infected with Schistosoma mansoni, which revealed the presence of both IgM and IgG antibody responses to HRP glycopeptides. This simple approach to glycopeptide purification and conjugation allows for the development of natural glycopeptide microarrays without the need to remove and derivatize glycans and potentially compromise their reducing end determinants.

Sensitive Determination of Volatile Organic Compounds and Aldehydes in Tattoo Inks.

PubMed

Lim, Hyun-Hee; Shin, Ho-Sang

2017-02-01

As the popularity of body art including tattoo ink has increased, the safety associated with it has become an important interest. In this study, twenty volatile organic compounds (VOCs) and two aldehydes in tattoo inks were identified and quantified. Headspace and gas chromatography-mass spectrometry (HS GC-MS) for the VOCs and HS GC-MS based on derivatization with 2,2,2-trifluoroethylhydrazine (TFEH) for aldehydes was developed. Benzene, chloroform, toluene, ethylbenzene, m-xylene, p-xylene, o-xylene, propylbenzene, chlorobenzene, tert-butylbenzene, 1,3,5-trimethylbenzene, styrene, 1,2,4-trimethylbenzene, 2-chlorotoluene, 4-chlorotoluene, 1,3-dichlorobenzene, 1,4-dichlorobenzene, 1,2-dichlorobenzene, 1,2,4-trichlorobenzene and isopropyl alcohol were detected with the concentration range of 0.02-207,000 mg/kg in 16 different tattoo inks. Formaldehyde and acetaldehyde were detected with the concentration range of 0.4-308 mg/kg in the same samples. Our analytical results represent solvents used intentionally or non-intentionally in tattoo inks, and thus they may provide important information for national regulation. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Development of a fast and simple gas chromatographic protocol based on the combined use of alkyl chloroformate and solid phase microextraction for the assay of polyamines in human urine.

PubMed

Naccarato, Attilio; Elliani, Rosangela; Cavaliere, Brunella; Sindona, Giovanni; Tagarelli, Antonio

2018-05-11

Polyamines are aliphatic amines with low molecular weight that are widely recognized as one of the most important cancer biomarkers for early diagnosis and treatment. The goal of the work herein presented is the development of a rapid and simple method for the quantification of free polyamines (i.e., putrescine, cadaverine, spermidine, spermine) and N-monoacetylated polyamines (i.e., N 1 -Acetylspermidine, N 8 -Acetylspermidine, and N 1 -Acetylspermine) in human urine. A preliminary derivatization with propyl chloroformate combined with the use of solid phase microextraction (SPME) allowed for an easy and automatable protocol involving minimal sample handling and no consumption of organic solvents. The affinity of the analytes toward five commercial SPME coatings was evaluated in univariate mode, and the best result in terms of analyte extraction was achieved using the divinylbenzene/carboxen/polydimethylsiloxane fiber. The variables affecting the performance of SPME analysis were optimized by the multivariate approach of experimental design and, in particular, using a central composite design (CCD). The optimal working conditions in terms of response values are the following: extraction temperature 40 °C, extraction time of 15 min and no addition of NaCl. Analyses were carried out by gas chromatography-triple quadrupole mass spectrometry (GC-QqQ-MS) in selected reaction monitoring (SRM) acquisition mode. The developed method was validated according to the guidelines issued by the Food and Drug Administration (FDA). The satisfactory performances reached in terms of linearity, sensitivity (LOQs between 0.01 and 0.1 μg/mL), matrix effect (68-121%), accuracy, and precision (inter-day values between -24% and +16% and in the range 3.3-28.4%, respectively) make the proposed protocol suitable to be adopted for quantification of these important biomarkers in urine samples. Copyright © 2018 Elsevier B.V. All rights reserved.

Molecular complexes of some anthraquinone anti-cancer drugs: experimental and computational study

NASA Astrophysics Data System (ADS)

El-Gogary, Tarek M.

2003-03-01

It is known that anti-cancer drugs target DNA in the cell. The mechanism of interaction of anti-cancer drugs with DNA is not fully understood. It is thought that the forces of interaction have some contribution from charge-transfer (CT) binding. The ability of some anthraquinones (AQs) anti-cancer drugs to form CT complexes with well-known electron donor molecules was investigated by NMR. The NMR spectroscopy has indicated the formation of CT complexes between 1,4-bis{[2-(dimethylamino) ethyl]amino}-5,8-dihydroxyanthracene-9,10-dione, (AQ4), and its des-hydroxylated equivalent 1,4-bis{[2-(dimethylamino) ethyl]amino}anthracene-9,10-dione, (AQ4H), as electron acceptors and pyrene (PY) and hexamethylbenzene (HMB) as electron donors. Association constants of the formed CT complexes were determined from the NMR data. AQ4 showed weaker electron accepting power than AQ4H, which could be easily explained on the basis of the electron donating nature of the two-hydroxyl groups. AQ4 and AQ4H have higher stability constant with PY than with HMB. This reflects the weaker interaction of the AQs with the latter, which is a direct effect of the six bulky methyl groups. Electronic absorption spectroscopy of the studied system was performed in chloroform and showed the absence of new absorption bands. The extent of interaction between AQs and donors has been computed using molecular mechanics and quantum mechanics. The computed values were compared with the experimental results of association constants.

The Scope of Direct Alkylation of Gold Surface with Solutions of C1-C4 n-Alkylstannanes.

PubMed

Kaletová, Eva; Kohutová, Anna; Hajduch, Jan; Kaleta, Jiří; Bastl, Zdeněk; Pospíšil, Lubomír; Stibor, Ivan; Magnera, Thomas F; Michl, Josef

2015-09-23

Treatment of cleaned gold surfaces with dilute tetrahydrofuran or chloroform solutions of tetraalkylstannanes (alkyl = methyl, ethyl, n-propyl, n-butyl) or di-n-butylmethylstannyl tosylate under ambient conditions causes a self-limited growth of disordered monolayers consisting of alkyls and tin oxide. Extensive use of deuterium labeling showed that the alkyls originate from the stannane and not from ambient impurities, and that trialkylstannyl groups are absent in the monolayers, contrary to previous proposals. Methyl groups attached to the Sn atom are not transferred to the surface. Ethyl groups are transferred slowly, and propyl and butyl rapidly. In all cases, tin oxide is codeposited in submonolayer amounts. The monolayers were characterized by ellipsometry, contact angle goniometry, polarization modulated IR reflection absorption spectroscopy, X-ray photoelectron spectroscopy, and electrochemical impedance spectroscopy with ferrocyanide/ferricyanide, which revealed a very low charge-transfer resistance. The thermal stability of the monolayers and their resistance to solvents are comparable with those of an n-octadecanethiol monolayer. A preliminary examination of the kinetics of monolayer deposition from a THF solution of tetra-n-butylstannane revealed an approximately half-order dependence on the bulk solution concentration of the stannane, hinting that more than one alkyl can be transferred from a single stannane molecule. A detailed structure of the attachment of the alkyl groups is not known, and it is proposed that it involves direct single or multiple bonding of one or more C atoms to one or more Au atoms.

Dissolution DNP-NMR spectroscopy using galvinoxyl as a polarizing agent

NASA Astrophysics Data System (ADS)

Lumata, Lloyd L.; Merritt, Matthew E.; Malloy, Craig R.; Sherry, A. Dean; van Tol, Johan; Song, Likai; Kovacs, Zoltan

2013-02-01

The goal of this work was to test feasibility of using galvinoxyl (2,6-di-tert-butyl-α-(3,5-di-tert-butyl-4-oxo-2,5-cyclohexadien-1-ylidene)-p-tolyloxy) as a polarizing agent for dissolution dynamic nuclear polarization (DNP) NMR spectroscopy. We have found that galvinoxyl is reasonably soluble in ethyl acetate, chloroform, or acetone and the solutions formed good glasses when mixed together or with other solvents such as dimethyl sulfoxide. W-band electron spin resonance (ESR) measurements revealed that galvinoxyl has an ESR linewidth D intermediate between that of carbon-centered free radical trityl OX063 and the nitroxide-based 4-oxo-TEMPO, thus the DNP with galvinoxyl for nuclei with low gyromagnetic ratio γ such as 13C and 15N is expected to proceed predominantly via the thermal mixing process. The optimum radical concentration that would afford the highest 13C nuclear polarization (approximately 6% for [1-13C]ethyl acetate) at 3.35 T and 1.4 K was found to be around 40 mM. After dissolution, large liquid-state NMR enhancements were achieved for a number of 13C and 15N compounds with long spin-lattice relaxation time T1. In addition, the hydrophobic galvinoxyl free radical can be easily filtered out from the dissolution liquid when water is used as the solvent. These results indicate that galvinoxyl can be considered as an easily available free radical polarizing agent for routine dissolution DNP-NMR spectroscopy.

Phytochemical analysis, hepatoprotective and antioxidant activity of Alchornea cordifolia methanol leaf extract on carbon tetrachloride-induced hepatic damage in rats.

PubMed

Osadebe, Patience O; Okoye, Festus B C; Uzor, Philip F; Nnamani, Nneka R; Adiele, Ijeoma E; Obiano, Nkemakonam C

2012-04-01

To investigate the hepatoprotective and antioxidant activities of Alchornea cordifolia (A. cordifolia) leaf extract. Various solvent fractions of the methanol extract of the leaf of the plant A. cordifolia Mull. Arg (Fam: Euphorbiaceae) were evaluated for hepatoprotective activity by carbon tetrachloride-induced liver damage in rats. The degree of protection was measured by using biochemical parameters such as serum glutamate oxalate transaminase (SGOT/AST), serum glutamate pyruvate transaminase (SGPT/ALT), alkaline phosphatase (ALP) and total bilirubin. The in vitro antioxidant activity of the extract was also evaluated by the 1, 1-diphenyl- 2-picrylhydrazyl (DPPH) free radical scavenging assay. The extract was subjected to preliminary phytochemical screening. The ethyl acetate and chloroform fractions, at a dose of 300 mg/kg, produced significant (P<0.05) hepatoprotection by decreasing the activities of the serum enzymes and bilirubin while there were marked scavenging of the DPPH free radicals by the fractions. The effects were comparable to those of the standard drugs used for the respective experiments, silymarin and ascorbic acid. Alkaloids, flavonoids, saponins and tannins were detected in the phytochemical screening. From this study, it was concluded that the plant of A. cordifolia possesses hepatoprotective as well as antioxidant activities and these activities reside mainly in the ethyl acetate and acetone fractions of methanol leaf extract. Copyright © 2012 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Phytochemical screening and evaluation of in vitro antioxidant and antimicrobial activities of the indigenous medicinal plant Albizia odoratissima.

PubMed

Banothu, Venkanna; Neelagiri, Chandrasekharnath; Adepally, Uma; Lingam, Jayalakshmi; Bommareddy, Kesavaharshini

2017-12-01

Albizia odoratissima (L. f.) Benth has been used in Indian folk medicine to treat numerous inflammatory pathologies, such as leprosy, ulcers, burns and asthma. To evaluate the antioxidant and antimicrobial properties of A. odoratissima. Dried leaves of A. odoratissima were extracted in organic solvents (hexane, chloroform, ethyl acetate, and methanol). The total phenolic content (TPC) and total flavonoid content (TFC) were determined using the Folin-Ciocalteu and aluminum chloride colorimetric methods, respectively. The antioxidant activity was examined using 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydrogen peroxide (H 2 O 2 ), 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS), and ferric reducing antioxidant power (FRAP) assays. The antibacterial activity was examined using minimum inhibitory concentration (MIC) and the minimum bacterial concentration (MBC), determined by broth microdilution method against Gram-negative bacteria (Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, and Proteus vulgaris) and Gram-positive bacterium (Staphylococcus aureus). The TPC ranged from 4.40 ± 1.06 to 1166.66 ± 31.85 mg GAE/g of dry weight (DW), and the TFC ranged from 48.35 ± 3.62 to 109.74 ± 1.84 mg QE/g of DW. The IC 50 values of the ethyl acetate extract for DPPH, ABTS, and H 2 O 2 were 10.96 ± 0.40, 4.35 ± 0.07, and 163.82 ± 1.52 μg/mL, respectively. Both methanol and ethyl acetate extracts demonstrated effective antibacterial activity with MICs and MBCs values ranging 136-546 μg/mL and 273-1093 μg/mL, respectively, against the tested pathogenic species. The leaves of A. odoratissima showed potent free radical scavenging property and antimicrobial activity.

Synthesis of some glycoside analogs and related compounds from 9-amino-6-(methylthio)-9H-purine.

PubMed

Temple, C; Kussner, C L; Montgomery, J A

1975-12-01

Additional information on the anticancer activity of 9-amino-9H-purine-6(1H)-thione and its derivatives was sought by the synthesis of some 9-(substituted amino)-6-(methylthio)-9H-purines in which the 9-substituent contained functional groups capable of either reversible or irreversible binding with an enzymatic site. Condensation of 9-amino-6-(methylthio)-9H-purine (1) with some carbonyl compounds followed by hydride reduction of the azomethine linkage in the intermediates leads to the 2-pyrrolylmethyl (8), 2,3,4-trihydroxybutyl (10), and the 1,5-dihydroxy-2- and 3-pentyl (11 and 12) compounds. A 4-hydroxybutyl derivative (13) was obtained by alkylation of 18, the 9-acetyl derivative of 1, with 4-chlorobutyl acetate followed by saponification. The cyclization of 13 and 11 with a sulfonyl chloride gave the 9-pyrrolidin-1-yl (27) and the 9-[2-(tosyloxymethyl)pyrrolidin-1-yl] (28), respectively. Acylation of 1 with ethyl L-2-pyrrolidine-5-carboxylate and ethyl 1-methyl-5-pyrrolidone-3-carboxylate, respectively, in Me2SO containing NaH gave the corresponding amides 15 and 17. Alkylation of 18 with 1-bromo-2-chloroethane and epichlorohydrin gave the N-(2-chloroethyl) and N-(1,2-epoxy-3-propyl) derivatives 19 and 20. The chloro group of the chlorobutyl derivative of 18 was displaced with KSCN and NaN3, respectively, to give the thiocyanate and azido derivatives 23 and 24. Hydrogenation of the latter gave the amine (25), which was acylated with ethyl chloroformate to give the (ethoxycarbonyl)amino compound 26. None of these compounds showed activity against L1210 leukemia cells implanted ip in mice on a single-dose schedule, suggesting that the activity observed in the simpler 9-aminopurines resulted from cleavage of the hydrazino linkage to give pH-purine-6(1H)-thione.

In vitro evaluation of the comprehensive antimicrobial and antioxidant properties of Curtisia dentata (Burm.f) C.A. Sm: toxicological effect on the Human embryonic kidney (HEK293) and Human hepatocellular carcinoma (HepG2) cell lines

PubMed Central

Fadipe, VO; Mongalo, NI; Opoku, AR

2015-01-01

Curtisia dentata is used in African traditional medicine to treat variety of infections. C. dentata leaves were collected from Buffelskloof Nature Reserve, South Africa. The ethanol, chloroform, ethyl acetate and acetone extracts were evaluated for antimicrobial activity using micro dilution assay against Escherichia coli, Pseudomonas aeruginosa, Mycobacterium smegmatis, Mycoplasma hominis, Candida albicans and some clinical isolates of Moraxella catarrhalis, Proteus mirabilis and Staphylococcus aureus isolated from HIV patient. Acetone extract exhibited lowest MIC of 0.01 mg/ml against Candida albicans compared to other extracts. Besides lupeol, betulinic acid and ursolic acid, β-sitosterol was isolated for the first time from C. dentata leaves and exhibited antimicrobial activity with MIC values ranging from 0.20 to 6.25 mg/ml. Furthermore, the ethanol extract and the four isolated compounds revealed microbicidal effect, with MIC index of less than 4. Ethanol extract revealed the best total activity of 2400 ml/g against Mycoplasma hominis. Cytotoxicity of the isolated compounds was further investigated against the Human embryonic kidney (HEK293) and Human hepatocellular carcinoma (HepG2) cell lines using the MTT assay. Ursolic acid exhibited the lowest LD50 of 122.4 µg/ml against HEK293 cell line while lupeol exhibited LD50 of 278.8 and 289.4 µg/ml against HEK293 and HepG2 respectively. Lupeol exhibited low selectivity index. Ethyl acetate and acetone extracts were further investigated for antioxidant activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH). The acetone extract exhibited potent inhibition of DPPH compared to ethyl acetate extract. The findings of the current work validate the use of the plant species in the treatment of various human infections. PMID:27065768

DOE Office of Scientific and Technical Information (OSTI.GOV)

McDonald, M.N.; Vire, D.E.

This article reviews the role chloroform has played in dentistry and describes an occupational health clinical investigation into the possible hazards of chloroform use in the operatory. Due to a Food and Drug Administration ban on drugs and cosmetics containing chloroform, there has been some confusion as to whether the use of chloroform in the practice of dentistry is considered unsafe or has been prohibited. Utilizing common endodontic treatment methods employing chloroform, this study reports no negative health effects to the dentist or assistant and air vapor levels well below Occupational Health and Safety Administration mandated maximum levels. The reportmore » concludes that, with careful and controlled use, chloroform can be a useful adjunct in the practice of dentistry. The Food and Drug Administration has no jurisdiction over a dentist's use of chloroform in clinical practice and has not proven that chloroform is a human carcinogen.« less

Mechanism of chloroform-induced renal toxicity: Non-involvement of hepatic cytochrome P450-dependent metabolism

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fang Cheng; Behr, Melissa; Xie Fang

2008-02-15

Chloroform causes hepatic and renal toxicity in a number of species. In vitro studies have indicated that chloroform can be metabolized by P450 enzymes in the kidney to nephrotoxic intermediate, although direct in vivo evidence for the role of renal P450 in the nephrotoxicity has not been reported. This study was to determine whether chloroform renal toxicity persists in a mouse model with a liver-specific deletion of the P450 reductase (Cpr) gene (liver-Cpr-null). Chloroform-induced renal toxicity and chloroform tissue levels were compared between the liver-Cpr-null and wild-type mice at 24 h following differing doses of chloroform. At a chloroform dosemore » of 150 mg/kg, the levels of blood urea nitrogen (BUN) were five times higher in the exposed group than in the vehicle-treated one for the liver-Cpr-null mice, but they were only slightly higher in the exposed group than in the vehicle-treated group for the wild-type mice. Severe lesions were found in the kidney of the liver-Cpr-null mice, while only mild lesions were found in the wild-type mice. At a chloroform dose of 300 mg/kg, severe kidney lesions were observed in both strains, yet the BUN levels were still higher in the liver-Cpr-null than in the wild-type mice. Higher chloroform levels were found in the tissues of the liver-Cpr-null mice. These findings indicated that loss of hepatic P450-dependent chloroform metabolism does not protect against chloroform-induced renal toxicity, suggesting that renal P450 enzymes play an essential role in chloroform renal toxicity.« less

Applicability of solid-phase microextraction combined with gas chromatography atomic emission detection (GC-MIP AED) for the determination of butyltin compounds in sediment samples.

PubMed

Carpinteiro, J; Rodríguez, I; Cela, R

2004-11-01

The performance of solid-phase microextraction (SPME) applied to the determination of butyltin compounds in sediment samples is systematically evaluated. Matrix effects and influence of blank signals on the detection limits of the method are studied in detail. The interval of linear response is also evaluated in order to assess the applicability of the method to sediments polluted with butyltin compounds over a large range of concentrations. Advantages and drawbacks of including an SPME step, instead of the classic liquid-liquid extraction of the derivatized analytes, in the determination of butyltin compounds in sediment samples are considered in terms of achieved detection limits and experimental effort. Analytes were extracted from the samples by sonication using glacial acetic acid. An aliquot of the centrifuged extract was placed on a vial where compounds were ethylated and concentrated on a PDMS fiber using the headspace mode. Determinations were carried out using GC-MIP AED.

Development of a test method for carbonyl compounds from stationary source emissions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhihua Fan; Peterson, M.R.; Jayanty, R.K.M.

1997-12-31

Carbonyl compounds have received increasing attention because of their important role in ground-level ozone formation. The common method used for the measurement of aldehydes and ketones is 2,4-dinitrophenylhydrazine (DNPH) derivatization followed by high performance liquid chromatography and ultra violet (HPLC-UV) analysis. One of the problems associated with this method is the low recovery for certain compounds such as acrolein. This paper presents a study in the development of a test method for the collection and measurement of carbonyl compounds from stationary source emissions. This method involves collection of carbonyl compounds in impingers, conversion of carbonyl compounds to a stable derivativemore » with O-2,3,4,5,6-pentafluorobenzyl hydroxylamine hydrochloride (PFBHA), and separation and measurement by electron capture gas chromatography (GC-ECD). Eight compounds were selected for the evaluation of this method: formaldehyde, acetaldehyde, acrolein, acetone, butanal, methyl ethyl ketone (MEK), methyl isobutyl ketone (MIBK), and hexanal.« less

Production of trichothecene mycotoxins by Australian Fusarium species.

PubMed

McLachlan, A; Shaw, K J; Hocking, A D; Pitt, J I; Nguyen, T H

1992-01-01

Australian isolates of Fusarium species were grown on potato dextrose agar. Trichothecenes produced by these species were extracted by ethyl acetate followed by methanol and a silica gel column was used to clean-up the extract. The extracted samples were derivatized by acetylation with trifluoroacetic anhydride and the derivatives analysed by gas chromatography/mass spectrometry (GC/MS). Multiple ion detection was used to trace ions characteristic of the trichothecenes expected to be present. Quantitation of those found was based on a known mass of pentabromophenol that was added as an internal standard. Eight species of Fusarium (nineteen strains) were surveyed, of which three species, F. acuminatum, F. equiseti and F. sporotrichioides, produced the trichothecenes scirpentriol, diacetoxyscirpenol, neosolaniol, HT-2 toxin, T-2 toxin, T-2 tetraol and deoxynivalenol. Wheat samples were inoculated with four different species of Fusarium, F. acuminatum, F. equiseti, F. graminearum and F. sporotrichioides, and in these samples diacetoxyscirpenol, neosolaniol, HT-2 toxin and T-2 toxin were found.

Identification of a dicaffeoylquinic acid isomer from Arctium lappa with a potent anti-ulcer activity.

PubMed

Carlotto, Juliane; da Silva, Luisa M; Dartora, Nessana; Maria-Ferreira, Daniele; Sabry, Diego de A; Filho, Arquimedes P S; de Paula Werner, Maria F; Sassaki, Guilherme L; Gorin, Philip A J; Iacomini, Marcello; Cipriani, Thales R; de Souza, Lauro M

2015-04-01

Leaves of Arctium lappa contain several mono- and dicaffeoylquinic acids, as evaluated by liquid chromatography-mass spectrometry. In order to investigate the protection on gastric mucosa against ulcers, rats were treated with fractions from leaf extract prior to ethanol-induced ulcers. The original fraction obtained as ethanol soluble fraction from hot aqueous extract was able to protect de gastric mucosa, and this effect was retained in the ethyl acetate fraction, obtained from liquid/liquid fractionation. The main compound in this fraction was isolated and chemically characterized by nuclear magnetic resonance and mass spectrometry, assisted by isopropylidene derivatization which gave rise a mass increment of 40 units. Therefore, the underivatized compound that had m/z 515.119 [M-H](-) was shifted to m/z 555.151, being confirmed as 1,3-O-dicaffeoylquinic acid, which presented an ED50 of 57 µg kg(-1) on gastric protection, lesser than the therapeutic concentration of omeprazole (40 mg kg(-1)). Copyright © 2014 Elsevier B.V. All rights reserved.

In Vitro Antimicrobial Bioassays, DPPH Radical Scavenging Activity, and FTIR Spectroscopy Analysis of Heliotropium bacciferum.

PubMed

Ahmad, Sohail; AbdEl-Salam, Naser M; Ullah, Riaz

2016-01-01

The present study deals with the antimicrobial, antioxidant, and functional group analysis of Heliotropium bacciferum extracts. Disc diffusion susceptibility method was followed for antimicrobial assessment. Noteworthy antimicrobial activities were recorded by various plant extracts against antibiotic resistant microorganisms. Plant flower extracts antioxidant activity was investigated against 2, 2-diphenyl-1-picryl hydrazyl radical by ultraviolet spectrophotometer (517 nm). Plant extracts displayed noteworthy radical scavenging activities at all concentrations (25-225 μg/mL). Notable activities were recorded by crude, chloroform and ethyl acetate extracts up to 88.27% at 225 μg/mL concentration. Compounds functional groups were examined by Fourier transform infrared spectroscopic studies. Alkanes, alkenes, alkyl halides, amines, carboxylic acids, amides, esters, alcohols, phenols, nitrocompounds, and aromatic compounds were identified by FTIR analysis. Thin layer chromatography bioautography was carried out for all plant extracts. Different bands were separated by various solvent systems. The results of the current study justify the use of Heliotropium bacciferum in traditional remedial herbal medicines.

Evaluation of medicinal plant extracts and isolated compound epicatechin from Ricinus communis against Paramphistomum cervi.

PubMed

Zahir, Abdul Abduz; Rahuman, Abdul Abdul; Bagavan, Asokan; Geetha, Kannappan; Kamaraj, Chinnaperumal; Elango, Gandhi

2012-10-01

The purpose of this study is to determine the efficacies of hexane, chloroform, ethyl acetate, acetone, and methanol leaf extracts of Euphorbia hirta L., Psidium guajava L., Ricinus communis L., Solanum trilobatum L., and Tridax procumbens L. against sheep fluke Paramphistomum cervi (Digenea: Paramphistomatidae). All plant extracts showed moderate effects after 24 h of exposure; however, the highest parasite mortality was found in the methanol extract of R. communis. In the present study, bioassay-guided fractionation of methanol extract of R. communis led to the separation and identification of epicatechin as a potential new compound (LC(50) = 31.2; LC(90) = 105.0 ppm) against P. cervi. The structures were established from infrared, ultraviolet, (1)H-nuclear magnetic resonance (NMR), (13)C-NMR, and mass spectral data which confirmed the identification of the compound epicatechin from R. communis. Results of this study showed that the methanol extract of R. communis may be considered as a potent source and epicatechin as a new natural parasitic agent.

Calix[4]arene coated QCM sensors for detection of VOC emissions: Methylene chloride sensing studies.

PubMed

Temel, Farabi; Tabakci, Mustafa

2016-06-01

This paper describes the sensing studies of QCM sensors with coated some calixarene derivatives bearing different functional groups for some selected Volatile Organic Compounds (VOCs) such as acetone, acetonitrile, carbon tetrachloride, chloroform, methylene chloride (MC), N,N-dimethylformamide, 1,4-dioxane, ethanol, ethyl acetate, xylene, methanol, n-hexane and toluene. The initial experiments have revealed that whole the calix[4]arene modified QCM sensors exhibited strongest sensing ability to MC emissions. Thus, the detailed studies were performed for only MC emissions after the determination of relatively more effective calix-coated QCM sensors for MC emissions in aqueous media. The results demonstrated that QCM sensor coated with calix-7 bearing both amino and imidazole groups was most useful sensor for MC emissions with 54.1ppm of detection limit. Moreover, it was understood that cyclic structures, H-bonding capabilities and also good preorganization properties of calixarene derivatives played an important role in VOC sensing processes. Copyright © 2016 Elsevier B.V. All rights reserved.

A Review on the Pharmacological Activities and Phytochemicals of Alpinia officinarum (Galangal) Extracts Derived from Bioassay-Guided Fractionation and Isolation

PubMed Central

Basri, Aida Maryam; Taha, Hussein; Ahmad, Norhayati

2017-01-01

The rhizomes of Alpinia officinarum Hance have been used conventionally for the treatment of various ailments, triggering a wide interest from the scientific research community on this ethnomedicinal plant. This review summarizes the phytochemical and pharmacological properties of the extracts and fractions from A. officinarum, a plant species of the Zingiberaceae family. Different parts of the plant – leaves, roots, rhizomes, and aerial parts – have been extracted in various solvents – methanol, ethanol, ethyl acetate, hexane, dichloromethane, aqueous, chloroform, and petroleum ether, using various techniques – Soxhlet extraction, maceration, ultrasonication, and soaking, whereas fractionation of the plant extracts involves the solvent–solvent partition method. The extracts, fractions, and isolated compounds have been studied for their biological activities – antioxidant, antibacterial, anti-inflammatory, anticancer, antiproliferative, inhibition of enzymes, as well as the inhibition of nitric oxide production. More findings on A. officinarum are certainly important to further develop potential bioactive drug compounds. PMID:28503054

A new ester coumarin from Ferula Persica wild, indigenous to Iran.

PubMed

Razavi, Seyed Mehdi; Janani, Mehrnoush

2015-01-01

Ferula persica wild (Apiaceae) is a perennial herb indigenous to Iran. It has been used in folk medicine for treatment of diabetes, lowering of blood pressure and for antispasmodic, carminative, laxative and expectorant effects in central Iran. Dried ground roots of F. persica (150 g) were extracted sequentially with n-hexane, dichloromethane and methanol (MeOH), 500 ml each, using a Soxhlet apparatus. The n-hexane extract of the roots (3 g) was subjected to vacuum liquid chromatography on silica gel, eluting with solvent mixtures of increasing polarity: 100% n-hexane-ethyl acetate (EtOAc), to yield a number of fractions, Fraction 4 (80% EtOAc in n-hexane) was further analysed by preparative TLC (mobile phase was 12% acetone in chloroform) to yield a coumarin ester (10.1 mg, Rf = 0.31, blue florescent). The structure of the isolated compound was elucidated by spectroscopic means. The compound is 7-O-(4,8,12 -trihydroxy-4,8,12-trimethyl-tridecanoyl)-coumarin, named, ferulone C as a new natural product.

Sister chromatid exchanges induced by inhaled anesthetics

DOE Office of Scientific and Technical Information (OSTI.GOV)

White,A.E.; Takehisa, S.; Eger II, E.I.

1970-05-01

There is sufficient evidence that anesthetics may cause cancer to justify a test of their carcinogenic potential. Baden et al., using the Ames test, a rapid and inexpensive genetic indicator of carcinogenicity, have shown that among currently used anesthetics fluorxene alone caused bacterial mutations. The authors used the sister chromatid exchange (SCE) technique, another rapid assay of mutagenic-carcinogenic potential. The frequency of sister chromatid exchanges in Chinese hamster ovary cells increases when the cell cultures are exposed to mutagen-carcinogens, particulary in the presence of a metabolic activating system. With this test system a one-hour exposure to 1 MAC nitrous oxide,more » diethyl ether, trichloroethylene, halothane, enflurane, isoflurane, methoxyflurane, or chloroform did not increase SCE values. Divinyl ether, fluroxene and ethyl vinyl ether increased SCE values in the same circumstances. Results of this study of mammalian cells suggest that no currently used anesthetic is a mutagen-carcinogen. The results also suggest that anesthetics containing a vinyl moiety may be mutagen-carcinogens.« less

In vitro antibacterial and antifungal activities of Rhus tripartitum used as antidiarrhoeal in Tunisian folk medicine.

PubMed

Abbassi, Feten; Hani, Khaled

2012-01-01

Rhus tripartitum (African sumac) is a plant commonly used in Tunisian traditional medicine to treat diarrhoea and dysentery. In this study, we have demonstrated that R. tripartitum extracts exhibited a significant broad spectrum activity against one or more of the test microorganisms with a zone size ranging from 8 to 28 mm in diameter. These diameters are much higher than those obtained with standard antibiotics. The chloroformic extracts were found to be effective against bacterial and fungal strains tested, with MIC values ranging between 0.07 and 0.62 mg mL(-1) against Staphylococcus aureus and Candida albicans. However, ethyl-acetate/methanol fractions showed a selective activity only against bacterial microorganisms with low MIC values between 0.07 and 0.15 mg mL(-1). The overall results suggested that the traditional use of R. tripartitum for the treatment of diarrhoea tract infections was attributed to the presence of antibacterial agents.

Evaluation of the Wound Healing Potential of Achillea biebersteinii Afan. (Asteraceae) by In Vivo Excision and Incision Models

PubMed Central

Akkol, Esra Küpeli; Koca, Ufuk; Pesin, Ipek; Yilmazer, Demet

2011-01-01

Achillea species are widely used for diarrhea, abdominal pain, stomachache and healing of wounds in folk medicine. To evaluate the wound healing activity of the plant, extracts were prepared with different solvents; hexane, chloroform, ethyl acetate and methanol, respectively from the roots of Achillea biebersteinii. Linear incision by using tensiometer and circular excision wound models were employed on mice and rats. The wound healing effect was comparatively evaluated with the standard skin ointment Madecassol. The n-hexane extract treated groups of animals showed 84.2% contraction, which was close to contraction value of the reference drug Madecassol (100%). On the other hand the same extract on incision wound model demonstrated a significant increase (40.1%) in wound tensile strength as compared to other groups. The results of histoptological examination supported the outcome of linear incision and circular excision wound models as well. The experimental data demonstrated that A. biebersteinii displayed remarkable wound healing activity. PMID:19546149

[Determination of solubility parameters of high density polyethylene by inverse gas chromatography].

PubMed

Wang, Qiang; Chen, Yali; Liu, Ruiting; Shi, Yuge; Zhang, Zhengfang; Tang, Jun

2011-11-01

Inverse gas chromatographic (IGC) technology was used to determine the solubility parameters of high density polyethylene (HDPE) at the absolute temperatures from 303.15 to 343.15 K. Six solvents were applied as test probes including hexane (n-C6), heptane (n-C7), octane (n-C8), nonane (n-C9), chloroform (CHCl3) and ethyl acetate (EtAc). Some thermodynamic parameters were obtained by IGC data analysis such as the specific retention volumes of the solvents (V(0)(g)), the molar enthalpy of sorption (delta H(S)(1)), the partial molar enthalpy of mixing at infinite dilution (delta H(1)(infinity)), the molar enthalpy of vaporization (delta H(v)), the activity coefficients at infinite dilution (omega (1)(infinity)), and Flow-Huggins interaction parameters (X(1,2)(infinity)) between HDPE and probe solvents. The results showed that the above six probes are poor solvents for HDPE. The solubility parameter of HDPE at room temperature (298.15 K) was also derived as 19.00 (J/cm3)(0.5).

Aprotic solvents effect on the UV-visible absorption spectra of bixin.

PubMed

Rahmalia, Winda; Fabre, Jean-François; Usman, Thamrin; Mouloungui, Zéphirin

2014-10-15

We describe here the effects of aprotic solvents on the spectroscopic characteristics of bixin. Bixin was dissolved in dimethyl sulfoxide, acetone, dichloromethane, ethyl acetate, chloroform, dimethyl carbonate, cyclohexane and hexane, separately, and its spectra in the resulting solutions were determined by UV-visible spectrophotometry at normal pressure and room temperature. We analyzed the effect of aprotic solvents on λmax according to Onsager cavity model and Hansen theory, and determined the approximate absorption coefficient with the Beer-Lambert law. We found that the UV-visible absorption spectra of bixin were found to be solvent dependent. The S0→S2 transition energy of bixin in solution was dependent principally on the refractive index of the solvents and the bixin-solvent dispersion interaction. There was a small influence of the solvents dielectric constant, permanent dipole interaction and hydrogen bonding occurred between bixin and solvents. The absorbance of bixin in various solvents, with the exception of hexane, increased linearly with concentration. Copyright © 2014 Elsevier B.V. All rights reserved.

Aprotic solvents effect on the UV-visible absorption spectra of bixin

NASA Astrophysics Data System (ADS)

Rahmalia, Winda; Fabre, Jean-François; Usman, Thamrin; Mouloungui, Zéphirin

2014-10-01

We describe here the effects of aprotic solvents on the spectroscopic characteristics of bixin. Bixin was dissolved in dimethyl sulfoxide, acetone, dichloromethane, ethyl acetate, chloroform, dimethyl carbonate, cyclohexane and hexane, separately, and its spectra in the resulting solutions were determined by UV-visible spectrophotometry at normal pressure and room temperature. We analyzed the effect of aprotic solvents on λmax according to Onsager cavity model and Hansen theory, and determined the approximate absorption coefficient with the Beer-Lambert law. We found that the UV-visible absorption spectra of bixin were found to be solvent dependent. The S0 → S2 transition energy of bixin in solution was dependent principally on the refractive index of the solvents and the bixin-solvent dispersion interaction. There was a small influence of the solvents dielectric constant, permanent dipole interaction and hydrogen bonding occurred between bixin and solvents. The absorbance of bixin in various solvents, with the exception of hexane, increased linearly with concentration.

In vitro antifungal activity of extracts obtained from Hypericum perforatum adventitious roots cultured in a mist bioreactor against planktonic cells and biofilm of Malassezia furfur.

PubMed

Simonetti, Giovanna; Tocci, Noemi; Valletta, Alessio; Brasili, Elisa; D'Auria, Felicia Diodata; Idoux, Alicia; Pasqua, Gabriella

2016-01-01

Xanthone-rich extracts from Hypericum perforatum root cultures grown in a Mist Bioreactor as antifungal agents against Malassezia furfur. Extracts of Hypericum perforatum roots grown in a bioreactor showed activity against planktonic cells and biofilm of Malassezia furfur. Dried biomass, obtained from roots grown under controlled conditions in a ROOTec mist bioreactor, has been extracted with solvents of increasing polarity (i.e. chloroform, ethyl acetate and methanol). The methanolic fraction was the richest in xanthones (2.86 ± 0.43 mg g(-1) DW) as revealed by HPLC. The minimal inhibitory concentration of the methanol extract against M. furfur planktonic cells was 16 μg mL(-1). The inhibition percentage of biofilm formation, at a concentration of 16 μg mL(-1), ranged from 14% to 39%. The results show that H. perforatum root extracts could be used as new antifungal agents in the treatment of Malassezia infections.

Leaf extracts from Moricandia arvensis promote antiproliferation of human cancer cells, induce apoptosis, and enhance antioxidant activity.

PubMed

Skandrani, Ines; Boubaker, Jihed; Bhouri, Wissem; Limem, Ilef; Kilani, Soumaya; Ben Sghaier, Mohamed; Neffati, Aicha; Bouhlel, Ines; Ghedira, Kamel; Chekir-Ghedira, Leila

2010-01-01

The in vitro antiproliferative, apoptotic, and antioxidant activities from leaf extracts of Moricandia arvensis, which are used in traditional cooking and medicines, were investigated. The MTT assay revealed that only TOF (total oligomer flavonoids), ethyl acetate (EA), chloroform (Chl), and petroleum ether (PE) extracts inhibited the proliferation of K562 cells. Apoptosis plays a very important role in the treatment of cancer by promoting the apoptosis of cancer cells and limiting the concurrent death of normal cells. Thus, the possible effects of M. arvensis extracts on the induction of apoptosis in human leukemic cells (K562 cells) were investigated. The electrophoretic analysis of DNA fragmentation confirms that TOF, Chl, PE, and EA extracts provoke DNA fragmentation. Using the lipid peroxidation inhibitory assay, the antioxidant capacity of M. arvensis extracts was evaluated by the ability of each extract to inhibit malondialdehyde formation. It was revealed that EA and TOF extracts are the most active in scavenging the hydroxyl radicals.

Isolation, characterization and chromatography based purification of antibacterial compound isolated from rare endophytic actinomycetes Micrococcus yunnanensis.

PubMed

Ranjan, Ravi; Jadeja, Vasantba

2017-10-01

Endophytic actinomycetes are considered as one of the relatively unexplored potential sources in search of antibiotic producer against antibiotic resistant pathogens. A potent strain isolated from Catharanthus roseus that displays antibacterial potential against antibiotic resistant human pathogen Staphylococcus aureus was characterized and designated as Micrococcus yunnanensis strain rsk5. Rsk5 is capable of producing optimum antibacterial metabolites on starch casein medium at 30 °C, pH 5 and 2% NaCl condition. The crude antibacterial agent was extracted from fermentation broth by ethyl acetate and separated by TLC using chloroform-methanol (24:1, v/v) solvent system with R f value of 0.26. It was partially purified by flash chromatography, followed by HPLC and analyzed by ultraviolet visible spectrophotometer to get absorption maxima at 208.4 nm. The ESI-MS spectra showed molecular ion peaks at m / z 472.4 [M-H], which does not match with any known antibacterial compound.

Phytochemicals from Mangifera pajang Kosterm and their biological activities.

PubMed

Ahmad, Sadikah; Sukari, Mohd Aspollah; Ismail, Nurussaadah; Ismail, Intan Safinar; Abdul, Ahmad Bustamam; Abu Bakar, Mohd Fadzelly; Kifli, Nurolaini; Ee, Gwendoline C L

2015-03-26

Mangifera pajang Kosterm is a plant species from the mango family (Anacardiaceae). The fruits are edible and have been reported to have high antioxidant content. However, the detailed phytochemical studies of the plant have not been reported previously. This study investigates the phytochemicals and biological activities of different parts of Mangifera pajang. The plant samples were extracted with solvents of different polarity to obtain the crude extracts. The isolated compounds were characterized using spectroscopic methods. The extracts and isolated compounds were subjected to cytotoxicity tests using human breast cancer (MCF-7), human cervical cancer (HeLa) and human colon cancer (HT-29) cells. The free radical scavenging activity test was conducted using the DPPH assay. Antimicrobial activity tests were carried out by using the disc diffusion method. Phytochemical investigation on the kernel, stem bark and leaves of Mangifera pajang led to the isolation of methyl gallate (1), mixture of benzaldehyde (2) and benzyl alcohol (3), mangiferonic acid (4), 3β-hydroxy-cycloart-24-ene-26-oic acid (5), 3β,23-dihydroxy-cycloart-24-ene-26-oic acid (6), lupeol(7) lupenone(8), β-sitosterol(9), stigmasterol(10), trans-sobrerol(11) and quercitrin (12). Crude ethyl acetate and methanol extracts from the kernel indicated strong cytotoxic activity towards MCF-7 and HeLa cells with IC50 values of less than 10 μg/mL, while petroleum ether, chloroform and ethyl acetate extracts of the stem bark showed strong to moderate activity against MCF-7, HeLa and HT-29 cancer cell lines with IC50 values ranging from 5 to 30 μg/mL. As for the antimicrobial assays, only the ethyl acetate and methanol extracts from the kernel displayed some inhibition against the microbes in the antibacterial assays. The kernel extracts showed highest free radical scavenging activity with IC50 values of less than 10 μg/mL, while the ethyl acetate and methanol extracts of leaves displayed only weak activity in the DPPH assays. Phytochemical investigations on various parts of Mangifera pajang have identified terpenoids and a flavonol derivative as major constituents. Bioassay studies have indicated that the crude extracts and isolated compounds have potential as naturally-derived anticancer and antimicrobial agents, besides possess high free radical scavenging activity.

Solute-solvent complex switching dynamics of chloroform between acetone and dimethylsulfoxide-two-dimensional IR chemical exchange spectroscopy.

PubMed

Kwak, Kyungwon; Rosenfeld, Daniel E; Chung, Jean K; Fayer, Michael D

2008-11-06

Hydrogen bonds formed between C-H and various hydrogen bond acceptors play important roles in the structure of proteins and organic crystals, and the mechanisms of C-H bond cleavage reactions. Chloroform, a C-H hydrogen bond donor, can form weak hydrogen-bonded complexes with acetone and with dimethylsulfoxide (DMSO). When chloroform is dissolved in a mixed solvent consisting of acetone and DMSO, both types of hydrogen-bonded complexes exist. The two complexes, chloroform-acetone and chloroform-DMSO, are in equilibrium, and they rapidly interconvert by chloroform exchanging hydrogen bond acceptors. This fast hydrogen bond acceptor substitution reaction is probed using ultrafast two-dimensional infrared (2D-IR) vibrational echo chemical exchange spectroscopy. Deuterated chloroform is used in the experiments, and the 2D-IR spectrum of the C-D stretching mode is measured. The chemical exchange of the chloroform hydrogen bonding partners is tracked by observing the time-dependent growth of off-diagonal peaks in the 2D-IR spectra. The measured substitution rate is 1/30 ps for an acetone molecule to replace a DMSO molecule in a chloroform-DMSO complex and 1/45 ps for a DMSO molecule to replace an acetone molecule in a chloroform-acetone complex. Free chloroform exists in the mixed solvent, and it acts as a reactive intermediate in the substitution reaction, analogous to a SN1 type reaction. From the measured rates and the equilibrium concentrations of acetone and DMSO, the dissociation rates for the chloroform-DMSO and chloroform-acetone complexes are found to be 1/24 ps and 1/5.5 ps, respectively. The difference between the measured rate for the complete substitution reaction and the rate for complex dissociation corresponds to the diffusion limited rate. The estimated diffusion limited rate agrees well with the result from a Smoluchowski treatment of diffusive reactions.

1-(3-aminopropyl)-3-butylimidazolium bromide for carboxyl group derivatization: potential applications in high sensitivity peptide identification by mass spectrometry.

PubMed

Qiao, Xiaoqiang; Zhou, Yuan; Hou, Chunyan; Zhang, Xiaodan; Yang, Kaiguang; Zhang, Lihua; Zhang, Yukui

2013-03-01

The cationic reagent 1-(3-aminopropyl)-3-butylimidazolium bromide (BAPI) was exploited for the derivatization of carboxyl groups on peptides. Nearly 100% derivatization efficiency was achieved with the synthetic peptide RVYVHPI (RI-7). Furthermore, the peptide derivative was stable in a 0.1% TFA/water solution or a 0.1% (v/v) TFA/acetonitrile/water solution for at least one week. The effect of BAPI derivatization on the ionization of the peptide RI-7 was further investigated, and the detection sensitivity was improved >42-fold via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), thus outperforming the commercial piperazine derivatization approach. Moreover, the charge states of the peptide were largely increased via BAPI derivatization by electrospray ionization (ESI) MS. The results indicate the potential merits of BAPI derivatization for high sensitivity peptide analysis by MS.

Quantitative Silylation Speciations of Primary Phenylalkyl Amines, Including Amphetamine and 3,4-Methylenedioxyamphetamine Prior to Their Analysis by GC/MS.

PubMed

Molnár, Borbála; Fodor, Blanka; Boldizsár, Imre; Molnár-Perl, Ibolya

2015-10-20

A novel, quantitative trimethylsilylation approach derivatizing 11 primary phenylalkyl amines (PPAAs), including amphetamine (A) and 3,4-methylenedioxyamphetamine (MDA), was noted. Triggering the fully derivatized ditrimethylsilyl (diTMS) species with the N-methyl-N-(trimethylsilyl)-trifluoroacetamide (MSTFA) reagent, a new principle was recognized followed by GC/MS. In the course of method optimization, the complementary impact of solvents (acetonitrile, ACN; ethyl acetate, ETAC; pyridine, PYR) and catalysts (trimethylchlorosilane, TMCS; trimethyliodosilane, TMIS) was studied: the role of solvent and catalyst proved to be equally crucial. Optimum, proportional, huge responses were obtained with the MSTFA/PYR = 2/1-9/1 (v/v) reagent applying catalysts; A and MDA needed the TMIS, while the rest of PPAAs provided the diTMS products also with TMCS. Similar to derivatives generated with hexamethyldisilazane and perfluorocarboxylic acid (HMDS and PFCA) ( Molnár et al. Anal. Chem. 2015 , 87 , 848 - 852 ), the fully silylated PPAAs offer several advantages. Both of our methods save time and cost by allowing for direct injection of analytes into the column; this is in stark contrast with the requirement to evaporate acid anhydrides by nitrogen prior to their injection. Efficiences of the novel catalyzed trimethylsilylation (MSTFA) and our recently introduced (now, for A and MDA extended) acylation principle were contrasted. Catalyzed trimethylsilylation led to diTMS derivatives resulting in on average a 1.7 times larger response compared to the corresponding acylated species. Catalyzed trimethylsilylation of PPAAs, A, and MDA were characterized with retention, mass fragmentation, and analytical performance properties (R(2), LOQ values). The practical utility of ditrimethylsilyation was shown by analyzing A in urine and mescaline (MSC) in cactus samples.

Mapping of contact sites in complex formation between transducin and light-activated rhodopsin by covalent crosslinking: Use of a photoactivatable reagent

PubMed Central

Cai, Kewen; Itoh, Yoshiki; Khorana, H. Gobind

2001-01-01

Interaction of light-activated rhodopsin with transducin (T) is the first event in visual signal transduction. We use covalent crosslinking approaches to map the contact sites in interaction between the two proteins. Here we use a photoactivatable reagent, N-[(2-pyridyldithio)-ethyl], 4-azido salicylamide. The reagent is attached to the SH group of cytoplasmic monocysteine rhodopsin mutants by a disulfide-exchange reaction with the pyridylthio group, and the derivatized rhodopsin then is complexed with T by illumination at λ >495 nm. Subsequent irradiation of the complex at λ310 nm generates covalent crosslinks between the two proteins. Crosslinking was demonstrated between T and a number of single cysteine rhodopsin mutants. However, sites of crosslinks were investigated in detail only between T and the rhodopsin mutant S240C (cytoplasmic loop V-VI). Crosslinking occurred predominantly with Tα. For identification of the sites of crosslinks in Tα, the strategy used involved: (i) derivatization of all of the free cysteines in the crosslinked proteins with N-ethylmaleimide; (ii) reduction of the disulfide bond linking the two proteins and isolation of all of the Tα species carrying the crosslinked moiety with a free SH group; (iii) adduct formation of the latter with the N-maleimide moiety of the reagent, maleimido-butyryl-biocytin, containing a biotinyl group; (iv) trypsin degradation of the resulting Tα derivatives and isolation of Tα peptides carrying maleimido-butyryl-biocytin by avidin-agarose chromatography; and (v) identification of the isolated peptides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. We found that crosslinking occurred mainly to two C-terminal peptides in Tα containing the amino acid sequences 310–313 and 342–345. PMID:11320237

Determination of orthophthalaldehyde in air using 2,4-dinitrophenylhydrazine-impregnated silica cartridge and high-performance liquid chromatography.

PubMed

Uchiyama, Shigehisa; Matsushima, Erika; Tokunaga, Hiroshi; Otsubo, Yasufumi; Ando, Masanori

2006-05-26

A new method is described for the determination of orthophthalaldehyde in air which is used for the disinfection of various instruments (e.g. endoscopes) in hospital. Orthophthalaldehyde in air was collected with a silica gel cartridge impregnated with acidified 2,4-dinitrophenylhydrazine (DNPH-cartridge) and derivatives were analyzed by high-performance liquid chromatography (HPLC). In this study, the derivatization was examined by comparing the process with three phthalaldehyde isomers (ortho-, iso- and tere-). In the case of iso- and tere-phthalaldehyde, derivatives synthesized with excess of aldehyde consisted mainly of mono-derivatives, and derivatives synthesized with excess of DNPH consisted mainly of bis-derivative. In the case of orthophthalaldehyde, derivative consisted of only bis-derivative and mono-derivative was never observed under any conditions. Orthophthalaldehyde was completely retained by the DNPH-cartridge during air sampling, however, the derivatization reaction was incomplete and unreacted orthophthalaldehyde was flushed from the cartridge during the subsequent solvent extraction process. Unreacted orthophthalaldehyde and DNPH reacted again in the extraction solvent solution. Immediately after the solvent extraction, both mono- and bis-DNPhydrazone derivatives of orthophthalaldehyde were present in the solution. However, over time, the mono-derivative decreased and bis-derivative increased until only the bis-derivative was left allowing accurate determination of the orthophthalaldehyde concentration. The transformation of mono-derivative to bis-derivative was faster in polar aprotic solvents such as acetonitrile, dimethyl sulfoxide and ethyl acetate. Transformation was found to occur most quickly in acetonitrile solvent and was completed in 4 h in this case. It was possible to measure orthophthalaldehyde in air as bis-derivative using a DNPH impregnated silica cartridge and HPLC analysis.

Apparent pollution of groundwater caused by natural formation of chloroform in forest soils

NASA Astrophysics Data System (ADS)

Jacobsen, O.; Laier, T.; Albers, C. N.; Hunkeler, D.

2011-12-01

Halogenated compounds are known to be formed in natural environments. Many of these compounds are similar to industrially produced compounds and are toxic or carcinogenic. High concentration of chloroform in groundwater is usually attributed to anthropogenic input, but we have found that the groundwater beneath some pristine areas contained chloroform exceeding 1 μg/L. We investigated four coniferous forests over a period of several years in order to measure the net-formation of chloroform. Field measurements of atmospheric and soil air concentrations of chloroform were monitored. Analyses of soil air at 40 cm depth in different parts of the forests and adjacent areas revealed an extremely large variation in chloroform concentration exceeding two orders of magnitude. Up to 100 ppbv was found in soil air under the spruce forest, to be compared to an ambient atmospheric concentration of 0.02 ppbv. The concentration of chloroform in soil air showed seasonal variation similar to that of CO2. Chloroform formation during incubation of undisturbed top-soil samples was found to be largest in soils from dense conifers stands with well-developed humus layers, while low chloroform formation occurred in soils from beech forest and agricultural grassland. We suggest that the mechanism behind the formation of chloroform is an unspecific chlorination of organic matter, caused by microbial activity in the soil. The aquifers are in fluvio-glacial sands with few layers of silt and a groundwater table from 4 to 7 m below the surface. In the shallowest parts of the aquifer, the groundwater has chloroform concentrations of 0.1 to 5 μg/L, and the groundwater is oxic with an age from 5 to 45 years using CFC-dating. Analyses of oxic groundwater > 40 years showed that it still contained chloroform at concentrations of 1 μg/L. Stable carbon isotopic analyses of chloroform from the uppermost groundwater in different parts of the forests and from soil water showed values from δ13C = -13 % to -27 %, corresponding to the ratio in natural organic materials and quite different from those of industrial products and from contaminated groundwater (δ13C = -46 % to -63 %). The isotopic ratio showed a minor decrease with depth due to a decomposition of chloroform. Measurements in a groundwater transect in one of the forest areas indicated that anoxic conditions in the groundwater depleted chloroform totally.

Magnetoliposomes Loaded with Poly-Unsaturated Fatty Acids as Novel Theranostic Anti-Inflammatory Formulations

PubMed Central

Calle, Daniel; Negri, Viviana; Ballesteros, Paloma; Cerdán, Sebastián

2015-01-01

We describe the preparation, physico-chemical characterization and anti-inflammatory properties of liposomes containing the superparamagnetic nanoparticle Nanotex, the fluorescent dye Rhodamine-100 and omega-3 polyunsaturated fatty acid ethyl ester (ω-3 PUFA-EE), as theranostic anti-inflammatory agents. Liposomes were prepared after drying chloroform suspensions of egg phosphatidylcholine, hydration of the lipid film with aqueous phases containing or not Nanotex, Rhodamine-100 dye or ω-3 PUFA-EE, and eleven extrusion steps through nanometric membrane filters. This resulted in uniform preparations of liposomes of approximately 200 nm diameter. Extraliposomal contents were removed from the preparation by gel filtration chromatography. High Resolution Magic Angle Spinning 1H NMR Spectroscopy of the liposomal preparations containing ω-3 PUFA-EE revealed well resolved 1H resonances from highly mobile ω-3 PUFA-EE, suggesting the formation of very small (ca. 10 nm) ω-3 PUFA-EE nanogoticules, tumbling fast in the NMR timescale. Chloroform extraction of the liposomal preparations revealed additionally the incorporation of ω-3 PUFA-EE within the membrane domain. Water diffusion weighted spectra, indicated that the goticules of ω-3 PUFA-EE or its insertion in the membrane did not affect the average translational diffusion coefficient of water, suggesting an intraliposomal localization, that was confirmed by ultrafiltration. The therapeutic efficacy of these preparations was tested in two different models of inflammatory disease as inflammatory colitis or the inflammatory component associated to glioma development. Results indicate that the magnetoliposomes loaded with ω-3 PUFA-EE allowed MRI visualization in vivo and improved the outcome of inflammatory disease in both animal models, decreasing significantly colonic inflammation and delaying, or even reversing, glioma development. Together, our results indicate that magnetoliposomes loaded with ω-3 PUFA-EE may become useful anti-inflammatory agents for image guided drug delivery. PMID:25767616

Anticholinesterase, antioxidant, analgesic and anti-inflammatory activity assessment of Xeranthemum annuum L. and isolation of two cyanogenic compounds.

PubMed

Orhan, Ilkay Erdogan; Gulyurdu, Fulya; Kupeli Akkol, Esra; Senol, Fatma Sezer; Arabaci Anul, Serap; Tatli, Iffet Irem

2016-11-01

Xeranthemum annuum L. (Asteraceae) (XA) is an ornamental and medicinal species with limited bioactivity and phytochemical data. Identification of anticholinesterase, antioxidant, anti-inflammatory and analgesic effects of the flower and root-stem (R-S) extracts of XA. Anticholinesterase (at 100 μg mL -1 ) and antioxidant (at 1000 μg mL -1 ) effects of various extracts were evaluated via microtiter assays, while anti-inflammatory and analgesic effects of the R-S extracts were tested using carrageenan-induced hind paw oedema (100 and 200 mg kg -1 ) and p-benzoquinone (PBQ) writhing models (200 mg kg -1 ) in male Swiss albino mice. The R-S ethanol extract of XA was subjected to isolation studies using conventional chromatographic methods. Most of the extracts showed inhibition over 85% against butyrylcholinesterase and no inhibition towards acetylcholinesterase. The flower chloroform and the R-S ethyl acetate extracts were most effective (97.85 ± 0.94% and 96.89 ± 1.09%, respectively). The R-S ethanol extract displayed a remarkable scavenging activity against DPPH (77.33 ± 1.99%) and in FRAP assay, while the hexane extract of the R-S parts possessed the highest metal-chelating capacity (72.79 ± 0.33%). The chloroform extract of the R-S caused a significant analgesic effect (24.4%) in PBQ writhing model. No anti-inflammatory effect was observed. Isolation of zierin and zierin xyloside, which were inactive in anticholinesterase assays, was achieved from the R-S ethanol extract. This is the first report of anticholinesterase, antioxidant, analgesic and anti-inflammatory activities and isolation of zierin and zierin xyloside from XA. Therefore, XA seems to contain antioxidant and BChE-inhibiting compounds.

Anti-acetylcholinesterase activity and antioxidant properties of extracts and fractions of Carpolobia lutea.

PubMed

Nwidu, Lucky Legbosi; Elmorsy, Ekramy; Thornton, Jack; Wijamunige, Buddhika; Wijesekara, Anusha; Tarbox, Rebecca; Warren, Averil; Carter, Wayne Grant

2017-12-01

There is an unmet need to discover new treatments for Alzheimer's disease. This study determined the anti-acetylcholinesterase (AChE) activity, DPPH free radical scavenging and antioxidant properties of Carpolobia lutea G. Don (Polygalaceae). The objective of this study is to quantify C. lutea anti-AChE, DPPH free radical scavenging, and antioxidant activities and cell cytotoxicity. Plant stem, leaves and roots were subjected to sequential solvent extractions, and screened for anti-AChE activity across a concentration range of 0.02-200 μg/mL. Plant DPPH radical scavenging activity, reducing power, and total phenolic and flavonoid contents were determined, and cytotoxicity evaluated using human hepatocytes. Carpolobia lutea exhibited concentration-dependent anti-AChE activity. The most potent inhibitory activity for the stem was the crude ethanol extract and hexane stem fraction oil (IC 50  = 140 μg/mL); for the leaves, the chloroform leaf fraction (IC 50  = 60 μg/mL); and for roots, the methanol, ethyl acetate and aqueous root fractions (IC 50  = 0.3-3 μg/mL). Dose-dependent free radical scavenging activity and reducing power were observed with increasing stem, leaf or root concentration. Total phenolic contents were the highest in the stem: ∼632 mg gallic acid equivalents/g for a hexane stem fraction oil. Total flavonoid content was the highest in the leaves: ∼297 mg quercetin equivalents/g for a chloroform leaf fraction. At 1 μg/mL, only the crude ethanol extract oil was significantly cytotoxic to hepatocytes. Carpolobia lutea possesses anti-AChE activity and beneficial antioxidant capacity indicative of its potential development as a treatment of Alzheimer's and other diseases characterized by a cholinergic deficit.

Photochemical properties of squarylium cyanine dyes.

PubMed

Ferreira, D P; Conceição, D S; Ferreira, V R A; Graça, V C; Santos, P F; Vieira Ferreira, L F

2013-11-01

This study presents several new squarylium dyes derived from benzothiazole and benzoselenazole with several structural variations, namely the nature of the heteroaromatic ring and the length of the N,N'-dialkyl groups. Before being investigated in connection with their effect on living cells and/or tissues, these novel compounds were characterized, namely with respect to the determination of their main photophysical parameters. Therefore, a study of the ground state absorption, fluorescence emission (quantum yields and lifetimes) and singlet oxygen generation quantum yields was performed for all the compounds synthesized in order to evaluate their efficiency as photosensitizers. An increase of the alkyl chain length from ethyl to hexyl did not produce a clear change in the fluorescence quantum yields, showing no influence on the photoisomerization process. Heavy atom inclusion (Se instead of S) enhanced the singlet oxygen generation efficiency and decreased the intensity of the fluorescence emission. The external heavy atom effect (I(-) as a counterion instead of CF3SO3(-)) produced a significant increase in the singlet oxygen formation quantum yield (about 20%). Transient absorption studies in aerated and oxygen free samples revealed that the photoisomerization process, which could compete with the triplet state formation for all dyes in solution, is a negligible pathway for the excited state deactivation, in accordance with the rigidity introduced by the squaric ring into the polymethine chain of the dye, both in chloroform and ethanol. However, in the case of the chloroform solution a new transient was detected in air equilibrated solutions, resulting from a reaction of the excited squarylium dye in the singlet state with CHCl3˙, and assigned to the radical cation (SQ(+)˙) of the dye.

Extraction Optimization for Phenolic- and Withanolide-Rich Fractions from Withania somnifera Roots: Identification and Quantification of Withaferin A, 12-Deoxywithastromonolide, and Withanolide A in Plant Materials and Marketed Formulations Using a Reversed-Phase HPLC-Photodiode Array Detection Method.

PubMed

Kumar, Satyanshu; Singh, Raghuraj; Gajbhiye, Narendra; Dhanani, Tushar

2018-06-26

Background : Both the roots and leaves of Withania somnifera are products of commerce. They contain active compounds of therapeutic value and mostly different withanolides. Several pharmacological activities of W. somnifera have links to one or more withanolides. The presence of phenolic compounds in extracts could play a vital role in the reduction of blood glucose levels in diabetic subjects. Objective : The present study was carried out for the selection of a solvent to prepare extracts rich in phenolics, withaferin A (WA), 12-deoxywithastromonolide (12WD), and withanolide A (WDA). A simple, rapid HPLC method was also developed for the identification and quantification of WA, 12WD, and WDA. Methods : The extraction efficiency of aqueous alcoholic solvents including hexane, chloroform, ethyl acetate, and methanol were compared for three selected withanolides and total phenolic content. The contents of WA, 12WD, and WDA and total phenolics were determined in the extracts. The quality of nine formulations containing W. sominfera were also compared in terms of the content of WA, 12WD, and WDA and total phenolics. Results : The maximum extract yield and the total withanolide and phenolic content were obtained from aqueous alcoholic compositions at 50:50 (v/v), 70:30 (v/v), and 100:0 (v/v), respectively. In the case of organic solvents, chloroform and ethyl acetate yielded the highest concentrations of phenolics and three withanolides, respectively. The total phenolic content in formulations was in the range of 1.84-3.13%, and total withanolide content showed wide variability. Conclusions : The outcome of the present investigation could be utilized for the selection of extraction solvents to prepare W. somnifera -enriched extracts and their quality monitoring by using the developed and validated HPLC-Photodiode array detection method. Highlights : A process for preparation of phenolics and withanolides (withaferin A, 12-deoxywithastramonolide and withanolide A) enriched extracts of Withania somnifera . Simple and rapid HPLC method was also developed and validated as per the ICH guidelines for identification and quantification of three major withanolides. The developed HPLC method was applied to analyze the quality of extracts and marketed herbal products (mono, as well as poly constituents). Optimized extraction process could be utilized for upscaling process development in preparation of enriched extracts from Withania somnifera , crop improvement, bio-prospection studies and quality control.

Sensitive and background-free determination of thiols from wastewater samples by MOF-5 extraction coupled with high-performance liquid chromatography with fluorescence detection using a novel fluorescence probe of carbazole-9-ethyl-2-maleimide.

PubMed

Lv, Zhengxian; Sun, Zhiwei; Song, Cuihua; Lu, Shuaimin; Chen, Guang; You, Jinmao

2016-12-01

A sensitive and background-free pre-column derivatization method for the determination of thiol compounds using metal-organic framework material (MOF-5) as dispersive solid-phase extraction (DSPE) adsorbent followed by high-performance liquid chromatography fluorescence detection (HPLC-FLD) has been developed. In this paper, a novel labeling reagent, carbazole-9-ethyl-2-maleimide(CAEM), was synthesized and reacted with thiols at 40°C for 10min in the presence of PBS buffer (0.02mol/L, pH 7.5). Interestingly, CAEM itself had no fluorescence, while its derivatives exhibited intense fluorescence with an excitation maximum at λ ex 274nm and an emission maximum at λ em 363nm, which greatly reduced the background interference and improved the sensitivity of the method. Furthermore, the MOF-5 was prepared and used as DSPE adsorbent for the selective adsorption of thiols from wastewater sample. Under the optimized experimental conditions, an excellent linearity for all analytes over their concentration ranges of 0.01-1.0μmol/L (R 2 >0.9986)were obtained with the limit of detection (LOD) ranging from 8 to 17.1pmol/L for nine tested thiols. The feasibility of this method for the determination of thiols in wastewater samples had been evaluated and satisfactory average recoveries (n=3) were achieved with the range of 86.6-98.5%. Copyright © 2016 Elsevier B.V. All rights reserved.

Comparison of two derivatization methods for the analysis of short chain fatty acids in the ambient aerosol using GC-MS

NASA Astrophysics Data System (ADS)

Kim, G.; Jeon, S.

2016-12-01

Fatty acids are one of the important compound classes in the polar organic fraction of ambient aerosols. Among them, short chain fatty acids play a significant role in the atmospheric transformation processes. For short-chain acids, the bottleneck of analysis has been the difficulty of sample preparation due to the high solubility and volatility. To overcome this problem, derivatization of polar organic fraction is widely used with silylation reagents to increase the resolution and sensitivity. Two different derivatization procedures; (1) using the tertbutyldimethylsilyl (TBDMS) derivatization and (2) the headspace-solid phase microextraction (HS-SPME) with in-fiber derivatization are compared using gas chromatography-mass spectrometry (GC-MS). At the second method, simultaneous derivatization and extraction were performed by a poly acrylate (PA) coated fiber doped with pyrenyldiazomethane (PDAM). We investigated the chromatographic property and relative sensitivities of each individual short chain acids according to two different derivatization procedures. For the method validation, the linearity, recovery and method detection limit (MDL) were compared. Also, two derivatization methods were applied to the ambient aerosol samples and evaluated with respect to the effectiveness.

Subchronic chloroform priming protects mice from a subsequently administered lethal dose of chloroform

DOE Office of Scientific and Technical Information (OSTI.GOV)

Philip, Binu K.; Anand, Sathanandam S.; Palkar, Prajakta S.

2006-10-01

Protection offered by pre-exposure priming with a small dose of a toxicant against the toxic and lethal effects of a subsequently administered high dose of the same toxicant is autoprotection. Although autoprotection has been extensively studied with diverse toxicants in acute exposure regimen, not much is known about autoprotection after priming with repeated exposure. The objective of this study was to investigate this concept following repeated exposure to a common water contaminant, chloroform. Swiss Webster (SW) mice, exposed continuously to either vehicle (5% Emulphor, unprimed) or chloroform (150 mg/kg/day po, primed) for 30 days, were challenged with a normally lethalmore » dose of chloroform (750 mg chloroform/kg po) 24 h after the last exposure. As expected, 90% of the unprimed mice died between 48 and 96 h after administration of the lethal dose in contrast to 100% survival of mice primed with chloroform. Time course studies indicated lower hepato- and nephrotoxicity in primed mice as compared to unprimed mice. Hepatic CYP2E1, glutathione levels (GSH), and covalent binding of {sup 14}C-chloroform-derived radiolabel did not differ between livers of unprimed and primed mice after lethal dose exposure, indicating that protection in liver is neither due to decreased bioactivation nor increased detoxification. Kidney GSH and glutathione reductase activity were upregulated, with a concomitant reduction in oxidized glutathione in the primed mice following lethal dose challenge, leading to decreased renal covalent binding of {sup 14}C-chloroform-derived radiolabel, in the absence of any change in CYP2E1 levels. Buthionine sulfoximine (BSO) intervention led to 70% mortality in primed mice challenged with lethal dose. These data suggest that higher detoxification may play a role in the lower initiation of kidney injury observed in primed mice. Exposure of primed mice to a lethal dose of chloroform led to 40% lower chloroform levels (AUC{sub 15-360min}) in the systemic circulation. Exhalation of {sup 14}C-chloroform was unchanged in primed as compared to unprimed mice (AUC{sub 1-6h}). Urinary excretion of {sup 14}C-chloroform was higher in primed mice after administration of the lethal dose. However, neither slightly higher urinary elimination nor unchanged expiration can account for the difference in systemic levels of chloroform. Liver and kidney regeneration was inhibited by the lethal dose in unprimed mice leading to progressive injury, organ failure, and 90% mortality. In contrast, sustained and highly stimulated compensatory hepato- and nephrogenic repair prevented the progression of injury resulting in 100% survival of primed mice challenged with the lethal dose. These findings affirm the critical role of tissue regeneration and favorable detoxification (only in kidney) of the lethal dose of chloroform in subchronic chloroform priming-induced autoprotection.« less

Phosphopeptide Enrichment by Covalent Chromatography after Derivatization of Protein Digests Immobilized on Reversed-Phase Supports

PubMed Central

Nika, Heinz; Nieves, Edward; Hawke, David H.; Angeletti, Ruth Hogue

2013-01-01

A rugged sample-preparation method for comprehensive affinity enrichment of phosphopeptides from protein digests has been developed. The method uses a series of chemical reactions to incorporate efficiently and specifically a thiol-functionalized affinity tag into the analyte by barium hydroxide catalyzed β-elimination with Michael addition using 2-aminoethanethiol as nucleophile and subsequent thiolation of the resulting amino group with sulfosuccinimidyl-2-(biotinamido) ethyl-1,3-dithiopropionate. Gentle oxidation of cysteine residues, followed by acetylation of α- and ε-amino groups before these reactions, ensured selectivity of reversible capture of the modified phosphopeptides by covalent chromatography on activated thiol sepharose. The use of C18 reversed-phase supports as a miniaturized reaction bed facilitated optimization of the individual modification steps for throughput and completeness of derivatization. Reagents were exchanged directly on the supports, eliminating sample transfer between the reaction steps and thus, allowing the immobilized analyte to be carried through the multistep reaction scheme with minimal sample loss. The use of this sample-preparation method for phosphopeptide enrichment was demonstrated with low-level amounts of in-gel-digested protein. As applied to tryptic digests of α-S1- and β-casein, the method enabled the enrichment and detection of the phosphorylated peptides contained in the mixture, including the tetraphosphorylated species of β-casein, which has escaped chemical procedures reported previously. The isolates proved highly suitable for mapping the sites of phosphorylation by collisionally induced dissociation. β-Elimination, with consecutive Michael addition, expanded the use of the solid-phase-based enrichment strategy to phosphothreonyl peptides and to phosphoseryl/phosphothreonyl peptides derived from proline-directed kinase substrates and to their O-sulfono- and O-linked β-N-acetylglucosamine (O-GlcNAc)-modified counterparts. Solid-phase enzymatic dephosphorylation proved to be a viable tool to condition O-GlcNAcylated peptide in mixtures with phosphopeptides for selective affinity purification. Acetylation, as an integral step of the sample-preparation method, precluded reduction in recovery of the thiolation substrate caused by intrapeptide lysine-dehydroalanine cross-link formation. The solid-phase analytical platform provides robustness and simplicity of operation using equipment readily available in most biological laboratories and is expected to accommodate additional chemistries to expand the scope of solid-phase serial derivatization for protein structural characterization. PMID:23997662

Metabolomics relative quantitation with mass spectrometry using chemical derivatization and isotope labeling

DOE PAGES

O'Maille, Grace; Go, Eden P.; Hoang, Linh; ...

2008-01-01

Comprehensive detection and quantitation of metabolites from a biological source constitute the major challenges of current metabolomics research. Two chemical derivatization methodologies, butylation and amination, were applied to human serum for ionization enhancement of a broad spectrum of metabolite classes, including steroids and amino acids. LC-ESI-MS analysis of the derivatized serum samples provided a significant signal elevation across the total ion chromatogram to over a 100-fold increase in ionization efficiency. It was also demonstrated that derivatization combined with isotopically labeled reagents facilitated the relative quantitation of derivatized metabolites from individual as well as pooled samples.

Antioxidant activity and protective effect of banana peel against oxidative hemolysis of human erythrocyte at different stages of ripening.

PubMed

Sundaram, Shanthy; Anjum, Shadma; Dwivedi, Priyanka; Rai, Gyanendra Kumar

2011-08-01

Phytochemicals such as polyphenols and carotenoids are gaining importance because of their contribution to human health and their multiple biological effects such as antioxidant, antimutagenic, anticarcinogenic, and cytoprotective activities and their therapeutic properties. Banana peel is a major by-product in pulp industry and it contains various bioactive compounds like polyphenols, carotenoids, and others. In the present study, effect of ripening, solvent polarity on the content of bioactive compounds of crude banana peel and the protective effect of peel extracts of unripe, ripe, and leaky ripe banana fruit on hydrogen peroxide-induced hemolysis and their antioxidant capacity were investigated. Banana (Musa paradisica) peel at different stages of ripening (unripe, ripe, leaky ripe) were treated with 70% acetone, which were partitioned in order of polarity with water, ethyl acetate, chloroform (CHCl₃), and hexane sequentially. The antioxidant activity of the samples was evaluated by the red cell hemolysis assay, free radical scavenging (1,1-diphenyl-2-picrylhydrazyl free radical elimination) and superoxide dismutase activities. The Folin-Ciocalteu's reagent assay was used to estimate the phenolic content of extracts. The findings of this investigation suggest that the unripe banana peel sample had higher antioxidant potency than ripe and leaky ripe. Further on fractionation, ethyl acetate and water soluble fractions of unripe peel displayed high antioxidant activity than CHCl₃ and hexane fraction, respectively. A positive correlation between free radical scavenging capacity and the content of phenolic compound were found in unripe, ripe, and leaky ripe stages of banana peel.

Essential oils chemical composition, antioxidant activities and total phenols of Astrodaucus persicus.

PubMed

Goodarzi, Saeid; Hadjiakhoondi, Abbas; Yassa, Narguess; Khanavi, Mahnaz; Tofighi, Zahra

2016-02-01

Astrodaucus persicus, Apiaceae, is used as vegetable or food additive in some parts of Iran. The essential oils of different parts of Astrodaucus persicus from Kordestan province were analyzed for the first time and compared with other regions. In this study, antioxidant activities and total phenols determination of aerial parts essential oils and root fractions of A. persicus were investigated. The essential oils were obtained by hydro-distillation from flowers/fruits, leaves/stems, ripe fruits and roots of plant and analyzed by GC-MS. Crude root extract was fractionated with hexane, chloroform, ethyl acetate and methanol. Antioxidant activities by DPPH and FRAP methods and total phenols by Folin-ciocalteu assay were measured. The abundant compounds of flowers/fruits blue essential oil were α-thujene, β-pinene and α-pinene. The predominant components of blue leaves/stems essential oil were α-thujene, α-pinene and α-fenchene. The major volatiles of ripe fruits blue essential oil were β-pinene, α-thujene and α-pinene. The chief compounds of root yellow essential oil were trans-caryophyllene, bicycogermacrene and germacrene-D. Total root extract and ethyl acetate fraction showed potent antioxidant activities and high amount of total phenols in comparison to other samples. Among volatile oils, the flowers/fruits essential oil showed potent reducing capacity. The major compounds of aerial parts essential oils were hydrocarbon monoterpenes while the chief percentage of roots essential oil constituents were hydrocarbon sesquiterpenes. α-Eudesmol and β-eudesmol were identified as responsible for creation of blue color in aerial parts essential oils. A. persicus was known as a potent antioxidant among Apiaceae.

Quantitation of polyamines in cultured cells and tissue homogenates by reversed-phase high-performance liquid chromatography of their benzoyl derivatives.

PubMed

Verkoelen, C F; Romijn, J C; Schroeder, F H; van Schalkwijk, W P; Splinter, T A

1988-04-08

A rapid and simple method, originally described by Redmond and Tseng [J. Chromatogr., 170 (1979) 479] was applied to the analysis of di- and polyamines in cultured human tumour cells and human tumour xenografts. Optimization of the procedures and evaluation of the characteristic features of the assay are described. The (modified) procedure employs precolumn derivatization with benzoyl chloride, extraction of the derivatives by chloroform, separation by reversed-phase high-performance liquid chromatography under isocratic conditions and detection by ultraviolet absorbance measurement at 229 nm. The complete analysis was accomplished within 10 min per sample. The detection limit was ca. 1 pmol. The intra- and inter-assay coefficients of variation were 2.5-4.4% and 3.4-13.1%, respectively. The presence of well known inhibitors of polyamine biosynthesis, such as DL-alpha-difluoromethylornithine and methylglyoxal bis(guanylhydrazone), did not interfere with the assay, and disturbance by cyclohexylamine could be avoided by changing the polarity of the mobile phase. The method proved to be very suitable because it is rapid, simple, requires a minimum of sample pretreatment, and still provides sufficient sensitivity to quantitate polyamines in relatively small amounts of cells (10(5) cells) or tumour tissues (less than 1 mg), even after treatment with inhibitors of polyamine biosynthesis.

Physicochemical and biological characteristics of DEAE-derivatized PS7 biopolymer of Beijerinckia indica.

PubMed

Lee, Kyung Hee; Yoo, Sang-Ho; Baek, Seung Hee; Lee, Hyeon Gyu

2007-07-01

Physicochemical and biological characteristics of the exopolysaccharide, PS7, produced from Beijerinckia indica were investigated. The PS7 weight fractions of Glc and GlcUA were 0.45 and 0.25, respectively, and the molar ratio of Glc:Rha:GalUA was approximately 5:1:1.3. The PS7 was chemically derivatized with diethylaminoethyl chloride-HCl (DEAE-HCl), and the resulting modified PS7 contained both positive and negative charges. The elemental and IR analyses were conducted to confirm the successful incorporation of DEAE groups into PS7. Large increase in nitrogen fraction was observed from the derivatized PS7 by elemental analysis. The characteristic CH(3) and CH(2) peaks originated from DEAE group were detected in (1)H NMR spectrum of the derivatized PS7 as well. Solubility of native PS7 was improved almost twice from 40 to 75% after DEAE-derivatization, while water holding capacity (WHC) drastically decreased from 10,026 to 245%. Oil binding capacity (OBC) of PS7 also significantly dropped from 1528 to 331% after the derivatization. The [eta] values of native and derivatized PS7 were 27.6 and 0.31 dL/g at 25 degrees C, respectively, which means that the DEAE-derivatization significantly decreased the [eta] of PS7. The bile acid binding capacity of PS7 was indirectly determined by measuring the holding capability of cholic acid inside the dialysis membrane. When PS7 was DEAE-derivatized, there was substantial decrease in the cholic acid retardation index (CRI). Up to 8-9h of dialysis, the derivatized PS7 hold 8.6% less of cholic acid compared to native one.

Bacterial Cellular Materials as Precursors of Chloroform

NASA Astrophysics Data System (ADS)

Wang, J.; Ng, T.; Zhang, Q.; Chow, A. T.; Wong, P.

2011-12-01

The environmental sources of chloroform and other halocarbons have been intensively investigated because their effects of stratospheric ozone destruction and environmental toxicity. It has been demonstrated that microorganisms could facilitate the biotic generation of chloroform from natural organic matters in soil, but whether the cellular materials itself also serves as an important precursor due to photo-disinfection is poorly known. Herein, seven common pure bacterial cultures (Acinetobacter junii, Aeromonas hydrophila, Bacillus cereus, Bacillus substilis, Escherichia coli, Shigella sonnei, Staphylococcus sciuri) were chlorinated to evaluate the yields of chloroform, dibromochloromethane, dichlorobromomethane, and bromoform. The effects of bromide on these chemical productions and speciations were also investigated. Results showed that, on average, 5.64-36.42 μg-chloroform /mg-C were generated during the bacterial chlorination, in similar order of magnitude to that generated by humic acid (previously reported as 78 μg-chloroform/mg-C). However, unlike humic acid in water chlorination, chloroform concentration did not simply increase with the total organic carbon in water mixture. In the presence of bromide, the yield of brominated species responded linearly to the bromide concentration. This study provides useful information to understand the contributions of chloroform from photodisinfection processes in coastal environments.

A comparison of the effectiveness of chloroform and eucalyptus oil in dissolving root canal sealers.

PubMed

Schäfer, Edgar; Zandbiglari, Tannaz

2002-05-01

The solubility of 8 different root canal sealers in chloroform and in eucalyptus oil was compared. For standardized samples (n=12), ring molds were filled with mixed sealers based on epoxy resin, silicone, calcium hydroxide, zinc oxide-eugenol, glass ionomer, and polyketone. These samples were immersed in chloroform or eucalyptus oil for 30 seconds, 1 minute, 2 minutes, 5 minutes, 10 minutes, and 20 minutes. Then, the mean weight loss was determined and statistically analyzed. With the exception of the silicone, all the sealers showed significantly higher solubilities (P <.05) in chloroform than in eucalyptus oil. Epoxy resin was the most soluble sealer in chloroform. In eucalyptus oil, calcium hydroxide, and zinc oxide-eugenol showed the highest solubility. Under the conditions of this study, chloroform was a far more effective solvent of root canal sealers than eucalyptus oil. Because of the potential hazards of chloroform, further studies on the dissolution of root canal sealers in different solvents seem to be necessary.

Acute chloroform ingestion successfully treated with intravenously administered N-acetylcysteine.

PubMed

Dell'Aglio, Damon M; Sutter, Mark E; Schwartz, Michael D; Koch, David D; Algren, D A; Morgan, Brent W

2010-06-01

Chloroform, a halogenated hydrocarbon, causes central nervous system depression, cardiac arrhythmias, and hepatotoxicity. We describe a case of chloroform ingestion with a confirmatory serum level and resultant hepatotoxicity successfully treated with intravenously administered N-acetylcysteine (NAC). A 19-year-old man attempting suicide ingested approximately 75 mL of chloroform. He was unresponsive and intubated upon arrival. Intravenously administered NAC was started after initial stabilization was complete. His vital signs were normal. Admission laboratory values revealed normal serum electrolytes, AST, ALT, PT, BUN, creatinine, and bilirubin. Serum ethanol level was 15 mg/dL, and aspirin and acetaminophen were undetectable. The patient was extubated but developed liver function abnormalities with a peak AST of 224 IU/L, ALT of 583 IU/L, and bilirubin level reaching 16.3 mg/dL. NAC was continued through hospital day 6. Serum chloroform level obtained on admission was 91 μg/mL. The patient was discharged to psychiatry without known sequelae and normal liver function tests. The average serum chloroform level in fatal cases of inhalational chloroform poisoning was 64 μg/mL, significantly lower than our patient. The toxicity is believed to be similar in both inhalation and ingestion routes of exposure, with mortality predominantly resulting from anoxia secondary to central nervous system depression. Hepatocellular toxicity is thought to result from free radical-induced oxidative damage. Previous reports describe survival after treatment with orally administered NAC, we report the first use of intravenously administered NAC for chloroform ingestion. Acute oral ingestion of chloroform is extremely rare. Our case illustrates that with appropriate supportive care, patients can recover from chloroform ingestion, and intravenously administered NAC may be of benefit in such cases.

Microbial reductive dehalogenation of trihalomethanes by a Dehalobacter-containing co-culture.

PubMed

Zhao, Siyan; Rogers, Matthew J; He, Jianzhong

2017-07-01

Trihalomethanes such as chloroform and bromoform, although well-known as a prominent class of disinfection by-products, are ubiquitously distributed in the environment due to widespread industrial usage in the past decades. Chloroform and bromoform are particularly concerning, of high concentrations detected and with long half-lives up to several hundred days in soils and groundwater. In this study, we report a Dehalobacter- and Desulfovibrio-containing co-culture that exhibits dehalogenation of chloroform (~0.61 mM) to dichloromethane and bromoform (~0.67 mM) to dibromomethane within 10-15 days. This co-culture was further found to dechlorinate 1,1,1-trichloroethane (1,1,1-TCA) (~0.65 mM) to 1,1-dichloroethane within 12 days. The Dehalobacter species present in this co-culture, designated Dehalobacter sp. THM1, was found to couple growth with dehalogenation of chloroform, bromoform, and 1,1,1-TCA. Strain THM1 harbors a newly identified reductive dehalogenase (RDase), ThmA, which catalyzes chloroform, bromoform, and 1,1,1-TCA dehalogenation. Additionally, based on the sequences of thmA and other identified chloroform RDase genes, ctrA, cfrA, and tmrA, a pair of chloroform RDase gene-specific primers were designed and successfully applied to investigate the chloroform dechlorinating potential of microbial communities. The comparative analysis of chloroform RDases with tetrachloroethene RDases suggests a possible approach in predicting the substrate specificity of uncharacterized RDases in the future.

Cobra venom factor immunoconjugates: effects of carbohydrate-directed versus amino group-directed conjugation.

PubMed

Zara, J; Pomato, N; McCabe, R P; Bredehorst, R; Vogel, C W

1995-01-01

Human IgM monoclonal antibody 16-88, derived from patients immunized with autologous colon carcinoma cells, was derivatized with two different cross-linkers, S-(2-thiopyridyl)-L-cysteine hydrazide (TPCH), which is carbohydrate-directed, and N-succinimidyl-3-(2- pyridyldithio)propionate (SPDP), which is amino group-directed. Two antibody functions, antigen binding and complement activation, were assayed upon derivatization with TPCH and SPDP. TPCH allowed for extensive modification (up to 17 TPCH molecules per antibody) without impairment of antigen binding activity, while this function was significantly compromised upon derivatization with SPDP. Antibody molecules derivatized with 16 SPDP residues showed almost complete loss of their antigen binding function. The complement activating ability of antibody 16-88 was significantly decreased after derivatization with TPCH or SPDP. In the case of SPDP derivatization, this decrease of the complement activating ability is predominantly a consequence of the impaired binding function. Upon conjugation of cobra venom factor (CVF), a nontoxic 137-kDa glycoprotein which is capable of activating the alternative pathway of complement, the antigen binding activity of SPDP-derivatized antibody was further compromised, whereas that of TPCH-derivatized antibody remained unaffected even after attachment of three or four CVF molecules per antibody. In both conjugates CVF retained good functional activity. CVF was slightly more active when attached to SPDP-derivatized antibody, suggesting a better accessibility of amino group-coupled CVF for its interaction with other complement proteins. These results indicate that carbohydrate-directed conjugation compromises the antibody function of complement activation, but allows for the generation of immunoconjugates with unimpaired antigen binding capability.(ABSTRACT TRUNCATED AT 250 WORDS)

Chloroform-induced insanity defence confounds lawyer Lincoln.

PubMed

Spiegel, A D; Suskind, P B

1997-12-01

During an 1857 trial, the defence claimed that the accused should be absolved of wilful murder because an overdose of chloroform during surgery induced insanity. In a rare appearance as a prosecutor, Abraham Lincoln tried the case for the State of Illinois. Expert medical witnesses testified about the side effects of chloroform and chloroform-induced insanity. Significantly, Lincoln was not knowledgeable about medical jurisprudence and overlooked potential sources of evidence and expert witnesses. Defence lawyers presented an impressive array of physicians to testify about insanity, about chloroform and about the results of an overdosage during anaesthesia. Considering the state of scientific knowledge at the time, the trial was notable.

Derivatization of phytochelatins from Silene vulgaris, induced upon exposure to arsenate and cadmium: comparison of derivatization with Ellman's reagent and monobromobimane.

PubMed

Sneller, F E; van Heerwaarden, L M; Koevoets, P L; Vooijs, R; Schat, H; Verkleij, J A

2000-09-01

Phytochelatins (PCs) are a family of thiol-rich peptides, with the general structure (gamma-Glu-Cys)(n)()-Gly, with n = 2-11, induced in plants upon exposure to excessive amounts of heavy metals and some metalloids, such as arsenic. Two types of PC analyses are currently used, i.e., acid extraction and separation on HPLC with either precolumn derivatization (pH 8.2) with monobromobimane (mBBr) or postcolumn derivatization (pH 7.8) with Ellman's reagent [5, 5'-dithiobis(2-nitrobenzoic acid), DTNB]. Although both methods were satisfactory for analysis of Cd-induced PCs, formation of (RS)(3)-As complexes during extraction of As-induced PCs rendered the DTNB method useless. This paper shows that precolumn derivatization with mBBr, during which the (RS)(3)-As complexes are disrupted, provides a qualitative and quantitative analysis of both Cd- and As-induced PCs. In addition, derivatization efficiencies of both methods for the oligomers with n = 2-4 (PC(2)(-)(4)) are compared. Derivatization efficiency decreased from 71.8% and 81.4% for mBBr and DTNB derivatization, respectively, for PC(2) to 27.4% and 50.2% for PC(4). This decrease is most likely due to steric hindrance. Correction of measured thiol concentration is therefore advised for better quantification of PC concentrations in plant material.

Derivatization of Dextran for Multiply Charged Ion Formation and Electrospray Ionization Time-of-Flight Mass Spectrometric Analysis

NASA Astrophysics Data System (ADS)

Tapia, Jesus B.; Hibbard, Hailey A. J.; Reynolds, Melissa M.

2017-10-01

We present the use of a simple, one-pot derivatization to allow the polysaccharide dextran to carry multiple positive charges, shifting its molecular weight distribution to a lower m/ z range. We performed this derivatization because molecular weight measurements of polysaccharides by mass spectrometry are challenging because of their lack of readily ionizable groups. The absence of ionizable groups limits proton abstraction and suppresses proton adduction during the ionization process, producing mass spectra with predominantly singly charged metal adduct ions, thereby limiting the detection of large polysaccharides. To address this challenge, we derivatized dextran T1 (approximately 1 kDa) by attaching ethylenediamine, giving dextran readily ionizable, terminal amine functional groups. The attached ethylenediamine groups facilitated proton adduction during the ionization process in positive ion mode. Using the low molecular weight dextran T1, we tracked the number of ethylenediamine attachments by measuring the mass shift from underivatized to derivatized dextran T1. Using electrospray ionization time-of-flight mass spectrometry, we observed derivatized dextran chains ranging from two to nine glucose residues with between one and four attachments/charges. Our success in shifting derivatized dextran T1 toward the low m/ z range suggests potential for this derivatization as a viable route for analysis of high molecular weight polysaccharides using electrospray ionization time-of-flight mass spectrometry. [Figure not available: see fulltext.

Measurement of oxidative DNA damage by gas chromatography-mass spectrometry: ethanethiol prevents artifactual generation of oxidized DNA bases.

PubMed Central

Jenner, A; England, T G; Aruoma, O I; Halliwell, B

1998-01-01

Analysis of oxidative damage to DNA bases by GC-MS enables identification of a range of base oxidation products, but requires a derivatization procedure. However, derivatization at high temperature in the presence of air can cause 'artifactual' oxidation of some undamaged bases, leading to an overestimation of their oxidation products, including 8-hydroxyguanine. Therefore derivatization conditions that could minimize this problem were investigated. Decreasing derivatization temperature to 23 degrees C lowered levels of 8-hydroxyguanine, 8-hydroxyadenine, 5-hydroxycytosine and 5-(hydroxymethyl)uracil measured by GC-MS in hydrolysed calf thymus DNA. Addition of the reducing agent ethanethiol (5%, v/v) to DNA samples during trimethylsilylation at 90 degrees C also decreased levels of these four oxidized DNA bases as well as 5-hydroxyuracil. Removal of guanine from hydrolysed DNA samples by treatment with guanase, prior to derivatization, resulted in 8-hydroxyguanine levels (54-59 pmol/mg of DNA) that were significantly lower than samples not pretreated with guanase, independent of the derivatization conditions used. Only hydrolysed DNA samples that were derivatized at 23 degrees C in the presence of ethanethiol produced 8-hydroxyguanine levels (56+/-8 pmol/mg of DNA) that were as low as those of guanase-pretreated samples. Levels of other oxidized bases were similar to samples derivatized at 23 degrees C without ethanethiol, except for 5-hydroxycytosine and 5-hydroxyuracil, which were further decreased by ethanethiol. Levels of 8-hydroxyguanine, 8-hydroxyadenine and 5-hydroxycytosine measured in hydrolysed calf thymus DNA by the improved procedures described here were comparable with those reported previously by HPLC with electrochemical detection and by GC-MS with prepurification to remove undamaged base. We conclude that artifactual oxidation of DNA bases during derivatization can be prevented by decreasing the temperature to 23 degrees C, removing air from the derivatization reaction and adding ethanethiol. PMID:9531471

Use of Linear Free Energy Relationships (LFERs) to Elucidate the Mechanisms of Reaction of a γ-Methyl-β-alkynyl and an ortho-Substituted Aryl Chloroformate Ester

PubMed Central

D’Souza, Malcolm J.; Knapp, Jaci A.; Fernandez-Bueno, Gabriel A.; Kevill, Dennis N.

2012-01-01

The specific rates of solvolysis of 2-butyn-1-yl-chloroformate (1) and 2-methoxyphenyl chloroformate (2) are studied at 25.0 °C in a series of binary aqueousorganic mixtures. The rates of reaction obtained are then analyzed using the extended Grunwald-Winstein (G-W) equation and the results are compared to previously published G-W analyses for phenyl chloroformate (3), propargyl chloroformate (4), p-methoxyphenyl choroformate (5), and p-nitrophenyl chloroformate (6). For 1, the results indicate that dual side-by-side addition-elimination and ionization pathways are occurring in some highly ionizing solvents due to the presence of the electron-donating γ-methyl group. For 2, the analyses indicate that the dominant mechanism is a bimolecular one where the formation of a tetrahedral intermediate is rate-determining. PMID:22312278

Extended Grunwald-Winstein Analysis - LFER Used to Gauge Solvent Effects in p-Nitrophenyl Chloroformate Solvolysis

PubMed Central

D’Souza, Malcolm J.; Shuman, Kevin E.; Carter, Shannon E.; Kevill, Dennis N.

2008-01-01

Specific rates of solvolysis at 25 °C for p-nitrophenyl chloroformate (1) are analyzed using the extended (two-term) Grunwald-Winstein equation. For 39 solvents, the sensitivities (l = 1.68±0.06 and m = 0.46±0.04) towards changes in solvent nucleophilicity (l) and solvent ionizing power (m) obtained, are similar to those previously observed for phenyl chloroformate (2) and p-methoxyphenyl chloroformate (3). The observations incorporating new kinetic data in several fluoroalcohol-containing mixtures, are rationalized in terms of the reaction being sensitive to substituent effects and the mechanism of reaction involving the addition (association) step of an addition-elimination (association-dissociation) pathway being rate-determining. The l/m ratios obtained for 1, 2, and 3, are also compared to the previously published l/m ratios for benzyl chloroformate (4) and p-nitrobenzyl chloroformate (5). PMID:19330071

Characterization of hot spots for natural chloroform formation: Relevance for groundwater quality

NASA Astrophysics Data System (ADS)

Jacobsen, Ole S.; Albers, Christian N.; Laier, Troels

2015-04-01

Chloroform soil hot spot may deteriorate groundwater quality and may even result in chloroform concentration exceeding the Danish maximum limit of 1 µg/L in groundwater for potable use. In order to characterize the soil properties important for the chloroform production, various ecosystems were examined with respect to soil air chloroform and soil organic matter type and content. Coniferous forest areas, responsible for highest chloroform concentrations, were examined on widely different scales from km to cm scale. Furthermore, regular soil gas measurements including chloroform were performed during 4 seasons at various depths, together with various meteorological measurements and soil temperature recordings. Laboratory incubation experiments were also performed on undisturbed soil samples in order to examine the role of various microbiota, fungi and bacteria. To identify hot spots responsible for the natural contamination we have measured the production of chloroform in the upper soil from different terrestrial systems. Field measurements of chloroform in top soil air were used as production indicators. The production was however not evenly distributed at any scale. The ecosystems seem to have quite different net-productions of chloroform from very low in grassland to very high in some coniferous forests. Within the forest ecosystem we found large variation in chloroform concentrations depending on vegetation. In beech forest we found the lowest values, somewhat higher in an open pine forest, but the highest concentrations were detected in spruce forest without any vegetation beneath. Within this ecotype, it appeared that the variation was also large; hot spots with 2-4 decades higher production than the surrounding area. These hot spots were not in any way visually different from the surroundings and were of variable size from 3 to 20 meters in diameter. Besides this, measurements within a seemingly homogenous hot spot showed that there was still high variability at 10 cm level. We suggest that the mechanism behind the formation of chloroform is an unspecific chlorination of organic matter, caused by microbial activity in the soil forming trichloroacetyl compounds. Laboratory measurements on intact soil cores have identified that the F and H horizons in the forest soil are the main producers of chloroform. Despite various attempts to identify the mechanisms responsible for the variability within a visually and chemically homogeneous area we have not yet succeeded. Parameters like soil respiration, inorganic and total organic chlorine, organic matter and soil structure were studied without any significant difference in favour of hot spots. By the use of 13C-isotopes we could identify the natural origin of the chloroform, and over a three years period we could conclude that the hot spots were permanent on the sites. At the same time a significant seasonal variation were measured depending on temperature and soil moisture.

Separation of thiol and cyanide hydrolysis products of chemical warfare agents by capillary electrophoresis.

PubMed

Copper, Christine L; Collins, Greg E

2004-03-01

The fluorescence derivatizing agent, o-phthalaldehyde (OPA), has been applied to the separation and detection of cyanide and several structurally similar thiols by capillary electrophoresis (CE)-laser induced fluorescence (LIF). Of particular interest to this investigation was the separation of 2-dimethylaminoethanethiol, 2-diethylaminoethanethiol, and cyanide, each of which are hydrolysis products or hydrolysis product simulants of the chemical warfare (CW) agents O-ethyl S-2-diisopropylaminoethyl methylphosphonothiolate (VX), O-isobutyl S-2-diethylaminoethyl methylphosphonothiolate (R-VX), and tabun (GA). Other structurally similar thiols simultaneously resolved by this method include 1-pentanethiol and 2-mercaptoethanol. Instrumental parameters were probed and optimum values for capillary length (50 cm) and inner diameter (75 microm), injection time (30 s) and field strength (15 kV) were determined. Sample stacking methods enabled detection limits of 9.3 microg/L for cyanide, 1.8 microg/L for 2-diethylaminoethanethiol, 35 microg/L for 2-dimethylaminoethanethiol, 15 microg/L for 2-mercaptoethanol, and 89 microg/L for 1-pentanethiol. The linearity of the method was verified over an order of magnitude and the reproducibility was found to be 3.0%.

Transformation of tributyltin in zebrafish eleutheroembryos (Danio rerio).

PubMed

Borges, Aline Rocha; López-Serrano Oliver, Ana; Gallego-Gallegos, Mercedes; Muñoz-Olivas, Riansares; Rodrigues Vale, Maria Goreti; Cámara, Carmen

2014-12-01

Organotin compounds are highly versatile group of organometallic chemicals used in industrial and agricultural applications. Their endocrine-disrupting effects are well known and their extensive uses as biocide materials, e.g., in antifouling paints, for many years have led to serious environmental problems. So far, attention has mainly been given to tributyltin pollution in water, sediments, and marine organisms because of its highly toxic effects and high accumulation levels at very low concentrations. In this study, we will focus on the conversion of tributyltin after it is absorbed by zebrafish eleutheroembryos, presented here as an alternative model to adult fish for describing bioconcentration. A simplified analytical extraction procedure based on the use of an assisted ultrasonic probe and derivatization by ethylation, followed by gas chromatography with a flame photometric detector (GC-FPD) is proposed. This classical methodology for organotin determination has been validated by inductively coupled plasma mass spectrometry (ICP-MS) and Zeeman graphite furnace atomic absorption spectrometry (ZGF-AAS) in terms of total tin content. The speciation analysis results show that zebrafish eleutheroembryos absorb high amounts of tributyltin and convert it into monobutyltin and likely in inorganic tin.

Development and validation of a method for the determination of low-ppb levels of macrocyclic lactones in butter, using HPLC-fluorescence.

PubMed

Macedo, Fabio; Marsico, Eliane Teixeira; Conte-Júnior, Carlos Adam; de Resende, Michele Fabri; Brasil, Taila Figueiredo; Pereira Netto, Annibal Duarte

2015-07-15

An analytical method was developed and validated for the simultaneous determination of four macrocyclic lactones (ML) (abamectin, doramectin, ivermectin and moxidectin) in butter, using liquid chromatography with fluorescence detection. The method employed heated liquid-liquid extraction and a mixture of acetonitrile, ethyl acetate and water, with preconcentration and derivatization, to produce stable fluorescent derivatives. The chromatographic run time was <12.5 min, with excellent separation. The method validation followed international guidelines and employed fortified butter samples. The figures of merit obtained, e.g. recovery (72.4-106.5%), repeatability (8.8%), within-laboratory reproducibility (15.7%) and limits of quantification (0.09-0.16 μg kg(-1)) were satisfactory for the desired application. The application of the method to real samples showed that ML residues were present in six of the ten samples evaluated. The method proved to be simple, easy and appropriate for simultaneous determination of ML residues in butter. To our knowledge, this is the first method described for the evaluation of ML in butter. Copyright © 2015. Published by Elsevier Ltd.

Efficient encapsulation of chloroform with cryptophane-M and the formation of exciplex studied by fluorescence spectroscopy.

PubMed

Shi, Yanqi; Li, Xueming; Yang, Jianchun; Gao, Fang; Tao, Chuanyi

2011-03-01

Efficient encapsulation of small molecules with supermolecules is one of significantly important subjects due to strong application potentials. This article presents the interaction between cryptophane-M and chloroform by fluorescence spectroscopy. The sonicated cryptophane-M solution exhibits light green color in chloroform, and the solid obtained from the evaporation of chloroform also has different color from that of cryptophane-M. In contrast, the sonicated cryptophane-M solutions in other solvents are colorless, and the solid obtained from the evaporation of these solvents has the same color as that of cryptophane-M. Furthermore, the freshly prepared cryptophane-M solution in different solvents is almost colorless, and the solid obtained from the evaporation of these solvents displays the same color as that of cryptophane-M. Although the sonicated cryptophane-M solutions in different solvents have very similar absorption spectra, they exhibit quite different emission spectra in chloroform. In contrast, the freshly-prepared cryptophane-M solutions show similar absorption and emission spectroscopy in various solvents. The variation of the fluorescence spectroscopy in binary solvents with the increasing chloroform ratio suggests that cryptophane-M and chloroform form a 1:1 exciplex, and the binding constant is estimated to be 292.95 M(-1). Although all solvents are able to enter into the cavity of cryptophane-M, only chloroform can stay in the cavity of cryptophane-M for a while, which is mostly due to the strong intermolecular interaction between cryptophane-M and chloroform, and this results in the formation of the exciplex between them. © Springer Science+Business Media, LLC 2010

Retreatability of two endodontic sealers, EndoSequence BC Sealer and AH Plus: a micro-computed tomographic comparison.

PubMed

Oltra, Enrique; Cox, Timothy C; LaCourse, Matthew R; Johnson, James D; Paranjpe, Avina

2017-02-01

Recently, bioceramic sealers like EndoSequence BC Sealer (BC Sealer) have been introduced and are being used in endodontic practice. However, this sealer has limited research related to its retreatability. Hence, the aim of this study was to evaluate the retreatability of two sealers, BC Sealer as compared with AH Plus using micro-computed tomographic (micro-CT) analysis. Fifty-six extracted human maxillary incisors were instrumented and randomly divided into 4 groups of 14 teeth: 1A, gutta-percha, AH Plus retreated with chloroform; 1B, gutta-percha, AH Plus retreated without chloroform; 2A, gutta-percha, EndoSequence BC Sealer retreated with chloroform; 2B, gutta-percha, EndoSequence BC Sealer retreated without chloroform. Micro-CT scans were taken before and after obturation and retreatment and analyzed for the volume of residual material. The specimens were longitudinally sectioned and digitized images were taken with the dental operating microscope. Data was analyzed using an ANOVA and a post-hoc Tukey test. Fisher exact tests were performed to analyze the ability to regain patency. There was significantly less residual root canal filling material in the AH Plus groups retreated with chloroform as compared to the others. The BC Sealer samples retreated with chloroform had better results than those retreated without chloroform. Furthermore, patency could be re-established in only 14% of teeth in the BC Sealer without chloroform group. The results of this study demonstrate that the BC Sealer group had significantly more residual filling material than the AH Plus group regardless of whether or not both sealers were retreated with chloroform.

IRIS Toxicological Review of Chloroform (Final Report)

EPA Science Inventory

EPA is announcing the release of the final report, Toxicological Review of Chloroform: in support of the Integrated Risk Information System (IRIS). The updated Summary for Chloroform and accompanying Quickview have also been added to the IRIS Database.

Ethyl acetate extract from Panax ginseng C.A. Meyer and its main constituents inhibit α-melanocyte-stimulating hormone-induced melanogenesis by suppressing oxidative stress in B16 mouse melanoma cells.

PubMed

Jiang, Rui; Xu, Xiao-Hao; Wang, Ke; Yang, Xin-Zhao; Bi, Ying-Fei; Yan, Yao; Liu, Jian-Zeng; Chen, Xue-Nan; Wang, Zhen-Zhong; Guo, Xiao-Li; Zhao, Da-Qing; Sun, Li-Wei

2017-08-17

Hyperpigmentation disease involves darkening of the skin color due to melanin overproduction. Panax ginseng C.A. Meyer is a well-known traditional Chinese medicine and has a long history of use as a skin lightener to inhibit melanin formation in China, Korea and some other Asian countries. However, the constituents and the molecular mechanisms by which they affect melanogenesis are not fully clear. The purpose of this study was to identify the active ingredient in Panax ginseng C.A. Meyer extract that inhibits mushroom tyrosinase activity and to investigate the antioxidative capacity and molecular mechanisms of the effective extract on melanogenesis in B16 mouse melanoma cells. Aqueous extracts of Panax ginseng C.A. Meyer were successively fractionated with an equal volume of chloroform, ethyl acetate, and n-butyl alcohol to determine the effects by examining the activity of mushroom tyrosinase. The effective fraction was analyzed using HPLC and LC-MS. The antioxidative capacity and the inhibitory effects on melanin content, cell intracellular tyrosinase activity, and melanogenesis protein levels were determined in α-melanocyte-stimulating hormone (α-MSH)-treated B16 mouse melanoma cells. The ethyl acetate extract from Panax ginseng C.A. Meyer (PG-2) had the highest inhibiting effect on mushroom tyrosinase, mainly contained phenolic acids, including protocatechuic acid, vanillic acid, p-coumaric acid, salicylic acid, and caffeic acid, and exhibited apparent antioxidant activity in vitro. PG-2 and its main constituents significantly decreased melanin content, suppressed cellular tyrosinase activity, and reduced expression of tyrosinase protein to inhibit B16 cells melanogenesis induced by α-MSH, and no cytotoxic effects were observed. They also inhibited cellular reactive oxygen species (ROS) generation, increased superoxide dismutase (SOD) activity and glutathione (GSH) level in α-MSH-treated B16 cells effectively. And those activities of its main constituents could reach more than 80% of PG-2. The ROS scavengers N-acetyl-L-cysteine (NAC) had a similar inhibitory effect on melanogenesis. These results suggest that ethyl acetate extract from Panax ginseng C.A. Meyer has the highest effect on inhibiting melanogenesis, and that its main components are polyphenolic compounds, which may inhibit melanogenesis by suppressing oxidative stress. This work provides new insight into the active constituents and molecular mechanisms underlying skin-lightening effect of Panax ginseng C.A. Meyer. Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.

Dynamic real-time monitoring of chloroform in an indoor swimming pool air using open-path Fourier transform infrared spectroscopy.

PubMed

Chen, M-J; Duh, J-M; Shie, R-H; Weng, J-H; Hsu, H-T

2016-06-01

This study used open-path Fourier transform infrared (OP-FTIR) spectroscopy to continuously assess the variation in chloroform concentrations in the air of an indoor swimming pool. Variables affecting the concentrations of chloroform in air were also monitored. The results showed that chloroform concentrations in air varied significantly during the time of operation of the swimming pool and that there were two peaks in chloroform concentration during the time of operation of the pool. The highest concentration was at 17:30, which is coincident with the time with the highest number of swimmers in the pool in a day. The swimmer load was one of the most important factors influencing the chloroform concentration in the air. When the number of swimmers surpassed 40, the concentrations of chloroform were on average 4.4 times higher than the concentration measured without swimmers in the pool. According to the results of this study, we suggest that those who swim regularly should avoid times with highest number of swimmers, in order to decrease the risk of exposure to high concentrations of chloroform. It is also recommended that an automatic mechanical ventilation system is installed to increase the ventilation rate during times of high swimmer load. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Piperazines for peptide carboxyl group derivatization: effect of derivatization reagents and properties of peptides on signal enhancement in matrix-assisted laser desorption/ionization mass spectrometry.

PubMed

Qiao, Xiaoqiang; Sun, Liangliang; Chen, Lingfan; Zhou, Yuan; Yang, Kaiguang; Liang, Zhen; Zhang, Lihua; Zhang, Yukui

2011-03-15

Piperazine-based derivatives, including 1-(2-pyridyl)piperazine (2-PP), 1-(2-pyrimidyl)piperazine (2-PMP), 1-(4-pyridyl)piperazine (4-PP), and 1-(1-methyl-4-piperidinyl)piperazine (M-PP), were used for the derivatization of carboxyl groups on peptides with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and 1-hydroxy-7-azabenzotriazole (HOAt) as coupling reagents, and trifluoroacetic acid (TFA) as activator. Taking synthetic peptides RVYVHPI (RI-7) and APGDRIYVHPF (AF-11) as samples, the yields of derivatized peptides by 2-PP, 2-PMP and 4-PP were higher than 94%. The effect of piperazine derivatives on the signals of tryptic digests of α-transferrin and bovine serum albumin (BSA) was investigated, and it was found that peptides derivatized by 2-PP and 2-PMP exhibited obviously improved ionization efficiency. Furthermore, comparison of identified peptides before and after derivatization showed that peptides with low molecular weight (MW) and high pI value were preferably detected after derivatization. In addition, after derivatization with 2-PP and 2-PMP, protein myelin basic protein S, 20 kDa protein, and histone H were confidently identified from the tryptic digests of two fractions of rat brain protein separated by reversed-phase high-performance liquid chromatography (HPLC), indicating the potential application of 2-PP and 2-PMP for the highly sensitive determination of peptides in comprehensive proteome analysis. Copyright © 2011 John Wiley & Sons, Ltd.

Accurate determination of aldehydes in amine catalysts or amines by 2,4-dinitrophenylhydrazine derivatization.

PubMed

Barman, Bhajendra N

2014-01-31

Carbonyl compounds, specifically aldehydes, present in amine catalysts or amines are determined by reversed-phase liquid chromatography using ultraviolet detection of their corresponding 2,4-dinitrophenylhydrazones. The primary focus has been to establish optimum conditions for determining aldehydes accurately because these add exposure concerns when the amine catalysts are used to manufacture polyurethane products. Concentrations of aldehydes determined by this method are found to vary with the pH of the aqueous amine solution and the derivatization time, the latter being problematic when the derivatization reaction proceeds slowly and not to completion in neutral and basic media. Accurate determination of aldehydes in amines through derivatization can be carried out at an effective solution pH of about 2 and with derivatization time of 20min. Hydrochloric acid has been used for neutralization of an amine. For complete derivatization, it is essential to protonate all nitrogen atoms in the amine. An approach for the determination of an adequate amount of acid needed for complete derivatization has been described. Several 0.2M buffer solutions varying in pH from 4 to 8 have also been used to make amine solutions for carrying out derivatization of aldehydes. These solutions have effective pHs of 10 or higher and provide much lower aldehyde concentrations compared to their true values. Mechanisms for the formation of 2,4-dinitrophenylhydrazones in both acidic and basic media are discussed. Copyright © 2014 Elsevier B.V. All rights reserved.

In Vitro Antimicrobial Bioassays, DPPH Radical Scavenging Activity, and FTIR Spectroscopy Analysis of Heliotropium bacciferum

PubMed Central

2016-01-01

The present study deals with the antimicrobial, antioxidant, and functional group analysis of Heliotropium bacciferum extracts. Disc diffusion susceptibility method was followed for antimicrobial assessment. Noteworthy antimicrobial activities were recorded by various plant extracts against antibiotic resistant microorganisms. Plant flower extracts antioxidant activity was investigated against 2, 2-diphenyl-1-picryl hydrazyl radical by ultraviolet spectrophotometer (517 nm). Plant extracts displayed noteworthy radical scavenging activities at all concentrations (25–225 μg/mL). Notable activities were recorded by crude, chloroform and ethyl acetate extracts up to 88.27% at 225 μg/mL concentration. Compounds functional groups were examined by Fourier transform infrared spectroscopic studies. Alkanes, alkenes, alkyl halides, amines, carboxylic acids, amides, esters, alcohols, phenols, nitrocompounds, and aromatic compounds were identified by FTIR analysis. Thin layer chromatography bioautography was carried out for all plant extracts. Different bands were separated by various solvent systems. The results of the current study justify the use of Heliotropium bacciferum in traditional remedial herbal medicines. PMID:27597961

Evaluation of the anticonvulsant effect of Centella asiatica (gotu kola) in pentylenetetrazol-induced seizures with respect to cholinergic neurotransmission.

PubMed

Visweswari, Gopalreddygari; Prasad, Kanchi Siva; Chetan, Pandanaboina Sahitya; Lokanatha, Valluru; Rajendra, Wudayagiri

2010-03-01

The study described here was carried out to investigate the anticonvulsant effect of different extracts of Centella asiatica with respect to cholinergic activity on pentylenetetrazol (PTZ)-induced seizures. Rats were randomly divided into eight groups of six rats each: nonepileptic rats treated with saline; PTZ (60 mg/kg, IP)-induced seizure rats treated with saline; PTZ-induced seizure rats pretreated with n-hexane, chloroform, ethyl acetate, n-butanol, and water extracts of C. asiatica; and PTZ-induced seizure rats pretreated with diazepam (2mg/kg body wt). The seized rats pretreated with different extracts were administered a dose of 200mg/kg body wt orally for 1 week before induction of epilepsy. Increased acetylcholine content and decreased acetylcholinesterase activity were recorded in different brain regions during PTZ-induced seizures. Pretreatment with C. asiatica extracts caused recovery of the levels of acetylcholine and acetylcholinesterase. These findings suggest that C. asiatica causes perceptible changes in the cholinergic system as one of the facets of its anticonvulsant activity. (c) 2010 Elsevier Inc. All rights reserved.

The antiepileptic effect of Centella asiatica on the activities of Na+/K+, Mg2+ and Ca2+-ATPases in rat brain during pentylenetetrazol–induced epilepsy

PubMed Central

G., Visweswari; K., Siva Prasad; V., Lokanatha; Rajendra, W.

2010-01-01

Background: To study the anticonvulsant effect of different extracts of Centella asiatica (CA) in male albino rats with reference to Na+/K+, Mg2+ and Ca2+-ATPase activities. Materials and Methods: Male Wistar rats (150±25 g b.w.) were divided into seven groups of six each i.e. (a) control rats treated with saline, (b) pentylenetetrazol (PTZ)-induced epileptic group (60 mg/kg, i.p.), (c) epileptic group pretreated with n-hexane extract (n-HE), (d) epileptic group pretreated with chloroform extract (CE), (e) epileptic group pretreated with ethyl acetate extract (EAE), (f) epileptic group pretreated with n-butanol extract (n-BE), and (g) epileptic group pretreated with aqueous extract (AE). Results: The activities of three ATPases were decreased in different regions of brain during PTZ-induced epilepsy and were increased in epileptic rats pretreated with different extracts of CA except AE. Conclusion: The extracts of C. asiatica, except AE, possess anticonvulsant and neuroprotective activity and thus can be used for effective management in treatment of epileptic seizures. PMID:20711371

Study of total phenol, flavonoids contents and phytochemical screening of various leaves crude extracts of locally grown Thymus vulgaris.

PubMed

Hossain, Mohammad Amzad; AL-Raqmi, Khulood Ahmed Salim; AL-Mijizy, Zawan Hamood; Weli, Afaf Mohammed; Al-Riyami, Qasim

2013-09-01

To prepare various crude extracts using different polarities of solvent and to quantitatively evaluate their total phenol, flavonoids contents and phytochemical screening of Thymus vulgaris collected from Al Jabal Al Akhdar, Nizwa, Sultanate of Oman. The leave sample was extracted with methanol and evaporated. Then it was defatted with water and extracted with different polarities organic solvents with increasing polarities. The prepare hexane, chloroform, ethyl acetate, butanol and methanol crude extracts were used for their evaluation of total phenol, flavonoids contents and phytochemical screening study. The established conventional methods were used for quantitative determination of total phenol, flavonoids contents and phytochemical screening. Phytochemical screening for various crude extracts were tested and shown positive result for flavonoids, saponins and steroids compounds. The result for total phenol content was the highest in butanol and the lowest in methanol crude extract whereas the total flavonoids contents was the highest in methanol and the lowest hexane crude extract. The crude extracts from locally grown Thymus vulgaris showed high concentration of flavonoids and it could be used as antibiotics for different curable and uncurable diseases.

Retrospective cohort mortality study of workers at an aircraft maintenance facility. II. Exposures and their assessment.

PubMed Central

Stewart, P A; Lee, J S; Marano, D E; Spirtas, R; Forbes, C D; Blair, A

1991-01-01

Methods are presented that were used for assessing exposures in a cohort mortality study of 15,000 employees who held 150,000 jobs at an Air Force base from 1939 to 1982. Standardisation of the word order and spelling of the job titles identified 43,000 unique job title organisation combinations. Walkthrough surveys were conducted, long term employees were interviewed, and available industrial hygiene data were collected to evaluate historic exposures. Because of difficulties linking air monitoring data and use of specific chemicals to the departments identified in the work histories, position descriptions were used to identify the tasks in each job. From knowledge of the tasks and the chemicals used in those tasks the presence or absence of 23 chemicals or groups of chemicals were designated for each job organisation combination. Also, estimates of levels of exposure were made for trichloroethylene and for mixed solvents, a category comprising several solvents including trichloroethylene, Stoddard solvent, carbon tetrachloride, JP4 gasoline, freon, alcohols, 1,1,1-trichloroethane, acetone, toluene, methyl ethyl ketone, methylene chloride, o-dichlorobenzene, perchloroethylene, chloroform, styrene, and xylene. PMID:1878309

Anti-diarrhoea and analgesic activities of the methanol extract and its fractions of Jasminum amplexicaule Buch.-Ham. (Oleaceae).

PubMed

Jia, Qiang; Su, Weiwei; Peng, Wei; Li, Peibo; Wang, Yonggang

2008-09-26

Jasminum amplexicaule Buch.-Ham. (Oleaceae) has been commonly used in the traditional medicine in dysentery, diarrhoea and bellyache in China. In the present work, the methanol extract of Jasminum amplexicaule and different fractions of this extract were studied for anti-diarrhoea and analgesic activities. The anti-diarrhoea activities were investigated using castor oil-induced, magnesium sulphate-induced diarrhoea models, antienteropooling assay and gastrointestinal motility models in mice. The analgesic activities were studied using hot-plate, writhing and formalin models in mice. At the doses of 100, 200 and 400mg/kg, the methanol extract (ME) showed significant and dose-dependent anti-diarrhoea and analgesic activity in these models. The chloroform fraction (CHF), ethyl acetate fraction (EAF) and the residual methanol fraction (RMF) exhibited similar activity using a dose of 200mg/kg in these models. The pharmacological activities of the n-butanol fraction (BUF) were lesser than the ME extract and other fractions. These results may support the fact that this plant is traditionally used to cure diarrhoea and pain.

Comparative study on the antioxidant activities of extracts of Coreopsis tinctoria flowering tops from Kunlun Mountains, Xinjiang, north-western China.

PubMed

Yao, Xincheng; Gu, Chengzhi; Tian, Liping; Wang, Xinbing; Tang, Hui

2016-01-01

Coreopsis tinctoria flowering tops (CTFs) from the Kunlun Mountains in Xinjing (north-western China) have been used for tea production for about a century. This study aims to assess the antioxidant activities and total phenolic, flavonoid and proanthocyanidin contents of various solvent extracts of CTF. CTF was extracted using n-hexane, chloroform, ethyl acetate, n-butanol, 75% aqueous ethanol (AEE) and water. The antioxidant activities of the CTF extracts were investigated through DPPH, ABTS, *OH, *O2(-), total antioxidant capacity and reducing power assays. The results showed that n-butanol extract showed the highest contents of total phenols and flavonoids, with DPPH, ABTS and *OH radical-scavenging activities with IC50 values of 134, 90.72 and 13.8 μg mL(-1), respectively. The AEE demonstrated the strongest DPPH and ABTS radical-scavenging activities, with IC50 values of 103 and 75.16 μg mL(-1), respectively. Given its high antioxidant effect, CTF is a good source of natural antioxidants or functional food materials.

Toxicity of amorphigenin from the seeds of Amorpha fruticosa against the larvae of Culex pipiens pallens (Diptera: Culicidae).

PubMed

Liang, Yaping; Li, Xiuwei; Gu, Zumin; Qin, Peiwen; Ji, Mingshan

2015-02-16

The larvicidal activity of the crude petroleum ether, ethyl acetate, acetone, chloroform and ethanol extracts of Amorpha fruticosa seeds was individually assayed for toxicity against the early fourth-instar larva of the mosquito, Culex pipiens pallens after 24 h exposure. Of the tested extracts, the ethanol one exhibited the highest larvicidal activity (LC50 = 22.69 mg/L). Amorphigenin (8'-hydroxyrotenone), a rotenoid compound which exhibits a strong larvicidal activity with LC50 and LC90 values of 4.29 and 11.27 mg/L, respectively, was isolated from the ethanol extract by column chromatograpy. Its structure was elucidated by 1H-NMR, UV and IR spectral data. Furthermore, investigation of amorphigenin's effects on mitochondrial complex I activity and protein synthesis in C. pipiens pallens larvae reveals that amorphigenin decreases mitochondrial complex I activities to 65.73% at 10.45 μmol/L, compared to the control, when NADH were used as the substrate. Meanwhile, amorphigenin at 10.45 μmol/L also caused a 1.98-fold decrease in protein content, compared to the control larvae treated with acetone only.

Preliminary phytochemical analysis, Antioxidant and cytotoxicity test of Carissa edulis Vahl dried fruits

NASA Astrophysics Data System (ADS)

Fowsiya, J.; Madhumitha, G.

2017-11-01

Plants are the main source of medicine which is used in traditional as well as modern medicine in recent years for curing many diseases. Carissa edulis Vahl is one of the traditional plants which have healing property on diarrhea, toothache and chest pain. The present work aims on phytochemical, antioxidant and in vitro cytotoxicity test of C. edulis dried fruits. The different solvent extracts obtained from petroleum ether, ethyl acetate, chloroform, ethanol and water have been evaluated the presence of phytochemicals. Several assays were carried out like total antioxidant, DPPH, reducing power and thiobarbituric acid to investigate the free radical scavenging property. In addition, the cytotoxicity study also carried out on human lung cancer cells (A549). Among different solvent extract, ethanol exhibited strong antioxidant activity. Additionally, the in vitro cytotoxicity test of C. edulis on human lung cancer cell (A549) showed IC50 value 405.704 ± 2.42 μg/mL. Therefore, C. edulis could be useful as a potential preventive intervention for free radicals mediated diseases as well as an antioxidant drug in the pharmaceutical industry.

Solubility and surface thermodynamics of conducting polymers by inverse gas chromatography. III: polypyrrole chloride.

PubMed

Duaij, Omar K; Alghamdi, Ali; Al-Saigh, Zeki Y

2013-05-24

Inverse gas chromatography, IGC, was applied to characterize conducting polypyrrole chloride (PPyCl) using twenty three solvents. IGC is able to reveal the change in the morphology, the strength of solvent-PPyCl interactions, thermodynamics parameters (χ12, Ω1(∞)), solvent and polymer solubility parameters, and molar heats of sorption, mixing and evaporation (ΔH1(s), ΔH1(∞), ΔH1(v)). The following solvents showed stronger interactions than others; yet, none of these solvents are good solvents for PPyCl: dodecane among the alkane family, tetrahydrofuran and methyl ethyl ketone among the oxy and keto group, dichloromethane among the chlorinated group up to 120°C and chloroform at 180°C, and toluene among the cyclic and aromatic group. Overall, the groups showed higher affinities to PPyCl are: acetates, oxy and cyclic, and chlorinated groups. Comprehensive solvents and PPyCl solubility parameters are obtained. The latter showed that PPyCl is not soluble in any solvent used. Copyright © 2013 Elsevier B.V. All rights reserved.

Evaluation of phenolic contents and antioxidant activity of various solvent extracts of Sonchus asper (L.) Hill

PubMed Central

2012-01-01

Background Sonchus asper (SA) is traditionally used for the treatment of various ailments associated with liver, lungs and kidneys. This study was aimed to investigate the therapeutic potential of nonpolar (hexane, SAHE; ethyl acetate, SAEE and chloroform, SACE) and polar (methanol, SAME) crude extracts of the whole plant. Methods To achieve these goals, several parameters including free-radical (DPPH•, ABTS•+, H2O2 and •OH) scavenging, iron chelating activity, scavenging of superoxide radicals, total flavonoids and total phenolic content (TPC) were examined. Results The SA extracts presented a remarkable capacity to scavenge all the tested reactive species with IC50 values being found at the μg ⁄ ml level. The SAME was shown to have the highest TPCs while lowest IC50 values for the DPPH•, ABTS•+ radical scavenging capacities and iron chelating scavenging efficiency, moreover, SAME had best activities in scavenging of superoxide radicals and hydrogen peroxide as well as potently scavenged the hydroxyl radicals. Conclusion These results suggest the potential of S. asper as a medicine against free-radical-associated oxidative damage. PMID:22305477

Mycochemical Investigation of the Turkey Tail Medicinal Mushroom Trametes versicolor (Higher Basidiomycetes): A Potential Application of the Isolated Compounds in Documented Pharmacological Studies.

PubMed

Habibi, Emran; Sadat-Ebrahimi, Seyed Esmaeel; Mousazadeh, Saeed Ali; Amanzadeh, Yaghoub

2015-01-01

The purpose of this study was to elucidate the chemical properties of the n-hexane, chloroform, and ethyl acetate extracts of the fruiting body of the medicinal mushroom Trametes versicolor. The study led to the isolation of 5 sterols, 2 triterpene derivatives, 1 hydroquinone-derived aromatic compound, and, finally, 1 cerebroside and 1 triglyceride derivative. These compounds were identified for first time in T. versicolor and were named as follows: 4-isobutoxyphenyl palmitate (5), N-D-2'-hydroxyheptanoic-1-O-β-D-glucopyranosyl-9-methyl-4,8-sphinga-dienine(cerebroside) (6), 3β-linoleyloxyergosta-7,22-diene (7), 3β-linoleyloxyergosta-7-ene (8), and betulinic acid (9). Other compounds elucidated in our study were ergosterol (1), ergosterol peroxide (2), trilinolein (3), ergosta-7, 22-dien-3β-ol (4), and betuline (10). These compounds were obtained via column or thin-layer chromatography before being identified by nuclear magnetic resonance spectroscopic analyses and infrared data. In addition, the beneficial pharmacological effects of the compounds are described here.

Quantitative analysis of the endogenous GHB level in the hair of the Chinese population using GC/MS/MS.

PubMed

Shi, Yan; Cui, Xiaopei; Shen, Min; Xiang, Ping

2016-04-01

Endogenous production complicates interpretation when gamma-hydroxybutyrate (GHB) is measured in hair for forensic purposes. A method capable of quantifying the endogenous concentration of GHB in human head hair was developed and validated using GC/MS/MS. Hair was digested under alkaline conditions (1 mol/L NaOH, 90 °C 10 min), and GHB-d6 was used as an internal standard. Before derivatization with BSTFA and ethyl acetate, a liquid-liquid extraction with ethyl acetate under acidic conditions was performed. GHB-TMS derivatives were detected using GC/MS/MS in the multiple-reaction monitoring mode. This method exhibited good linearity (y = 0.018x + 0.038, R(2) = 0.9998), and the limit of detection was 0.02 ng/mg. The extraction recoveries were more than 60%, and the inter-day and intra-day relative standard deviations (RSD) were less than 15%. This method has been applied for the analysis of the endogenous GHB in hair samples from 66 drug-free Chinese donors. The mean measured concentration for 0-3 cm hair was 1.93 ± 1.40 ng/mg (n = 66), and extreme values were in the range of 0.28-4.91 ng/mg. The mean male endogenous GHB level was 2.95 ng/mg (0.92-4.91 ng/mg, n = 35), while the mean female level was 0.77 ng/mg (0.28-1.95 ng/mg, n = 31). This method was applied to a forensic case for the determination of GHB in hair samples but it is hard to make a reasonable "cut off" in hair. The solution is to use each subject as his own control. Copyright © 2016 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved.

CANCER RISK ASSESSMENT FOR CHLOROFORM

EPA Science Inventory

Chloroform is a common chlorination by-product in drinking water. EPA has regulated chloroform as a probable human carcinogen under the Safe Drinking Water Act. The cancer risk estimate via ingestion was based on the 1985 Jorgenson study identifying kidney tumors in male Osborne ...

CONTROLLED, SHORT-TERM DERMAL AND INHALATION EXPOSURE TO CHLOROFORM

EPA Science Inventory

Studies were conducted to determine the uptake by humans of chloroform as a result of controlled short-term dermal and inhalation exposures. The approach used continuous real-time breath analysis to determine exhaled-breath profiles and evaluate chloroform kinetics in the huma...

Synthesis, structural characterization, and thermal stability studies of heteroleptic cadmium(II) dithiocarbamate with different pyridyl groups

NASA Astrophysics Data System (ADS)

Onwudiwe, Damian C.; Hosten, Eric C.

2018-01-01

The synthesis, characterization and crystal structures of three chloroform solvated adducts of cadmium with mixed ligands of N-alkyl-N-phenyldithiocarbamate and pyridine, 2,2-bipyridine and 1, 10 phenanthroline represented as [CdL1L2 (py)2]·CHCl3(1), [CdL1L2bpy]•CHCl3(2), and [CdL1L2phen]•CHCl3(3) (LI = N-methyl-N-phenyldithiocarbamate, L2 = N-ethyl-N-phenyldithiocarbamate, py = pyridine, bpy = 2,2-bipyridine and phen = 1,10-phenanthroline) respectively are reported. Complex 1, which crystallized in the monoclinic space group P-1, is a centrosymmetric dimeric structure where each Cd center is bonded to two monodentate pyridine, a bidentate terminal dithiocarbamate, and another bidentate bridging dithiocarbamate to form a four-membered ring. Complex 2 crystallized in the monoclinic space group P21/c, with four discrete monomeric molecules in the asymmetric unit. The structure presents a cadmium atom coordinated by two sulphur atoms of a dithiocarbamate ligand and two nitrogen atoms of the 2,2‧-bipyridine to form a CdS4N2 fragment, thus giving the structure around the Cd atom a distorted trigonal prism geometry. Complex 3 contains two discrete monomeric molecules of (phenanthroline) (N, N-methyl phenyl-N, N-ethyl phenyl dithiocarbamato)cadmium (II) per unit cell, and the complex crystallized in the triclinic space group P-1. The structure showed that the Cd atom is bonded to two bidentate dithiocarbamate ligands and to one bidentate phenanthroline ligand in a distorted trigonal prism geometry. All the compounds resulted in CdS as residue upon thermal decomposition process conducted under inert atmosphere.

Antioxidant Activity of Phenolic Compounds from Fava Bean Sprouts.

PubMed

Okumura, Koharu; Hosoya, Takahiro; Kawarazaki, Kai; Izawa, Norihiko; Kumazawa, Shigenori

2016-06-01

Fava beans are eaten all over the world and recently, marketing for their sprouts began in Japan. Fava bean sprouts contain more polyphenols and l-3,4-dihydroxyphenylalanine (l-DOPA) than the bean itself. Our antioxidant screening program has shown that fava bean sprouts also possess a higher antioxidant activity than other commercially available sprouts and mature beans. However, the individual constituents of fava bean sprouts are not entirely known. In the present study, we investigated the phenolic compounds of fava bean sprouts and their antioxidant activity. Air-dried fava bean sprouts were treated with 80% methanol and the extract was partitioned in water with chloroform and ethyl acetate. HPLC analysis had shown that the ethyl acetate-soluble parts contained phenolic compounds, separated by preparative HPLC to yield 5 compounds (1-5). Structural analysis using NMR and MS revealed that the compounds isolated were kaempferol glycosides. All isolated compounds had an α-rhamnose at the C-7 position with different sugars attached at the C-3 position. Compounds 1-5 had β-galactose, β-glucose, α-rhamnose, 6-acetyl-β-galactose and 6-acetyl-β-glucose, respectively, at the C-3 position. The amount of l-DOPA in fava bean sprouts was determined by the quantitative (1) H NMR technique. The l-DOPA content was 550.45 mg ± 11.34 /100 g of the raw sprouts. The antioxidant activities of compounds 2-5 and l-DOPA were evaluated using the 2,2-diphenyl-1-picrylhydrazyl scavenging assay. l-DOPA showed high antioxidant activity, but the isolated kaempferol glycosides showed weak activity. Therefore, it can be suggested that l-DOPA contributed to the antioxidant activity of fava bean sprouts. © 2016 Institute of Food Technologists®

Bio-prospecting of coral (Porites lutea) mucus associated bacteria, Palk Bay reefs, Southeast coast of India.

PubMed

Ahila, N K; Prakash, S; Manikandan, B; Ravindran, J; Prabhu, N M; Kannapiran, E

2017-12-01

Coral mucus is one of the key localization in the coral holobiont, as this serves as an energy rich substrate for a wide range of abundant, diverse and multifunctional microbiota. However, very little is known about the functional role of bacterial communities in their associations with corals. In the present study, a total of 48 isolates were obtained from Porites lutea wherein the genus of Bacillus sp. and Vibrio sp. were predominant. Bio-prospecting the coral mucus revealed the existence of (10.42%) antagonistic bacteria against the tested bacterial pathogens. Molecular taxonomy (16S rRNA) proved the identity of these antagonistic bacteria belong to Enterobacter cloacae (CM1), Bacillus subtilis (CM2), Bacillus sp. (CM11) and Bacillus marisflavi (CM12). The secondary screening emphasized that the ethyl acetate extract of B. subtilis showed strong antagonistic effect, followed by the chloroform extract of E. cloacae and ethyl acetate extract of B. marisflavi. The antagonistic activity was statistically confirmed by Principal Component Analysis and Hierarchical Cluster Analysis. The privileged coral mucus associated bacterial (CMAB) solvent extracts inhibited the bacterial pathogens at 100 μg/ml (MIC) and ceased the growth at 200 μg/ml (MBC). The hemolytic and brine shrimp lethality assays disclosed the non-toxic nature of solvent extracts of CMAB. Altogether, the present investigation brought out the diversity of bacteria associated with the mucus of P. lutea. In addition, bio-prospecting corroborated the CMAB as the potential source of pharmacologically important bioactive compounds against a wide range of bacterial pathogens. Copyright © 2017 Elsevier Ltd. All rights reserved.

Radiochemical studies of 99mTc complexes of modified cysteine ligands and bifunctional chelating agents.

PubMed

Pillai, M R; Kothari, K; Banerjee, S; Samuel, G; Suresh, M; Sarma, H D; Jurisson, S

1999-07-01

The synthesis of four novel ligands using the amino-acid cysteine and its ethyl carboxylate derivative is described. The synthetic method involves a two-step procedure, wherein the intermediate Schiff base formed by the condensation of the amino group of the cysteine substrate and salicylaldehyde is reduced to give the target ligands. The intermediates and the final products were characterized by high resolution nuclear magnetic resonance spectroscopy. Complexation studies of the ligands with 99mTc were optimized using stannous tartrate as the reducing agent under varying reaction conditions. The complexes were characterized using standard quality control techniques such as thin layer chromatography, paper electrophoresis, and paper chromatography. Lipophilicities of the complexes were estimated by solvent extraction into chloroform. Substantial changes in net charge and lipophilicity of the 99mTc complexes were observed on substituting the carboxylic acid functionality in ligands I and II with the ethyl carboxylate groups (ligands II and IV). All the ligands formed 99mTc complexes in high yield. Whereas the complexes with ligands I and II were observed to be hydrophilic in nature and not extractable into CHCl3, ligands III and IV resulted in neutral and lipophilic 99mTc complexes. The 99mTc complex with ligand II was not stable and on storage formed a hydrophilic and nonextractable species. The biodistribution of the complexes of ligands I and II showed that they cleared predominantly through the kidneys, whereas the complexes with ligands III and IV were excreted primarily through the hepatobiliary system. No significant brain uptake was observed with the 99mTc complexes with ligands III and IV despite their favorable properties of neutrality, lipophilicity, and conversion into a hydrophilic species. These ligands offer potential for use as bifunctional chelating agents.

Spectroscopic investigations (FT-IR, UV, 1H and 13C NMR) and DFT/TD-DFT calculations of potential analgesic drug 2-[2-(dimethylamino)ethyl]-6-methoxy-4-(pyridin-2-yl)-1(2H)-phthalazinone

NASA Astrophysics Data System (ADS)

Sroczyński, Dariusz; Malinowski, Zbigniew

2017-12-01

The theoretical molecular geometry and the IR, UV, 1H and 13C NMR spectroscopic properties of 2-[2-(dimethylamino)ethyl]-6-methoxy-4-(pyridin-2-yl)-1(2H)-phthalazinone with the previously demonstrated in vivo analgesic activity were characterized. The conformational analysis, performed using the molecular mechanics method with the General AMBER Force Field (GAFF) and the Density Functional Theory (DFT) approach with the B3LYP hybrid functional and the 6-31 + g(d) basis sets, allowed to determine the most stable rotamer. The theoretical molecular geometry of this conformer was then calculated at the B3LYP/6-311++g(d,p) level of theory, and its phthalazinone core was compared with the experimental geometry of 1(2H)-phthalazinone. The calculated vibrational frequencies and the potential energy distribution enabled to assign the theoretical vibrational modes to the experimental FT-IR bands. The UV spectrum calculated with the Time-Dependent Density Functional Theory (TD-DFT) method in methanol identified the main electronic transitions and their character. 1H and 13C NMR chemical shifts simulated by the Gauge-Independent Atomic Orbital (GIAO) method in chloroform confirmed the previous assignment of the experimental resonance signals. The stability of the molecule was considered taking into account the hyperconjugation and electron density delocalization effects evaluated by the Natural Bond Orbital (NBO) method. The calculated spatial distribution of molecular electrostatic potential made possible to estimate the regions with nucleophilic and electrophilic properties. The results of the potentiodynamic polarization measurements were also indicated the corrosion inhibition activity of the title compound on 100Cr6 bearing steel in 1 mol dm-3 HCl solution.

Derivatization of amino acids with N,N-dimethyl-2,4-dinitro-5-fluorobenzylamine for liquid chromatography/electrospray ionization mass spectrometry.

PubMed

Liu, Zhongfa; Minkler, Paul E; Lin, De; Sayre, Lawrence M

2004-01-01

Derivatization, separation and identification of amino acids with a novel compound, N,N-dimethyl-2,4-dinitro-5-fluorobenzylamine (DMDNFB), using high-performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS) was demonstrated. Compared to derivatization with 2,4-dinitrofluorobenzene (DNFB), DMDNFB-derivatized amino acids and dipeptides exhibit much larger ion current signals in the commonly used ESI positive mode, which was attributed to the introduction of the N,N-dimethylaminomethyl protonatable site. Copyright 2004 John Wiley & Sons, Ltd.

Derivatization reagents in liquid chromatography/electrospray ionization tandem mass spectrometry.

PubMed

Santa, Tomofumi

2011-01-01

Liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) is one of the most prominent analytical techniques owing to its inherent selectivity and sensitivity. In LC/ESI-MS/MS, chemical derivatization is often used to enhance the detection sensitivity. Derivatization improves the chromatographic separation, and enhances the mass spectrometric ionization efficiency and MS/MS detectability. In this review, an overview of the derivatization reagents which have been applied to LC/ESI-MS/MS is presented, focusing on the applications to low molecular weight compounds. 2010 John Wiley & Sons, Ltd.

Simultaneous determination of diastereoisomeric and enantiomeric impurities in SSS-octahydroindole-2-carboxylic acid by chiral high-performance liquid chromatography with pre-column derivatization.

PubMed

Wang, Jin Zhao; Zeng, Su; Hu, Gong Yun; Wang, Dan Hua

2009-04-10

SSS-Octahydroindole-2-carboxylic acid (SSS-Oic) is a key intermediate used in the synthesis of some angiotensin-converting enzyme (ACE) inhibitors. The separation of diastereoisomers and enantiomers of Oic was performed using a pre-column derivatization chiral HPLC method. Phenyl isothiocyanate (PITC) was used as the derivatization reagent. Three PITC derivatives of Oic stereoisomers were separated on an Ultron ES-OVM chiral column (150 mm x 4.6 mm, 5 microm). Derivatization conditions such as reaction temperature, reaction time and derivatization reagent concentration were investigated. The chromatographic conditions for separation of the three PITC-Oic derivatives were optimized. The method was successfully applied in the diastereoisomeric and enantiomeric purity test of SSS-Oic.

Determination of volatile organic acids in oriental tobacco by needle-based derivatization headspace liquid-phase microextraction coupled to gas chromatography/mass spectrometry.

PubMed

Sun, Shi-Hao; Xie, Jian-Ping; Xie, Fu-Wei; Zong, Yong-Li

2008-02-01

A method coupling needle-based derivatization headspace liquid-phase microextraction with gas chromatography-mass spectrometry (HS-LPME/GC-MS) was developed to determine volatile organic acids in tobacco. The mixture of N,O-bis(trimethylsilyl)trifluoroacetamide and decane was utilized as the solvent for HS-LPME, resulting that extraction and derivatization were simultaneously completed in one step. The solvent served two purposes. First, it pre-concentrated volatile organic acids in the headspace of tobacco sample. Second, the volatile organic acids extracted were derivatized to form silyl derivatives in the drop. The main parameters affecting needle-based derivatization HS-LPME procedure such as extraction and derivatization reagent, microdrop volume, extraction and derivatization time, and preheating temperature and preheating time were optimized. The standard addition approach was essential to obtain accurate measurements by minimizing matrix effects. Good linearity (R(2)> or =0.9804) and good repeatability (RSDs< or =15.3%, n=5) for 16 analytes in spiked standard analytes sample were achieved. The method has the additional advantages that at the same time it is simple, fast, effective, sensitive, selective, and provides an overall profile of volatile organic acids in the oriental tobacco. This paper does offer an alternative approach to determine volatile organic acids in tobacco.

Retreatability of two endodontic sealers, EndoSequence BC Sealer and AH Plus: a micro-computed tomographic comparison

PubMed Central

Oltra, Enrique; Cox, Timothy C.; LaCourse, Matthew R.; Johnson, James D.

2017-01-01

Objectives Recently, bioceramic sealers like EndoSequence BC Sealer (BC Sealer) have been introduced and are being used in endodontic practice. However, this sealer has limited research related to its retreatability. Hence, the aim of this study was to evaluate the retreatability of two sealers, BC Sealer as compared with AH Plus using micro-computed tomographic (micro-CT) analysis. Materials and Methods Fifty-six extracted human maxillary incisors were instrumented and randomly divided into 4 groups of 14 teeth: 1A, gutta-percha, AH Plus retreated with chloroform; 1B, gutta-percha, AH Plus retreated without chloroform; 2A, gutta-percha, EndoSequence BC Sealer retreated with chloroform; 2B, gutta-percha, EndoSequence BC Sealer retreated without chloroform. Micro-CT scans were taken before and after obturation and retreatment and analyzed for the volume of residual material. The specimens were longitudinally sectioned and digitized images were taken with the dental operating microscope. Data was analyzed using an ANOVA and a post-hoc Tukey test. Fisher exact tests were performed to analyze the ability to regain patency. Results There was significantly less residual root canal filling material in the AH Plus groups retreated with chloroform as compared to the others. The BC Sealer samples retreated with chloroform had better results than those retreated without chloroform. Furthermore, patency could be re-established in only 14% of teeth in the BC Sealer without chloroform group. Conclusion The results of this study demonstrate that the BC Sealer group had significantly more residual filling material than the AH Plus group regardless of whether or not both sealers were retreated with chloroform. PMID:28194360

Chloroform Hydrodechlorination over Palladium–Gold Catalysts: A First-Principles DFT Study

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xu, Lang; Yao, Xiaoqian; Khan, Ahmad

2016-04-20

Hydrodechlorination is a promising method for treating toxic chlorocarbon compounds. Pd is among the most effective catalysts for chloroform hydrodechlorination, and experiments have shown that the Pd–Au alloy catalyst yields superior catalytic performance over pure Pd. In this paper, we examine the chloroform hydrodechlorination mechanism over Pd(1 1 1) and Pd ML/Au(1 1 1) surfaces using periodic, self-consistent density functional theory calculations (DFT, GGA–PW91) and maximum rate analysis. We suggest that the reaction occurs on both surfaces through complete dechlorination of chloroform followed by hydrogenation of CH* to methane, and that the initial dechlorination step is likely the rate-limiting step.more » Finally, on Pd(1 1 1), the chloroform dechlorination barrier is 0.24 eV higher than the desorption barrier, whereas on Pd ML/Au(1 1 1), the chloroform dechlorination barrier is 0.07 eV lower than the desorption barrier, which can explain the higher hydrodechlorination activity of the Pd–Au alloy catalyst.« less

Investigating chloroperoxidase-catalyzed formation of chloroform from humic substances using stable chlorine isotope analysis.

PubMed

Breider, Florian; Hunkeler, Daniel

2014-01-01

Chloroperoxidase (CPO) is suspected to play an important role in the biosynthesis of natural chloroform. The aims of the present study are to evaluate the variability of the δ(37)Cl value of naturally produced chloroform and to better understand the reaction steps that control the chlorine isotope signature of chloroform. The isotope analyses have shown that the chlorination of the humic substances (HS) in the presence of high H3O(+) and Cl(-) concentrations induces a large apparent kinetic isotope effect (AKIE = 1.010-1.018) likely associated with the transfer of chlorine between two heavy atoms, whereas in the presence of low H3O(+) and Cl(-) concentrations, the formation of chloroform induces a smaller AKIE (1.005-1.006) likely associated with the formation of an HOCl-ferriprotoporphyrin IX intermediate. As the concentration of H3O(+) and Cl(-) in soils are generally at submillimolar levels, the formation of the HOCl-ferriprotoporphyrin IX intermediate is likely rate-limiting in a terrestrial environment. Given that the δ(37)Cl values of naturally occurring chloride tend to range between -1 and +1‰, the δ(37)Cl value of natural chloroform should vary between -5‰ and -8‰. As the median δ(37)Cl value of industrial chloroform is -3.0‰, the present study suggests that chlorine isotopic composition of chloroform might be used to discriminate industrial and natural sources in the environment.

A microfluidic device for the automated derivatization of free fatty acids to fatty acid methyl esters.

PubMed

Duong, Cindy T; Roper, Michael G

2012-02-21

Free fatty acid (FFA) compositions are examined in feedstock for biodiesel production, as source-specific markers in soil, and because of their role in cellular signaling. However, sample preparation of FFAs for gas chromatography-mass spectrometry (GC-MS) analysis can be time and labor intensive. Therefore, to increase sample preparation throughput, a glass microfluidic device was developed to automate derivatization of FFAs to fatty acid methyl esters (FAMEs). FFAs were delivered to one input of the device and methanolic-HCl was delivered to a second input. FAME products were produced as the reagents traversed a 29 μL reaction channel held at 55 °C. A Design of Experiment protocol was used to determine the combination of derivatization time (T(der)) and ratio of methanolic-HCl:FFA (R(der)) that maximized the derivatization efficiencies of tridecanoic acid and stearic acid to their methyl ester forms. The combination of T(der) = 0.8 min and R(der) = 4.9 that produced optimal derivatization conditions for both FFAs within a 5 min total sample preparation time was determined. This combination of T(der) and R(der) was used to derivatize 12 FFAs with a range of derivatization efficiencies from 18% to 93% with efficiencies of 61% for tridecanoic acid and 84% for stearic acid. As compared to a conventional macroscale derivatization of FFA to FAME, the microfluidic device decreased the volume of methanolic-HCl and FFA by 20- and 1300-fold, respectively. The developed microfluidic device can be used for automated preparation of FAMEs to analyze the FFA compositions of volume-limited samples.

Tandem MS Analysis of Selenamide-Derivatized Peptide Ions

NASA Astrophysics Data System (ADS)

Zhang, Yun; Zhang, Hao; Cui, Weidong; Chen, Hao

2011-09-01

Our previous study showed that selenamide reagents such as ebselen and N-(phenylseleno)phthalimide (NPSP) can be used for selective and rapid derivatization of protein/peptide thiols in high conversion yield. This paper reports the systematic investigation of MS/MS dissociation behaviors of selenamide-derivatized peptide ions upon collision induced dissociation (CID) and electron transfer dissociation (ETD). In the positive ion mode, derivatized peptide ions exhibit tag-dependent CID dissociation pathways. For instance, ebselen-derivatized peptide ions preferentially undergo Se-S bond cleavage upon CID to produce a characteristic fragment ion, the protonated ebselen ( m/z 276), which allows selective identification of thiol peptides from protein digest as well as selective detection of thiol proteins from protein mixture using precursor ion scan (PIS). In contrast, NPSP-derivatized peptide ions retain their phenylselenenyl tags during CID, which is useful in sequencing peptides and locating cysteine residues. In the negative ion CID mode, both types of tags are preferentially lost via the Se-S cleavage, analogous to the S-S bond cleavage during CID of disulfide-containing peptide anions. In consideration of the convenience in preparing selenamide-derivatized peptides and the similarity of Se-S of the tag to the S-S bond, we also examined ETD of the derivatized peptide ions to probe the mechanism for electron-based ion dissociation. Interestingly, facile cleavage of Se-S bond occurs to the peptide ions carrying either protons or alkali metal ions, while backbone cleavage to form c/z ions is severely inhibited. These results are in agreement with the Utah-Washington mechanism proposed for depicting electron-based ion dissociation processes.

Biofiltration of Chloroform in a Trickle Bed Air Biofilter Under Acidic Conditions

EPA Science Inventory

In this paper, the application of biofiltration is investigated for controlled removal of gas phase chloroform through cometabolic degradation with ethanol. A trickle bed air biofilter (TBAB) operated under acidic pH 4 is subjected to aerobic biodegradation of chloroform and etha...

Tyrosinase Inhibitory Activities of Carissa opaca Stapf ex Haines Roots Extracts and Their Phytochemical Analysis

PubMed Central

Malik, Wajeeha; Ahmed, Dildar; Izhar, Sania

2017-01-01

Objective: Carissa opaca is a medicinal plant with rich folkloric applications. The present research was conducted to explore the tyrosinase inhibitory potential of aqueous decoction (AD) and methanolic extract (ME) of roots of C. opaca and its fractions in various solvents and their phytochemical analysis. Materials and Methods: AD of the dried powdered roots of C. opaca was prepared by boiling in water. ME was prepared by cold maceration. Its fractions were obtained in solvents of increasing polarity, i.e., hexane, chloroform, ethyl acetate, n-butanol, and water. The biomass left after extraction with methanol was boiled in water to get its decoction Biomass aqueous decoction (BAD). Tyrosinase inhibitory activities of the samples were studied according to a reported method. Chemical compounds in the samples were identified by gas chromatography-mass spectrometry (GC-MS). Results: The AD, BAD, and ME and its fractions displayed remarkable tyrosinase inhibitory activity. The IC50 of AD was 23.33 μg/mL as compared to 15.80 μg/mL of the standard arbutin and that of BAD was 21.24 μg/mL. The IC50 of ME was 34.76 μg/mL while that of hexane, chloroform, ethyl acetate, n-butanolic, and aqueous fractions was 21.0, 44.73, 43.40, 27.66, and 25.06 μg/mL, respectively. The hexane fraction was thus most potent followed by aqueous fraction. By phytochemical analysis, campesterol, stigmasterol, gamma-sitosterol, alpha-amyrin, 9,19-cyclolanostan-3-ol, 24-methylene-,(3 β)-, lupeol, lup-20(29)-en-3-one, lup-20(29)-en-3-ol, acetate,(3 β)-, 2(1H) naphthalenone, 3,5,6,7,8,8a-hexahydro-4,8a-dimethyl-6-(1-methylethenyl)-, and 2,3,3-trimethyl-2-(3-methylbuta-1,3-dienyl)-6-methylenecyclohexanone were identified in the extracts by GC-MS. Other compounds included fatty acids and their esters. Some of these compounds are being first time reported here from this plant. Conclusions: The roots extracts exhibited considerable tyrosinase inhibitory activities, alluding to a possible application of the plant in cosmetic as whitening agent subject to further pharmacological studies. SUMMARY The present study aimed to explore the tyrosinase inhibitory potential of aqueous decoction and methanolic extract of roots of Carissa opaca and its fractions in various solvents and their phytochemical constituents. GCMS analysis was conducted to identify the phytochemicals. The extracts and fractions of C. opaca roots showed remarkable anti-tyrosinase activities alluding to their possible application to treat disorders related to overproduction of melanin. Abbreviations used: AD: Aqueous decoction; ME: Methanolic extract; BAD: Biomass aqueous decoction; GC-MS: Gas chromatography-mass spectrometry. PMID:29142412

Enhancement of Drug Detection and Identification by Use of Various Derivatizing Reagents on GC-FTIR Analysis.

DTIC Science & Technology

1992-07-01

pseudoephedrine and L- studied the effects of five different phenylpropanolamine ephedrine, which are stereoisomers of each other, infrared...FIGURE 2. IR spectrum of D- pseudoephedrine . dard because it does not derivatize with any of the derivatizing agents chosen for this study and it has a

Development of a compound-specific isotope analysis method for acetone via 2,4-dinitrophenylhydrazine derivatization.

PubMed

Wen, Sheng; Feng, Yanli; Wang, Xinming; Sheng, Guoying; Fu, Jiamo; Bi, Xinhui

2004-01-01

A novel method has been developed for compound-specific isotope analysis for acetone via DNPH (2,4-dinitrophenylhydrazine) derivatization together with combined gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). Acetone reagents were used to assess delta13C fractionation during the DNPH derivatization process. Reduplicate delta13C analyses were designed to evaluate the reproducibility of the derivatization, with an average error (1 standard deviation) of 0.17 +/- 0.05 per thousand, and average analytical error of 0.28 +/- 0.09 per thousand. The derivatization process introduces no isotopic fractionation for acetone (the average difference between the predicted and analytical delta13C values was 0.09 +/- 0.20 per thousand, within the precision limits of the GC/C/IRMS measurements), which permits computation of the delta13C values for the original underivatized acetone through a mass balance equation. Together with further studies of the carbon isotopic effect during the atmospheric acetone-sampling procedure, it will be possible to use DNPH derivatization for carbon isotope analysis of atmospheric acetone. Copyright (c) 2004 John Wiley & Sons, Ltd.

Extraction and derivatization of polar herbicides for GC-MS analyses.

PubMed

Ranz, Andreas; Maier, Eveline; Motter, Herbert; Lankmayr, Ernst

2008-09-01

A sample preparation procedure including a simultaneous microwave-assisted (MA) extraction and derivatization for the determination of chlorophenoxy acids in soil samples is presented. For a selective and sensitive measurement, an analytical technique such as GC coupled with MS needs to be adopted. For GC analyses, chlorophenoxy acids have to be converted into more volatile and thermally stable derivatives. Derivatization by means of microwave radiation offers new alternatives in terms of shorter derivatization time and reduces susceptibility for the formation of artefacts. Extraction and derivatization into methyl esters (ME) were performed with sulphuric acid and methanol. Due to the novelty of the simultaneous extraction and derivatization assisted by means of microwave radiation, a careful investigation and optimization of influential reaction parameters was necessary. It could be shown that the combination of sulphuric acid and methanol provides a fast sample preparation including an efficient clean up procedure. The data obtained by the described method are in good agreement with those published for the reference material. Finally, compared to conventional heating and also to the standard procedure of the EPA, the sample preparation time could be considerably shortened.

Chemical derivatization for enhancing sensitivity during LC/ESI-MS/MS quantification of steroids in biological samples: a review.

PubMed

Higashi, Tatsuya; Ogawa, Shoujiro

2016-09-01

Sensitive and specific methods for the detection, characterization and quantification of endogenous steroids in body fluids or tissues are necessary for the diagnosis, pathological analysis and treatment of many diseases. Recently, liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) has been widely used for these purposes due to its specificity and versatility. However, the ESI efficiency and fragmentation behavior of some steroids are poor, which lead to a low sensitivity. Chemical derivatization is one of the most effective methods to improve the detection characteristics of steroids in ESI-MS/MS. Based on this background, this article reviews the recent advances in chemical derivatization for the trace quantification of steroids in biological samples by LC/ESI-MS/MS. The derivatization in ESI-MS/MS is based on tagging a proton-affinitive or permanently charged moiety on the target steroid. Introduction/formation of a fragmentable moiety suitable for the selected reaction monitoring by the derivatization also enhances the sensitivity. The stable isotope-coded derivatization procedures for the steroid analysis are also described. Copyright © 2015 Elsevier Ltd. All rights reserved.

Physicochemical properties and biological activities of DEAE-derivatized Sphingomonas gellan.

PubMed

Yoo, Sang-Ho; Lee, Kyung Hee; Lee, Ji-Soo; Cha, Jaeho; Park, Cheon Seok; Lee, Hyeon Gyu

2005-08-10

Physicochemical characteristics and biological activities of Sphingomonas gellan (S-gellan) were investigated. The S-gellan weight fractions of Glc and GlcUA were 0.45 and 0.25, respectively, and the molar ratio of Glc:Rha:GlcUA was approximately 4:2:3. The S-gellan was chemically derivatized with diethylaminoethyl chloride-HCl (DEAE-HCl), and the resulting modified S-gellan contained both positive and negative charges. The elemental and IR analyses were conducted to confirm the successful incorporation of DEAE groups into S-gellan. A large increase in nitrogen fraction was observed from the derivatized S-gellan by elemental analysis. The IR absorption bands induced by C-H, C-N, and C-O-C stretching were noticeable at 2950, 1310-1380, and 1000-1150 cm(-1), respectively, resulting from the DEAE substitution. The characteristic CH3 and CH2 peaks originated from the DEAE group were detected in the 1H NMR spectrum of the derivatized S-gellan as well. The solubility of native S-gellan was improved almost twice from 40% to 75% after DEAE derivatization, while water holding capacity (WHC) drastically decreased from 10026% to 245%. Oil binding capacity (OBC) of S-gellan also significantly dropped from 1528% to 331% after the derivatization. The bile acid binding capacity of S-gellan was indirectly determined by measuring the holding capability of cholic acid inside the dialysis membrane (MWCO 12,000-14,000 Da). Once S-gellan was DEAE derivatized, there was substantial increase in the cholic acid retardation index (CRI). Up to 9 h of dialysis, the derivatized S-gellan released 29.3% less of cholic acid compared to the control group that did not contain S-gellan. From these results of the improved water solubility and stronger bile acid binding capacity, it would be suggested that the DEAE-derivatized S-gellan has more advantages than gellan itself for functional food applications.

THE ENHANCEMENT OF CHLOROFORM-INDUCED PLASMA PROTEOLYTIC ACTIVITY BY EPSILON AMINOCAPROIC ACID

PubMed Central

Donaldson, Virginia H.; Ratnoff, Oscar D.

1962-01-01

The proteolytic activity in chloroform-treated plasma euglobulins has been attributed to plasmin. Plasmin can digest both casein and fibrin. Epsilon aminocaproic acid, which inhibits the activation of plasminogen, the precursor of plasmin, by streptokinase, urokinase, and tissue activators enhanced the development of casein hydrolytic activity in a mixture of chloroform and plasma euglobulins. Fibrinolytic activity was also enhanced, but this was evident only if the epsilon aminocaproic acid was removed from the chloroform-treated euglobulins prior to assay. The reasons for the paradoxical enhancement of chloroform-induced casein hydrolysis by euglobulins containing epsilon aminocaproic acid are unclear. However, studies of optimal pH, heat stability, and the effect of ionic strength on the activation of the precursor of this proteolytic enzyme do not differentiate it from plasminogen. PMID:13887179

A new agent for derivatizing carbonyl species used to investigate limonene ozonolysis

NASA Astrophysics Data System (ADS)

Wells, J. R.; Ham, Jason E.

2014-12-01

A new method for derivatizing carbonyl compounds is presented. The conversion of a series of dicarbonyls to oximes in aqueous solution and from gas-phase sampling was achieved using O-tert-butylhydroxylamine hydrochloride (TBOX). Some advantages of using this derivatization agent include: aqueous reactions, lower molecular weight oximes, and shortened oxime-formation reaction time. Additionally, the TBOX derivatization technique was used to investigate the carbonyl reaction products from limonene ozonolysis. With ozone (O3) as the limiting reagent, four carbonyl compounds were detected: 7-hydroxy-6-oxo-3-(prop-1-en-2-yl)heptanal; 3-Isopropenyl-6-oxoheptanal (IPOH), 3-acetyl-6-oxoheptanal (3A6O) and one carbonyl of unknown structure. Using cyclohexane as a hydroxyl (OHrad) radical scavenger, the relative yields (peak area) of the unknown carbonyl, IPOH, and 3A6O were reduced indicating the influence secondary OH radicals have on limonene ozonolysis products. The relative yield of the hydroxy-dicarbonyl based on the chromatogram was unchanged suggesting it is only made by the limonene + O3 reaction. The detection of 3A6O using TBOX highlights the advantages of a smaller molecular weight derivatization agent for the detection of multi-carbonyl compounds. The use of TBOX derivatization if combined with other derivatization agents may address a recurring need to simply and accurately detect multi-functional oxygenated species in air.

Rapid analysis of aflatoxins B1, B2, and ochratoxin A in rice samples using dispersive liquid-liquid microextraction combined with HPLC.

PubMed

Lai, Xian-Wen; Sun, Dai-Li; Ruan, Chun-Qiang; Zhang, He; Liu, Cheng-Lan

2014-01-01

A novel, simple, and rapid method is presented for the analysis of aflatoxin B1, aflatoxin B2, and ochratoxin A in rice samples by dispersive liquid-liquid microextraction combined with LC and fluorescence detection. After extraction of the rice samples with a mixture of acetonitrile/water/acetic acid, mycotoxins were rapidly partitioned into a small volume of organic solvent (chloroform) by dispersive liquid-liquid microextraction. The three mycotoxins were simultaneously determined by LC with fluorescence detection after precolumn derivatization for aflatoxin B1 and B2. Parameters affecting both extraction and dispersive liquid-liquid microextraction procedures, including the extraction solvent, the type and volume of extractant, the volume of dispersive solvent, the addition of salt, the pH and the extraction time, were optimized. The optimized protocol provided an enrichment factor of approximately 1.25 and with detection of limits (0.06-0.5 μg/kg) below the maximum levels imposed by current regulations for aflatoxins and ochratoxin A. The mean recovery of three mycotoxins ranged from 82.9-112%, with a RSD less than 7.9% in all cases. The method was successfully applied to measure mycotoxins in commercial rice samples collected from local supermarkets in China. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Surface derivatization strategy for combinatorial analysis of cell response to mixtures of protein domains.

PubMed

Chiang, Chunyi; Karuri, Stella W; Kshatriya, Pradnya P; Schwartz, Jeffrey; Schwarzbauer, Jean E; Karuri, Nancy W

2012-01-10

We report a robust strategy for conjugating mixtures of two or more protein domains to nonfouling polyurethane surfaces. In our strategy, the carbamate groups of polyurethane are reacted with zirconium alkoxide from the vapor phase to give a surface-bound oxide that serves as a chemical layer that can be used to bond organics to the polymer substrate. A hydroxyalkylphosphonate monolayer was synthesized on this layer, which was then used to covalently bind primary amine groups in protein domains using chloroformate-derived cross-linking. The effectiveness of this synthesis strategy was gauged by using an ELISA to measure competitive, covalent bonding of cell-binding (III(9-10)) and fibronectin-binding (III(1-2)) domains of the cell adhesion protein fibronectin. Cell adhesion, spreading, and fibronectin matrix assembly were examined on surfaces conjugated with single domains, a 1:1 surface mixture of III(1-2) and III(9-10), and a recombinant protein "duplex" containing both domains in one fusion protein. The mixture performed as well as or better than the other surfaces in these assays. Our surface activation strategy is amenable to a wide range of polymer substrates and free amino group-containing protein fragments. As such, this technique may be used to create biologically specific materials through the immobilization of specific protein groups or mixtures thereof on a substrate surface.

Determination of Carvedilol Enantiomers in Pharmaceutical Dosages by SBSE-HPLC Based on Diastereomer Formation.

PubMed

Taraji, Maryam; Talebpour, Zahra; Adib, Nuoshin; Karimi, Shima; Haghighi, Farideh; Aboul-Enein, Hassan Y

2015-09-01

A sensitive, selective and simple method for the simultaneous determination of carvedilol enantiomers in aqueous solution has been developed using stir bar sorptive extraction (SBSE) followed by high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection. This method is based on the reaction of carvedilol enantiomers with (-)-menthyl chloroformate (MCF) after extraction by the SBSE method to produce diastereomeric derivatives. The separation was achieved by use of a C18 analytical column and the influence of mobile phase composition on the enantioseparation of carvedilol was studied. The applicability of two sorptive phases, poly(methyl methacrylate/ethyleneglycol dimethacrylate) (PA-EG) and polydimethylsiloxane, were tested for extraction of carvedilol enantiomers from aqueous samples. The obtained results showed excellent linear dynamic ranges and precisions for each of them. The least limit of detection for (S)- and (R)-carvedilol obtained 8 and 11 µg L(-1), respectively, using the PA-EG sorptive phase. Inter- and intra-mean recoveries were also satisfactory, ranging from 98 to 103%, with coefficient of variation in the range of 1-5% at three fortified levels using a PA-EG coated stir bar. The proposed SBSE (PA-EG)-MCF derivatization-HPLC-UV method was successfully applied to enantioselective analysis of carvedilol in water and pharmaceutical dosages, confirming the application of this method. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Synthesis and photochemical properties of a novel iron-sulfur-nitrosyl cluster derivatized with the pendant chromophore protoporphyrin IX.

PubMed

Conrado, Christa L; Wecksler, Stephen; Egler, Christian; Magde, Douglas; Ford, Peter C

2004-09-06

The novel Roussin red-salt ester (PPIX-RSE) with a pendant porphyrin chromophore was prepared and investigated as a precursor for the photochemical generation of nitric oxide. PPIX-RSE has the general formula Fe(2)(NO)(4)[(mu-S,mu-S')P] (where (S,S')P is the bis(2-thiolatoethyl) diester of protoporphyrin IX. The photoexcitation of PPIX-RSE with 436- or 546-nm light in an aerated chloroform solution led to the photodecomposition of the cluster with the respective quantum yields (5.2 +/- 0.7) x 10(-4) and (2.5 +/- 0.5 x 10(-4)) and the concomitant release of NO. PPIX-RSE is a significantly more effective NO generator at longer wavelength excitation than are other Fe(2)(mu-SR)(2)(NO)(4) esters for which R is a simple alkyl group such as CH(3)CH(2)- because of the much higher absorptivity of the pendant PPIX chromophore at these wavelengths and a modestly higher quantum yield. Fluorescence intensity and lifetime data indicate that the photoexcited porphyrin of PPIX-RSE is largely quenched by the energy transfer to the Fe(2)S(2)(NO)(4) cluster's core. However, a small fraction of this emission is not quenched, and it is proposed that PPIX-RSE may exist in solution as two conformers.

Amino acids implicated in plant defense are higher in Candidatus Liberibacter asiaticus-tolerant citrus varieties

PubMed Central

Killiny, Nabil; Hijaz, Faraj

2016-01-01

ABSTRACT Citrus Huanglongbing (HLB), also known as citrus greening, has been threatening the citrus industry since the early 1900's and up to this date there are no effective cures for this disease. Field observations and greenhouse controlled studies demonstrated that some citrus genotypes are more tolerant to Candidatus Liberibacter asiaticus (CLas) pathogen than others. However, the mechanisms underpinning tolerance has not been determined yet. The phloem sap composition of CLas-tolerant and sensitive citrus varieties was studied to identify metabolites that could be responsible for their tolerance to CLas. The citrus phloem sap was collected by centrifugation and was analyzed with gas chromatography-mass spectrometry after methyl chloroformate derivatization. Thirty-three metabolites were detected in the phloem sap of the studied varieties: twenty 20 amino acids, eight 8 organic acids, and five 5 fatty acids. Interestingly, the levels of most amino acids, especially those implicated in plantdefense to pathogens such as phenylalanine, tyrosine, tryptophan, lysine, and asparagine were higher in tolerant varieties. Although the level of organic acids varied between cultivars, this variation was not correlated with citrus resistance to CLas and could be cultivar specific. The fatty acids were found in trace amounts and in most cases their levels were not significantly different among varieties. Better understanding of the mechanisms underpinning citrus tolerance to CLas will help in developing economically tolerant varieties. PMID:27057814

Amino acids implicated in plant defense are higher in Candidatus Liberibacter asiaticus-tolerant citrus varieties.

PubMed

Killiny, Nabil; Hijaz, Faraj

2016-01-01

Citrus Huanglongbing (HLB), also known as citrus greening, has been threatening the citrus industry since the early 1900's and up to this date there are no effective cures for this disease. Field observations and greenhouse controlled studies demonstrated that some citrus genotypes are more tolerant to Candidatus Liberibacter asiaticus (CLas) pathogen than others. However, the mechanisms underpinning tolerance has not been determined yet. The phloem sap composition of CLas-tolerant and sensitive citrus varieties was studied to identify metabolites that could be responsible for their tolerance to CLas. The citrus phloem sap was collected by centrifugation and was analyzed with gas chromatography-mass spectrometry after methyl chloroformate derivatization. Thirty-three metabolites were detected in the phloem sap of the studied varieties: twenty 20 amino acids, eight 8 organic acids, and five 5 fatty acids. Interestingly, the levels of most amino acids, especially those implicated in plantdefense to pathogens such as phenylalanine, tyrosine, tryptophan, lysine, and asparagine were higher in tolerant varieties. Although the level of organic acids varied between cultivars, this variation was not correlated with citrus resistance to CLas and could be cultivar specific. The fatty acids were found in trace amounts and in most cases their levels were not significantly different among varieties. Better understanding of the mechanisms underpinning citrus tolerance to CLas will help in developing economically tolerant varieties.

Method to simultaneously determine the sphingosine 1-phosphate breakdown product (2E)-hexadecenal and its fatty acid derivatives using isotope-dilution HPLC-electrospray ionization-quadrupole/time-of-flight mass spectrometry.

PubMed

Neuber, Corinna; Schumacher, Fabian; Gulbins, Erich; Kleuser, Burkhard

2014-09-16

Sphingosine 1-phosphate (S1P), a bioactive lipid involved in various physiological processes, can be irreversibly degraded by the membrane-bound S1P lyase (S1PL) yielding (2E)-hexadecenal and phosphoethanolamine. It is discussed that (2E)-hexadecenal is further oxidized to (2E)-hexadecenoic acid by the long-chain fatty aldehyde dehydrogenase ALDH3A2 (also known as FALDH) prior to activation via coupling to coenzyme A (CoA). Inhibition or defects in these enzymes, S1PL or FALDH, result in severe immunological disorders or the Sjögren-Larsson syndrome, respectively. Hence, it is of enormous importance to simultaneously determine the S1P breakdown product (2E)-hexadecenal and its fatty acid metabolites in biological samples. However, no method is available so far. Here, we present a sensitive and selective isotope-dilution high performance liquid chromatography-electrospray ionization-quadrupole/time-of-flight mass spectrometry method for simultaneous quantification of (2E)-hexadecenal and its fatty acid metabolites following derivatization with 2-diphenylacetyl-1,3-indandione-1-hydrazone and 1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide. Optimized conditions for sample derivatization, chromatographic separation, and MS/MS detection are presented as well as an extensive method validation. Finally, our method was successfully applied to biological samples. We found that (2E)-hexadecenal is almost quantitatively oxidized to (2E)-hexadecenoic acid, that is further activated as verified by cotreatment of HepG2 cell lysates with (2E)-hexadecenal and the acyl-CoA synthetase inhibitor triacsin C. Moreover, incubations of cell lysates with deuterated (2E)-hexadecenal revealed that no hexadecanoic acid is formed from the aldehyde. Thus, our method provides new insights into the sphingolipid metabolism and will be useful to investigate diseases known for abnormalities in long-chain fatty acid metabolism, e.g., the Sjögren-Larsson syndrome, in more detail.

Validated Spectrophotometric and RP-HPLC-DAD Methods for the Determination of Ursodeoxycholic Acid Based on Derivatization with 2-Nitrophenylhydrazine.

PubMed

El-Kafrawy, Dina S; Belal, Tarek S; Mahrous, Mohamed S; Abdel-Khalek, Magdi M; Abo-Gharam, Amira H

2017-05-01

This work describes the development, validation, and application of two simple, accurate, and reliable methods for the determination of ursodeoxycholic acid (UDCA) in bulk powder and in pharmaceutical dosage forms. The carboxylic acid group in UDCA was exploited for the development of these novel methods. Method 1 is the colorimetric determination of the drug based on its reaction with 2-nitrophenylhydrazine hydrochloride in the presence of a water-soluble carbodiimide coupler [1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride] and pyridine to produce an acid hydrazide derivative, which ionizes to yield an intense violet color with maximum absorption at 553 nm. Method 2 uses reversed-phase HPLC with diode-array detection for the determination of UDCA after precolumn derivatization using the same reaction mentioned above. The acid hydrazide reaction product was separated using a Pinnacle DB C8 column (4.6 × 150 mm, 5 μm particle size) and a mobile phase consisting of 0.01 M acetate buffer (pH 3)-methanol-acetonitrile (30 + 30 + 40, v/v/v) isocratically pumped at a flow rate of 1 mL/min. Ibuprofen was used as the internal standard (IS). The peaks of the reaction product and IS were monitored at 400 nm. Different experimental parameters for both methods were carefully optimized. Analytical performance of the developed methods were statistically validated for linearity, range, precision, accuracy, specificity, robustness, LOD, and LOQ. Calibration curves showed good linear relationships for concentration ranges 32-192 and 60-600 μg/mL for methods 1 and 2, respectively. The proposed methods were successfully applied for the assay of UDCA in bulk form, capsules, and oral suspension with good accuracy and precision. Assay results were statistically compared with a reference pharmacopeial HPLC method, and no significant differences were observed between proposed and reference methods.

A Low-Cost Liquid-Chromatography System Using a Spectronic 20-Based Detector.

ERIC Educational Resources Information Center

Jezorek, John R.; And Others

1986-01-01

Describes the design and evaluation of a Spectronic 20-based detector as well as a simple system for postcolumn derivatization useful for metal-ion chromatographic detection. Both detection and derivatization can be performed in the ultra-violet (UV) mode using a low-cost UV-visible spectrophotometer and UV-region derivatization reagents. (JN)

Matrix effect in matrix-assisted laser desorption/ionization mass spectra of derivatized oligomeric polyols.

PubMed

Borisov, Roman S; Polovkov, Nikolai Yu; Zhilyaev, Dmitry I; Zaikin, Vladimir G

2013-01-30

Herein we describe a strong matrix effect observed in the matrix-assisted laser desorption/ionization time-of-flight (MALDI-ToF) mass spectra of silylated glycerol alkoxylates and manifested in the loss of the silyl groups in the presence of carboxyl-containing matrices. Commercially available glycerol alkoxylates containing three end OH groups as well as three matrices - 2,5-dihydroxybenzoic acid (DHB), 3-indoleacrylic acid (IAA) and 1,8,9-anthracenetriol (dithranol) - were chosen for the investigation. N,O-Bis(trimethylsilyl)trifluoroacetamide containing 1% trimethylchlorosilane, acetic anhydride and a formylation mixture (formic acid/acetyl chloride) were used for derivatization. Initial oligomers and derivatized products were analyzed by MALDI-ToF-mass spectrometry (MS) on an Autoflex II instrument, equipped with a nitrogen laser (λ 337 nm), in positive ion reflectron mode. Only [M + Na](+) ions were observed for underivatized polymers and for completely derivatized polymers in the presence of DHB and dithranol, respectively. In the case of IAA the mass spectra revealed sets of peaks for underivatized, and for partially and completely derivatized oligomers. No similar 'matrix effect' was observed in the case of acylated glycerol alkoxylates (acyl = formyl, acetyl): only peaks for completely derivatized oligomers were obtained in all matrices: DHB, IAA and dithranol. Using 1,9-nonandiol, we showed that the 'matrix effect' was due to trans-silylation of carboxyl-containing matrices (DHB and IAA) during co-crystallization of silylated oligomers and matrices. The obtained results show that matrix molecules can participate as reactive species in MALDI-ToF-MS experiments. The matrix should be carefully chosen when a derivatization approach is applied because the analysis of spectra of the completely derivatized products is particularly desirable in the quantitative determination of functional end-groups. Copyright © 2012 John Wiley & Sons, Ltd.

Analysis of Organic Molecules Extracted from Mars Analogues and Influence of Their Mineralogy Using N-Methyl-N-(tert-butyldimethylsilyl)Trifluoroacetamide Derivatization Coupled with Gas Chromatography Mass Spectrometry in Preparation for the Sample Analysis at Mars Derivatization Experiment on the Mars Science Laboratory Mission

NASA Technical Reports Server (NTRS)

Stalport, F.; Glavin, D. P.; Eigenbrode, J. L.; Bish, D.; Blake, D.; Coll, P.; Szopa, C.; Buch, A.; McAdam, A.; Dworkin, J. P.;

Injection port derivatization following ion-pair hollow fiber-protected liquid-phase microextraction for determining acidic herbicides by gas chromatography/mass spectrometry.

PubMed

Wu, Jingming; Lee, Hian Kee

2006-10-15

Injection port derivatization following ion-pair hollow fiber-protected liquid-phase microextraction (LPME) for the trace determination of acidic herbicides (2,4-dichlorobenzoic acid, 2,4-dichlorophenoxyacetic acid, 2-(2,4-dichlorophenoxy)propionic acid, 3,5-dichlorobenzoic acid, 2-(2,4,5-trichlorophenoxy)propionic acid) in aqueous samples by gas chromatography/mass spectrometry (GC/MS) was developed. Prior to GC injection port derivatization, acidic herbicides were converted into their ion-pair complexes with tetrabutylammonium chloride in aqueous samples and then extracted by 1-octanol impregnated in the hollow fiber. Upon injection, ion pairs of acidic herbicides were quantitatively derivatized to their butyl esters in the GC injection port. Thus, several parameters related to the derivatization process (i.e., injection temperature, purge-off time) were evaluated, and main parameters affecting the hollow fiber-protected LPME procedure such as extraction organic solvent, ion-pair reagent type, pH of aqueous medium, concentration of ion-pair reagent, sodium chloride concentration added to the aqueous medium, stirring speed, and extraction time profile, optimized. At the selected extraction and derivatization conditions, no matrix effects were observed. This method proved good repeatability (RSDs <12.3%, n = 6) and good linearity (r2 > or = 0.9939) for spiked deionized water samples for five analytes. The limits of detection were in the range of 0.51-13.7 ng x L(-1) (S/N =3) under GC/MS selected ion monitoring mode. The results demonstrated that injection port derivatization following ion-pair hollow fiber-protected LPME was a simple, rapid, and accurate method for the determination of trace acidic herbicides from aqueous samples. In addition, this method proved to be environmentally friendly since it completely avoided open derivatization with potentially hazardous reagents.

USE OF BIOLOGICALLY BASED COMPUTATIONAL MODELING IN MODE OF ACTION-BASED RISK ASSESSMENT – AN EXAMPLE OF CHLOROFORM

EPA Science Inventory

The objective of current work is to develop a new cancer dose-response assessment for chloroform using a physiologically based pharmacokinetic/pharmacodynamic (PBPK/PD) model. The PBPK/PD model is based on a mode of action in which the cytolethality of chloroform occurs when the ...

Selective chloroform sensor using thiol functionalized reduced graphene oxide at room temperature

NASA Astrophysics Data System (ADS)

Midya, Anupam; Mukherjee, Subhrajit; Roy, Shreyasee; Santra, Sumita; Manna, Nilotpal; Ray, Samit K.

2018-02-01

This paper presents a highly selective chloroform sensor using functionalised reduced graphene oxide (RGO) as a sensing layer. Thiol group is covalently attached on the basal plan of RGO film by a simple one-step aryl diazonium chemistry to improve its selectivity. Several spectroscopic techniques like X-ray photoelectron, Raman and Fourier transform infrared spectroscopy confirm successful thiol functionalization of RGO. Finally, the fabricated chemiresistor type sensor is exposed to chloroform in the concentration range 200-800 ppm (parts per million). The sensor shows a 4.3% of response towards 800 ppm chloroform. The selectivity of the sensor is analyzed using various volatile organic compounds as well. The devices show enhanced response and faster recovery attributed to the physiosorption of chloroform onto thiol functionalized graphene making them attractive for 2D materials based sensing applications.

Antioxidant, anticholinesterase and antibacterial activities of Stachys guyoniana and Mentha aquatica.

PubMed

Ferhat, Maria; Erol, Ebru; Beladjila, Khadidja Aya; Çetintaş, Yunus; Duru, Mehmet Emin; Öztürk, Mehmet; Kabouche, Ahmed; Kabouche, Zahia

2017-12-01

Stachys guyoniana Noë ex. Batt. and Mentha aquatica L. are two Algerian Lamiaceae used in folk medicine. To investigate their antioxidant, anticholinesterase and antibacterial activities. n-Butanol (BESG), ethyl acetate (EESG) and chloroform (CESG) extracts of S. guyoniana and methanol (MEMA) and chloroform (CEMA) aerial part extracts of M. aquatica and methanol (MERMA) and acetone (AERMA) roots extracts of M. aquatica were evaluated for their antioxidant activity by the β-carotene-linoleic acid, DPPH • and ABTS • +  scavenging, CUPRAC and metal chelating assays. The anticholinesterase activity was tested against AChE and BChE. The antibacterial activity was assessed by MICs determination against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella heidelberg, Klebsiella pneumoniae, Enterobacter aerogenes and Morganella morganii strains. In the β-carotene test, the CESG (IC 50 : 2.3 ± 1.27 μg/mL) exhibited the highest activity. The BESG was the best scavenger of DPPH • (IC 50 : 2.91 ± 0.14 μg/mL). In the ABTS test, AERMA was the most active (IC 50 : 4.21 ± 0.28 μg/mL). However, with the CUPRAC, the BESG exhibited the best activity (A 0.50 : 0.15 ± 0.05 μg/mL) and was active in metal chelating assay with 48% inhibition at 100 μg/mL. The BESG was the best AChE inhibitor (IC 50 : 5.78 ± 0.01 μg/mL) however, the AERMA showed the highest BChE inhibitory activity (IC 50 : 19.23 ± 1.42 μg/mL). The tested extracts exhibited a good antibacterial activity. This study demonstrated good antioxidant, anticholinesterase and antibacterial potential of S. guyoniana and M. aquatica, which fits in well with their use in folk medicine.

Inter-species comparative antioxidant assay and HPTLC analysis of sakuranetin in the chloroform and ethanol extracts of aerial parts of Rhus retinorrhoea and Rhus tripartita.

PubMed

Alam, Perwez; Parvez, Mohammad Khalid; Arbab, Ahmed Hassan; Siddiqui, Nasir Ali; Al-Dosary, Mohammed Salem; Al-Rehaily, Adnan Jathlan; Ahmed, Sarfaraz; Kalam, Mohd Abul; Ahmad, Mohammad Shamim

2017-12-01

Extensive research on Rhus (Anacardiaceae) shows their antioxidant potential, which warrants further evaluation of its other species. To perform a comparative antioxidant assay on extracts of R. retinorrhoea and R. tripartita, including sakuranetin quantification by a validated HPTLC method. In vitro antioxidant assay was performed on chloroform and ethanol extracts of R. retinorrhoea Steud. ex Oliv. (RRCE and RREE) and R. tripartita (Ucria) Grande (RTCE and RTEE) by DPPH radical scavenging (at 31.25, 62.5, 125, 250 and 500 μg/mL concentrations) and β-carotene-linoleic acid bleaching methods at 500 μg/mL concentration. Densitometric HPTLC method was developed and validated using toluene: ethyl acetate: methanol (8:2:0.2; v/v/v) as mobile phase, executed on glass-backed silica gel F 254 plate and scanned at 292 nm. Antioxidant activity of Rhus extracts tested by the two methods (DPPH/BCB) was found in order of RTEE > RREE > RTCE > RRCE with IC 50 118.67/256.26, 315.75/82.35, 827.92/380.0 and 443.69/292.75, respectively. Scanning of the HPTLC plate provided an intense peak of sakuranetin at R f = 0.59. The estimated sakuranetin content in the dry weight of the extracts was highest in RREE (27.95 μg/mg) followed by RRCE (25.22 μg/mg), RTEE (0.487 μg/mg) and RTCE (0.0 μg/mg). Presence of sakuranetin in RREE, RRCE and RTEE supported the highest antioxidant property of the two Rhus species. Nonetheless, low sakuratenin in R. tripartita indicated the presence of other bioactive constituents responsible for synergistic antioxidant activity. The developed HPTLC method therefore guarantees its application in quality control of commercialized herbal drugs and formulations containing sakuranetin.

Bioactive plants from Argentina and Bolivia.

PubMed

Bardón, Alicia; Borkosky, Susana; Ybarra, María I; Montanaro, Susana; Cartagena, Elena

2007-04-01

Antibacterial and molluscicidal activities of methanol and chloroform extracts of 16 plant species belonging to the families Compositae and Melastomataceae were evaluated. The chloroform extract of Vernonanthura tweediana and the methanol extract of Senecio santelisis resulted to be very toxic to brine shrimp nauplii (LC(50)=1 microg/ml). Chloroform extracts of S. santelisis and Senecio leucostachys as well as the methanol extract of Wedelia subvaginata displayed molluscicidal effects on Biomphalaria peregrina showing LC(100)<100 microg/ml. Moderate antibacterial action was produced by the chloroform extracts of Flaveria bidentis, Grindelia scorzonerifolia and Vernonia incana against two strains of Staphylococcus aureus.

Self-Diffusion and Heteroassociation in an Acetone-Chloroform Mixture at 298 K

NASA Astrophysics Data System (ADS)

Golubev, V. A.; Gurina, D. L.; Kumeev, R. S.

2018-01-01

The self-diffusion coefficients of acetone and chloroform in a binary acetone-chloroform mixture at 298 K are determined via pulsed field gradient NMR spectroscopy. It is estimated that the hydrodynamic radii of the mixture's components, calculated using the Stokes-Einstein equation, grow as the concentrations of the components fall. It is shown that such behavior of hydrodynamic radii is due to acetone-chloroform heteroassociation. The hydrodynamic radii of monomers and heteroassociates in a 1: 1 ratio are determined along with the constant of heteroassociation, using the proposed model of an associated solution.

Modified Method for Detection of Benzoylecgonine in Human Urine by GC-MS: Derivatization Using Pentafluoropropanol/Acetic Anhydride.

PubMed

Serafin, Michelle C; Paulemon, Kasandra M; Fuller, Zachary J; Bronner, William E

2017-05-01

An existing GC-MS method for detecting benzoylecgonine (BZE) in urine was modified by changing derivatizing reagents. This method modification presents a cost-effective alternative derivatization procedure for the detection of BZE in urine by GC-MS. The combination of pentafluoropropanol and acetic anhydride was found to produce the same reaction product for BZE as pentafluoropropanol with pentafluoropropionic anhydride, while reducing reagent cost. With no anhydride present, derivatization of BZE by pentafluoropropanol did not occur. Published by Oxford University Press 2017. This work is written by (a) US Government employee(s) and is in the public domain in the US.

USE OF SENSITIVITY ANALYSIS ON A PHYSIOLOGICALLY-BASED PHARMACOKINETIC (PBPK) MODEL FOR CHLOROFORM IN RATS TO DETERMINE AGE-RELATED TOXICITY

EPA Science Inventory

USE OF SENSITIVITY ANALYSIS ON A PHYSIOLOGICALLY BASED PHARMACOKINETIC (PBPK) MODEL FOR CHLOROFORM IN RATS TO DETERMINE AGE-RELATED TOXICITY.
CR Eklund, MV Evans, and JE Simmons. US EPA, ORD, NHEERL, ETD,PKB, Research Triangle Park, NC.

Chloroform (CHCl3) is a disinfec...

Immunotoxicological Profile of Chloroform in Female B6c3f1 Mice When Administered In Drinking Water

EPA Science Inventory

Chloroform can be formed as a disinfection by-product during water chlorination, one of the primary modalities for purifying municipal water supplies for human consumption. The goal of this study was to characterize the immunotoxic effects of chloroform in female B6C3F1 mice when...

Preserving ground water samples with hydrochloric acid does not result in the formation of chloroform

USGS Publications Warehouse

Squillace, Paul J.; Pankow, James F.; Barbash, Jack E.; Price, Curtis V.; Zogorski, John S.

1999-01-01

Water samples collected for the determination of volatile organic compounds (VOCs) are often preserved with hydrochloric acid (HCl) to inhibit the biotransformation of the analytes of interest until the chemical analyses can he performed. However, it is theoretically possible that residual free chlorine in the HCl can react with dissolved organic carbon (DOC) to form chloroform via the haloform reaction. Analyses of 1501 ground water samples preserved with HCl from the U.S. Geological Survey's National Water-Quality Assessment Program indicate that chloroform was the most commonly detected VOC among 60 VOCs monitored. The DOC concentrations were not significantly larger in samples with detectable chloroform than in those with no delectable chloroform, nor was there any correlation between the concentrations of chloroform and DOC. Furthermore, chloroform was detected more frequently in shallow ground water in urban areas (28.5% of the wells sampled) than in agricultural areas (1.6% of the wells sampled), which indicates that its detection was more related to urban land-use activities than to sample acidification. These data provide strong evidence that acidification with HCl does not lead to the production of significant amounts of chloroform in ground water samples. To verify these results, an acidification study was designed to measure the concentrations of all trihalomethanes (THMs) that can form as a result of HCl preservation in ground water samples and to determine if ascorbic acid (C6H8O6) could inhibit this reaction if it did occur. This study showed that no THMs were formed as a result of HCl acidification, and that ascorbic acid had no discernible effect on the concentrations of THMs measured.

Theodore Roosevelt Chloroforming Uncle Sam "In the Hopeless Ward".

PubMed

Drew, Benjamin A; Bause, George S

2016-10-01

In March of 1905 in Judge magazine, Louis Dalrymple published his political cartoon of Theodore Roosevelt chloroforming "Uncle Sam." Having sampled a host of Democratic remedies, the 125-year-old Sam can expect that Roosevelt's chloroform will either cure him with major Republican surgery or kill him with Osler-linked euthanasia. Copyright © 2016 Elsevier Inc. All rights reserved.

THE FAILURE OF CHLOROFORM ADMINISTERED IN THE DRINKING WATER TO INDUCE RENAL TUBULAR CELL NEOPLASIA IN MALE F344/N RATS

EPA Science Inventory

The failure of chloroform administered in drinking water to induce renal tubular cell neoplasia in male F344/N rats

Chloroform (TCM) has been demonstrated to be a renal carcinogen in the male Osborne-
Mendel rat when administered either by corn oil gavage or in drin...

Kinetic Studies that Evaluate the Solvolytic Mechanisms of Allyl and Vinyl Chloroformate Esters

PubMed Central

D’Souza, Malcolm J.; Givens, Aaron F.; Lorchak, Peter A.; Greenwood, Abigail E.; Gottschall, Stacey L.; Carter, Shannon E.; Kevill, Dennis N.

2013-01-01

At 25.0 °C the specific rates of solvolysis for allyl and vinyl chloroformates have been determined in a wide mix of pure and aqueous organic mixtures. In all the solvents studied, vinyl chloroformate was found to react significantly faster than allyl chloroformate. Multiple correlation analyses of these rates are completed using the extended (two-term) Grunwald-Winstein equation with incorporation of literature values for solvent nucleophilicity (NT) and solvent ionizing power (YCl). Both substrates were found to solvolyze by similar dual bimolecular carbonyl-addition and unimolecular ionization channels, each heavily dependent upon the solvents nucleophilicity and ionizing ability. PMID:23549265

Simultaneous Solid Phase Extraction and Derivatization of Aliphatic Primary Amines Prior to Separation and UV-Absorbance Detection

PubMed Central

Felhofer, Jessica L.; Scida, Karen; Penick, Mark; Willis, Peter A.; Garcia, Carlos D.

2013-01-01

To overcome the problem of poor sensitivity of capillary electrophoresis-UV absorbance for the detection of aliphatic amines, a solid phase extraction and derivatization scheme was developed. This work demonstrates successful coupling of amines to a chromophore immobilized on a solid phase and subsequent cleavage and analysis. Although the analysis of many types of amines is relevant for myriad applications, this paper focuses on the derivatization and separation of amines with environmental relevance. This work aims to provide the foundations for future developments of an integrated sample preparation microreactor capable of performing simultaneous derivatization, preconcentration, and sample cleanup for sensitive analysis of primary amines. PMID:24054648

Applications of derivatization reactions to trace organic compounds during sample preparation based on pressurized liquid extraction.

PubMed

Carro, Antonia M; González, Paula; Lorenzo, Rosa A

2013-06-28

Pressurized liquid extraction (PLE) is an exhaustive technique used for the extraction of analytes from solid samples. Temperature, pressure, solvent type and volume, and the addition of other reagents notably influence the efficiency of the extraction. The analytical applications of this technique can be improved by coupling with appropriate derivatization reactions. The aim of this review is to discuss the recent applications of the sequential combination of PLE with derivatization and the approaches that involve simultaneous extraction and in situ derivatization. The potential of the latest developments to the trace analysis of environmental, food and biological samples is also analyzed. Copyright © 2013 Elsevier B.V. All rights reserved.

Selective determination of arginine-containing and tyrosine-containing peptides using capillary electrophoresis and laser-induced fluorescence detection.

PubMed

Cobb, K A; Novotny, M V

1992-01-01

The use of two different amino acid-selective fluorogenic reagents for the derivatization of peptides is investigated. One such scheme utilizes a selective reaction of benzoin with the guanidine moiety to derivatize arginine residues occurring in a peptide. The second scheme involves the formylation of tyrosine, followed by reaction with 4-methoxy-1,2-phenylenediamine. The use of capillary electrophoresis and laser-induced fluorescence detection allows enhanced efficiencies and sensitivities to be obtained for the separations of either arginine- or tyrosine-containing peptides. A helium-cadmium laser (325 nm) is ideally suited for the laser-based detection system due to a close match of the excitation maxima of derivatized peptides from both reactions. A detection limit of 270 amol is achieved for model arginine-containing peptides, while the detection limit for model tyrosine-containing peptides is measured at 390 amol. Both derivatization reactions are found to be useful for high-sensitivity peptide mapping applications in which only the peptides containing the derivatized amino acids are detected.

15N and13C NMR investigation of hydroxylamine-derivatized humic substances

USGS Publications Warehouse

Thorn, K.A.; Arterburn, J.B.; Mikita, M.A.

1992-01-01

Five fulvic and humic acid samples of diverse origins were derivatized with 15N-labeled hydroxylamine and analyzed by liquid-phase 15N NMR spectrometry. The 15N NMR spectra indicated that hydroxylamine reacted similarly with all samples and could discriminate among carbonyl functional groups. Oximes were the major derivatives; resonances attributable to hydroxamic acids, the reaction products of hydroxylamine with esters, and resonances attributable to the tautomeric equilibrium position between the nitrosophenol and monoxime derivatives of quinones, the first direct spectroscopic evidence for quinones, also were evident. The 15N NMR spectra also suggested the presence of nitriles, oxazoles, oxazolines, isocyanides, amides, and lactams, which may all be explained in terms of Beckmann reactions of the initial oxime derivatives. INEPT and ACOUSTIC 15N NMR spectra provided complementary information on the derivatized samples. 13C NMR spectra of derivatized samples indicated that the ketone/quinone functionality is incompletely derivatized with hydroxylamine. ?? 1991 American Chemical Society.

Separation-oriented derivatization of native fluorescent compounds through fluorous labeling followed by liquid chromatography with fluorous-phase.

PubMed

Sakaguchi, Yohei; Yoshida, Hideyuki; Todoroki, Kenichiro; Nohta, Hitoshi; Yamaguchi, Masatoshi

2009-06-15

We have developed a new and simple method based on "fluorous derivatization" for LC of native fluorescent compounds. This method involves the use of a column with a fluorous stationary phase. Native fluorescent analytes with target functional groups are precolumn derivatized with a nonfluorescent fluorous tag, and the fluorous-labeled analytes are retained in the column, whereas underivatized substances are not. Only the retained fluorescent analytes are detected fluorometrically at appropriate retention times, and retained substrates without fluorophores are not detected. In this study, biologically important carboxylic acids (homovanillic acid, vanillylmandelic acid, and 5-hydroxyindoleacetic acid) and drugs (naproxen, felbinac, flurbiprofen, and etodolac) were used as model native fluorescent compounds. Experimental results indicate that the fluorous-phase column can selectively retain fluorous compounds including fluorous-labeled analytes on the basis of fluorous separation. We believe that separation-oriented derivatization presented here is the first step toward the introduction of fluorous derivatization in quantitative LC analysis.

Quantitative extraction and concentration of synthetic water-soluble acid dyes from aqueous media using a quinine-chloroform solution

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kobayashi, F.; Ozawa, N.; Hanai, J.

Twenty-one water-soluble acid dyes, including eleven azo, five triphenylmethane four xanthene, one naphthol derivatives, used at practical concentrations for food coloration, were quantitatively extracted from water and various carbonated beverages into a 0.1 M quinine-chloroform solution in the presence of 0.5 M boric acid by brief shaking. Quantitative extraction of these dyes was also accomplished by the 0.1 M quinine-chloroform solution made conveniently from chloroform, quinine hydrochloride, and sodium hydroxide added successively to water or beverages containing boric acid. Quinine acted as a countercation on the dyes having sulfonic and/or carboxylic acid group(s) to form chloroform-soluble ion-pair complexes. The diacidicmore » base alkaloid interacted with each acid group of mono-, di-, tri-, and tetrasulfonic acid dyes approximately in the ratio 0.8-0.9 to 1. The dyes in the chloroform solution were quantitatively concentrated into a small volume of sodium hydroxide solution also by brief shaking. The convenient quinine-chloroform method was applicable to the quantitative extraction of a mixture of 12 dyes from carbonated beverages, which are all currently used for food coloration. A high-pressure liquid chromatographic method is also presented for the systematic separation and determination of these 12 dyes following their concentration into the aqueous alkaline solution. The chromatogram was monitored by double-wavelength absorptiometry in the visible and ultraviolet ray regions.« less

Ionic liquid-based microwave-assisted dispersive liquid-liquid microextraction and derivatization of sulfonamides in river water, honey, milk, and animal plasma.

PubMed

Xu, Xu; Su, Rui; Zhao, Xin; Liu, Zhuang; Zhang, Yupu; Li, Dan; Li, Xueyuan; Zhang, Hanqi; Wang, Ziming

2011-11-30

The ionic liquid-based microwave-assisted dispersive liquid-liquid microextraction (IL-based MADLLME) and derivatization was applied for the pretreatment of six sulfonamides (SAs) prior to the determination by high-performance liquid chromatography (HPLC). By adding methanol (disperser), fluorescamine solution (derivatization reagent) and ionic liquid (extraction solvent) into sample, extraction, derivatization, and preconcentration were continuously performed. Several experimental parameters, such as the type and volume of extraction solvent, the type and volume of disperser, amount of derivatization reagent, microwave power, microwave irradiation time, pH of sample solution, and ionic strength were investigated and optimized. When the microwave power was 240 W, the analytes could be derivatized and extracted simultaneously within 90 s. The proposed method was applied to the analysis of river water, honey, milk, and pig plasma samples, and the recoveries of analytes obtained were in the range of 95.0-110.8, 95.4-106.3, 95.0-108.3, and 95.7-107.7, respectively. The relative standard deviations varied between 1.5% and 7.3% (n=5). The results showed that the proposed method was a rapid, convenient and feasible method for the determination of SAs in liquid samples. Copyright © 2011 Elsevier B.V. All rights reserved.

Resonance energy transfer between the active sites of creatine kinase from rabbit brain.

PubMed

Grossman, S H

1990-09-03

Resonance energy transfer was measured between the active site domains of the brain isozyme of creatine kinase (CK-BB). The reactive thiol near the active sites, one on each subunit of the dimeric protein, was derivatized using 5-[2-[iodoacetyl)amino)ethyl]aminonaphthalene-1-sulfonic acid (AED), 2-[4'-iodoacetamidoanilino]naphthalene-6-sulfonic acid (AANS) and 5-iodoacetamidofluorescein (AF). Suitable donor/acceptor protein conjugated hybrids were prepared by controlled kinetics producing CK-BB-AED/AF and CK-BB-AANS/AF. Transfer efficiencies, measured from the quenching of the donor lifetime and steady-state sensitized acceptor emission, ranged from 0.10 to 0.17. From determination of the donor/acceptor overlap integrals, donor quantum yields and attempts to delimit the orientation factor using steady-state and phase-resolved anisotropy measurements, it was found that a suitable estimate of the range between the active sites was between 45 and 57 A. This range is similar to that reported previously for the muscle isozyme of creatine kinase (Grossman, S.H. (1989) Biochemistry 28, 4894-4902) but is a significantly greater distance than detected for the hybrid, myocardial specific isozyme (Grossman, S.H. (1983) Biochemistry 22, 5369-5375).

Structures of asparagine-linked oligosaccharides from hen egg-yolk antibody (IgY). Occurrence of unusual glucosylated oligo-mannose type oligosaccharides in a mature glycoprotein.

PubMed

Ohta, M; Hamako, J; Yamamoto, S; Hatta, H; Kim, M; Yamamoto, T; Oka, S; Mizuochi, T; Matsuura, F

1991-10-01

Asparagine-linked oligosaccharides present on hen egg-yolk immunoglobulin, termed IgY, were liberated from the protein by hydrazinolysis. After N-acetylation, the oligosaccharides were labelled with a UV-absorbing compound, p-aminobenzoic acid ethyl ester (ABEE). The ABEE-derivatized oligosaccharides were fractionated by anion exchange, normal phase and reversed phase HPLC, and their structures were determined by a combination of sugar composition analysis, methylation analysis, negative ion FAB-MS, 500 MHz 1H-NMR and sequential exoglycosidase digestions. IgY contained monoglucosylated oligomannose type oligosaccharides with structures of Glc alpha 1-3Man7-9-GlcNAc-GlcNAc, oligomannose type oligosaccharides with the size range of Man5-9GlcNAc-GlcNAc, and biantennary complex type oligosaccharides with core region structure of Man alpha 1-6(+/- GlcNAc beta 1-4)(Man alpha 1-3)Man beta 1-4GlcNAc beta 1-4(+/- Fuc alpha 1-6)GlcNAc. The glucosylated oligosaccharides, Glc1Man8GlcNAc2 and Glc1Man7GlcNAc2, have not previously been reported in mature glycoproteins from any source.

HPTLC-FLD-SERS as a facile and reliable screening tool: Exemplarily shown with tyramine in cheese.

PubMed

Wang, Liao; Xu, Xue-Ming; Chen, Yi-Sheng; Ren, Jie; Liu, Yun-Tao

2018-04-01

The serious cytotoxicity of tyramine attracted marked attention as it induced necrosis of human intestinal cells. This paper presented a novel and facile high performance thin-layer chromatography (HPTLC) method tailored for screening tyramine in cheese. Separation was performed on glass backed silica gel plates, using methanol/ethyl acetate/ammonia (6/4/1 v/v/v) as the mobile phase. Special efforts were focused on optimizing conditions (substrate preparation, laser wavelength, salt types and concentrations) of surface enhanced Raman spectroscopy (SERS) measurements directly on plates after derivatization, which enabled molecule-specific identification of targeted bands. In parallel, fluorescent densitometry (FLD) scanning at 380

The Influence of Mineralogy on Recovering Organic Acids from Mars Analogue Materials Using the One-Pot Derivatization Experiment on the Sample Analysis at Mars(SAM) Instrument Suite

NASA Technical Reports Server (NTRS)

Stalport, Fabien; Glavin, Daniel P.; Eigenbrode, J. L.; Bish, D.; Blake, D.; Coll, P.; Szopa, C.; Buch, A.; McAdam, A.; Dworkin, J. P.;

Microwave-assisted on-spot derivatization for gas chromatography-mass spectrometry based determination of polar low molecular weight compounds in dried blood spots.

PubMed

Sadones, Nele; Van Bever, Elien; Archer, John R H; Wood, David M; Dargan, Paul I; Van Bortel, Luc; Lambert, Willy E; Stove, Christophe P

2016-09-23

Dried blood spot (DBS) sampling and analysis is increasingly being applied in bioanalysis. Although the use of DBS has many advantages, it is also associated with some challenges. E.g. given the limited amount of available material, highly sensitive detection techniques are often required to attain sufficient sensitivity. In gas chromatography coupled to mass spectrometry (GC-MS), derivatization can be helpful to achieve adequate sensitivity. Because this additional sample preparation step is considered as time-consuming, we introduce a new derivatization procedure, i.e. "microwave-assisted on-spot derivatization", to minimize sample preparation of DBS. In this approach the derivatization reagents are directly applied onto the DBS and derivatization takes place in a microwave instead of via conventional heating. In this manuscript we evaluated the applicability of this new concept of derivatization for the determination of two polar low molecular weight molecules, gamma-hydroxybutyric acid (GHB) and gabapentin, in DBS using a standard GC-MS configuration. The method was successfully validated for both compounds, with imprecision and bias values within acceptance criteria (<20% at LLOQ, <15% at 3 other QC levels). Calibration lines were linear over the 10-100μg/mL and 1-30μg/mL range for GHB and gabapentin, respectively. Stability studies revealed no significant decrease of gabapentin and GHB in DBS upon storage at room temperature for at least 84 days. Furthermore, DBS-specific parameters, including hematocrit and volume spotted, were evaluated. As demonstrated by the analysis of GHB and gabapentin positive samples, "microwave-assisted on-spot derivatization" proved to be reliable, fast and applicable in routine toxicology. Moreover, other polar low molecular weight compounds of interest in clinical and/or forensic toxicology, including vigabatrin, beta-hydroxybutyric acid, propylene glycol, diethylene glycol, 1,4-butanediol and 1,2-butanediol, can also be detected using this method. Copyright © 2016 Elsevier B.V. All rights reserved.

Determination of NH2 concentration on 3-aminopropyl tri-ethoxy silane layers and cyclopropylamine plasma polymers by liquid-phase derivatization with 5-iodo 2-furaldehyde

NASA Astrophysics Data System (ADS)

Manakhov, Anton; Čechal, Jan; Michlíček, Miroslav; Shtansky, Dmitry V.

2017-08-01

The quantification of concentration of primary amines, e.g. in plasma polymerized layers is a very important task for surface analysis. However, the commonly used procedure, such as gas phase derivatization with benzaldehydes, shows several drawbacks, the most important of which are the side reaction effects. In the present study we propose and validate a liquid phase derivatization using 5-iodo 2-furaldehyde (IFA). It was demonstrated that the content of NH2 groups can be determined from the atomic concentrations measured by X-ray photoelectron spectroscopy (XPS), in particular from the ratio of I 3d and N 1s peak intensities. First, we demonstrate the method on a prototypical system such as 3-aminopropyl tri-ethoxy silane (APTES) layer. Here the XPS analysis carried out after reaction of APTES layer with IFA gives the fraction of primary amines (NH2/N) of 38.3 ± 7.9%. Comparing this value with that obtained by N 1s curve fitting of APTES layer giving 40.9 ± 9.5% of amine groups, it can be concluded that all primary amines were derivatized by reaction with IFA. The second system to demonstrate the method comprises cyclopropylamine (CPA) plasma polymers that were free from conjugated imines. In this case the method gives the NH2 fraction ∼8.5%. This value is closely matching the NH2/N ratio estimated by 4-trifluoromethyl benzaldehyde (TFBA) derivatization. The reaction of IFA with CPA plasma polymer exhibiting high density of conjugated imines revealed the NH2/N fraction of ∼10.8%. This value was significantly lower compared to 17.3% estimated by TFBA derivatization. As the overestimated density of primary amines measured by TFBA derivatization is probably related to the side reaction of benzaldehydes with conjugated imines, the proposed IFA derivatization of primary amines can be an alternative procedure for the quantification of surface amine groups.

A high selective cataluminescence sensor for the determination of tetrahydrofuran vapor

NASA Astrophysics Data System (ADS)

Cao, Xiaoan; Dai, Huimei; Chen, Suilin; Zeng, Jiayi; Zhang, Keke; Sun, Yan

2013-02-01

A novel tetrahydrofuran (THF) vapor sensor was designed based on the cataluminescence (CTL) of THF on nanosized γ-Al2O3/MgO (mol ratio = 1.5:1). SEM and XRD were applied for its characterization. We found that the CTL was strongly produced when THF vapor flowed through a nanosized Al-Mg mixed-metal oxide surface, while the CTL was weakly generated when THF vapor flowed through a single nanosized γ-Al2O3 or MgO surface. Quantitative analysis was performed at an optimal temperature of 279 °C, a wavelength of 460 nm and a flow rate of 360 mL min-1. The linear range of the CTL intensity versus concentrations of THF vapor was 1.0-3000 mL m-3 with a detection limit of 0.67 mL m-3. No (or only very low) interference was observed by formaldehyde, methanol, ethanol, benzene, toluene, ethyl acetate, ammonia, cyclohexane, chloroform, glycol armour ether, glycol ether, isopropyl ether and n-butyl ether or acetic acid. Since the response of the sensor was rapid and the system was easy to handle, we believe that the sensor has great potential for real-world use.

Antioxidant lipoxygenase inhibitors from the leaf extracts of Simmondsia chinensis.

PubMed

Abdel-Mageed, Wael Mostafa; Bayoumi, Soad Abdel Latief Hassan; Salama, Awwad Abdoh Radwan; Salem-Bekhit, Mounir Mohamed; Abd-Alrahman, Sherif Hussein; Sayed, Hanaa Mohamed

2014-09-01

To isolate and identify chemical constituents with antioxidant and lipoxygenase inhibitory effects of the ethanolic extract of Simmondsia chinensis (Jojoba) leaves. The alcoholic extract was subjected to successive solvent fractionation. The antioxidant active fractions (chloroform, ethyl acetate and aqueous fractions) were subjected to a combination of different chromatographic techniques guided by the antioxidant assay with DPPH. The structures of the isolated compounds were elucidated on the basis of spectroscopic evidences and correlated with known compounds. The antioxidant activity was assessed quantitively using DPPH and β-carotene methods. The inhibitory potential against enzyme lipoxygenase was assessed on soybean lipoxygenase enzyme. Ten flavonoids and four lignans were isolated. Flavonoid aglycones showed stronger antioxidant and lipoxygenase inhibitory effects than their glycosides. Lignoid glycosides showed moderate to weak antioxidant and lipoxygenase inhibitory effects. A total of 14 compounds were isolated and identified from Simmondsia chinensis; 12 of them were isolated for the first time. This is the first report that highlights deeply on the phenolic content of jojoba and their potential biological activities and shows the importance of this plant as a good source of phenolics in particular the flavonoid content. Copyright © 2014 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Qualitative and quantitative analysis of four species of Curcuma rhizomes using twice development thin layer chromatography.

PubMed

Zhang, J S; Guan, J; Yang, F Q; Liu, H G; Cheng, X J; Li, S P

2008-11-04

The rhizomes of Curcuma phaeocaulis, Curcuma kwangsiensis, Curcuma wenyujin and Curcuma longa are used as Ezhu or Jianghuang in traditional Chinese medicine for a long time. Due to their similar morphological characters, it is difficult to distinguish their origins of raw materials used in clinic. In this study, a simple, rapid and reliable twice development TLC method was developed for qualitative and quantitative analysis of the four species of Curcuma rhizomes. The chromatography was performed on silica gel 60F(254) plate with chloroform-methanol-formic acid (80:4:0.8, v/v/v) and petroleum ether-ethyl acetate (90:10, v/v) as mobile phase for twice development. The TLC markers were colorized with 1% vanillin-H(2)SO(4) solution. The four species of Curcuma were easily discriminated based on their characteristic TLC profiles, and simultaneous quantification of eight compounds, including bisdemethoxycurcumin, demethoxycurcumin, curcumine, curcumenol, curcumol, curdione, furanodienone and curzerene, in Curcuma were also performed densitometrically at lambda(scan)=518nm and lambda(reference)=800 nm. The investigated compounds had good linearity (r(2)>0.9905) within test ranges. Therefore, the developed TLC method can be used for quality control of Curcuma rhizomes.

Water fraction of edible medicinal fern Stenochlaena palustris is a potent α-glucosidase inhibitor with concurrent antioxidant activity.

PubMed

Chai, Tsun-Thai; Kwek, Meng-Tee; Ong, Hean-Chooi; Wong, Fai-Chu

2015-11-01

This study aimed to isolate a potent antiglucosidase and antioxidant fraction from Stenochlaena palustris. Extraction was performed with hexane, chloroform, ethyl acetate, methanol, and water. Antiglucosidase, 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging and ferric reducing antioxidant power (FRAP) assays found methanol extract (ME) to be the most active. Water fraction (WF) of ME was a stronger α-glucosidase inhibitor (EC50 2.9 μg/mL) than quercetin, with weak antiamylase activity. WF was a competitive α-glucosidase inhibitor. DPPH scavenging activity of WF (EC50 7.7 μg/mL) was weaker than quercetin. WF (EC50 364 μg/mL) was a stronger hydrogen peroxide scavenger than gallic acid (EC50 838 μg/mL) and was equally strong as quercetin in scavenging superoxide. WF possessed moderate copper chelating activity. WF was enriched in total phenolics (TP) and hydroxycinnamic acids (THC). TP correlated with antioxidant activity (R(2) > 0.76). Only THC correlated with antiglucosidase activity (R(2) = 0.86). Overall, WF demonstrated concurrent, potent antiglucosidase and antioxidant activities. Copyright © 2015 Elsevier Ltd. All rights reserved.

Antidiabetic potential of Caesalpinia sumatrana, a medicinal herbs traditionally used by local tribe in East Kalimantan

NASA Astrophysics Data System (ADS)

Wicaksono, D. A.; Rosamah, E.; Kusuma, I. W.

2018-04-01

The aims of the research was to analyze the content of phytochemicals, to examine the antioxidant and antidiabeticpotentials of n-hexane, chloroform, ethyl acetate, and ethanol extracts of Caesalpinia sumatrana. Method to measure antioxidant capacity of sample involves the use of the free radical, 1,1-diphenyl-2-picrylhydrazyl (DPPH) which is widely used to test the ability of compounds to act as free radical. Analysis the potential of antidiabeticactivity of the extracts was determined by α-glucosidase and α-amylase inhibitory assay. Of all extracts obtained by successive maceration, ethanol maceration gave the highest extract by 2.63% of extract on the dry weigh basis. The result of phytochemicals showed that all extracts contain alkaloid and flavonoid. The highest antioxidant activity was 82.32% with IC50 value of 5.00 µg/ml obtained by ethanol extract. The results of enzyme inhibitory assay of α-glucosidase showed that ethanol extract of C. sumatrana had IC50 value 17.16 µg/mL to inhibit ɑ-glucosidase activity and IC50 value 16.78 µg/mL for ɑ-amylase. The present result displayed potential of the plant to be developed as natural antidiabetic and antioxidant agents.

Determination of six pesticides in the medicinal herb Cordia salicifolia by matrix solid-phase dispersion and gas chromatography/mass spectrometry.

PubMed

de Carvalho, Pedro Henrique Viana; Prata, Vanessa de Menezes; Alves, Péricles Barreto; Navickiene, Sandro

2009-01-01

A simple and effective extraction method based on matrix solid-phase dispersion was developed for acephate, chlorpropham, pyrimicarb, bifenthrin, tetradifon, and phosalone in leaves of the medicinal plant Cordia salicifolia, whose extracts are commercialized in Brazil as diuretic, appetite suppressant, and weight loss products. The determination method was GC/MS with selected-ion monitoring. Different parameters of the method were evaluated, such as type of solid phase (C18, alumina, silica gel, and Florisil) and the amount of solid phase and eluent (dichloromethane, ethyl acetate, chloroform, and cyclohexane). The best results were obtained using 0.5 g herb sample, 0.5 g neutral alumina as the dispersant sorbent, 0.5 g C18 as the cleanup sorbent, and cyclohexane-dichloromethane (3 + 1, v/v) as the eluting solvent. The method was validated using herb samples fortified with pesticides at different concentration levels (0.3, 0.5, and 1.0 mg/kg). Average recoveries (seven replicates) ranged from 67.7 to 129.9%, with relative standard deviations between 6.3 and 26%. Detection and quantitation limits for the herb ranged from 0.10 to 0.15 and 0.15 to 0.25 mg/kg, respectively.

Effects of deuteration on the metabolism of halogenated anesthetics in the rat

DOE Office of Scientific and Technical Information (OSTI.GOV)

McCarty, L.P.; Malek, R.S.; Larsen, E.R.

1979-08-01

The authors studied the effects of substituting deuterium for hydrogen in several volatile anesthetics on their metabolism in the Fischer rat. Substitution of deuterium in the ethyl portion of methoxyflurane increased the metabolic production of fluoride ion by 19 percent when administered at a concentration of 0.05%. Total replacement of hydrogen by deuterium resulted in a 29% decrease in the amount of fluoride produced, while deuteration of only the methoxyl group produced a 33% decrease in fluoride produced. Deuteration of halothane resulted in a 15 or 26% decrease in serum bromide at 0.75% or 1.0%, respectively. Deuteration in the ethylmore » portions of enflurane and two experimental agents, CF2HOCF2CFBrH and CF2HOCF2CCl2H resulted in 65, 76, and 29% decreases in urinary fluoride, respectively. Anesthesia with deuterated chloroform at a concentration of 0.36% produced a 35% decrease in serum glutamic pyruvic transaminase (SGPT). It is concluded that deuteration of volatile anesthetics changes their metabolism, in most cases producing decreases in metabolism. This effect may lessen the organ toxicity believed to occur with some of these anesthetics.« less

Urease inhibitory profile of extracts and chemical constituents of Pistacia atlantica ssp. cabulica Stocks.

PubMed

Uddin, Ghias; Ismail; Rauf, Abdur; Raza, Muslim; Khan, Haroon; Nasruddin; Khan, Majid; Farooq, Umar; Khan, Ajmal; Arifullah

2016-06-01

The current study was designed to evaluate the urease inhibitory profile of extract and fractions of Pistacia atlantica ssp. cabulica Stocks followed by bioactivity-guided isolated compounds. The crude extract was found significantly active with urease inhibitor (95.40% at 0.2 mg/mL) with IC50 values of 32.0 ± 0.28 μg/mL. Upon fractionation, ethyl acetate fraction displayed 100% urease inhibition with IC50 values of 19.9 ± 0.51 μg/mL at 0.2 mg/mL. However, n-hexane and chloroform fractions exhibited insignificant urease inhibition. Similarly, the isolated compound, transilitin (1) and dihydro luteolin (2) demonstrated marked urease attenuation with 95 and 98% respectively, at 0.15 mg/mL. Both the isolated compounds showed marked potency with IC50 values of 8.54 ± 0.54 and 9.58 ± 2.22 μg/mL, respectively. In short, both the extract and fractions and isolated compounds showed marked urease inhibition and thus a useful natural source of urease inhibition.

Effect of binder liquid type on spherical crystallization.

PubMed

Maghsoodi, Maryam; Hajipour, Ali

2014-11-01

Spherical crystallization is a process of formation of agglomerates of crystals held together by binder liquid. This research focused on understanding the effect of type of solvents used as binder liquid on the agglomeration of crystals. Carbamazepine and ethanol/water were used respectively as a model drug and crystallization system. Eight solvents as binder liquid including chloroform, dichloromethane, isopropyl acetate, ethyl acetate, n-hexane, dimethyl aniline, benzene and toluene were examined to better understand the relationship between the physical properties of the binder liquid and its ability to bring about the formation of the agglomerates. Moreover, the agglomerates obtained from effective solvents as binder liquid were evaluated in term of size, apparent particle density and compressive strength. In this study the clear trend was observed experimentally in the agglomerate formation as a function of physical properties of the binder liquid such as miscibility with crystallization system. Furthermore, the properties of obtained agglomerates such as size, apparent particle density and compressive strength were directly related to physical properties of effective binder liquids. RESULTS of this study offer a useful starting point for a conceptual framework to guide the selection of solvent systems for spherical crystallization.

Repellent properties of Cardiospermum halicacabum Linn. (Family: Sapindaceae) plant leaf extracts against three important vector mosquitoes

PubMed Central

Govindarajan, M; Sivakumar, R

2012-01-01

Objective To determine repellent activity of hexane, ethyl acetate, benzene, chloroform and methanol extract of Cardiospermum halicacabum (C. halicacabum) against Culex quinquefasciatus (Cx. quinquefasciatus), Aedes aegypti (Ae. aegypti) and Anopheles stephensi (An. stephensi). Methods Evaluation was carried out in a net cage (45 cm×30 cm×25 cm) containing 100 blood starved female mosquitoes of three mosquito species and were assayed in the laboratory condition by using the protocol of WHO 2005; The plant leaf crude extracts of C. halicacabum was applied at 1.0, 2.5, and 5.0 mg/cm2 separately in the exposed area of the fore arm. Only ethanol served as control. Results In this observation, the plant crude extracts gave protection against mosquito bites without any allergic reaction to the test person, and also, the repellent activity was dependent on the strength of the plant extracts. The tested plant crude extracts had exerted promising repellent against all the three mosquitoes. Conclusions From the results it can be concluded the crude extract of C. halicacabum was potential for controlling Cx. quinquefasciatus, Ae. aegypti and An. stephensi mosquitoes. PMID:23569979

NITRIC OXIDE INHIBITORY ACTIVITY OF STRYCHNOS SPINOSA (LOGANIACEAE) LEAF EXTRACTS AND FRACTIONS

PubMed Central

AI, Isa; JP, Dzoyem; SA, Adebayo; MM, Suleiman; JN, Eloff

2016-01-01

Background: The study was aimed at determining the anti-inflammatory activity of fractions and extracts obtained from Strychnos spinosa leaves on a mediator of inflammation nitric oxide (NO). Materials and Methods: Leaves were extracted with acetone and separated into fractions with different polarities by solvent-solvent fractionation. The Griess assay was used to determine the nitric oxide (NO) inhibitory activity. Cellular toxicity was determined by “using the MTT reduction assay”. Results: With the exception of the ethyl acetate fraction which had an IC50 >750 μg/mL, all extracts and fractions had significant nitric oxide-inhibitory activity. The most active being the water fraction, chloroform fraction and the dichloromethane/methanol extracts with IC50 values of 88.43 μg/mL, 96.72 μg/mL and 115.62 μg/mL, respectively. The extracts and fractions had low cytotoxicity on macrophage U937 cell lines. Conclusion: Extracts and fractions of Strychnos spinosa leaves may be promising sources of natural anti-inflammatory agents. Findings obtained from this study showed that Strychnos spinosa leaves possess promising anti-inflammatory action and could be used in the treatment of inflammation-related conditions. PMID:28480356

NITRIC OXIDE INHIBITORY ACTIVITY OF STRYCHNOS SPINOSA (LOGANIACEAE) LEAF EXTRACTS AND FRACTIONS.

PubMed

Ai, Isa; Jp, Dzoyem; Sa, Adebayo; Mm, Suleiman; Jn, Eloff

2016-01-01

The study was aimed at determining the anti-inflammatory activity of fractions and extracts obtained from Strychnos spinosa leaves on a mediator of inflammation nitric oxide (NO). Leaves were extracted with acetone and separated into fractions with different polarities by solvent-solvent fractionation. The Griess assay was used to determine the nitric oxide (NO) inhibitory activity. Cellular toxicity was determined by "using the MTT reduction assay". With the exception of the ethyl acetate fraction which had an IC 50 >750 μg/mL, all extracts and fractions had significant nitric oxide-inhibitory activity. The most active being the water fraction, chloroform fraction and the dichloromethane/methanol extracts with IC 50 values of 88.43 μg/mL, 96.72 μg/mL and 115.62 μg/mL, respectively. The extracts and fractions had low cytotoxicity on macrophage U937 cell lines. Extracts and fractions of Strychnos spinosa leaves may be promising sources of natural anti-inflammatory agents. Findings obtained from this study showed that Strychnos spinosa leaves possess promising anti-inflammatory action and could be used in the treatment of inflammation-related conditions.

Determination of flavonoids from Orthosiphon stamineus in plasma using a simple HPLC method with ultraviolet detection.

PubMed

Loon, Yit Hong; Wong, Jia Woei; Yap, Siew Ping; Yuen, Kah Hay

2005-02-25

A simple liquid chromatographic method was developed for the simultaneous determination of flavonoids from Orthosiphon stamineus Benth, namely sinensitin, eupatorin and 3'-hydroxy-5,6,7,4'-tetramethoxyflavone, in plasma. Prior to analysis, the flavonoids and the internal standard (naproxen) were extracted from plasma samples using a 1:1 mixture of ethyl acetate and chloroform. The detection and quantification limits for the three flavonoids were similar being 3 and 5 ng/ml, respectively. The within-day and between-day accuracy values, expressed as percentage of true values, for the three flavonoids were between 95 and 107%, while the corresponding precision, expressed as coefficients of variation, for the three flavonoids were less than 14%. In addition, the mean recovery values of the extraction procedure for all the flavonoids were between 92 and 114%. The calibration curves were linear over a concentration range of 5-4000 ng/ml. The present method was applied to analyse plasma samples obtained from a pilot study using rats in which the mean absolute oral bioavailability values for sinensitin, eupatorin and 3'-hydroxy-5,6,7,4'-tetramethoxyflavone was 9.4, 1.0 and 1.5%, respectively.

Moricandia arvensis extracts protect against DNA damage, mutagenesis in bacteria system and scavenge the superoxide anion.

PubMed

Skandrani, Ines; Bouhlel, Ines; Limem, Ilef; Boubaker, Jihed; Bhouri, Wissem; Neffati, Aicha; Ben Sghaier, Mohamed; Kilani, Soumaya; Ghedira, Kamel; Ghedira-Chekir, Leila

2009-02-01

The mutagenic potential of total aqueous, total oligomers flavonoids (TOF), ethyl acetate (EA), chloroform (Chl), petroleum ether (PE) and methanol (MeOH) extracts from aerial parts of Moricandia arvensis was assessed using Ames Salmonella tester strains TA100 and TA1535 with and without metabolic activation (S9), and using plasmid pBluescript DNA assay. None of the different extracts produced a mutagenic effect, except aqueous extract when incubated with Salmonella typhimurium TA100 after metabolic activation. Likewise, the antimutagenicity of the same extracts was tested using the "Ames test". Our results showed that M. arvensis extracts possess antimutagenic effects against sodium azide (SA) in the two tested Salmonella assay systems, except metabolized aqueous and PE extracts when tested with S. typhimurium TA100 assay system. Different extracts were also found to be effective in protecting plasmid DNA against the strand breakage induced by hydroxyl radicals, except PE and aqueous extracts. Antioxidant capacity of the tested extracts was evaluated using the enzymatic (xanthine/xanthine oxidase assay) (X/XOD) and the non enzymatic (NBT/Riboflavine assay) systems. TOF extract was the more effective one in inhibiting both xanthine oxidase activity and NBT reduction.

Seeds of Peganum Harmala L. chemical analysis, antimalarial and antioxidant activities, and cytotoxicity against human breast cancer cells.

PubMed

Chabir, Naziha; Ibrahim, Hany; Romdhane, Hany; Valentin, Alexis; Moukarzel, Beatrice; Mars, Mohamed; Bouajila, Jalloul

2014-01-01

The present study evaluated the levels of total phenolics, flavonoids, tannins and anthocyanins from Peganum harmala L. seeds and determined their antioxidant, antiplasmodial and anticancer potentials. Antioxidant activity was determined by DPPH and ABTS assays. Extracts of P. harmala seeds from Oudref and Djerba (two places in Tunisia) were obtained by successive extraction solvents: petroleum ether, chloroform, ethyl acetate, ethanol and water. Their composition was evaluated for phenolics (gallic acid equivalent 2.48 to 72.52 g/kg), tannins (catechin equivalent 0 to 25.27 g/kg), anthocyanins (cyanidin equivalent 0 to 20.56 mg/kg) and flavonoids (quercetin equivalent 0 to 3.12 g/kg). Ethanolic extract exerted the highest activities against a chloroquine-resistant strain of Plasmodium falciparum (IC₅₀=23 mg/L), against human breast cancer cells MCF7 (IC₅₀=32 mg/L) and against free radical (IC₅₀=19.09±3.07 mg/L). Correlations were studied between each chemical family and the three activities. Total phenolics content exhibited the highest correlation with antiplasmodial activity (R²=0.92) and with anticancer activity (R²=0.86), respectively.

Anti-inflammatory and analgesic activities of Tunisian Citrullus colocynthis Schrad. immature fruit and seed organic extracts.

PubMed

Marzouk, B; Marzouk, Z; Fenina, N; Bouraoui, A; Aouni, M

2011-06-01

Inflammations and immune-related diseases including rheumatoid arthritis are widespread in the entire globe. The treatment of these illnesses is mainly based on the use of synthetic and biotechnological drugs, in recent years. Tunisian traditional medicine is a potential source of new remedies namely Citrullus colocynthis Schrad. (Cucurbitaceae): endemic in southern Tunisia and used in folk medicine to treat many inflammation disorders. Our goal was to assess the in vivo analgesic and anti-inflammatory activities of Tunisian Citrullus colocynthis immature fruit and seed organic extracts (petroleum ether, chloroform, ethyl acetate, acetone and finely methanol extract). Yields of prepared organic extracts are gravimetrically determined. For the analgesic and anti-inflammatory activities, we have used respectively, the acetic acid writhing test in mice and the carrageenan-induced paw edema assay in rats. All extracts displayed an important analgesic and anti-inflammatory activities at different doses without inducing any side effects. This study has demonstrated the analgesic and anti-inflammatory activities of Citrullus colocynthis immature fruit and seed extracts. Experiment results provide scientific insight into the ancient practice of utilizing Citrullus colocynthis Schrad. as analgesic and as anti-inflammatory agents.

High Performance Liquid Chromatography-mass Spectrometry Analysis of High Antioxidant Australian Fruits with Antiproliferative Activity Against Cancer Cells.

PubMed

Sirdaarta, Joseph; Maen, Anton; Rayan, Paran; Matthews, Ben; Cock, Ian Edwin

2016-05-01

High antioxidant capacities have been linked to the treatment and prevention of several cancers. Recent reports have identified several native Australian fruits with high antioxidant capacities. Despite this, several of these species are yet to be tested for anticancer activity. Solvent extracts prepared from high antioxidant native Australian fruits were analyzed for antioxidant capacity by the di (phenyl)-(2,4,6-trinitrophenyl) iminoazanium free radical scavenging assay. Antiproliferative activities against CaCo2 and HeLa cancer cells were determined by a multicellular tumor spheroid-based cell proliferation assay. Toxicity was determined by Artemia franciscana bioassay. Methanolic extracts of all plant species displayed high antioxidant contents (equivalent to approximately 7-16 mg of vitamin C per gram of fruit extracted). Most aqueous extracts also contained relatively high antioxidant capacities. In contrast, the ethyl acetate, chloroform, and hexane extracts of most species (except lemon aspen and bush tomato) had lower antioxidant contents (below 1.5 mg of vitamin C equivalents per gram of plant material extracted). The antioxidant contents correlated with the ability of the extracts to inhibit proliferation of CaCo2 and HeLa cancer cell lines. The high antioxidant methanolic extracts of all species were potent inhibitors of cell proliferation. The methanolic lemon aspen extract was particularly effective, with IC50 values of 480 and 769 μg/mL against HeLa and CaCo2 cells, respectively. In contrast, the lower antioxidant ethyl acetate and hexane extracts (except the lemon aspen ethyl acetate extract) generally did not inhibit cancer cell proliferation or inhibited to only a minor degree. Indeed, most of the ethyl acetate and hexane extracts induced potent cell proliferation. The native tamarind ethyl acetate extract displayed low-moderate toxicity in the A. franciscana bioassay (LC50 values below 1000 μg/mL). All other extracts were nontoxic. A total of 145 unique mass signals were detected in the lemon aspen methanolic and aqueous extracts by nonbiased high-performance liquid chromatography-mass spectrometry analysis. Of these, 20 compounds were identified as being of particular interest due to their reported antioxidant and/or anticancer activities. The lack of toxicity and antiproliferative activity of the high antioxidant plant extracts against HeLa and CaCo2 cancer cell lines indicates their potential in the treatment and prevention of some cancers. Australian fruit extracts with high antioxidant contents were potent inhibitors of CaCo2 and HeLa carcinoma cell proliferationMethanolic lemon aspen extract was particularly potent, with IC50 values of 480 μg/mL (HeLa) and 769 μg/mL (CaCo2)High-performance liquid chromatography-mass spectrometry-quadrupole time-of-flight analysis highlighted and putatively identified 20 compounds in the antiproliferative lemon aspen extractsIn contrast, lower antioxidant content extracts stimulated carcinoma cell proliferationAll extracts with antiproliferative activity were nontoxic in the Artemia nauplii assay. Abbreviations used: DPPH: di (phenyl)- (2,4,6-trinitrophenyl) iminoazanium, HPLC: High-performance liquid chromatography, IC50: The concentration required to inhibit by 50%, LC50: The concentration required to achieve 50% mortality, MS: Mass spectrometry.

Competition between roots and microorganisms for phosphorus: A novel 33P labeling approach

NASA Astrophysics Data System (ADS)

Zilla, Thomas; Kuzyakov, Yakov; Zavišiæ, Aljoša; Polle, Andrea

2015-04-01

While organic N mineralization exhibits clear seasonal uptake dynamics, knowledge about seasonal variation in microbial P uptake and mineralization is scarce. We hypothesize that the dynamics of P uptake and mineralization by microorganisms in temperate forest soils exhibit a seasonality anti-cyclic to plant P uptake. Therefore, the ratio of microbial P to labile P increases by the transition from acquiring ecosystems (in spring) to recycling ones (in fall). To investigate this, intact soil-plant mesocosms containing Ah horizon with 1 year old F. sylvatica were removed from the P-rich field site Bad Brueckenau and the P-depleted field site Luess in Germany. During incubation under controlled conditions, seasonal pulse labeling by 33P-orthophosphate was performed at 5 time points over the course of one year. 33P recovery in microbial compounds of organic and mineral soil horizons was determined 7 and 30 days after the labeling. This procedure will account for temporal changes in P allocation and also considers the rather slow P transport from the mycorrhiza into the plants and other microorganisms. For the first time we analyzed the 33P incorporation into total PLFA and consequently provide a new technique for the analysis of P uptake by microorganisms, which has clear advantages compared to P quantification after chloroform fumigation. Polar lipids are hereby extracted with a Frostegård-modified Bligh-and-Dyer buffer, i.e. a single phase mixture of chloroform, methanol and citrate buffer (0.8:1:2, v:v:v). Phospholipids (PLFA) are isolated and purified by solid phase extraction via a silica gel column chromatography. Subsequently, PLFA are hydrolyzed and the resulting fatty acids derivatized by methylation. The fatty acid methyl esters were extracted with n-hexane and measured by GC/MS to investigate the composition of the microbial community. The remaining extract, containing head groups, phosphate units and glycerol backbones, was used to determine 33P activity and recovery in the microbial membrane lipids with a multi-purpose scintillation counter. This approach offers the unique possibility to quantify P fluxes through the microbial network. For the first time, P cycling can be linked to changes in microbial community structure and activity in soils in situ.

Drawing the battle lines: tracing the "Science War" in the construction of the chloroform and human health risks debate.

PubMed

Driedger, S Michelle; Eyles, John

2003-04-01

The United States Environmental Protection Agency (US EPA) and the Chlorine Chemistry Council, the Chemical Manufacturers Association, and others have been embroiled in a legal challenge concerning the US EPA's "reversal" regarding the scientific assessment of chloroform's carcinogenicity. This issue arose during the US EPA's November 1998 promulgation of a Maximum Contaminant Level Goal for chloroform in the Stage 1 Final Rules for Disinfectants and Disinfection Byproducts in drinking water. In this paper we adopt a claimsmaking approach: to trace the development and outcome of the chloroform court challenge in the USA, to examine the construction of scientific knowledge claims concerning chloroform risk assessments, and to investigate how different interpretations of scientific uncertainties regarding the evidence are contested when such uncertainties are brought into a regulatory and judicial arena. This "science war" (Chlorine Chemistry Council and others v. US EPA and others) took place in the US Court of Appeals for the District of Columbia Circuit. The scientific "authority" in the construction of scientific claims in this dispute is based on the International Life Sciences Institute expert panel report on chloroform. Examining these science wars is important because they signal critical shifts in science policy agendas. The regulatory outcome of the chloroform science war in the United States can have profound implications for the construction and acceptance of scientific claims regarding drinking water in other jurisdictions (e.g., Canada). In this challenge, we argue that the actors involved in the dispute constructed "boundaries" around accepted and credible scientific claims.

Methyl Chloroform Elimination from the Production of Space Shuttle Sold Rocket Motors

NASA Technical Reports Server (NTRS)

Golde, Rick P.; Burt, Rick; Key, Leigh

1997-01-01

Thiokol Space Operations manufactures the Reusable Solid Rocket Motors used to launch America's fleet of Space Shuttles. In 1989, Thiokol used more than 1.4 Mlb of methyl chloroform to produce rocket motors. The ban placed by the Environmental Protection Agency on the sale of methyl chloroform had a significant effect on future Reusable Solid Rocket Motor production. As a result, changes in the materials and processes became necessary. A multiphased plan was established by Thiokol in partnership with NASA's Marshall Space Flight Center to eliminate the use of methyl chloroform in the Reusable Solid Rocket Motor production process. Because of the extensive scope of this effort, the plan was phased to target the elimination of the majority of methyl chloroform use (90 percent) by January 1, 1996, the 3 Environmental Protection Agency deadline. Referred to as Phase I, this effort includes the elimination of two large vapor degreasers, grease diluent processes, and propellant tooling handcleaning using methyl chloroform. Meanwhile, a request was made for an essential use exemption to allow the continued use of the remaining 10 percent of methyl chloroform after the 1996 deadline, while total elimination was pursued for this final, critical phase (Phase II). This paper provides an update to three previous presentations prepared for the 1993, 1994, and 1995 CFC/Halon Alternative Conferences, and will outline the overall Ozone Depleting Compounds Elimination Program from the initial phases through the final testing and implementation phases, including facility and equipment development. Processes and materials to be discussed include low-pressure aqueous wash systems, high-pressure water blast systems- environmental shipping containers, aqueous and semi-aqueous cleaning solutions, and bond integrity and inspection criteria. Progress toward completion of facility implementation and lessons learned during the scope of the program, as well as the current development efforts and basic requirements of future methyl chloroform handcleaning elimination, will also be outlined.

The role of renal proximal tubule P450 enzymes in chloroform-induced nephrotoxicity: Utility of renal specific P450 reductase knockout mouse models

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, Senyan; Wadsworth Center, New York State Department of Health, and School of Public Health, State University of New York, Albany, NY 12201; Yao, Yunyi

The kidney is a primary target for numerous toxic compounds. Cytochrome P450 enzymes (P450) are responsible for the metabolic activation of various chemical compounds, and in the kidney are predominantly expressed in proximal tubules. The aim of this study was to test the hypothesis that renal proximal tubular P450s are critical for nephrotoxicity caused by chemicals such as chloroform. We developed two new mouse models, one having proximal tubule-specific deletion of the cytochrome P450 reductase (Cpr) gene (the enzyme required for all microsomal P450 activities), designated proximal tubule-Cpr-null (PTCN), and the other having proximal tubule-specific rescue of CPR activity withmore » the global suppression of CPR activity in all extra-proximal tubular tissues, designated extra-proximal tubule-Cpr-low (XPT-CL). The PTCN, XPT-CL, Cpr-low (CL), and wild-type (WT) mice were treated with a single oral dose of chloroform at 200 mg/kg. Blood, liver and kidney samples were obtained at 24 h after the treatment. Renal toxicity was assessed by measuring BUN and creatinine levels, and by pathological examination. The blood and tissue levels of chloroform were determined. The severity of toxicity was less in PTCN and CL mice, compared with that of WT and XPT-CL mice. There were no significant differences in chloroform levels in the blood, liver, or kidney, between PTCN and WT mice, or between XPT-CL and CL mice. These findings indicate that local P450-dependent activities play an important role in the nephrotoxicity induced by chloroform. Our results also demonstrate the usefulness of these novel mouse models for studies of chemical-induced kidney toxicity. - Highlights: • New mouse models were developed with varying P450 activities in the proximal tubule. • These mouse models were treated with chloroform, a nephrotoxicant. • Studies showed the importance of local P450s in chloroform-induced nephrotoxicity.« less

Development and validation of a general derivatization HPLC method for the trace analysis of acyl chlorides in lipophilic drug substances.

PubMed

Zheng, Xiangyuan; Luo, Lan; Zhou, Jie; Ruan, Xiaoling; Liu, Wenyuan; Zheng, Feng

2017-06-05

Acyl chlorides are important acylating agents in the synthesis of active pharmaceutical ingredients. Determining the residual acyl chlorides in drug substances is a challenge due to their high reactivity and the matrix interferences from drug substances and their related impurities. This paper describes a general derivatization HPLC method for the determination of aromatic and aliphatic acyl chlorides in lipophilic drug substances. Since most drug substances have weak absorptions in the visible range (above 380nm), the nitro-substituted anilines and nitro-substituted phenylhydrazines were selected as the derivatization reagents due to their weak basicity and red-shift of UV absorption spectra. The maximum wavelength and absorption intensity of nitro-substituted anilines decreased after derivatization with acyl chlorides, whereas the derivatization products of nitro-substituted phenylhydrazines showed the slight increases of maximum wavelength and absorbance intensity. Hence, 2-nitrophenylhydrazine was selected as the suitable derivatization reagent because the derivatives have the maximum UV wavelength absorbance at 395nm, which could largely minimize the matrix interferences. The optimization of the concentration of 2-nitrophenylhydrazine is important for the sensitivity and stability of derivatives. Other reaction conditions including reaction temperature, time and the influence of three competitive solvents (water, methanol and ethanol) on the reaction efficiency were also studied. After derivatization with 100μgmL -1 2-nitrophenylhydrazine at room temperature for 30min, the method was validated for high specificity and sensitivity with the detection limits in the range of 0.01-0.03μgmL -1 . The proposed method was applied as a generic method to determine the residual acyl chlorides in lipophilic drug substances. Copyright © 2017 Elsevier B.V. All rights reserved.

Underivatized oxysterols and nanoLC-ESI-MS: A mismatch.

PubMed

Roberg-Larsen, Hanne; Vesterdal, Caroline; Wilson, Steven Ray; Lundanes, Elsa

2015-07-01

Due to their non-charged character, liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) measurements of oxysterols are often performed after derivatization with e.g. charged Girard reagents. However, derivatization reactions are time-consuming and may require numerous steps to remove excess reagent. In addition, extensive sample handling can be associated with cholesterol autoxidation, resulting in analyte artifacts and hence false positives. Nano scale liquid chromatography in combination with electrospray-mass spectrometry (nanoLC-ESI-MS) is a powerful tool for analyzing limited samples, due to substantially increased sensitivity compared to conventional LC-ESI-MS. The signal enhancement may compensate for the poor ionization of the oxysterols; hence we have explored the possibility to quantify oxysterols without derivatization using nanoLC-ESI-MS. Non-derivatized oxysterols and nanoLC were however not compatible, due to persistent and large carry-over. This was attributed to the extended contribution of surface to volume ratio in such miniaturized systems and interactions with the materials of the nanoLC instrumentation (e.g. adsorption to the fused silica tubing). Two contemporary MS instruments (Q-Exactive™ hybrid quadrupole-Orbitrap and TSQ Quantiva™ triple quadrupole) were used. However, both the MS and MS/MS spectra of non-derivatized oxysterols were ambiguous and/or unrepeatable for both of the instruments employed. Derivatizing oxysterols is more cumbersome, but provides more selective and reliable results, and Girard derivatization+nanoLC-ESI-MS continues to be our recommended choice for measuring oxysterols in very limited samples. These investigations also indicate that extra care should be taken to remove lipids prior to nanoLC of other analytes, as adsorbed oxysterols, etc. can compromise analysis. Copyright © 2015 Elsevier Inc. All rights reserved.

Review of in situ derivatization techniques for enhanced bioanalysis using liquid chromatography with mass spectrometry.

PubMed

Baghdady, Yehia Z; Schug, Kevin A

2016-01-01

Accurate and specific analysis of target molecules in complex biological matrices remains a significant challenge, especially when ultra-trace detection limits are required. Liquid chromatography with mass spectrometry is often the method of choice for bioanalysis. Conventional sample preparation and clean-up methods prior to the analysis of biological fluids such as liquid-liquid extraction, solid-phase extraction, or protein precipitation are time-consuming, tedious, and can negatively affect target recovery and detection sensitivity. An alternative or complementary strategy is the use of an off-line or on-line in situ derivatization technique. In situ derivatization can be incorporated to directly derivatize target analytes in their native biological matrices, without any prior sample clean-up methods, to substitute or even enhance the extraction and preconcentration efficiency of these traditional sample preparation methods. Designed appropriately, it can reduce the number of sample preparation steps necessary prior to analysis. Moreover, in situ derivatization can be used to enhance the performance of the developed liquid chromatography with mass spectrometry-based bioanalysis methods regarding stability, chromatographic separation, selectivity, and ionization efficiency. This review presents an overview of the commonly used in situ derivatization techniques coupled to liquid chromatography with mass spectrometry-based bioanalysis to guide and to stimulate future research. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A Derivatization and Validation Strategy for Determining the Spatial Localization of Endogenous Amine Metabolites in Tissues using MALDI Imaging Mass Spectrometry

PubMed Central

Manier, M. Lisa; Spraggins, Jeffrey M.; Reyzer, Michelle L.; Norris, Jeremy L.; Caprioli, Richard M.

2014-01-01

Imaging mass spectrometry (IMS) studies increasingly focus on endogenous small molecular weight metabolites and consequently bring special analytical challenges. Since analytical tissue blanks do not exist for endogenous metabolites, careful consideration must be given to confirm molecular identity. Here we present approaches for the improvement in detection of endogenous amine metabolites such as amino acids and neurotransmitters in tissues through chemical derivatization and matrix-assisted laser desorption/ionization (MALDI) IMS. Chemical derivatization with 4-hydroxy-3-methoxycinnamaldehyde (CA) was used to improve sensitivity and specificity. CA was applied to the tissue via MALDI sample targets precoated with a mixture of derivatization reagent and ferulic acid (FA) as a MALDI matrix. Spatial distributions of chemically derivatized endogenous metabolites in tissue were determined by high-mass resolution and MSn imaging mass spectrometry. We highlight an analytical strategy for metabolite validation whereby tissue extracts are analyzed by high-performance liquid chromatography (HPLC)-MS/MS to unambiguously identify metabolites and distinguish them from isobaric compounds. PMID:25044893

Purification of Derivatized Oligosaccharides by Solid Phase Extraction for Glycomic Analysis

PubMed Central

Zhang, Qiwei; Li, Henghui; Feng, Xiaojun; Liu, Bi-Feng; Liu, Xin

2014-01-01

Profiling of glycans released from proteins is very complex and important. To enhance the detection sensitivity, chemical derivatization is required for the analysis of carbohydrates. Due to the interference of excess reagents, a simple and reliable purification method is usually necessary for the derivatized oligosaccharides. Various SPE based methods have been applied for the clean-up process. To demonstrate the differences among these methods, seven types of self-packed SPE cartridges were systematically compared in this study. The optimized conditions were determined for each type of cartridge and it was found that microcrystalline cellulose was the most appropriate SPE material for the purification of derivatized oligosaccharide. Normal phase HPLC analysis of the derivatized maltoheptaose was realized with a detection limit of 0.12 pmol (S N−1 = 3) and a recovery over 70%. With the optimized SPE method, relative quantification analysis of N-glycans from model glycoproteins were carried out accurately and over 40 N-glycans from human serum samples were determined regardless of the isomers. Due to the high stability and sensitivity, microcrystalline cellulose cartridge showed potential applications in glycomics analysis. PMID:24705408

Effect of AOT Microemulsion Composition on the Hydrodynamic Diameter and Electrophoretic Mobility of Titanium Oxide Nanoparticles

NASA Astrophysics Data System (ADS)

Shaparenko, N. O.; Beketova, D. I.; Demidova, M. G.; Bulavchenko, A. I.

2018-05-01

The hydrodynamic diameter and electrophoretic mobility of titania nanoparticles in AOT microemulsions are studied depending on their water content (from 0 to 1.5 vol %), chloroform content in n-decane-chloroform mixture (from 0 to 30 vol %) and temperature (from 0 to 60°C). Considerable changes in diameter (from 20 to 400 nm) are detected upon adding water to the microemulsion. The electrophoretic mobility grows by 2-3 times upon adding chloroform, or as the temperature falls. The observed features allow us to halve the time of electrophoretic concentration for 140 nm TiO2 nanoparticles, and to concentrate 14 nm nanoparticles that do not exhibit electrophoretic mobility in the absence of chloroform.

Ophthalmic acid accumulation in an Escherichia coli mutant lacking the conserved pyridoxal 5'-phosphate-binding protein YggS.

PubMed

Ito, Tomokazu; Yamauchi, Ayako; Hemmi, Hisashi; Yoshimura, Tohru

2016-12-01

Escherichia coli YggS is a highly conserved pyridoxal 5'-phosphate (PLP)-binding protein whose biochemical function is currently unknown. A previous study with a yggS-deficient E. coli strain (ΔyggS) demonstrated that YggS controls l-Ile- and l-Val-metabolism by modulating 2-ketobutyrate (2-KB), l-2-aminobutyrate (l-2-AB), and/or coenzyme A (CoA) availability in a PLP-dependent fashion. In this study, we found that ΔyggS accumulates an unknown metabolite as judged by amino acid analyses. LC/MS and MS/MS analyses of the compound with propyl chloroformate derivatization, and co-chromatography analysis identified this compound as γ-l-glutamyl-l-2-aminobutyryl-glycine (ophthalmic acid), a glutathione (GSH) analogue in which the l-Cys moiety is replaced by l-2-AB. We also determine the metabolic consequence of the yggS mutation. Absence of YggS initially increases l-2-AB availability, and then causes ophthalmic acid accumulation and CoA limitation in the cell. The expression of a γ-glutamylcysteine synthetase and a glutathione synthetase in a ΔyggS background causes high-level accumulation of ophthalmic acid in the cells (∼1.2 nmol/mg cells) in a minimal synthetic medium. This opens the possibility of a first fermentative production of ophthalmic acid. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Liquid chromatography/electrospray ionization/isotopic dilution mass spectrometry analysis of n-(phosphonomethyl) glycine and mass spectrometry analysis of aminomethyl phosphonic acid in environmental water and vegetation matrixes.

PubMed

Grey, L; Nguyen, B; Yang, P

2001-01-01

A liquid chromatography/electrospray/mass spectrometry (LC/ES/MS) method was developed for the analysis of glyphosate (n-phosphonomethyl glycine) and its metabolite, aminomethyl phosphonic acid (AMPA) using isotope-labelled glyphosate as a method surrogate. Optimized parameters were achieved to derivatize glyphosate and AMPA using 9-fluorenylmethyl chloroformate (FMOC-Cl) in borate buffer prior to a reversed-phase LC analysis. Method spike recovery data obtained using laboratory and real world sample matrixes indicated an excellent correlation between the recovery of the native and isotope-labelled glyphosate. Hence, the first performance-based, isotope dilution MS method with superior precision, accuracy, and data quality was developed for the analysis of glyphosate. There was, however, no observable correlation between the isotope-labelled glyphosate and AMPA. Thus, the use of this procedure for the accurate analysis of AMPA was not supported. Method detection limits established using standard U.S. Environmental Protection Agency protocol were 0.06 and 0.30 microg/L, respectively, for glyphosate and AMPA in water matrixes and 0.11 and 0.53 microg/g, respectively, in vegetation matrixes. Problems, solutions, and the method performance data related to the analysis of chlorine-treated drinking water samples are discussed. Applying this method to other environmental matrixes, e.g., soil, with minimum modifications is possible, assuring accurate, multimedia studies of glyphosate concentration in the environment and the delivery of useful multimedia information for regulatory applications.