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Sample records for 9-fluorenylmethyl chloroformate fmoc

  1. 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.

  2. Generation of a Natural Glycan Microarray Using 9-Fluorenylmethyl Chloroformate (FmocCl) as a Cleavable Fluorescent Tag

    PubMed Central

    Song, Xuezheng; Lasanajak, Yi; Rivera-Marrero, Carlos; Luyai, Anthony; Willard, Margaret; Smith, David F.; Cummings, Richard D.

    2009-01-01

    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 α1,3-fucose and core α1,2-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. PMID:19699706

  3. Studies on the interaction between 9-fluorenylmethyl chloroformate and Fe3+ and Cu2+ ions: spectroscopic and theoretical calculation approach.

    PubMed

    Gu, Zhenyan; Lei, Wu; Shi, Wenyan; Hao, Qingli; Si, Weimeng; Xia, Xifeng; Wang, Fengxiang

    2014-11-11

    The interaction between 9-fluorenylmethyl chloroformate (FMOC-Cl) and Fe3+ and Cu2+ ions was investigated using fluorescence, UV/Vis absorption spectroscopies and theoretical calculation. The optical property of FMOC-Cl was studied in detail in absence and presence of various transition metal ions with particular affinity to Fe3+ and Cu2+ ions. With the fluorescence characteristic band centered at 307 and 315 nm for FMOC-Cl, the introduction of Fe3+ or Cu2+ ions leads to the fluorescence quenching of FMOC-Cl with different shift and intensities of two fluorescent bands. It allows us to differentiate between FMOC-Cl and Fe3+ and Cu2+ ions interaction behavior. The study on fluorescent kinetics confirms that the fluorescence quenching of FMOC-Cl with Fe3+ and Cu2+ ions is based on the formation of non-fluorescent material, that is, static quenching. Further analyses of bond lengths, Mulliken atomic charges and the frontier orbital compositions for FMOC-Cl and its complexes with Fe3+ and Cu2+ ions were carried out. The theoretical calculations prove the fluorescence quenching originates from the formation of coordination bonds between the oxygen atom of the carbonyl group of FMOC-Cl and Fe3+ and Cu2+ ions. The commercially available FMOC-Cl can be used as excellent fluorescent probe toward Fe3+ and Cu2+ ions with high sensitivity.

  4. Studies on the interaction between 9-fluorenylmethyl chloroformate and Fe3+ and Cu2+ ions: Spectroscopic and theoretical calculation approach

    NASA Astrophysics Data System (ADS)

    Gu, Zhenyan; Lei, Wu; Shi, Wenyan; Hao, Qingli; Si, Weimeng; Xia, Xifeng; Wang, Fengxiang

    2014-11-01

    The interaction between 9-fluorenylmethyl chloroformate (FMOC-Cl) and Fe3+ and Cu2+ ions was investigated using fluorescence, UV/Vis absorption spectroscopies and theoretical calculation. The optical property of FMOC-Cl was studied in detail in absence and presence of various transition metal ions with particular affinity to Fe3+ and Cu2+ ions. With the fluorescence characteristic band centered at 307 and 315 nm for FMOC-Cl, the introduction of Fe3+ or Cu2+ ions leads to the fluorescence quenching of FMOC-Cl with different shift and intensities of two fluorescent bands. It allows us to differentiate between FMOC-Cl and Fe3+ and Cu2+ ions interaction behavior. The study on fluorescent kinetics confirms that the fluorescence quenching of FMOC-Cl with Fe3+ and Cu2+ ions is based on the formation of non-fluorescent material, that is, static quenching. Further analyses of bond lengths, Mulliken atomic charges and the frontier orbital compositions for FMOC-Cl and its complexes with Fe3+ and Cu2+ ions were carried out. The theoretical calculations prove the fluorescence quenching originates from the formation of coordination bonds between the oxygen atom of the carbonyl group of FMOC-Cl and Fe3+ and Cu2+ ions. The commercially available FMOC-Cl can be used as excellent fluorescent probe toward Fe3+ and Cu2+ ions with high sensitivity.

  5. Spectrofluorimetric determination of sertraline in dosage forms and human plasma through derivatization with 9-fluorenylmethyl chloroformate

    PubMed Central

    2011-01-01

    Background Sertraline is primarily used to treat major depression in adult outpatients as well as obsessive-compulsive, panic and social anxiety disorders in both adults and children. A survey of the literature reveals that most of the reported methods are either insufficiently sensitive or tedious and require highly sophisticated and dedicated instrumentation. The proposed method is considered to be specific for determination of SER in presence of its metabolite (deaminated form). Results A sensitive, simple and specific spectrofluorimetric method was developed for the determination of sertraline (SER) in pharmaceutical formulations and biological fluids. The method is based on its reaction with 9-fluorenylmethyl chloroformate (FMOC-Cl) in borate buffer of pH 8.0 to yield a highly fluorescent derivative peaking at 315 nm after excitation at 265 nm. The different experimental parameters affecting the development and stability of the reaction product were carefully studied and optimized. The fluorescence concentration plot was rectilinear over the range of 0.05-1.0 μg mL-1 with a lower detection limit of 5.34 × 10-3 μg mL-1 and limit of quantitation of 0.016 μg mL-1. Conclusions The proposed method was successfully applied to the analysis of commercial tablets and the results obtained were in good agreement with those obtained using the reference method. Furthermore, the method was applied for the determination of SER in spiked and real human plasma. The mean % recovery (n = 3) was 94.33 ± 1.53 and 92.00 ± 2.65, respectively. A proposal of the reaction pathway was postulated. PMID:21978386

  6. High performance liquid chromatography for the determination of glucosamine sulfate in human plasma after derivatization with 9-fluorenylmethyl chloroformate.

    PubMed

    Huang, Tao-min; Deng, Chun-Hui; Chen, Nian-zhu; Liu, Zhen; Duan, Geng-Li

    2006-10-01

    In this study, we developed a simple, rapid, sensitive, and reliable method for the determination of glucosamine sulfate in human plasma, which was based on derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) followed by reverse-phase HPLC-FLD. For the first time, FMOC-Cl was introduced into derivatization of glucosamine sulfate in human plasma. The amino groups of glucosamine sulfate and vertilmicin sulfate (the internal standard) were trapped with FMOC-Cl to form glucosamine-FMOC-Cl and vertilmicin-FMOC-Cl adducts, which can be very suitable for HPLC-FLD. Precipitation of plasma proteins by acetonitrile was followed by vortex mixing and centrifugation. Chromatographic separation was performed on a C18 column (DIAMONSIL 150 x 4 mm id, 5 microm) with a mobile phase gradient consisting of acetonitrile and water at a flow-rate of 1 mL/min. The retention times of glucosamine-FMOC-Cl and vertilmicin-FMOC-Cl adducts were 8.9 and 21.2 min, respectively. This method was shown to be selective and sensitive for glucosamine sulfate. The limit of detection was 15 ng/mL for glucosamine sulfate in plasma and the linear range was 0.1-10 mg/mL in plasma with a correlation coefficient (r) of 0.9999. The relative standard deviations (RSDs) of intra-day and inter-day assays were 5.2-8.1% and 6.1- 8.5%, respectively. Extraction recoveries of glucosamine sulfate in plasma were greater than 90%. The validated method was successfully applied to the determination of glucosamine sulfate in human plasma samples.

  7. Determination of sulphonamides in animal tissues by high performance liquid chromatography with pre-column derivatization of 9-fluorenylmethyl chloroformate.

    PubMed

    Zou, Qiong-Hui; Xie, Meng-Xia; Wang, Xiang-Feng; Liu, Yuan; Wang, Jin; Song, Jia; Gao, Hui; Han, Jie

    2007-11-01

    A novel approach for simultaneous determination of 12 sulphonamides (sulphadiazine, sulphamethazine, sulphathiazole, sulphadimethoxine, sulphamerazine, sulphapyridine, sulphamethoxazole, suphamethizole, sulphaquinoxaline, sulphameter, sulphamonomethoxine, and sulphachloropyridazine) in animal tissues (swine muscle and liver, chicken muscle, beef muscle) by HPLC with UV detection has been developed. A pre-column derivatization of the sulphonamide compounds with 9-fluorenylmethyl chloroformate (FMOC-Cl) has been proposed and the reaction conditions have been optimized. The FMOC-sulphonamide derivatives were purified by SPE with silica gel as solid support prior to HPLC separation. The limits of detection for the sulphonamide compounds were greatly improved after the derivatization and purification step for the derivatives. Sulphonamide residues in animal tissues were extracted by acetonitrile and purified by solid phase extraction with C(18) as the solid support. The method developed has high sensitivity and good repeatability, and the average recoveries for most of the sulphonamides at various spiking levels were above 70% with relative standard deviations below 13.7%. The limits of detection for most sulphonamides can reach 3-5 microg/kg.

  8. Measurement of histidinohydroxylysinonorleucine and hydroxyproline in skin collagen by reversed-phase high-performance liquid chromatography after 9-fluorenylmethyl chloroformate labeling.

    PubMed

    Kondo, A; Ishikawa, O; Okada, K; Miyachi, Y; Abe, S; Kuboki, Y

    1997-10-15

    A novel, highly sensitive method to quantify histidinohydroxylysinonorleucine (HHL), a trifunctional type of cross-link in skin collagen, was developed. HHL in skin hydrolysates labeled with 9-fluorenylmethyl chloroformate (FMOC-Cl) was separated by reversed-phase high-performance liquid chromatography. Mass spectrometric analysis revealed that two FMOCs were bound to two primary amino acid residues, histidine and hydroxylysine, but not to lysine residue in one HHL molecule. Hydroxyproline was simultaneously measured to express the molar ratio of HHL to collagen. The detection range of HHL was from 1 to 10 pmol and that of hydroxyproline from 1 to 50 pmol. A 6-mm punch-biopsied human skin sample contained 0.40 to 0.69 mol of HHL per one molecule of collagen. This sensitive method is useful as it is rapid and can be used to examine the aging process or the change of HHL content in skin collagens of various pathologic states.

  9. Determination of 1-deoxynojirimycin in Morus alba L. leaves by derivatization with 9-fluorenylmethyl chloroformate followed by reversed-phase high-performance liquid chromatography.

    PubMed

    Kim, Jin-Won; Kim, Soo-Un; Lee, Heui Sam; Kim, Iksoo; Ahn, Mi Young; Ryu, Kang Sun

    2003-06-20

    A rapid and reliable method suitable for assays of a large number of Morus alba leaves for 1-deoxynojirimycin (DNJ) has been developed. DNJ in 0.1 g of freeze-dried leaves was double-extracted in 10 mL of aqueous 0.05 M HCl by vortexing for 15 s at room temperature, derivatized with 9-fluorenylmethyl chloroformate (FMOC-Cl), and analyzed by reversed-phase high-performance liquid chromatography (RP-HPLC) equipped with a fluorescence detector. The double extraction recovered > 99% of extractable DNJ from the leaves. Stabilization of FMOC-derivatized DNJ (DNJ-FMOC) was achieved by diluting the reactant with aqueous acetic acid after derivatization. DNJ-FMOC was stable for at least 16 days under acidic conditions at room temperature (24 degrees C). Linearity ranged between 0.3 and 30 microg mL(-1). The intra- and inter-day precision for DNJ-spiked biological samples was between 0.6 and 1.8% and between 3.7 and 4.5%, respectively.

  10. Simultaneous analysis of amino acids and amines as their o-phthalaldehyde-ethanethiol-9-fluorenylmethyl chloroformate derivatives in cheese by high-performance liquid chromatography.

    PubMed

    Korös, A; Varga, Zs; Molnár-Perl, I

    2008-09-05

    A high-performance liquid chromatographic method (HPLC) with combined diode array and fluorescence detection of amino acids and amines in various cheese samples is described. The proposal is based on acidic deproteinization, derivatization and gradient optimization studies, resulting in the identification and quantification of 21 amino acids and 9 amines from a single solution, by one injection. The optimized, simple protocol consists of deproteinization (1M perchloric acid), centrifugation, filtration and the subsequent derivatization with the o-phthalaldehyde-ethanethiol-9-fluorenylmethyl chloroformate (OPA-ET-FMOC) reagent. The method can be characterized with a linearity of wide concentration range (6.25-1000 pM/injection), a good chromatographic reproducibility (average: 2.69% RSD) and an excellent recovery (average: 100.2%; average 3.84% RSD). The developed method was successfully applied in the determination of the amino acid and amine contents of port salut cheese, blue cheese and smoked cheese samples.

  11. Sensitive high-performance liquid chromatographic quantitation of gabapentin in human serum using liquid-liquid extraction and pre-column derivatization with 9-fluorenylmethyl chloroformate.

    PubMed

    Bahrami, Gholamreza; Kiani, Amir

    2006-05-01

    Most of the published methods for analysis of gabapentin, an antiepileptic agent, in human serum are based on the same approach, involving o-phthaldialdehyde derivatization of deproteinized serum samples. The present paper however, describes a new, simple and sensitive high-performance liquid chromatographic method for determination of gabapentin in human serum using liquid-liquid extraction and 9-fluorenylmethyl chloroformate (FMOC-Cl) as pre-column labeling agent. The drug and an internal standard (azithromycin) were extracted from serum by salting-out approach using a mixture of dichloromethane-2 propanol (1:1, v/v) as the extracting solvent. The extracted analytes were subjected to derivatization with FMOC-Cl in the presence of phosphate buffer (pH 7). A mobile phase consisting of methanol-0.05 M sodium phosphate buffer (73/27, v/v; pH of 3.9) containing 1 ml/l triethylamine was eluted and chromatographic separation was performed on a Shimpack CLC-C18 (150 mm x 4.6 mm) column. The standard curve was linear over the range of 0.03-20 microg/ml and limit of quantification was 0.03 microg/ml. The performance of analysis was studied and the validated method showed excellent performance in terms of selectivity, specificity, sensitivity, precision and accuracy. No interferences were found from commonly co-administered antiepileptic agents.

  12. Determination of clarithromycin in human serum by high-performance liquid chromatography after pre-column derivatization with 9-fluorenylmethyl chloroformate: application to a bioequivalence study.

    PubMed

    Bahrami, Gholamreza; Mohammadi, Bahareh

    2007-05-01

    A sensitive liquid chromatographic method for the analysis of clarithromycin, a macrolide antibiotic, in human serum using pre-column derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) is described. The method involved liquid-liquid extraction of the drug and an internal standard (amantadine) followed by pre-column derivatization of the analytes with FMOC-Cl. A mixture of 0.05 M phosphate buffer containing triethylamine (2 mL L(-1); pH 3.8) and methanol (17:83, v/v) was used as mobile phase and chromatographic separation was achieved on a Shimpack CLC-ODS column. The eluate was monitored by a fluorescence detector with respective excitation and emission wavelengths of 265 and 315 nm. The analytical method was linear over the concentration range of 0.025-10 microg mL(-1) of clarithromycin in human serum with a limit of quantification of 0.025 microg mL(-1). The assay is sensitive enough to measure drug levels obtained in human single dose studies. In the present method, sensitivity and run time of analysis have been improved, and successfully applied in a bioequivalence study of three different clarithromycin preparations in 12 healthy volunteers.

  13. Quantification of fudosteine in human plasma by high-performance liquid chromatography-electrospray ionization mass spectrometry employing precolumn derivatization with 9-fluorenylmethyl chloroformate.

    PubMed

    Xu, Fengguo; Zhang, Zunjian; Jiao, Haoyan; Tian, Yuan; Zhang, Beibei; Chen, Yun

    2006-05-01

    This paper describes a novel method for the sensitive and selective determination of fudosteine in human plasma. The method involves a derivatization step with 9-fluorenylmethyl chloroformate (FMOC-Cl) in borate buffer and detection based on high-performance liquid chromatography-electrospray ionization mass spectrometry (LC/ESI/MS). After acetonitrile-induced protein precipitation of plasma samples, fudosteine was derivatized with FMOC-Cl, then extracted by ethyl acetate, evaporated, reconstituted and injected using an LC/ESI/MS instrument. Separation was achieved using an ODS column and isocratic elution. Excellent linearity was obtained for the entire calibration range from 0.05 to 20 microg/ml. Validation assays of the lower limit of quantification (LLOQ) as well as for the intra- and inter-batch precision and accuracy met the international acceptance criteria for bioanalytical method validation. Using the developed analytical method, fudosteine could be detected for the first time in human plasma with a low limit of detection (LLOD) of 0.03 microg/ml. The proposed method has been successfully applied to study the pharmacokinetics of fudosteine in healthy Chinese volunteers after single and multiple oral administration.

  14. Sensitive determination of trimetazidine in spiked human plasma by HPLC with fluorescence detection after pre-column derivatization with 9-fluorenylmethyl chloroformate.

    PubMed

    Khedr, Alaa; Sheha, Mahmoud M; Darwish, Ibrahim A

    2007-09-01

    A high-performance liquid chromatographic method for the determination of trimetazidine dihydrochloride (TMZ) in spiked human plasma is described. The method is based on the pre-column derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) using the fluorimetric detection technique. Fluoxetine HCl (FLX) was used as internal standard. Both, TMZ and FLX were completely derivatized after heating at 50 degrees C for 20 min in borate buffer pH 8.0. Samples were analyzed by high performance liquid chromatography (HPLC) using Zorbax-TMS column (250 mm x 4.6 mm, i.d., 5 microm) and mobile phase consist of acetonitrile, methanol and 20 mM sodium acetate pH 4.7 (44:6:50; v/v/v). Fluorescence detector (FLD) was adjusted at excitation and emission wavelengths; 265 and 311 nm, respectively. The linearity of the method was in the range of 4.5-200 ng/ml. Limits of detection (LOD) and quantification (LOQ) were 1.5 and 4.5 ng/ml, respectively. Trimetazidine recovery was 96.5+/-1.3% (n=6; RSD=2.1%).

  15. Sensitive fluorimetric determination of gentamicin sulfate in biological matrices using solid-phase extraction, pre-column derivatization with 9-fluorenylmethyl chloroformate and reversed-phase high-performance liquid chromatography.

    PubMed

    Stead, D A; Richards, R M

    1996-01-26

    A high-performance liquid chromatographic method is described for the determination of gentamicin in bacterial culture medium or plasma with increased sensitivity and improved separation of the C1 component. Gentamicin was extracted from the biological matrix with high efficiency using carboxypropyl (CBA)-bonded silica. Derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) followed by C18 reversed-phase chromatography allowed the fluorimetric detection of gentamicins C1, C1a and C2. A fourth component, considered to be gentamicin C2a, was partially resolved from the C2 peak. Optimal conditions for the extraction and derivatization of gentamicin are described. The detection limit was below 50 micrograms/l, the assay was linear to 5 mg/l and showed good reproducibility. It is concluded that pre-column derivatization with FMOC-Cl substantially improves the analysis of gentamicin compared with present methods based on reaction with o-phthaldialdehyde.

  16. Derivatization and fluorescence detection of amino acids and peptides with 9-fluorenylmethyl chloroformate on the surface of a solid adsorbent.

    PubMed

    Shangguan, D; Zhao, Y; Han, H; Zhao, R; Liu, G

    2001-05-01

    An approach that exploits the surface of a solid adsorbent is proposed for precolumn FMOC derivatization of amino acids and peptides. Amino acids (Ser, Glu, GABA, Val, Phe, Lys) and two neuropeptides (substance P and Leuenkephalin) were adsorbed on alkaline silica gel cartridges. After drying, they were reacted with 9-fluorenyl-methyl chloroformate (FMOC-Cl) in toluene. After washing off the excess FMOC-Cl with ethyl acetate, the derivatives were eluted with aqueous eluant. The eluates were separated and detected by means of HPLC with fluorescence detection. Compared with the traditional derivatization in the liquid phase, the extent of formation of byproducts of FMOC-Cl with water was greatly decreased, and the excess FMOC-Cl was eliminated completely.

  17. Analysis of synephrine in da-cheng-qi decoction by HPLC employing precolumn derivatization with 9-fluorenylmethyl chloroformate.

    PubMed

    Liu, Ying; Xu, Fengguo; Zhang, Zunjian; Yang, Cheng; Song, Rui; Tian, Yuan

    2009-01-01

    A sensitive and reliable method has been developed to determine synephrine in a famous purgative Chinese medicinal prescription, da-cheng-qi decoction. After purified by solid-phase extraction (SPE), the decoction was derivatized with 9-fluorenylmethyl chloroformate in borate buffer and analyzed by reversed-phase high-performance liquid chromatography equipped with a diode-array detector. The chromatographic separation was carried out on a Shimadzu shim-pack VP-ODS column using acetonitrile-water as mobile phase. This method was validated in terms of linearity, recovery, precision, accuracy, and repeatability. Calibration curves were linear (r(2) > 0.99) over the concentration range of 0.78-100 microg/mL, and the mean recovery was 93.28%. The intra- and inter-batch precisions were less than 2% in terms of relative standard deviations. The accuracy and repeatability were well within the acceptable range. The limit of detection was 0.25 microg/mL. This validated method also can be used for assays of herbal products, fruits, dietary supplements, and biological samples containing synephrine.

  18. 9-fluorenylmethyl chloroformate as a fluorescence-labeling reagent for derivatization of carboxylic acid moiety of sodium valproate using liquid chromatography/tandem mass spectrometry for binding characterization: a human pharmacokinetic study.

    PubMed

    Mohammadi, Bahareh; Majnooni, Mohammad Bagher; Khatabi, Pyman Malek; Jalili, Ronak; Bahrami, Gholamreza

    2012-01-01

    In High Performance Liquid Chromatographic (HPLC) determination of chemicals with acidic functions, different labeling agents are used to improve sensitivity of the assay. 9-Fluorenylmethyl chloroformate (FMOC-Cl), on the other hand, is a suitable labeling agent, which reacts with both primary and secondary amines and less readily with hydroxyl groups in alkaline conditions. However, the reagent has not been applied in labeling of chemicals with acidic function yet. In this study which is the first report on application of FMOC-Cl in derivatization and analysis of a drug with acidic function, valproic acid (VPA), one of a series of fatty carboxylic acids with anticonvulsant activity, was derivatized using the reagent and quantified in serum samples by HPLC with fluorescence detection. In addition, to document the reaction between the labeling agent and carboxylic acid moiety of the drug, we developed a liquid chromatography-tandem MS/MS (LC-MS/MS) method. Following liquid-liquid extraction, derivatization of the drug and an internal standard was achieved in alkaline medium. The elute was monitored by a fluorescence detector with respective excitation and emission wavelengths of 265 and 315 nm. The present method is more sensitive comparing with other published HPLC procedures for analysis of VPA. The assay is sensitive enough to measure drug levels obtained in human single dose studies with a limit of quantification of 0.01 μg/mL. Also the method is linear over the concentrations range of 0.01-32 μg/mL of VPA in human serum using 100 μL serum sample and 5 μL injection. The coefficient variation values of both inter and intra day analysis were less than 12% and the percentage error was less than 4%. The method performance was studied and the validated procedure applied in a randomized cross-over bioequivalence study of two different VPA preparations in 24 healthy volunteers.

  19. Determination of glyphosate and aminomethylphosphonic acid in aqueous soil matrices: a critical analysis of the 9-fluorenylmethyl chloroformate derivatization reaction and application to adsorption studies.

    PubMed

    Báez, María E; Fuentes, Edwar; Espina, María José; Espinoza, Jeannette

    2014-11-01

    The assessment of the environmental fate of glyphosate and its degradation product (aminomethylphosphonic acid) is of great interest given the widespread use of the herbicide. Studies of adsorption-desorption and transport processes in soils require analytical methods with sensitivity, accuracy, and precision suitable for determining the analytes in aqueous equilibrium solutions of varied complexity. In this work, the effect of factors on the yield of the derivatization of both compounds with 9-fluorenylmethyl chloroformate for applying in aqueous solutions derived from soils was evaluated through factorial experimental designs. Interference effects coming from background electrolytes and soil matrices were established. The whole method had a linear response up to 640 ng/mL (R(2) > 0.999) under optimized conditions for high-performance liquid chromatography with fluorescence detection. Limits of detection were 0.6 and 0.4 ng/mL for glyphosate and aminomethylphosphonic acid, respectively. The relative standard deviation was 4.4% for glyphosate (20 ng/mL) and 5.9% for aminomethylphosphonic acid (10 ng/mL). Adsorption of compounds on four different soils was assessed. Isotherm data fitted well the Freundlich model (R(2) > 0.97). Kf constants varied between 93 ± 3.1 and 2045 ± 157 for glyphosate and between 99 ± 4.1 and 1517 ± 56 (μg(1-1/) (n)  mL(1/) (n) ( ) g(-1) ) for aminomethylphosphonic acid, showing the broad range of applicability of the proposed method.

  20. Amino acid analysis by reverse-phase high-performance liquid chromatography: improved derivatization and detection conditions with 9-fluorenylmethyl chloroformate.

    PubMed

    Bank, R A; Jansen, E J; Beekman, B; te Koppele, J M

    1996-09-05

    An improved method for the quantitative derivatization of amino acids with fluorenylmethyl chloroformate (FMOC-Cl) is described. Amino acids are derivatized in borate buffer at pH 11.4 for 40 min at ambient temperature. All amino acids resulted in stable derivatives. In particular, improved derivatization was obtained with the troublesome amino acids His and Tyr: exclusively monosubstituted His and disubstituted Tyr were formed, eluting as free peaks in the chromatogram. These derivatives show a higher fluorescence response than their disubstituted and monosubstituted counterparts, respectively, resulting from other protocols. Under the new conditions, considerable less of the hydrolysis product of FMOC-Cl is seen in the chromatograms. Baseline noise was substantially reduced at a higher emission wavelength (630 nm instead of 313 or 340 nm). With simple precautions, extensive adsorption of the disubstituted derivatives (Lys, Hyl, and Tyr) on plastic or glass surfaces could be prevented. Calibration curves were linear over a 10 to 300 molar ratio of FMOC-Cl to total amino acid. The detection limits are in the femtomole range and the derivatives are stable for more than 48 h, thus permitting automated analysis of multiple samples.

  1. A simple method for the assay of colistin in human plasma, using pre-column derivatization with 9-fluorenylmethyl chloroformate in solid-phase extraction cartridges and reversed-phase high-performance liquid chromatography.

    PubMed

    Li, J; Milne, R W; Nation, R L; Turnidge, J D; Coulthard, K; Johnson, D W

    2001-09-25

    A simple, selective and sensitive high-performance liquid chromatographic (HPLC) method is described for the determination of colistin in human plasma. Derivatization with 9-fluorenylmethyl chloroformate was performed in the same solid-phase extraction C18 cartridge used for sample pre-treatment, followed by reversed-phase HPLC with fluorimetric detection. Quantification was achieved using the ratio of the summed peak areas of colistin A and B derivatives to that of the derivative of netilmicin (internal standard). Linear calibration curves were obtained within the concentrations of colistin sulfate from 0.10 to 4.0 mg/l in plasma. Accuracy was within 10% and reproducibility (RSD) was less than 10%.

  2. Simultaneous analysis of N(ε) -(carboxymethyl)Lysine and N(ε) -(carboxyethyl)lysine in foods by ultra-performance liquid chromatography-mass spectrometry with derivatization by 9-fluorenylmethyl chloroformate.

    PubMed

    Zhou, Yanqiong; Lin, Qin; Jin, Cheng; Cheng, Lu; Zheng, Xiaoyan; Dai, Ming; Zhang, Ying

    2015-02-01

    Isotope dilution ultra-performance liquid chromatography-electrospray tandem mass spectrometry with derivatization by 9-fluorenylmethyl chloroformate was successfully applied to quantify N(ε) -(carboxymethyl)lysine (CML) and N(ε) -(carboxyethyl)lysine (CEL) in processed foods. We demonstrate that this analytical method is well validated for the determination of CML and CEL contents in processed foods. Relative standard deviations (RSD) indicate repeatability (RSD < 6% for CML and CEL) and reproducibility (RSD < 6% for CML and < 7% for CEL) in this method. Percent recovery is also good. We obtain recoveries of 102% to 112% for CML and 86% to 114% for CEL. CML levels detected in the samples vary from 2.29 to 480 mg/kg food, whereas CEL is detected in significantly lower concentrations ranging from 0.56 to 107 mg/kg food. These data could help consumers make better food choices by monitoring intake of advanced glycation end-products, which may pose a risk to human health.

  3. Capillary electrophoresis-electrospray ionization mass spectrometry for rapid and sensitive N-glycan analysis of glycoproteins as 9-fluorenylmethyl derivatives.

    PubMed

    Nakano, Miyako; Higo, Daisuke; Arai, Etsuo; Nakagawa, Takatoshi; Kakehi, Kazuaki; Taniguchi, Naoyuki; Kondo, Akihiro

    2009-02-01

    It is well known that most protein therapeutics such as monoclonal antibody pharmaceuticals and other biopharmaceuticals including cancer biomarkers are glycoproteins, and thus the development of high-throughput and sensitive analytical methods for glycans is essential in terms of their determination and quality control. We previously reported a novel alternative labeling method for glycans involving 9-fluorenylmethyl chloroformate (Fmoc-Cl) instead of the conventional reductive amination procedure. The derivatives were analyzed by high-performance liquid chromatography (HPLC) (Kamoda S, Nakano M, Ishikawa R, Suzuki S, Kakehi K. 2005. Rapid and sensitive screening of N-glycans as 9-fluorenylmethyl derivatives by high-performance liquid chromatography: A method which can recover free oligosaccharides after analysis. J Proteome Res. 4:146-152). This method was rapid and simple; however, it was time-consuming in terms of analysis by HPLC and did not provide so much information such as the detailed structures and mass numbers of glycans. Here we have developed a high-throughput and highly sensitive method. It comprises three steps, i.e., release of glycans, derivatization with Fmoc, and capillary electrophoresis-electrospray ionization mass spectrometry (CE-ESI MS) analysis. We analyzed several glycoproteins such as fetuin, alpha1 acid glycoprotein, IgG, and transferrin in order to validate this method. We were able to analyze the above glycoproteins with the three-step procedure within only 5 h, which provided detailed N-glycan patterns. Moreover, the MS/MS analysis allowed identification of the N-glycan structures. As novel applications, the method was employed for the analysis of N-glycans derived from monoclonal antibody pharmaceuticals and also from alpha-fetoprotein; the latter is known as one of the tumor markers of hepatocellular carcinomas. We were able to easily and rapidly determine the detailed structures of the N-glycans. The present method is very useful

  4. Rapid and sensitive screening of N-glycans as 9-fluorenylmethyl derivatives by high-performance liquid chromatography: a method which can recover free oligosaccharides after analysis.

    PubMed

    Kamoda, Satoru; Nakano, Miyako; Ishikawa, Rika; Suzuki, Shigeo; Kakehi, Kazuaki

    2005-01-01

    There are a large number of labeling methods for asparagine-type oligosaccharides with fluorogenic and chromophoric reagents. We have to choose the most appropriate labeling method based on the purposes such as mass spectrometry, high-performance liquid chromatography and capillary electrophoresis. Asparagine-type glycans are released from core proteins as N-glycosylamine at the initial step of the releasing reaction when glycoamidase F is employed as the enzyme. The N-glycosylamine-type oligosaccharides thus released by the enzyme are subjected to hydrolysis or mutarotation to form free-form oligosaccharides. In the detailed studies on the enzyme reaction, we found a condition in which the released N-glycosylamine-type oligosaccharides were exclusively present at least during the course of enzyme reaction, and developed a method for in situ derivatization of the glycosylamine-type oligosaccharides with 9-fluorenylmethyl chloroformate (Fmoc-Cl). The Fmoc labeled sialo- and asialo- (or high-mannose and hybrid) oligosaccharides were successfully analyzed on an amine-bonded polymer column and amide-silica column, respectively. The present method showed approximately 5 times higher sensitivities than that using 2-aminobenzoic acid (2-AA). The separation profile was similar to that observed using 2-AA method as examined by the analyses of carbohydrate chains derived from several glycoproteins including complex-type, high-mannose type and hybrid type of N-linked oligosaccharides. The labeled oligosaccharides were stable at least for several months when stored at -20 degrees C. Furthermore, it should be emphasized that the Fmoc-derivatized oligosaccharides could be easily recovered as free reducing oligosaccharides simply by incubation with morpholine in dimethylformamide solution. We obtained a pure triantennary oligosaccharide with 3 sialic acid residues as a free reducing form from fetuin in good yield after isolation of the corresponding Fmoc oligosaccharide followed

  5. Analysis of O-glycans as 9-fluorenylmethyl derivatives and its application to the studies on glycan array.

    PubMed

    Yamada, Keita; Hirabayashi, Jun; Kakehi, Kazuaki

    2013-03-19

    A method is proposed for the analysis of O-glycans as 9-fluorenylmethyl (Fmoc) derivatives. After releasing the O-glycans from the protein backbone in the presence of ammonia-based media, the glycosylamines thus formed are conveniently labeled with Fmoc-Cl and analyzed by HPLC and MALDI-TOF MS after easy purification. Fmoc labeled O-glycans showed 3.5 times higher sensitivities than those labeled with 2-aminobenzoic acid in fluorescent detection. Various types of O-glycans having sialic acids, fucose, and/or sulfate residues were successfully labeled with Fmoc and analyzed by HPLC and MALDI-TOF MS. The method was applied to the comprehensive analysis of O-glycans expressed on MKN45 cells (human gastric adenocarcinoma). In addition, Fmoc-derivatized O-glycans were easily converted to free hemiacetal or glycosylamine-form glycans that are available for fabrication of glycan array and neoglycoproteins. To demonstrate the availability of our methods, we fabricate the glycan array with Fmoc labeled glycans derived from mucin samples and cancer cells. The model studies using the glycan array showed clear interactions between immobilized glycans and some lectins.

  6. Micellular Electrokinetic Capillary Chromatography in the Undergraduate Curriculum: Separation and Identification of the Amino Acid Residues in an Unknown Dipeptide Using FMOC Derivatization

    NASA Astrophysics Data System (ADS)

    Strein, Timothy G.; Poechmann, James L.; Prudenti, Mark

    1999-06-01

    This manuscript describes our efforts to introduce biochemistry students to micellar electrokinetic capillary chromatography (MEKC), a mode of capillary electrophoresis that employs micelles in the operating buffer. Unlike free solution capillary electrophoresis, MEKC is capable of resolving both charged and uncharged analytes because the micellar pseudo stationary phase allows for the separation of uncharged species. The experiment described herein includes a comparison of MEKC, employing sodium dodecyl sulfate (SDS) as the micelle-forming species, with reverse-phase HPLC. Both methods are used to determine the amino acid residues in an unknown dipeptide. Advanced undergraduate chemistry, biochemistry, and biology majors perform this experiment in the Biochemical Methods course at Bucknell University. The students cleave the peptide bond, derivatize the resultant amino acids with 9-fluorenylmethyl chloroformate (FMOC), and separate the FMOC-amino acid derivatives using HPLC and MEKC. This manuscript details the analytical procedures for the MEKC separation and presents typical student data obtained using this relatively new method.

  7. 9-Fluorenylmethyl (Fm) Disulfides: Biomimetic Precursors for Persulfides

    SciTech Connect

    Park, Chung-Min; Johnson, Brett A.; Duan, Jicheng; Park, Jeong-Jin; Day, Jacob J.; Gang, David; Qian, Wei-Jun; Xian, Ming

    2016-03-04

    Protein S-sulfhydration has been recognized as an important post-translational modification that regulates H2S signals. However, the reactivity and biological implications of the products of S-sulfhydration, i.e. persulfides, are still unclear. This is mainly due to the instability of persulfides and difficulty to access these molecules. Under physiological conditions persulfides mainly exist in anionic forms because of their low pKa values. However, current methods do not allow for the direct generation of persulfide anions under biomimetic and non-H2S conditions. Herein we report the development of a functional disulfide, FmSSPy-A (Fm =9-fluorenylmethyl; Py = pyridinyl). This reagent can effectively convert both small molecule and protein thiols (-SH) to form –S-SFm adducts under mild conditions. It allows for a H2S-free and biomimetic protocol to generate highly reactive persulfides (in their anionic forms). We also demonstrated the high nucleophilicity of persulfides toward a number of thiol-blocking reagents. This method holds promise for further understanding the chemical biology of persulfides and S-sulfhydration.

  8. Analysis of fumonisin B1 in rodent feed by CE with fluorescence detection of the FMOC derivative.

    PubMed

    Holcomb, M; Thompson, H C

    1996-01-01

    A fast, simple, and environmentally friendly method has been developed for the analysis of fumonisin B1 (FB1) in rodent feed using CE. FB1 is the major fumonisin metabolite produced by the fungus Fusarium moniliforme and has been implicated in human and animal diseases. FB1 was extracted from rodent feed with acetonitrile/water (50/50) (vol/vol) and cleaned up with a C18 Sep-Pak Vac cartridge (Waters Corp., Milford, MA, U.S.A.). FB1 was quantitated after elution from the column using capillary electrophoresis with a 25-mM sodium borate buffer (pH 9.0) containing 10% acetonitrile and fluorescence detection of the (9-fluorenylmethyl) chloroformate (FMOC) derivative. The minimum detectable amount in rodent feed was 0.5 pmm. Recovery values in spiked rodent feed averaged more than 87% over the 2-20 ppm range.

  9. Statistical Methods for the Investigation of Solvolysis Mechanisms Illustrated by the Chlorides of the Carbomethoxy Protecting Groups NVOC and FMOC

    PubMed Central

    D'Souza, Malcolm J.; Deol, Jasbir K.; Pavey, Maryeah T.; Kevill, Dennis N.

    2015-01-01

    The solvolysis of 4,5-dimethoxy-2-nitrobenzyl chloroformate (NVOC-Cl, 1) is followed at 25.0°C in twenty hydroxylic solvents. A comparison with previously published rates for benzyl chloroformate and p-nitrobenzyl chloroformate indicates that the inductive effect of the nitro and the two methoxy groups strongly influences the rate of reaction. For 1, the specific rates of solvolysis are correlated using an extended Grunwald-Winstein (G-W) treatment. A direct comparison with the data for phenyl chloroformate (PhOCOCl) in identical solvents strongly suggests that the addition step within an addition-elimination mechanism is rate-determining for both substrates. A reevaluation of the kinetic data for 9-fluorenylmethyl chloroformate (FMOC-Cl, 2) involves a correlation of log⁡(k/ko)2 versus log⁡(k/ko)PhOCOCl. In this plot, deviations were observed in solvents rich in a hydrogen-bonding fluoroalcohol component. Omitting the aqueous fluoroalcohol rate measurements for 2 in an analysis using the extended G-W equation suggested the occurrence of dual pathways differing in the dependences upon the ionizing power and nucleophilicity of the solvent. In addition, the fluorenyl ring is rotated out of the plane containing the ether oxygen and, as a result, PhOCOCl is found to solvolyze 20 times faster than 2 in ethanol and methanol. PMID:25767738

  10. Statistical Methods for the Investigation of Solvolysis Mechanisms Illustrated by the Chlorides of the Carbomethoxy Protecting Groups NVOC and FMOC.

    PubMed

    D'Souza, Malcolm J; Deol, Jasbir K; Pavey, Maryeah T; Kevill, Dennis N

    2015-01-01

    The solvolysis of 4,5-dimethoxy-2-nitrobenzyl chloroformate (NVOC-Cl, 1) is followed at 25.0°C in twenty hydroxylic solvents. A comparison with previously published rates for benzyl chloroformate and p-nitrobenzyl chloroformate indicates that the inductive effect of the nitro and the two methoxy groups strongly influences the rate of reaction. For 1, the specific rates of solvolysis are correlated using an extended Grunwald-Winstein (G-W) treatment. A direct comparison with the data for phenyl chloroformate (PhOCOCl) in identical solvents strongly suggests that the addition step within an addition-elimination mechanism is rate-determining for both substrates. A reevaluation of the kinetic data for 9-fluorenylmethyl chloroformate (FMOC-Cl, 2) involves a correlation of log⁡(k/k o )2 versus log⁡(k/k o )PhOCOCl. In this plot, deviations were observed in solvents rich in a hydrogen-bonding fluoroalcohol component. Omitting the aqueous fluoroalcohol rate measurements for 2 in an analysis using the extended G-W equation suggested the occurrence of dual pathways differing in the dependences upon the ionizing power and nucleophilicity of the solvent. In addition, the fluorenyl ring is rotated out of the plane containing the ether oxygen and, as a result, PhOCOCl is found to solvolyze 20 times faster than 2 in ethanol and methanol.

  11. Chloroform

    Integrated Risk Information System (IRIS)

    EPA / 635 / R - 01 / 001 TOXICOLOGICAL REVIEW OF CHLOROFORM ( CAS No . 67 - 66 - 3 ) In Support of Summary Information on the Integrated Risk Information System ( IRIS ) October 2001 U.S . Environmental Protection Agency Washington , DC DISCLAIMER This document has been reviewed in accordance with U

  12. Characterization of Nα-Fmoc-protected dipeptide isomers by electrospray ionization tandem mass spectrometry (ESI-MS(n)): effect of protecting group on fragmentation of dipeptides.

    PubMed

    Ramesh, M; Raju, B; Srinivas, R; Sureshbabu, V V; Vishwanatha, T M; Hemantha, H P

    2011-07-30

    A series of positional isomeric pairs of Fmoc-protected dipeptides, Fmoc-Gly-Xxx-OY/Fmoc-Xxx-Gly-OY (Xxx=Ala, Val, Leu, Phe) and Fmoc-Ala-Xxx-OY/Fmoc-Xxx-Ala-OY (Xxx=Leu, Phe) (Fmoc=[(9-fluorenylmethyl)oxy]carbonyl) and Y=CH(3)/H), have been characterized and differentiated by both positive and negative ion electrospray ionization ion-trap tandem mass spectrometry (ESI-IT-MS(n)). In contrast to the behavior of reported unprotected dipeptide isomers which mainly produce y(1)(+) and/or a(1)(+) ions, the protonated Fmoc-Xxx-Gly-OY, Fmoc-Ala-Xxx-OY and Fmoc-Xxx-Ala-OY yield significant b(1)(+) ions. These ions are formed, presumably with stable protonated aziridinone structures. However, the peptides with Gly- at the N-terminus do not form b(1)(+) ions. The [M+H](+) ions of all the peptides undergo a McLafferty-type rearrangement followed by loss of CO(2) to form [M+H-Fmoc+H](+). The MS(3) collision-induced dissociation (CID) of these ions helps distinguish the pairs of isomeric dipeptides studied in this work. Further, negative ion MS(3) CID has also been found to be useful for differentiating these isomeric peptide acids. The MS(3) of [M-H-Fmoc+H](-) of isomeric peptide acids produce c(1)(-), z(1)(-) and y(1)(-) ions. Thus the present study of Fmoc-protected peptides provides additional information on mass spectral characterization of the dipeptides and distinguishes the positional isomers.

  13. Fmoc/Trt-amino acids: comparison to Fmoc/tBu-amino acids in peptide synthesis.

    PubMed

    Barlos, K; Gatos, D; Koutsogianni, S

    1998-03-01

    Model peptides containing the nucleophilic amino acids Trp and Met have been synthesized with the application of Fmoc/Trt- and Fmoc/tBu-amino acids, for comparison. The deprotection of the peptides synthesized using Fmoc/Trt-amino acids in all cases leads to crude peptides of higher purity than that of the same peptides synthesized using Fmoc/tBu-amino acids.

  14. Carcinogenicity of chloroform.

    PubMed Central

    Reuber, M D

    1979-01-01

    Chloroform is carcinogenic in rats, mice, and probably in dogs. Chloroform induced carcinomas of the liver and kidney and malignant tumors in other organs in rats and mice. Liver neoplasms have been described in three strains of mice. Carcinomas of the kidney were found in a first study in mice and in the repeat of that study. Dogs given chloroform developed neoplasms of the liver as well as in other organs. Rats given chloroform also developed toxic changes, particularly male rats, as a result of treatment. These lesions included interstitial fibrosis of the kidney; polyarteritis of the mesenteric, pancreatic, and other arterioles and arteries; and atrophy of the testes. These toxic changes may have interfered with the development of neoplasms in male rats. PMID:227668

  15. Investigation on enantiomeric separations of fluorenylmethoxycarbonyl amino acids and peptides by high-performance liquid chromatography using native cyclodextrins as chiral stationary phases.

    PubMed

    Tang, Y; Zukowski, J; Armstrong, D W

    1996-09-06

    A systematic study was carried out to investigate enantiomeric separations of fluorenylmethoxycarbonyl (FMOC) amino acids and their peptides. Twenty amino acids were derivatized by 9-fluorenylmethyl chloroformate (FMOC-Cl) and its analogues, FMOC-glycyl-Cl and FMOC-beta-alanyl-Cl. All derivatives were chromatographed on native beta- and gamma-cyclodextrin columns using acetonitrile as the main mobile phase component. The results indicated that glycyl and beta-alanyl groups between FMOC and amino acid moieties enhanced chiral selectivities of amino acid derivatives. The addition of modifiers, triethylamine, acetic acid and methanol, into the mobile phase caused alterations in retention, enantiorecognition and elution order. The structures of amino acids and the type of chiral stationary phase employed exhibited significant impacts on chiral resolutions. It is also found that the number and position of glycyl moieties affect the retentions and enantioselectivities of FMOC derivatized glycyl containing peptides.

  16. Advances in Fmoc solid‐phase peptide synthesis

    PubMed Central

    Behrendt, Raymond; White, Peter

    2016-01-01

    Today, Fmoc SPPS is the method of choice for peptide synthesis. Very‐high‐quality Fmoc building blocks are available at low cost because of the economies of scale arising from current multiton production of therapeutic peptides by Fmoc SPPS. Many modified derivatives are commercially available as Fmoc building blocks, making synthetic access to a broad range of peptide derivatives straightforward. The number of synthetic peptides entering clinical trials has grown continuously over the last decade, and recent advances in the Fmoc SPPS technology are a response to the growing demand from medicinal chemistry and pharmacology. Improvements are being continually reported for peptide quality, synthesis time and novel synthetic targets. Topical peptide research has contributed to a continuous improvement and expansion of Fmoc SPPS applications. Copyright © 2015 European Peptide Society and John Wiley & Sons, Ltd. PMID:26785684

  17. An additional fluorenylmethoxycarbonyl (Fmoc) moiety in di-Fmoc-functionalized L-lysine induces pH-controlled ambidextrous gelation with significant advantages.

    PubMed

    Reddy, Samala Murali Mohan; Shanmugam, Ganesh; Duraipandy, Natarajan; Kiran, Manikantan Syamala; Mandal, Asit Baran

    2015-11-07

    In recent years, several fluorenylmethoxycarbonyl (Fmoc)-functionalized amino acids and peptides have been used to construct hydrogels, which find a wide range of applications. Although several hydrogels have been prepared from mono Fmoc-functionalized amino acids, herein, we demonstrate the importance of an additional Fmoc-moiety in the hydrogelation of double Fmoc-functionalized L-lysine [Fmoc(Nα)-L-lysine(NεFmoc)-OH, (Fmoc-K(Fmoc))] as a low molecular weight gelator (LMWG). Unlike other Fmoc-functionalized amino acid gelators, Fmoc-K(Fmoc) exhibits pH-controlled ambidextrous gelation (hydrogelation at different pH values as well as organogelation), which is significant among the gelators. Distinct fibrous morphologies were observed for Fmoc-K(Fmoc) hydrogels formed at different pH values, which are different from organogels in which Fmoc-K(Fmoc) showed bundles of long fibers. In both hydrogels and organogels, the self-assembly of Fmoc-K(Fmoc) was driven by aromatic π-π stacking and hydrogen bonding interactions, as evidenced from spectroscopic analyses. Characterization of Fmoc-K(Fmoc) gels using several biophysical methods indicates that Fmoc-K(Fmoc) has several advantages and significant importance as a LMWG. The advantages of Fmoc-K(Fmoc) include pH-controlled ambidextrous gelation, pH stimulus response, high thermal stability (∼100 °C) even at low minimum hydrogelation concentration (0.1 wt%), thixotropic property, high kinetic and mechanical stability, dye removal properties, cell viability to the selected cell type, and as a drug carrier. While single Fmoc-functionalized L-lysine amino acids failed to exhibit gelation under similar experimental conditions, the pH-controlled ambidextrous gelation of Fmoc-K(Fmoc) demonstrates the benefit of a second Fmoc moiety in inducing gelation in a LMWG. We thus strongly believe that the current findings provide a lead to construct or design various new synthetic Fmoc-based LMW organic gelators for several

  18. A mild removal of Fmoc group using sodium azide.

    PubMed

    Chen, Chun-Chi; Rajagopal, Basker; Liu, Xuan Yu; Chen, Kuan Lin; Tyan, Yu-Chang; Lin, Fui; Lin, Po-Chiao

    2014-02-01

    A mild method for effectively removing the fluorenylmethoxycarbonyl (Fmoc) group using sodium azide was developed. Without base, sodium azide completely deprotected N (α)-Fmoc-amino acids in hours. The solvent-dependent conditions were carefully studied and then optimized by screening different sodium azide amounts and reaction temperatures. A variety of Fmoc-protected amino acids containing residues masked with different protecting groups were efficiently and selectively deprotected by the optimized reaction. Finally, a biologically significant hexapeptide, angiotensin IV, was successfully synthesized by solid phase peptide synthesis using the developed sodium azide method for all Fmoc removals. The base-free condition provides a complement method for Fmoc deprotection in peptide chemistry and modern organic synthesis.

  19. Chloroform in the endodontic operatory

    SciTech Connect

    McDonald, M.N.; Vire, D.E. )

    1992-06-01

    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 report 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.

  20. 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 ...

  1. Chloroform stripping from waste waters

    SciTech Connect

    Kolev, N.; Darakchiev, R.; Semkov, K.

    1997-01-01

    The problem treated in this paper is the purification of waste industrial waters from chloroform. An industrial installation with a stripping column is designed, and the results of its study and industrial tests are presented. It is shown that, in a column with 6400 mm total height of the used packing (Holpack), the chloroform concentration in the waste water decreases 150,000 times, approaching that of drinking water.

  2. Methyl chloroform and the atmosphere

    SciTech Connect

    Ravishankara, A.R.; Albritton, D.L.

    1995-07-14

    The atmospheric abundance of methyl chloroform, CH{sub 3}CCl{sub 3}, a compound of only anthropogenic origin, is actually decreasing because of emission reductions in compliance with the United Nations Montreal Protocol and its subsequent amendments. This observation, reported by Prinn and co-workers elsewhere in this issue, is based on data from surface-level monitoring stations. The observed trends in methyl chloroform abundance have a few straightforward scientific consequences and substantial policy relevance as discussed in this article. 6 refs., 1 fig.

  3. Application of matrix solid-phase dispersion and high-performance liquid chromatography for determination of sulfonamides in honey.

    PubMed

    Zou, Qiong-Hui; Wang, Jin; Wang, Xiang-Feng; Liu, Yuan; Han, Jie; Hou, Feier; Xie, Meng-Xia

    2008-01-01

    A novel method for simultaneous determination of 8 sulfonamide residues (sulfathiazole, sulfapyridine, sulfadiazine, sulfamerazine, sulfamonome-thoxine, sulfachloropyridazine, sulfamethoxazole, and sulfadimethoxine) in honey samples by high-performance liquid chromatography (HPLC) has been developed on the basis of precolumn derivatization with 9-fluorenylmethyl-chloroformate (FMOC-Cl). Sulfonamide residues in honey samples were extracted and purified by matrix solid-phase dispersion with C18 as the solid support. The residues were derivatized by FMOC-CI, and the FMOC-sulfonamide derivatives were further purified by solid-phase extraction with silica gel as the solid support prior to HPLC analysis. The average recoveries for most sulfonamide compounds at different spiking levels (from 10 to 250 microg/kg) were > 70% with relative standard deviations < 16%, and their limits of detection were 4.0 microg/kg. The established analytical method has high sensitivity and repeatability and can be applicable for determining the sulfonamide residues in various honey matrixes.

  4. Zinc-promoted simple synthesis of oligomer-free N(alpha)-Fmoc-amino acids using Fmoc-Cl as an acylating agent under neutral conditions.

    PubMed

    Gopi, H N; Suresh Babu, V V

    2000-04-01

    A range of N(alpha)-Fmoc-protected amino acids, including those that contain t-butyl moiety, have been synthesized by employing Fmoc-Cl utilizing the activated, commercial zinc dust-promoted synthesis of carbamates under neutral conditions. A general procedure is described that circumvents the oligomerization side reaction normally noticed in Schotten-Baumann conditions. It is a simple, convenient and clean method. Thus, Fmoc-amino acids are obtained in high yield (85-92%) and purity as checked by thin-layer chromatography, high-performance liquid chromatography and other physical methods.

  5. Applicability of LC-MS/MS to optimize derivatization of topiramate with FMOC-Cl using reacted/intact drug ratio.

    PubMed

    Mohammadi, Bahareh; Tammari, Esmail; Fakhri, Sajad; Bahrami, Gholamreza

    2013-06-01

    Topiramate is an antiepileptic agent, which is structurally different from the other anticonvulsants. The drug has no UV-Vis absorption or emits fluorescence. Thus for its analysis using high performance liquid chromatography (HPLC) with conventional UV or fluorescence detectors, the drug should be derivatized with a suitable reagent. In previous study using fluorenylmethyl chloroformate (FMOC-Cl) and HPLC coupled with fluorescence detector, we reported an analytical method for derivatization and analysis of the drug in human serum. In this method, several factors including time and temperature of the reaction, pH and concentration of the used buffer, ratio of organic phase in the medium and removal of the reagent excess by glycine should be optimized to obtain maximum yield of the product. In HPLC coupled with fluorescence detector, there is not any signal from intact topiramate and only the final product (FMOC-topiramate) is appeared. Thus to optimize the reaction conditions for obtaining the highest derived yield, intensity of the final product peak is considered as a criteria for progression of the reaction. In LC-MS/MS system however, both free and reacted topiramate are visible in observed spectra. In the present study reaction of the drug with FMOC-Cl was re-optimized using LC-MS/MS technique on the basis of reacted/free topiramate ratio as the new and more accurate index. The results showed that, ratio of organic/aqueous phase has a dominant effect on the reaction, the most efficient temperature is 70°C and the reaction is reversed following addition of the glycine.

  6. 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.

  7. The 2-(triphenylsilyl)ethoxycarbonyl-("Tpseoc"-) group: a new silicon-based, fluoride cleavable oxycarbonyl protecting group highly orthogonal to the Boc-, Fmoc- and Cbz-groups.

    PubMed

    Golkowski, Martin; Ziegler, Thomas

    2011-06-07

    Starting from 2-(triphenylsilyl)ethanol a new oxycarbonyl protecting group cleavable by fluoride ion induced Peterson-elimination has been developed. Known 2-(triphenylsilyl)ethanol has been prepared from commercially available triphenylvinyl-silane by a hydroboration-oxidation sequence using the sterically hindered borane reagent 9-BBN. The silyl alcohol was subsequently transformed into its chloroformate, imidazolylcarboxylic acid ester and p-nitrophenyl carbonate and used in standard protocols for the formation of carbamates and carbonates. The Tpseoc group proved to be highly resistant against acidic conditions applied in removal of tert-butyl esters and the t-Boc-group. It also withstood catalytic hydrogenation, treatment with morpholine, methylhydrazine and Pd-reagents/allyl-scavanger combinations, conditions required to cleave Cbz-, Fmoc-, phthalimide- and Alloc-groups. The Tpseoc-group is cleaved upon treatment with TBAF/CsF at 0 °C or r.t. with cleavage times reaching from.

  8. Rapid separation of phosphopeptides by microchip electrophoresis-electrospray ionization mass spectrometry.

    PubMed

    Ollikainen, Elisa; Bonabi, Ashkan; Nordman, Nina; Jokinen, Ville; Kotiaho, Tapio; Kostiainen, Risto; Sikanen, Tiina

    2016-04-01

    Protein phosphorylation is a significant biological process, but separation of phosphorylated peptide isomers is often challenging for many analytical techniques. We developed a microchip electrophoresis (MCE) method for rapid separation of phosphopeptides with on-chip electrospray ionization (ESI) facilitating online sample introduction to the mass spectrometer (MS). With the method, two monophosphorylated positional isomers of insulin receptor peptide (IR1A and IR1B) and a triply phosphorylated insulin receptor peptide (IR3), all with the same amino acid sequence, were separated from the nonphosphorylated peptide (IR0) in less than one minute. For efficient separation of the positional peptide isomers from each other derivatization with 9-fluorenylmethyl reagents (either chloroformate, Fmoc-Cl, or N-succinimidyl carbonate, Fmoc-OSu) was required before the analysis. The derivatization improved not only the separation of the monophosphorylated positional peptide isomers in MCE, but also identification of the phosphorylation site based on MS/MS.

  9. Fmoc-Cl fluorescent determination for amino groups of nanomaterial science.

    PubMed

    Zhang, Y; Chen, Y

    2012-06-01

    With the wide application of nanomaterials, the quantification of functional groups on nanomaterial surface becomes more and more necessary. A heterogeneous 9-fluorenylmethoxy carbonyl chloride (Fmoc-Cl) fluorescent method using an aqueous solution was established to determinate amino groups on nanomaterial surface. The effect factors of determination were investigated and the assay was optimised. The Fmoc fluorescent method is 200-fold more sensitive than the current UV assay using an organic solvent, and compared with chemical ninhydrin method and physical elemental analysis. Heterogeneous Fmoc-Cl fluorescent method can be used to determine amino groups on nanomaterials with big size, which is difficult to undergo a direct detection.

  10. Convenient and Scalable Synthesis of Fmoc-Protected Peptide Nucleic Acid Backbone

    PubMed Central

    Feagin, Trevor A.; Shah, Nirmal I.; Heemstra, Jennifer M.

    2012-01-01

    The peptide nucleic acid backbone Fmoc-AEG-OBn has been synthesized via a scalable and cost-effective route. Ethylenediamine is mono-Boc protected, then alkylated with benzyl bromoacetate. The Boc group is removed and replaced with an Fmoc group. The synthesis was performed starting with 50 g of Boc anhydride to give 31 g of product in 32% overall yield. The Fmoc-protected PNA backbone is a key intermediate in the synthesis of nucleobase-modified PNA monomers. Thus, improved access to this molecule is anticipated to facilitate future investigations into the chemical properties and applications of nucleobase-modified PNA. PMID:22848796

  11. 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.

  12. 2-Chlorotrityl chloride resin. Studies on anchoring of Fmoc-amino acids and peptide cleavage.

    PubMed

    Barlos, K; Chatzi, O; Gatos, D; Stavropoulos, G

    1991-06-01

    The esterification of 2-chlorotrityl chloride resin with Fmoc-amino acids in the presence of DIEA is studied under various conditions. High esterification yields are obtained using 0.6 equiv. Fmoc-amino acid/mmol resin in DCM or DCE, in 25 min, at room temperature. The reaction proceeds without by product formation even in the case of Fmoc-Asn and Fmoc-Gln. The quantitative and easy cleavage of amino acids and peptides from 2-chlorotrityl resin, by using AcOH/TFE/DCM mixtures, is accomplished within 15-60 min at room temperature, while t-butyl type protecting groups remain unaffected. Under these exceptionally mild conditions 2-chlorotrityl cations generated during the cleavage of amino acids and peptides from resin do not attack the nucleophilic side chains of Trp, Met, and Tyr.

  13. Ultrashort self-assembling Fmoc-peptide gelators for anti-infective biomaterial applications.

    PubMed

    McCloskey, Alice P; Draper, Emily R; Gilmore, Brendan F; Laverty, Garry

    2017-02-01

    Biomaterial-related infections have a significant impact on society and are a major contributor to the growing threat of antimicrobial resistance. Current licensed antibiotic classes struggle to breakdown or penetrate the exopolysaccharide biofilm barrier, resulting in sub-therapeutic concentrations of antibiotic at the surface of the biomaterial, treatment failure and increased spread of resistant isolates. This paper focuses for the first time on the ability of ultrashort Fmoc-peptide gelators to eradicate established bacterial biofilms implicated in a variety of medical device infections (Gram-positive: Staphylococcus aureus, Staphylococcus epidermidis and Gram-negative Escherichia coli, Pseudomonas aeruginosa). The effect of increasing the cationicity of FmocFF via addition of di-lysine and di-orntithine was also studied with regard to antibacterial activity. Our studies demonstrated that Fmoc-peptides (FmocFF, FmocFFKK, FmocFFFKK, FmocFFOO) formed surfactant-like soft gels at concentrations of 1% w/v and above using a method of glucono-δ-Lactone pH induction. The majority of Fmoc-peptides (0.5-2% w/v) demonstrated selective action against established (grown for 24 h) biofilms of Gram-positive and Gram-negative pathogens. These results are likely to increase the clinical translation of short-peptide gelator platforms within the area of anti-infective biomaterials including as wound dressings and coatings for prostheses, catheters, heart valves and surgical tubes. In the long term, this will lead to wider treatment choices for clinicians and patients involved in the management of medical device infections and reduce the burden of antimicrobial resistance. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.

  14. The effect of calcium chloride concentration on alginate/Fmoc-diphenylalanine hydrogel networks.

    PubMed

    Çelik, Ekin; Bayram, Cem; Akçapınar, Rümeysa; Türk, Mustafa; Denkbaş, Emir Baki

    2016-09-01

    Peptide based hydrogels gained a vast interest in the tissue engineering studies thanks to great superiorities such as biocompatibility, supramolecular organization without any need of additional crosslinker, injectability and tunable nature. Fmoc-diphenylalanine (FmocFF) is one of the earliest and widely used example of these small molecule gelators that have been utilized in biomedical studies. However, Fmoc-peptides are not feasible for long term use due to low stability and weak mechanical properties at neutral pH. In this study, Fmoc-FF dipeptides were mechanically enhanced by incorporation of alginate, a biocompatible and absorbable polysaccharide. The binary hydrogel is obtained via molecular self-assembly of FmocFF dipeptide in alginate solution followed by ionic crosslinking of alginate moieties with varying concentrations of calcium chloride. Hydrogel characterization was evaluated in terms of morphology, viscoelastic moduli and diffusional phenomena and the structures were tested as 3D scaffolds for bovine chondrocytes. In vitro evaluation of scaffolds lasted up to 14days and cell viability, sulphated glycosaminoglycan (sGAG) levels, collagen type II synthesis were determined. Our results showed that alginate incorporation into FmocFF hydrogels leads to better mechanical properties and higher stability with good biocompatibility.

  15. Fmoc-modified amino acids and short peptides: simple bio-inspired building blocks for the fabrication of functional materials.

    PubMed

    Tao, Kai; Levin, Aviad; Adler-Abramovich, Lihi; Gazit, Ehud

    2016-07-11

    Amino acids and short peptides modified with the 9-fluorenylmethyloxycarbonyl (Fmoc) group possess eminent self-assembly features and show distinct potential for applications due to the inherent hydrophobicity and aromaticity of the Fmoc moiety which can promote the association of building blocks. Given the extensive study and numerous publications in this field, it is necessary to summarize the recent progress concerning these important bio-inspired building blocks. Therefore, in this review, we explore the self-organization of this class of functional molecules from three aspects, i.e., Fmoc-modified individual amino acids, Fmoc-modified di- and tripeptides, and Fmoc-modified tetra- and pentapeptides. The relevant properties and applications related to cell cultivation, bio-templating, optical, drug delivery, catalytic, therapeutic and antibiotic properties are subsequently summarized. Finally, some existing questions impeding the development of Fmoc-modified simple biomolecules are discussed, and corresponding strategies and outlooks are suggested.

  16. CHARMM force field parameterization protocol for self-assembling peptide amphiphiles: the Fmoc moiety.

    PubMed

    Ramos Sasselli, I; Ulijn, R V; Tuttle, T

    2016-02-14

    Aromatic peptide amphiphiles are known to self-assemble into nanostructures but the molecular level structure and the mechanism of formation of these nanostructures is not yet understood in detail. Molecular dynamic simulations using the CHARMM force field have been applied to a wide variety of peptide-based systems to obtain molecular level details of processes that are inaccessible with experimental techniques. However, this force field does not include parameters for the aromatic moieties which dictate the self-assembly of these systems. The standard CHARMM force field parameterization protocol uses hydrophilic interactions for the non-bonding parameters evaluation. However, to effectively reproduce the self-assembling behaviour of these molecules, the balance between the hydrophilic and hydrophobic nature of the molecule is essential. In this work, a modified parameterization protocol for the CHARMM force field for these aromatic moieties is presented. This protocol is applied for the specific case of the Fmoc moiety. The resulting set of parameters satisfies the conformational and interactions analysis and is able to reproduce experimental results such as the Fmoc-S-OMe water/octanol partition free energy and the self-assembly of Fmoc-S-OH and Fmoc-Y-OH into spherical micelles and fibres, respectively, while also providing detailed information on the mechanism of these processes. The effectiveness of the parameters for the Fmoc moiety validates the protocol as a robust approach to paramterise this class of compounds.

  17. Investigating the effects of peptoid substitutions in self-assembly of Fmoc-diphenylalanine derivatives.

    PubMed

    Rajbhandary, Annada; Nilsson, Bradley L

    2017-03-01

    Low molecular weight agents that undergo self-assembly into fibril networks with hydrogel properties are promising biomaterials. Most low molecular weight hydrogelators are discovered empirically or serendipitously due to imperfect understanding of the mechanisms of self-assembly, the packing structure of self-assembled materials, and how the self-assembly process corresponds to emergent hydrogelation. Herein, the mechanisms of self-assembly and hydrogelation of N-fluorenylmethoxycarbonyl diphenylalanine (Fmoc-PhePhe), a well-studied low molecular weight hydrogelator, is probed by systematic comparison with derivatives in which Phe residues are replaced by corresponding N-benzyl glycine peptoid (Nphe) analogs. Peptoids are peptidomimetics that shift display of side chain functionality from the α-carbon to the terminal nitrogen. This alters the hydrogen bonding capacity, the side chain presentation geometry, amide cis/trans isomerization equilibrium, and β-sheet potential of the peptoid relative to the corresponding amino acid in the context of peptidic polymers. It was found that amino acid/peptoid hybrids Fmoc-Phe-Nphe and Fmoc-Nphe-Phe have altered fibril self-assembly propensity and reduced hydrogelation capacity relative to the parent dipeptide, and that fibril self-assembly of the dipeptoid, Fmoc-Nphe-Nphe, is completely curtailed. These findings provide insight into the potential of low molecular weight peptoids and peptide/peptoid hybrids as hydrogelation agents and illuminate the importance of hydrogen bonding and π-π interaction geometry in facilitating self-assembly of Fmoc-Phe-Phe.

  18. Chloroform uptake by gutta-percha and assessment of its concentration in air during the chloroform-dip technique.

    PubMed

    Margelos, J; Verdelis, K; Eliades, G

    1996-10-01

    The use of chloroform as an adjunct to the practice of endodontics has been a matter of debate. In the present study the chloroform uptake of gutta-percha cones was determined by a gravimetric assay for different times of chloroform dip. In conjunction with an assessment of the amount of gutta-percha dissolved during dip, this provided an estimate of the amount of chloroform that patients are exposed to in clinical conditions. An assay was also performed of the chloroform concentration in the air in a dental office. Chloroform uptake was shown to increase with an increasing dipping time. There also seems to be a difference in this uptake between pure chloroform and a chloroform preparation with colophonium. The concentration levels of chloroform evaporated during the practice of chloroform dip within a dental office do not exceed the safety limits.

  19. Study of the matrix effects and sample dilution influence on the LC-ESI-MS/MS analysis using four derivatization reagents.

    PubMed

    Oldekop, Maarja-Liisa; Herodes, Koit; Rebane, Riin

    2014-09-15

    For liquid chromatographic analysis of amino acids involving derivatization and mass-spectrometric detection, it becomes more important to evaluate the presence of matrix effects in complex samples. This is somewhat complicated for amino acid analysis where analyte free sample matrix is often unavailable. In this work, matrix effects were investigated using post-column infusion method for 9-fluorenylmethyl chloroformate (FMOC-Cl) derivatives of β-Ala, Gly and Phe and diethyl ethoxymethylenemalonate (DEEMM) derivative of β-Ala. While for DEEMM derivatives, the main signal suppression was due to the borate buffer, in case of FMOC-Cl, other FMOC-derivatives caused signal suppression. Analysis of amino acids in tea and honey with DEEMM, FMOC-Cl, p-N,N,N-trimethylammonioanilyl N'-hydroxysuccinimidyl carbamate iodide (TAHS) and dansyl chloride (DNS) showed that amino acid concentrations found with different reagents do not agree well. Sample dilution experiments indicated that the sample matrix affected the analysis results obtained with DEEMM the least, but with FMOC-Cl, TAHS and DNS, sample dilution had an influence on the results. When sample dilution and extrapolative dilution approach were applied on the latter results, an agreement of amino acid concentrations measured with different reagents was achieved within relative standard deviation (RSD) of 22% for most cases.

  20. Designing unconventional Fmoc-peptide-based biomaterials: structure and related properties.

    PubMed

    Chronopoulou, Laura; Sennato, Simona; Bordi, Federico; Giannella, Domenico; Di Nitto, Antonio; Barbetta, Andrea; Dentini, Mariella; Togna, Anna Rita; Togna, Giuseppina Ines; Moschini, Sabina; Palocci, Cleofe

    2014-03-28

    We have recently employed L-amino acids in the lipase-catalyzed biofabrication of a class of self-assembling Fmoc-peptides that form 3-dimensional nanofiber scaffolds. Here we report that using d-amino acids, the homochiral self-assembling peptide Fmoc-D-Phe3 (Fmoc-F*F*F*) also forms a 3-dimensional nanofiber scaffold that is substantially distinguishable from its L-peptide and heterochiral peptide (F*FF and FF*F*) counterparts on the basis of their physico-chemical properties. Such chiral peptides self-assemble into ordered nanofibers with well defined fibrillar motifs. Circular dichroism and atomic force microscopy have been employed to study in depth such fibrillar peptide structures. Dexamethasone release kinetics from PLGA and CS-PLGA nanoparticles entrapped within the peptidic hydrogel matrix encourage its use for applications in drug controlled release.

  1. Comparative dynamics of methionine side-chain in FMOC-methionine and in amyloid fibrils

    NASA Astrophysics Data System (ADS)

    Vugmeyster, Liliya; Ostrovsky, Dmitry

    2017-04-01

    We compared the dynamics of key methionine methyl groups in the water-accessible hydrophobic cavity of amyloid fibrils and Fluorenylmethyloxycarbonyl-Methionine (FMOC-Met), which renders general hydrophobicity to the environment without the complexity of the protein. Met35 in the hydrated cavity was recently found to undergo a dynamical cross-over from the dominance of methyl rotations at low temperatures to the dominance of diffusive motion of methyl axis at high temperatures. Current results indicate that in FMOC-Met this cross-over is suppressed, similar to what was observed for the dry fibrils, indicating that hydration of the cavity is driving the onset of the dynamical transition.

  2. Agglomeration and Sedimentation of MWCNTS in Chloroform

    NASA Astrophysics Data System (ADS)

    Eremin, Yu. S.; Kolesnikova, A. A.; Grekhov, A. M.

    The kinetics of agglomeration of multiwalled carbon nanotubes dispersed in chloroform has been studied by the methods of optical spectroscopy and dynamic light scattering. With the use of the models of the diffusion of cylindrical particles, the sizes of particles obtained by this method can be recalculated to the DLS data and the concentration at which the dispersion of individual МWCNTs occurs can be determined.

  3. SYNTHESIS OF PROTEINS BY NATIVE CHEMICAL LIGATION USING FMOC-BASED CHEMISTRY

    SciTech Connect

    Camarero, J A; Mitchell, A R

    2005-01-20

    C-terminal peptide {alpha}-thioesters are valuable intermediates in the synthesis/semisynthesis of proteins by native chemical ligation. They are prepared either by solid-phase peptide synthesis (SPPS) or biosynthetically by protein splicing techniques. The present paper reviews the different methods available for the chemical synthesis of peptide {alpha}-thioesters using Fmoc-based SPPS.

  4. Scope and Limitations of Fmoc Chemistry SPPS-Based Approaches to the Total Synthesis of Insulin Lispro via Ester Insulin.

    PubMed

    Dhayalan, Balamurugan; Mandal, Kalyaneswar; Rege, Nischay; Weiss, Michael A; Eitel, Simon H; Meier, Thomas; Schoenleber, Ralph O; Kent, Stephen B H

    2017-01-31

    We have systematically explored three approaches based on 9-fluorenylmethoxycarbonyl (Fmoc) chemistry solid phase peptide synthesis (SPPS) for the total chemical synthesis of the key depsipeptide intermediate for the efficient total chemical synthesis of insulin. The approaches used were: stepwise Fmoc chemistry SPPS; the "hybrid method", in which maximally protected peptide segments made by Fmoc chemistry SPPS are condensed in solution; and, native chemical ligation using peptide-thioester segments generated by Fmoc chemistry SPPS. A key building block in all three approaches was a Glu[O-β-(Thr)] ester-linked dipeptide equipped with a set of orthogonal protecting groups compatible with Fmoc chemistry SPPS. The most effective method for the preparation of the 51 residue ester-linked polypeptide chain of ester insulin was the use of unprotected peptide-thioester segments, prepared from peptide-hydrazides synthesized by Fmoc chemistry SPPS, and condensed by native chemical ligation. High-resolution X-ray crystallography confirmed the disulfide pairings and three-dimensional structure of synthetic insulin lispro prepared from ester insulin lispro by this route. Further optimization of these pilot studies could yield an efficient total chemical synthesis of insulin lispro (Humalog) based on peptide synthesis by Fmoc chemistry SPPS.

  5. A new on-line, in-tube pre-column derivatization technique for high performance liquid chromatographic determination of azithromycin in human serum.

    PubMed

    Bahrami, Gholamreza; Mohammadi, Bahareh

    2006-01-18

    Pre-column derivatization methods for high performance liquid chromatographic assay of specific pharmaceutical agents using 9-fluorenylmethyl chloroformate (FMOC-Cl) have received special attention because highly fluorescent and stable adducts are provided by these methods. However, unlike the post-column on-line techniques, long derivatization time is needed and the reaction cannot be well controlled. A new, sensitive and fast pre-column on-line derivatization technique coupled with high-performance liquid chromatography using FMOC-Cl as labeling agent is described and validated for determination of azithromycin in human serum. After extraction of the drug from serum, the residue was reconstituted in mixture of acetonitrile-phosphate buffer (3:1, v/v; pH 8.5) and directly injected onto the chromatographic system. Continuous on-line derivatization and analysis of the compounds were successfully performed using in-tube elution of FMOC-Cl. The total time needed for derivatization and chromatographic analysis of the drug was 13 min. The assay was reliable and reproducible, with limit of quantification of 10 ng/ml. The described technique may offer significant advantages over existing off-line derivatization methods using FMOC-Cl.

  6. Review of the mutagenicity of chloroform

    SciTech Connect

    Rosenthal, S.L.

    1987-01-01

    Although chloroform (CHCl/sub 3/) is metabolized in vivo and in vitro to a substance that covalently interacts with protein and lipid, its potential for binding to DNA is low. In addition, most of the assays for genotoxicity are negative. However, many of the genotoxicity results are inconclusive because of inadequacies in the experimental protocols. The types of genotoxicity tests this report is based on include bacterial, yeast, host-mediated, Drosophila sex-linked recessive lethal, mammalian cell mutagenicity, sperm head abnormality, cytogenetic, and DNA damage. On the basis of presently available information, no definitive conclusion on the mutagenic potential of CHCl/sub 3/ can be reached.

  7. Ultra-high pressure LC determination of glucosamine in shrimp by-products and migration tests of chitosan films.

    PubMed

    Sanches-Silva, Ana; Ribeiro, Tiago; Albuquerque, Tânia G; Paseiro, Perfecto; Sendón, Raquel; de Quirós, Ana Bernaldo; López-Cervantes, Jaime; Sánchez-Machado, Dalia I; Valdez, Herlinda Soto; Angulo, Inmaculada; Aurrekoetxea, Goizane P; Costa, Helena S

    2012-03-01

    Chitosan, a multiple applications molecule, was isolated from shrimp by-products by fermentation. The amount of chitosan in the solid fraction of the fermented extract was measured after its conversion in the respective glucosamine units. The procedure includes an acid hydrolysis (110 °C, 4 h with HCl 8 M) and a derivatization with 9-fluorenylmethyl chloroformate (Fmoc-Cl). Ultra-high-pressure liquid chromatography method was developed and optimized. Excellent peaks resolution was achieved in just 10 min. The method was evaluated in what concerns to validation parameters such as linearity, repeatability, quantification limit, and recovery. Migration tests of films prepared with chitosan were carried out in two simulants: ultrapure water and ethanol 95% (v/v).

  8. Determination of roxithromycin in human plasma by HPLC with fluorescence and UV absorbance detection: application to a pharmacokinetic study.

    PubMed

    Główka, Franciszek K; Karaźniewicz-Łada, Marta

    2007-06-01

    A selective HPLC method with fluorescence detection for the determination of roxithromycin (ROX) in human plasma was described. After solid-phase extraction (SPE), ROX and erythromycin (internal standard, I.S.) were derivatized by treatment with 9-fluorenylmethyl chloroformate (FMOC-Cl). Optimal resolution of fluorescence derivatives of ROX and I.S. was obtained during one analytical run using reversed phase, C(18) column. The mobile phase was composed of potassium dihydrogenphosphate solution, pH 7.5 and acetonitrile. Fluorescence of the compounds was measured at the maximum excitation, 255 nm and emission, 313 nm, of ROX derivatives. Validation parameters of the method were also established. After SPE, differences in recoveries of ROX and erythromycin from human plasma were observed. The linear range of the standard curve of ROX in plasma was 0.5-10.0 mg/l. The validated method was successfully applied for pharmacokinetic studies of ROX after administration of a single tablet of ROX.

  9. High-performance liquid chromatographic determination of vertilmicin in rat plasma using sensitive fluorometric derivatization.

    PubMed

    Liu, Zhen; Sha, Yunfei; Huang, Taomin; Yang, Bei; Duan, Geng-Li

    2005-12-15

    A sensitive and reliable high-performance liquid chromatographic method was developed for the determination of vertilmicin in rat plasma. Derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) followed by C(18) reversed-phase chromatography allowed the fluorimetric detection of vertilmicin. Optimal conditions for the derivatization of vertilmicin are described. The limit of quantification was 0.02 mg/L. The pharmacokinetics of vertilmicin was studied in 24 rats following intramuscular injection (i.m.) of different doses (4, 8, 16, 32 mg/kg of body weight). The pharmacokinetic parameter values were estimated by use of 3P97 program. In this study, we assessed the dose proportionality of vertilmicin after single intramuscular injection doses and obtained new information on the pharmacokinetics of the compound.

  10. High-performance liquid chromatographic determination of memantine hydrochloride in rat plasma using sensitive fluorometric derivatization.

    PubMed

    Xie, Mei-Fen; Zhou, Wei; Tong, Xin-Yi; Chen, Yi-Le; Cai, Yi; Li, Yan; Duan, Geng-Li

    2011-02-01

    In this study, we investigated a simple, sensitive and reliable liquid chromatography-fluorescence detection method for the determination of memantine hydrochloride in rat plasma which was based on derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl). For the first time, FMOC-Cl was introduced into derivatization of memantine hydrochloride in rat plasma. The amino groups of memantine hydrochloride and amantadine hydrochloride (internal standard) were trapped with FMOC-Cl to form memantine hydrochloride-FMOC-Cl and amantadine hydrochloride-FMOC-Cl compositions, which can be very compatible for LC-FLD. Precipitation of plasma proteins by acetonitrile was followed by vortex mixing and centrifugation. Chromatographic separation was performed on a C(18) column (DIAMONSIL 150 × 4.6 mm, id 5 μm) with a mobile phase consisting of acetonitrile and water at a flow rate of 1.0 mL/min. The retention times of memantine hydrochloride-FMOC-Cl and amantadine hydrochloride-FMOC-Cl compositions were 23.69 and 40.27 min, respectively. Optimal conditions for the derivatization of memantine hydrochloride were also described. The limit of quantification (LOQ) was 25 ng/mL for memantine hydrochloride in plasma, the linear range was 0.025-5.0 μg/mL in plasma with a correlation coefficient (r) of 0.9999. The relative standard deviations (RSDs) of intra-day and inter-day assays were 4.46-12.19 and 5.23-11.50%, respectively. The validated method was successfully applied to the determination of memantine hydrochloride in rat plasma samples.

  11. DBU-catalyzed transprotection of N-Fmoc-cysteine di- and tripeptides into S-Fm-cysteine di- and tripeptides.

    PubMed

    Katritzky, Alan R; Abo-Dya, Nader E; Abdelmajeid, Abdelmotaal; Tala, Srinivasa R; Amine, M S; El-Feky, Said A

    2011-01-21

    The transprotection of N-Fmoc-cysteine containing di- and tripeptides possessing a free SH group to produce the corresponding S-Fm-cysteine di- and tripeptides bearing a free amino group is accomplished efficiently with DBU in dry THF. The N-Fmoc to S-Fm transformation mechanism is discussed. S-Fm-Cysteine di- and tripeptides readily form amide bonds on coupling with N-(Pg-α-aminoacyl)benzotriazoles and N-(Pg-α-dipeptidoyl)benzotriazoles to give larger peptides.

  12. Supramolecular Hydrogels from Self-Assembly of di-Fmoc-L-lysine

    NASA Astrophysics Data System (ADS)

    Hashemnejad, Seyed Meysam; Naas, Kinsey; Kundu, Santanu

    Mechanical properties and nanostructure of a supramolecular hydrogel formed by self-assembly of di-fluorenylmethyloxycarbonyl-lysine (di-Fmoc-L-lysine) are reported here. Hydrogels were prepared by solvent switch technique in which water was added to a solution of di-Fmoc-L-lysine in dimethyl sulfoxide (DMSO). Mechanical properties of the gels were investigated using shear and cavitation rheology. The gels display strain-softening behavior at moderate strain values. Morphological investigations of the samples were conducted using FTIR and CD spectroscopy, electron microscopy, and atomic force microscopy (AFM). Self-assembled fibers with lateral dimensions ranging from 10 to 50 nm were captured in microscopy studies. FTIR results indicate β-sheet-like conformation of the peptides in the hydrogel.

  13. Preparation and characterization of a novel sodium alginate incorporated self-assembled Fmoc-FF composite hydrogel.

    PubMed

    Gong, Xiao; Branford-White, Christopher; Tao, Lei; Li, Shubai; Quan, Jing; Nie, Huali; Zhu, Limin

    2016-01-01

    Dipeptides and their derivatives have attracted tremendous attention owning to their excellent abilities of self-assemble assembling into various structures which have great potentials for applications in biology and/or nanotechnology. In the present study, we dedicate to fabricate a rigid and structure controllable Fmoc-FF/SA composite hydrogel. We found that the modified dipeptide, fluorenyl-9-methoxycarbonyl (Fmoc)-diphenylalanine (Phe-Phe) can self-assemble into rigid hydrogels with structures of nanowires, layered thin films or honeycombs as the change of sodium alginate (SA) concentration. Meanwhile, CD-spectroscopy demonstrated that SA appeared to control the process, but it did not change the arrangement of the Fmoc-FF peptide. Our results demonstrated that the formed hydrogel showed physical and chemical stability as well as possessing good biocompatibility. Rheological measurements showed that the addition of SA could improve the stability of the hydrogel. Cell viability assay revealed that the Fmoc-FF and Fmoc-FF/SA hydrogels are both beneficial for cell proliferation in-vitro. Our results indicated that the fabricated Fmoc-FF/SA composite hydrogels could be used in tissue engineering and drug delivery in the future.

  14. Chronic toxicity of chloroform to Japanese medaka fish.

    PubMed Central

    Toussaint, M W; Rosencrance, A B; Brennan, L M; Beaman, J R; Wolfe, M J; Hoffmann, F J; Gardner, H S

    2001-01-01

    Japanese medaka (Oryzias latipes) were continually exposed in a flow-through diluter system for 9 months to measured chloroform concentrations of 0.017, 0.151, or 1.463 mg/L. Parameters evaluated were hepatocarcinogenicity, hepatocellular proliferation, hematology, and intrahepatic chloroform concentration. Histopathology was evaluated at 6 and 9 months. Chloroform was not hepatocarcinogenic to the medaka at the concentrations tested. Chronic toxicity was evidenced at these time points by statistically significant ([alpha] = 0.05) levels of gallbladder lesions and bile duct abnormalities in medaka treated with 1.463 mg/L chloroform. We assessed hepatocellular proliferation by exposing test fish to 5-bromo-2'-deoxyuridine in the aquarium water for 72 hr after 4 and 20 days of chloroform exposure; we then quantified area-labeling indices of the livers using computer-assisted image analysis. We observed no treatment-related increases in cellular proliferation. We analyzed cells in circulating blood in medaka after 6 months of chloroform exposure. Hematocrit, leukocrit, cell viability, and cell counts of treated fish were not significantly different from those of control fish. Using gas chromatography (GC), we evaluated intrahepatic concentrations of chloroform in fish after 9 months of exposure. Livers from the 0.151 and 1.463 mg/L chloroform-treated fish had detectable amounts of chloroform, but these levels were always lower than the aquaria concentrations of chloroform. Thus, it appeared that chloroform did not bioaccumulate in the liver. Unidentified presumptive metabolite peaks were found in the GC tracings of these fish livers. PMID:11171522

  15. Determination of atmospheric amines by on-fiber derivatization solid-phase microextraction with 2,3,4,5,6-pentafluorobenzyl chloroformate and 9-fluorenylmethoxycarbonyl chloride.

    PubMed

    Parshintsev, Jevgeni; Rönkkö, Tuukka; Helin, Aku; Hartonen, Kari; Riekkola, Marja-Liisa

    2015-01-09

    Alkylamines play an important role in atmospheric chemistry and are of concern for human health. Determining them from the vapor phase is challenging owing to their high polarity and volatility, water solubility, low concentrations, and poor chromatographic properties. We propose on-fiber derivatization solid-phase microextraction (SPME) to increase sensitivity and selectivity for the determination of alkylamines in air samples. SPME fibers coated in head-space with 2,3,4,5,6-pentafluorobenzyl chloroformate (PFBCF, 10min) or 9-fluorenylmethoxycarbonyl (FMOC) chloride (5min) were exposed to the sample for 5-120min, after which the derivatized alkylamines were thermally desorbed in the GC injection port and analyzed by GC-MS. The specific focus of the research was dimethylamine (DMA) but, as well as secondary amines, both coating agents readily react with primary and tertiary amines and with ammonia at ambient temperatures. The fiber coating procedures, sampling times, and analytical conditions were optimized, and methods were tested with natural samples. PFBCF was more selective and almost an order of magnitude more sensitive than FMOC chloride. Both reagents are applicable, however, depending on the requirements. With scan mode and use of molecular ion for quantification, the limit of quantification for DMA was 0.17μgL(-1) when derivatized with PFBCF and 3.4μgL(-1) when derivatized with FMOC chloride. When selected ion monitoring was used with the most abundant ion, the limit of quantification for DMA was 2.8ngL(-1). Intermediate reproducibility expressed as relative standard deviation was around 30% with PFBCF and less than 20% with FMOC chloride. Fibers coated with PFBCF could be used at least up to 24h when stored at 4°C and for 5 to 7h when stored at room temperature. After sampling/derivatization, storage time before analysis should not exceed 48h at 4°C or 24h at room temperature. At maximum, the PFBCF-coated fiber can be used in 100 coating

  16. 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...

  17. 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.

  18. Biotrickling filtration of complex pharmaceutical VOC emissions along with chloroform.

    PubMed

    Balasubramanian, P; Philip, Ligy; Murty Bhallamudi, S

    2012-06-01

    Biodegradation of chloroform along with a mixture of VOCs (methanol, ethanol, acetone and toluene) commonly found in pharmaceutical emissions using a biotrickling filter (BTF) was evaluated. The performance of the BTF was evaluated for both steady and transient conditions, for different inlet loading rates (ILR), empty bed residence time (EBRT) and inlet chloroform concentrations. Among the VOCs studied before chloroform feeding, toluene removal was the least, under all the operating conditions. Complete removal of all pollutants was achieved up to a chloroform loading rate of 14.22 g/m(3)/h. Increase in loading rate of chloroform adversely affected the removal efficiency of toluene and declined the overall performance of BTF. The results suggest that biodegradation of VOCs is influenced by the inlet loading rate and complexity of pollutants in the inlet air stream. Results from studies on shock loading and starvation indicated that the system was highly resilient to transient operating conditions.

  19. Chloroform mineralization by toluene-oxidizing bacteria.

    PubMed Central

    McClay, K; Fox, B G; Steffan, R J

    1996-01-01

    Seven toluene-oxidizing bacterial strains (Pseudomonas mendocina KR1, Burkholderia cepacia G4, Pseudomonas putida F1, Pseudomonas pickettii PKO1, and Pseudomonas sp. strains ENVPC5, ENVBF1, and ENV113) were tested for their ability to degrade chloroform (CF). The greatest rate of CF oxidation was achieved with strain ENVBF1 (1.9 nmol/min/mg of cell protein). CF also was oxidized by P. mendocina KR1 (0.48 nmol/min/mg of cell protein), strain ENVPC5 (0.49 nmol/min/mg of cell protein), and Escherichia coli DH510B(pRS202), which contained cloned toluene 4-monooxygenase genes from P. mendocina KR1 (0.16 nmol/min/mg of cell protein). Degradation of [14C]CF and ion analysis of culture extracts revealed that CF was mineralized to CO2 (approximately 30 to 57% of the total products), soluble metabolites (approximately 15%), a total carbon fraction irreversibly bound to particulate cellular constituents (approximately 30%), and chloride ions (approximately 75% of the expected yield). CF oxidation by each strain was inhibited in the presence of trichloroethylene, and acetylene significantly inhibited trichloroethylene oxidation by P. mendocina KR1. Differences in the abilities of the CF-oxidizing strains to degrade other halogenated compounds were also identified. CF was not degraded by B. cepacia G4, P. putida F1, P. pickettii PKO1, Pseudomonas sp. strain ENV113, or P. mendocina KRMT, which contains a tmo mutation. PMID:8702263

  20. Koetjapic acid chloro-form hemisolvate.

    PubMed

    Nassar, Z D; Aisha, A F A; Majid, A M S Abdul; Yeap, Chin Sing; Fun, Hoong-Kun

    2010-05-08

    The asymmetric unit of the title compound, C(30)H(46)O(4)·0.5CHCl(3), consists of one koetjapic acid [systematic name: (3R,4aR,4bS,7S,8S,10bS,12aS)-7-(2-carboxy-ethyl)-3,4b,7,10b,12a-penta-methyl-8-(prop-1-en-2-yl)-1,2,3,4,4a,4b,5,6,7,8,9,10,10b,11,12,12a-hexa-deca-hydro-chrysene-3-carboxylic acid] mol-ecule and one half-mol-ecule of chloro-form solvent, which is disordered about a twofold rotation axis. The symmetry-independent component is further disordered over two sites, with occupancies of 0.30 and 0.20. The koetjapic acid contains a fused four-ring system, A/B/C/D. The A/B, B/C and C/D junctions adopt E/trans/cis configurations, respectively. The conformation of ring A is inter-mediate between envelope and half-chair and ring B adopts an envelope conformation whereas rings C and D adopt chair conformations. A weak intra-molecular C-H⋯O hydrogen bond is observed. The koetjapic acid mol-ecules are linked into dimers by two pairs of inter-molecular O-H⋯O hydrogen bonds. The dimers are stacked along the c axis.

  1. A one-pot procedure for the preparation of N-9-fluorenylmethyloxycarbonyl-α-amino diazoketones from α-amino acids.

    PubMed

    Siciliano, Carlo; De Marco, Rosaria; Guidi, Ludovica Evelin; Spinella, Mariagiovanna; Liguori, Angelo

    2012-12-07

    The study describes a new "one-pot" route to the synthesis of N-9-fluorenylmethyloxycarbonyl (Fmoc) α-amino diazoketones. The procedure was tested on a series of commercially available free or side-chain protected α-amino acids employed as precursors. The conversion into the title compounds was achieved by masking and activating the α-amino acids with a single reagent, namely, 9-fluorenylmethyl chloroformate (Fmoc-Cl). The resulting N-protected mixed anhydrides were reacted with diazomethane to lead to the α-amino diazoketones, which were isolated by flash column chromatography in very good to excellent overall yields. The versatility of the procedure was verified on lipophilic α-amino acids and further demonstrated by the preparation of N-Fmoc-α-amino diazoketones also from α-amino acids containing side-chain masking groups, which are orthogonal to the Fmoc one. The results confirmed that tert-butyloxycarbonyl (Boc), tert-butyl ((t)Bu), and 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl (Pbf), three acid-labile protecting groups mostly adopted in the solution and solid-phase peptide synthesis, are compatible to the adopted reaction conditions. In all cases, the formation of the corresponding C-methyl ester of the starting amino acid was not observed. Moreover, the proposed method respects the chirality of the starting α-amino acids. No racemization occurred when the procedure was applied to the synthesis of the respective N-Fmoc-protected α-amino diazoketones from L-isoleucine and L-threonine and to the preparation of a diastereomeric pair of N-Fmoc-protected dipeptidyl diazoketones.

  2. Optimized syntheses of Fmoc azido amino acids for the preparation of azidopeptides.

    PubMed

    Pícha, Jan; Buděšínský, Miloš; Macháčková, Kateřina; Collinsová, Michaela; Jiráček, Jiří

    2017-03-01

    The rise of CuI-catalyzed click chemistry has initiated an increased demand for azido and alkyne derivatives of amino acid as precursors for the synthesis of clicked peptides. However, the use of azido and alkyne amino acids in peptide chemistry is complicated by their high cost. For this reason, we investigated the possibility of the in-house preparation of a set of five Fmoc azido amino acids: β-azido l-alanine and d-alanine, γ-azido l-homoalanine, δ-azido l-ornithine and ω-azido l-lysine. We investigated several reaction pathways described in the literature, suggested several improvements and proposed several alternative routes for the synthesis of these compounds in high purity. Here, we demonstrate that multigram quantities of these Fmoc azido amino acids can be prepared within a week or two and at user-friendly costs. We also incorporated these azido amino acids into several model tripeptides, and we observed the formation of a new elimination product of the azido moiety upon conditions of prolonged couplings with 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate/DIPEA. We hope that our detailed synthetic protocols will inspire some peptide chemists to prepare these Fmoc azido acids in their laboratories and will assist them in avoiding the too extensive costs of azidopeptide syntheses. Experimental procedures and/or analytical data for compounds 3-5, 20, 25, 26, 30 and 43-47 are provided in the supporting information. © 2017 The Authors Journal of Peptide Science published by European Peptide Society and John Wiley & Sons Ltd.

  3. Optimized syntheses of Fmoc azido amino acids for the preparation of azidopeptides

    PubMed Central

    Pícha, Jan; Buděšínský, Miloš; Macháčková, Kateřina; Collinsová, Michaela

    2017-01-01

    The rise of CuI‐catalyzed click chemistry has initiated an increased demand for azido and alkyne derivatives of amino acid as precursors for the synthesis of clicked peptides. However, the use of azido and alkyne amino acids in peptide chemistry is complicated by their high cost. For this reason, we investigated the possibility of the in‐house preparation of a set of five Fmoc azido amino acids: β‐azido l‐alanine and d‐alanine, γ‐azido l‐homoalanine, δ‐azido l‐ornithine and ω‐azido l‐lysine. We investigated several reaction pathways described in the literature, suggested several improvements and proposed several alternative routes for the synthesis of these compounds in high purity. Here, we demonstrate that multigram quantities of these Fmoc azido amino acids can be prepared within a week or two and at user‐friendly costs. We also incorporated these azido amino acids into several model tripeptides, and we observed the formation of a new elimination product of the azido moiety upon conditions of prolonged couplings with 2‐(1H‐benzotriazol‐1‐yl)‐1,1,3,3‐tetramethyluronium hexafluorophosphate/DIPEA. We hope that our detailed synthetic protocols will inspire some peptide chemists to prepare these Fmoc azido acids in their laboratories and will assist them in avoiding the too extensive costs of azidopeptide syntheses. Experimental procedures and/or analytical data for compounds 3–5, 20, 25, 26, 30 and 43–47 are provided in the supporting information. © 2017 The Authors Journal of Peptide Science published by European Peptide Society and John Wiley & Sons Ltd. PMID:28120383

  4. Tissue Necrosis due to Chloroform: A Case Report

    PubMed Central

    Mohammadzadeh Akhlaghi, Nahid; Baradaran Mohajeri, Ladan; Fazlyab, Mahta

    2013-01-01

    For many years, gutta-percha has been the root canal filling material of choice. Chloroform is one of the most efficient solvents widely used for gutta-percha removal in retreatment cases, despite being toxic and carcinogenic. The present case report discusses a chloroform extrusion through an existing perforation to the surrounding periodontal ligament space and subsequent necrosis in supporting bone and tissues, during an endodontic retreatment visit for an addicted patient. Subsequently, the management and preventive options are reviewed. PMID:24790633

  5. Human respiratory uptake of chloroform and haloketones during showering.

    PubMed

    Xu, Xu; Weisel, Clifford P

    2005-01-01

    Inhalation is an important exposure route for volatile water contaminants, including disinfection by-products (DBPs). A controlled human study was conducted on six subjects to determine the respiratory uptake of haloketones (HKs) and chloroform, a reference compound, during showering. Breath and air concentrations of the DBPs were measured using gas chromatography and electron capture detector during and following the inhalation exposures. A lower percentage of the HKs (10%) is released from shower water to air than that of chloroform (56%) under the experiment conditions due to the lower volatility of the HKs. Breath concentrations of the DBPs were elevated during the inhalation exposure, while breath concentrations decreased rapidly after the exposure. Approximately 85-90% of the inhaled HKs were absorbed, whereas only 70% of the inhaled chloroform was absorbed for the experiment conditions used. The respiratory uptake of the DBPs was estimated using a linear one-compartment model coupled with a plug flow stream model for the shower system. The internal dose of chloroform normalized to its water concentration was approximately four times that of the HKs after a 30-min inhalation exposure. Approximately 0.3-0.4% of the absorbed HKs and 2-9% of the absorbed chloroform were expired through lung excretion after the 30-min exposure. The inhalation exposure from a typical 10-15 min shower contributes significantly to the total dose for chloroform in chlorinated drinking water but only to a moderate extent for HKs.

  6. Durability of organobentonite-amended liner for decelerating chloroform transport.

    PubMed

    He, Shichong; Zhu, Lizhong

    2016-04-01

    Chloroform is added to landfill for suppressing methane generation, which however may transport through landfill liners and lead to contamination of groundwater. To decelerate chloroform transport, the enhanced sorption ability of clay liners following organobentonite addition was tested. In this study, we used batch sorption to evaluate sorption capacity of chloroform to organobentonite, followed by column tests and model simulations for assessing durability of different liners. Results show that adding 10% CTMAB-bentonite (organobentonite synthesized using cetyltrimethylammonium bromide) increased the duration of a bentonite liner by 88.5%. CTMAB-bentonite consistently showed the highest sorption capacity (Qm) among six typical organobentonites under various environmental conditions. The removal rate of chloroform by CTMAB-bentonite was 3.6-23 times higher than that by natural soils. According to the results derived by model simulation, a 70-cm 10% CTMAB-bentonite liner exhibited much better durability than a 100-cm compact clay liner (CCL) and natural bentonite liner evidenced by the delayed and lower peak of eluent concentration. A minimum thickness of 65.8 cm of the 10% CTMAB-bentonite liner could completely sorb the chloroform in a 100-m-high landfill. The 10% CTMAB-bentonite liner exhibiting much better durability has the promise for reducing environmental risk of chloroform in landfill.

  7. Synthesis of various 3-substituted 1,2,4-oxadiazole-containing chiral beta 3- and alpha-amino acids from Fmoc-protected aspartic acid.

    PubMed

    Hamzé, Abdallah; Hernandez, Jean-François; Fulcrand, Pierre; Martinez, Jean

    2003-09-19

    Various 3-substituted chiral 1,2,4-oxadiazole-containing Fmoc-beta(3)- and -alpha-amino acids were synthesized from Fmoc-(l or d)-Asp(OtBu)-OH and Fmoc-l-Asp-OtBu, respectively, in three steps (i.e., condensation of an aspartyl derivative with differentially substituted amidoximes, formation of the 1,2,4-oxadiazole, and cleavage of the tert-butyl ester). These compounds represent new series of nonnatural amino acids, which could be used in combinatorial synthesis. A simple protocol has been developed to generate the 1,2,4-oxadiazole ring. Indeed, common methods resulted in cleavage of the Fmoc group or required long reaction times. We found that sodium acetate in refluxing ethanol/water (86 degrees C) was a convenient and efficient catalyst to promote conversion of Fmoc-amino acyl amidoximes to 1,2,4-oxadiazoles, and this procedure proved to be compatible with Fmoc protection. It is shown that these compounds can be prepared without significant loss of enantiomerical purity. Furthermore, the alkaline conditions used to cleave the Fmoc protecting group from these compounds did not induce epimerization of their chiral center.

  8. Chiral HPLC analysis of milnacipran and its FMOC-derivative on cellulose-based stationary phases.

    PubMed

    Patti, Angela; Pedotti, Sonia; Sanfilippo, Claudia

    2008-02-01

    The HPLC enantioseparation of the last generation antidepressive drug milnacipran (+/-)-1 was investigated on different cellulose-based chiral stationary phases (CSPs). On carbamate-type columns, Chiralcel OD and OD-H (+/-)-1 could be separated with alpha value about 1.20 but the resolution was quite low because of the tailing of the peaks. Direct determination of (+/-)-1 with high selectivity and resolution was obtained on Chiralcel OJ in normal phase mode elution. Precolumn derivatization of milnacipran with Fmoc-Cl gave compound (+/-)-2 which was enantioseparated on all the investigated CSPs and allowed enhanced UV or fluorimetric detection. The Chiralpak IB, that could be considered the immobilized version of Chiralcel OD-H, was found completely ineffective in the chiral recognition of (+/-)-1 and moderately efficient in the separation of (+/-)-2.

  9. Microwave-assisted solid-phase peptide synthesis based on the Fmoc protecting group strategy (CEM).

    PubMed

    Vanier, Grace S

    2013-01-01

    Microwave-assisted peptide synthesis has become one of the most widely used tools by peptide chemists for the synthesis of both routine and difficult peptide sequences. Microwave technology significantly reduces the synthesis time while also improving the quality of the peptides produced. Microwave energy allows most amino acid couplings to be completed in just 5 min. The Fmoc removal can also be accelerated in the microwave decreasing the reaction time from at least 15 min to only 3 min in most cases. Common side reactions such as racemization and aspartimide formation are easily controllable with optimized methods that can be applied routinely. This protocol outlines the detailed procedure for performing both manual and automated microwave-assisted peptide synthesis of two difficult peptide sequences, ACP (65-74) and β-amyloid, in high purity and yield.

  10. Appraisal of four pre-column derivatization methods for the high-performance liquid chromatographic determination of free amino acids in biological materials.

    PubMed

    Fürst, P; Pollack, L; Graser, T A; Godel, H; Stehle, P

    1990-01-19

    Reversed-phase high-performance liquid chromatography (RP-HPLC) is a powerful method for assaying physiological amino acid concentrations in biological fluids. Four pre-column derivatization methods, with o-phthaldialdehyde (OPA), 9-fluorenylmethyl chloroformate (FMOC-Cl), phenyl isothiocyanate (PITC) and 1-dimethylaminonaphthalene-5-sulphonyl chloride (dansyl-Cl), were assessed with respect to their applicability in biological research. The methods permit the measurement of 21-26 major amino acids in 13-40 min. The superior sensitivity favours the use of OPA, FMOC-Cl and dansyl-Cl techniques. Because of instability of the OPA adducts, automated on-line derivatization is required when using this method in general practice. Application of the PITC method, although less sensitive, is useful in clinical chemistry, where sample availability is rarely a problem. Cystine determination is not feasible when using OPA or FMOC-Cl and with PITC the reproducibility and linearity are poor, whereas the dansyl-Cl method allows reliable quantitation. The four methods are currently used to perform ca. 8000 OPA and 5000-6000 FMOC-Cl, PITC and dansyl-Cl analyses of biological samples per year. The results obtained with the RP-HPLC methods compare favourably with those derived from conventional ion-exchange amino acid analyses. When the guard column is regularly changed after 120 analyses, the separation remains satisfactory for at least 700 OPA, 800 FMOC-Cl, 150 PITC and 500 dansyl-Cl analyses. Careful control of factors and limitations inherent in the various methodologies is a prerequesite for proper identification and appropriate quantitation.

  11. Unusual Case of Drug-Facilitated Sexual Assault using Chloroform.

    PubMed

    Richeval, Camille; Allorge, Delphine; Lopez, Vincent; Boyer, Baptiste; Gaulier, Jean-Michel

    2016-11-22

    Despite its well-known sedative properties, chloroform is rarely used for drug-facilitated sexual assault (DFSA) as its administration cannot be achieved without the victim's knowledge: we report an unusual case of DFSA using this solvent. A 26-year-old woman declared that her partner get her to sleep using chloroform the previous night. When she waked up at 3 am, her hands were tied. She immediately suspected violence and sexual penetration. Toxicological blood screening using a liquid chromatography-electrospray coupled tandem mass spectrometry method highlighted the presence of bamifylline and theophylline, two therapeutics of asthma. A screening method for volatile substances using a headspace-gas chromatography coupled with mass spectrometry method showed (i) the presence of chloroform in blood at a concentration subsequently estimated at 580 µg/L using an external calibration, and (ii) chloroform traces on a piece of a scarf brought by the patient and suspected to have been used to put her to sleep. These results were consistent with an exposure to chloroform by inhalation and demonstrate that there is no limit in the use of chemical weapons in DFSA.

  12. Biofiltration of Chloroform in a Trickle Bed Air Biofilter Under ...

    EPA Pesticide Factsheets

    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 ethanol. The TBAB is composed of pelleted diatomaceous earth filter media inoculated with filamentous fungi species, which served as the principle biodegrading microorganism. The removal efficiencies of 5 ppmv of chloroform mixed with different ratios of ethanol as cometabolite (25, 50, 100, 150, and 200 ppmv) ranged between 69.9 and 80.9%. The removal efficiency, reaction rate kinetics, and the elimination capacity increased proportionately with an increase in the cometabolite concentration. The carbon recovery from the TBAB amounted to 69.6% of the total carbon input. It is postulated that the remaining carbon contributed to excess biomass yield within the system. Biomass control strategies such as starvation and stagnation were employed at different phases of the experiment. The chloroform removal kinetics provided a maximum reaction rate constant of 0.0018 s−1. The highest ratio of chemical oxygen demand (COD)removal/nitrogenutilization was observed at 14.5. This study provides significant evidence that the biodegradation of a highly chlorinated methane can be favored by cometabolism in a fungi-based TBAB. Chloroform is volatile hazardous chemical emitted from publicly owned treatment

  13. Chloroform alters interleaflet coupling in lipid bilayers: an entropic mechanism

    PubMed Central

    Reigada, Ramon; Sagués, Francesc

    2015-01-01

    The interaction of the two leaflets of the plasmatic cell membrane is conjectured to play an important role in many cell processes. Experimental and computational studies have investigated the mechanisms that modulate the interaction between the two membrane leaflets. Here, by means of coarse-grained molecular dynamics simulations, we show that the addition of a small and polar compound such as chloroform alters interleaflet coupling by promoting domain registration. This is interpreted in terms of an entropic gain that would favour frequent chloroform commuting between the two leaflets. The implication of this effect is discussed in relation to the general anaesthetic action. PMID:25833246

  14. Preventing aspartimide formation in Fmoc SPPS of Asp-Gly containing peptides--practical aspects of new trialkylcarbinol based protecting groups.

    PubMed

    Behrendt, Raymond; Huber, Simon; White, Peter

    2016-02-01

    In our efforts to develop a universal solution to the problem of aspartimide formation in Fmoc SPPS, we investigated the application of our new β-trialkylmethyl protected aspartic acid building blocks to the synthesis of peptides containing the Asp-Gly motif. The N(α)-Fmoc aspartic acid β-tri-(ethyl/propyl/butyl)methyl esters were used in the synthesis of the classic model peptide scorpion toxin II (VKDGYI), and their effectiveness in minimising aspartimide formation during extended piperidine treatments was evaluated. Furthermore, we compared their efficacy against that of the commonly used approach of adding acids to the Fmoc deprotection solution. Finally, we applied our aspartic acid building blocks to the stepwise Fmoc SPPS of teduglutide, a human GLP-2 analogue, whose synthesis is made challenging by extensive aspartimide formation. In all experiments, our approach led to almost complete reduction of aspartimide formation with accompanied suppression of aspartic acid epimerisation.

  15. β Sheets Not Required: Combined Experimental and Computational Studies of Self-Assembly and Gelation of the Ester-Containing Analogue of an Fmoc-Dipeptide Hydrogelator

    PubMed Central

    2015-01-01

    In our work toward developing ester-containing self-assembling peptides as soft biomaterials, we have found that a fluorenylmethoxycarbonyl (Fmoc)-conjugated alanine-lactic acid (Ala-Lac) sequence self-assembles into nanostructures that gel in water. This process occurs despite Fmoc-Ala-Lac’s inability to interact with other Fmoc-Ala-Lac molecules via β-sheet-like amide–amide hydrogen bonding, a condition previously thought to be crucial to the self-assembly of Fmoc-conjugated peptides. Experimental comparisons of Fmoc-Ala-Lac to its self-assembling peptide sequence analogue Fmoc-Ala-Ala using a variety of microscopic, spectroscopic, and bulk characterization techniques demonstrate distinct features of the two systems and show that while angstrom-scale self-assembled structures are similar, their nanometer-scale size and morphological properties diverge and give rise to different bulk mechanical properties. Molecular dynamics simulations were performed to gain more insight into the differences between the two systems. An analysis of the hydrogen-bonding and solvent-surface interface properties of the simulated fibrils revealed that Fmoc-Ala-Lac fibrils are stronger and less hydrophilic than Fmoc-Ala-Ala fibrils. We propose that this difference in fibril amphiphilicity gives rise to differences in the higher-order assembly of fibrils into nanostructures seen in TEM. Importantly, we confirm experimentally that β-sheet-type hydrogen bonding is not crucial to the self-assembly of short, conjugated peptides, and we demonstrate computationally that the amide bond in such systems may act mainly to mediate the solvation of the self-assembled single fibrils and therefore regulate a more extensive higher-order aggregation of fibrils. This work provides a basic understanding for future research in designing highly degradable self-assembling materials with peptide-like bioactivity for biomedical applications. PMID:24786493

  16. Optical sensors for the detection of trace chloroform.

    PubMed

    Fong, Jonathan K; Pena, Justin K; Xue, Zi-Ling; Alam, Maksudul M; Sampathkumaran, Uma; Goswami, Kisholoy

    2015-02-03

    Optical thin film sensors have been developed to detect chloroform in aqueous and nonaqueous solutions. These sensors utilize a modified Fujiwara reaction, one of the only known methods for detecting halogenated hydrocarbons in the visible spectrum. The modified Fujiwara reagents, 2,2'-dipyridyl and tetra-n-butyl ammonium hydroxide (n-Bu4NOH or TBAH), are encapsulated in an ethyl cellulose (EC) or sol-gel film. Upon exposure of the EC sensor film to HCCl3 in petroleum ether, a colored product is produced within the film, which is analyzed spectroscopically, yielding a detection limit of 0.830 ppm (parts per million v/v or μL/L hereinafter) and a quantification limit of 2.77 ppm. When the chloroform concentration in pentane is ≥5 ppm, the color change of the EC sensor is visible to the naked eye. In aqueous chloroform solution, reaction in the sol-gel sensor film turns the sensor from colorless to dark yellow/brown, also visible to the naked eye, with a detection limit of 500 ppm. This is well below the solubility of chloroform in water (ca. 5,800 ppm). To our knowledge, these are the first optical quality thin film sensors using Fujiwara reactions for halogenated hydrocarbon detection.

  17. Design and synthesis of an Fmoc-SPPS-compatible amino acid building block mimicking the transition state of phosphohistidine phosphatase.

    PubMed

    Eerland, Martijn F; Hedberg, Christian

    2012-02-17

    The synthesis of a sulfonamide-based transition-state (TS) analogue of enzymatic phosphohistidine dephosphorylation as an amino acid building block is presented, together with the proof-of-concept of its incorporation into peptides. Key features include final global acidolytic protective group removal as well as full compatibility with standard Fmoc solid-phase peptide synthesis (SPPS). The peptides are designed as inhibitors of phosphohistidine phosphatase and as a pull-down probe for identification of phosphohistidine phosphatases, respectively.

  18. Effect of water temperature on dermal exposure to chloroform.

    PubMed

    Gordon, S M; Wallace, L A; Callahan, P J; Kenny, D V; Brinkman, M C

    1998-06-01

    We have developed and applied a new measurement methodology to investigate dermal absorption of chloroform while bathing. Ten subjects bathed in chlorinated water while breathing pure air through a face mask. Their exhaled breath was delivered to a glow discharge source/ion trap mass spectrometer for continuous real-time measurement of chloroform in the breath. This new method provides abundant data compared to previous discrete time-integrated breath sampling methods. The method is particularly well suited to studying dermal exposure because the full face mask eliminates exposure to contaminated air. Seven of the 10 subjects bathed in water at two or three different temperatures between 30 degrees C and 40 degrees C. Subjects at the highest temperatures exhaled about 30 times more chloroform than the same subjects at the lowest temperatures. This probably results from a decline in blood flow to the skin at the lower temperatures as the body seeks to conserve heat forcing the chloroform to diffuse over a much greater path length before encountering the blood. These results suggest that pharmacokinetic models need to employ temperature-dependent parameters. Two existing models predict quite different times of about 12 min and 29 min for chloroform flux through the stratum corneum to reach equilibrium. At 40 degrees C, the time for the flux to reach a near steady-state value is 6-9 min. Although uptake and decay processes involve several body compartments, the complicating effect of the stratum corneum lag time made it difficult to fit multiexponential curves to the data; however, a single-compartment model gave a satisfactory fit.

  19. A Comparative Examination of two Fmoc Removal Reagents for Process Improvement to Produce Peptide Drugs

    NASA Astrophysics Data System (ADS)

    Srivastava, K.; Davis, M.

    The importance of peptides as therapeutics has been recognized since they were found responsible for a wide variety of biological functions. The recent approval of peptide drugs such as Byetta® (Amylin Pharmaceuticals, Inc.), Fuzeon® (Hoffman-LaRoche Inc.), Integrelin™ (CDR Therapeutics, Inc.), Natrecor® (SCIOS Inc.), Symlin® (Amylin), Teriparatide, and Ziconotide, etc., which demonstrated applications for treatment of such problems as bone metabolism disorders, cardiovascular diseases, diabetes, viral infections and severe chronic pain control, has further endorsed the growing interest in peptides as a potential drug. This growing trend for peptide drugs has drawn our attention for their production in a cost-effective manner. To do so, the improvement in the quality of crude peptides during synthesis, the most critical parameter in the process, is important to prevent yield losses during the more expensive purification step. To accomplish it, we decided to examine the efficacy of the commonly used nucleophilic base piperidine and non-neucleophilic base DBU (1,8-Diazabicyclo[5.4.0]undec-7-ene) for the complete removal of Fmoc group during the synthesis of peptides. According to our investigation, application of piperidine was found more effective than DBU in solid phase synthesis. Details of the investigation will be discussed.

  20. High-throughput sequencing of peptoids and peptide-peptoid hybrids by partial edman degradation and mass spectrometry.

    PubMed

    Thakkar, Amit; Cohen, Allison S; Connolly, Michael D; Zuckermann, Ronald N; Pei, Dehua

    2009-03-09

    A method for the rapid sequence determination of peptoids [oligo(N-substituted glycines)] and peptide-peptoid hybrids selected from one-bead-one-compound combinatorial libraries has been developed. In this method, beads carrying unique peptoid (or peptide-peptoid) sequences were subjected to multiple cycles of partial Edman degradation (PED) by treatment with a 1:3 (mol/mol) mixture of phenyl isothiocyanate (PITC) and 9-fluorenylmethyl chloroformate (Fmoc-Cl) to generate a series of N-terminal truncation products for each resin-bound peptoid. After PED, the Fmoc group was removed from the N-terminus and any reacted side chains via piperidine treatment. The resulting mixture of the full-length peptoid and its truncation products was analyzed by matrix-assisted laser desorption ionization (MALDI) mass spectrometry, to reveal the sequence of the full-length peptoid. With a slight modification, the method was also effective in the sequence determination of peptide-peptoid hybrids. This rapid, high-throughput, sensitive, and inexpensive sequencing method should greatly expand the utility of combinatorial peptoid libraries in biomedical and materials research.

  1. Comparison of amino acid derivatization reagents for LC-ESI-MS analysis. Introducing a novel phosphazene-based derivatization reagent.

    PubMed

    Rebane, Riin; Oldekop, Maarja-Liisa; Herodes, Koit

    2012-09-01

    Amino acid analysis with high performance liquid chromatography with electrospray ionization mass spectrometry (LC-ESI-MS) is an emerging method. For more sensitive analysis, derivatization is used and next to commercially available derivatization reagents such as dansyl chloride (DNS), 9-fluorenylmethyl chloroformate (FMOC-Cl) and diethyl ethoxymethylenemalonate (DEEMM), new derivatization reagents are designed specially for LC-ESI-MS, like p-N,N,N-trimethylammonioanilyl N'-hydroxysuccinimidyl carbamate iodide (TAHS) which provides very low limits of detection. In this work, a novel phosphazene based derivatization reagent (FOSF) that provides comparable limits of quantitation (LoQ) to TAHS is introduced. Moreover, a thorough comparison between FOSF, TAHS, DNS, FMOC-Cl and DEEMM is carried out for 7 different amino acids - Arg, Asp, Gly, β-Ala, Pro, Trp and Phe. This is a first time that thorough comparison is carried out on the same instrument for amino acid derivatization reagents. Results on the same instrument for five amino acid derivatization reagents show that novel reagents are sensitive with LoQ values around 80 fmol but have disadvantages such as problematic chromatographic separation. Next to novel reagents, DEEMM offers very good LoQ-s (average of 150 fmol) and wide dynamic linear range.

  2. Determination of memantine in plasma and vitreous humour by HPLC with precolumn derivatization and fluorescence detection.

    PubMed

    Puente, Belen; Hernandez, Esther; Perez, Susana; Pablo, Luis; Prieto, Esther; Garcia, Maria Angeles; Bregante, Miguel Angel

    2011-01-01

    A new HPLC procedure with precolumn derivatization and rimantadine as the internal standard for determining memantine, a candidate agent for the treatment of glaucoma in plasma and vitreous humour, has been developed and validated. Precolumn derivatization was performed with 9-fluorenylmethyl-chloroformate-chloride (FMOC-Cl) as the derivatization reagent and followed by a liquid-liquid extraction with n-hexane. Optimal conditions for derivatization were an FMOC-Cl concentration of 1.5 mM, a reaction time of 20 min, the temperature at 30°C, the borate buffer pH 8.5, and a borate buffer-acetonitrile ratio of 1:1. The derivatives were analyzed by isocratic HPLC with the fluorescence detector λex 260 nm λem 315 nm on a Novapack C(18) reversed-phase column with a mobile phase of acetonitrile-water (73:27, v/v), 40°C, and a flow rate of 1.2 mL/min. The linear range was 10-1000 ng/mL with a quantification limit of ∼ 10 ng/mL for both types of samples. This analytical method may be suitable for using in ocular availability studies.

  3. Biological Hydrogen Production Using Chloroform-treated Methanogenic Granules

    NASA Astrophysics Data System (ADS)

    Hu, Bo; Chen, Shulin

    In fermentative hydrogen production, the low-hydrogen-producing bacteria retention rate limits the suspended growth reactor productivity because of the long hydraulic retention time (HRT) required to maintain adequate bacteria population. Traditional bacteria immobilization methods such as calcium alginate entrapment have many application limitations in hydrogen fermentation, including limited duration time, bacteria leakage, cost, and so on. The use of chloroform-treated anaerobic granular sludge as immobilized hydrogen-producing bacteria in an immobilized hydrogen culture may be able to overcome the limitations of traditional immobilization methods. This paper reports the findings on the performance of fed-batch cultures and continuous cultures inoculated with chloroform-treated granules. The chloroform-treated granules were able to be reused over four fed-batch cultures, with pH adjustment. The upflow reactor packed with chloroform-treated granules was studied, and the HRT of the upflow reactor was found to be as low as 4 h without any decrease in hydrogen production yield. Initial pH and glucose concentration of the culture medium significantly influenced the performance of the reactor. The optimum initial pH of the culture medium was neutral, and the optimum glucose concentration of the culture medium was below 20 g chemical oxygen demand/L at HRT 4 h. This study also investigated the possibility of integrating immobilized hydrogen fermentation using chloroform-treated granules with immobilized methane production using untreated granular sludge. The results showed that the integrated batch cultures produced 1.01 mol hydrogen and 2 mol methane per mol glucose. Treating the methanogenic granules with chloroform and then using the treated granules as immobilized hydrogen-producing sludge demonstrated advantages over other immobilization methods because the treated granules provide hydrogen-producing bacteria with a protective niche, a long duration of an active

  4. Asphyxial suicide by inhalation of chloroform inside a plastic bag.

    PubMed

    Zorro, Andres Rodriguez

    2014-01-01

    Asphyxia suicide by placing a plastic bag over the head in addition with inhalation of gases or use of sedative substances is an unusual method of committing suicide, but frequently referenced by right to die groups in the Internet. This article reports 2 suicides in which chloroform was used to induce unconsciousness and subsequent asphyxia by placing the head in a plastic bag. Case histories of 2 males, ages 23 and 28, are described with special emphasis on characteristics death related to suffocation using plastic bags and chloroform. The final remarkable point in both cases is that the victims previously searched the WEB for instructions of suicide methods. The importance of the phenomenon of misuse of Internet by young people who commit suicide is stressed.

  5. Global transcriptional response of Nitrosomonas europaea to chloroform and chloromethane.

    PubMed

    Gvakharia, Barbara O; Permina, Elizabeth A; Gelfand, Mikhail S; Bottomley, Peter J; Sayavedra-Soto, Luis A; Arp, Daniel J

    2007-05-01

    Upon exposure of Nitrosomonas europaea to chloroform (7 microM, 1 h), transcripts for 175 of 2,460 genes were found at higher levels in treated cells than in untreated cells and transcripts for 501 genes were found at lower levels. With chloromethane (3.2 mM, 1 h), transcripts for 67 genes were at higher levels and transcripts for 148 genes were at lower levels. Transcripts for 37 genes were at higher levels following both treatments and included genes for heat shock proteins, sigma-factors of the extracytoplasmic function subfamily, and toxin-antitoxin loci. N. europaea has higher levels of transcripts for a variety of defense genes when exposed to chloroform or chloromethane.

  6. Graphene oxide and adsorption of chloroform: A density functional study

    NASA Astrophysics Data System (ADS)

    Kuisma, Elena; Hansson, C. Fredrik; Lindberg, Th. Benjamin; Gillberg, Christoffer A.; Idh, Sebastian; Schröder, Elsebeth

    2016-05-01

    Chlorinated hydrocarbon compounds are of environmental concerns, since they are toxic to humans and other mammals, and are widespread, and exposure is hard to avoid. Understanding and improving methods to reduce the amount of the substances are important. We present an atomic-scale calculational study of the adsorption of chlorine-based substance chloroform (CHCl3) on graphene oxide, as a step in estimating the capacity of graphene oxide for filtering out such substances, e.g., from drinking water. The calculations are based on density functional theory, and the recently developed consistent-exchange functional for the van der Waals density-functional method is employed. We obtain values of the chloroform adsorption energy varying from roughly 0.2 to 0.4 eV per molecule. This is comparable to previously found results for chloroform adsorbed directly on clean graphene, using similar calculations. In a wet environment, like filters for drinking water, the graphene will not stay clean and will likely oxidize, and thus adsorption onto graphene oxide, rather than clean graphene, is a more relevant process to study.

  7. Evaluation of Direct Aqueous Injection Method for Analysis of Chloroform in Drinking Water

    ERIC Educational Resources Information Center

    Pfaender, Frederic K.; And Others

    1978-01-01

    A direct aqueous injection (DAI) technique was compared with the purge method for chloroform measurement in drinking water. The DAI method gave consistently higher values for chloroform than the purge method. The results indicated the need for caution in the interpretation of chloroform and other trihalomethane values generated by DAI. (Author/MA)

  8. 9-Fluorenylmethoxycarbonyl (Fmoc)-glycine coupling of saccharide beta-glycosylamines for the fractionation of oligosaccharides and the formation of neoglycoconjugates.

    PubMed

    Arsequell, G; Dwek, R A; Wong, S Y

    1994-01-01

    We have devised a new saccharide derivatization scheme to provide not only the temporary attachment of a chromophore for detecting and facilitating the chromatographic separation of carbohydrates, but also the intermediates for further derivatization to produce neoglycoconjugates. Several neutral unprotected saccharide beta-glycosylamines were formed by the direct condensation of the reducing saccharides with aqueous ammonium bicarbonate. The beta-glycosylamine derivatives of N-acetylglucosamine, di-N-acetylchitobiose, and asialo-, digalactosylated biantennary complex oligosaccharide were N-acylated separately with excess 9-fluorenylmethoxycarbonyl (Fmoc)-glycine. The Fmoc-glycinamido beta-derivatives of these unprotected saccharides were well separated by normal-phase high-performance liquid chromatography and detected by ultraviolet absorption. Similar derivatization and fractionation of a partial acid hydrolyzate of chitin were equally successful resulting in the separation of Fmoc-glycinamido derivatives of di-N-acetylchitobiose to hepta-N-acetylchitoheptaose in the hydrolyzate. The reversibility of the Fmoc derivatization was demonstrated by treating the Fmoc-glycinamido derivative of N-acetylglucosamine with piperidine to generate its 1-N-glycyl-beta-saccharide derivative. The structure and stereochemistry of this product was confirmed by proton nuclear magnetic resonance spectroscopy. The 1-N-glycyl-beta-saccharide derivatives are stable intermediates for the formation of asparagine-linked neoglycoconjugates.

  9. Synthesis of the very acid-sensitive Fmoc-Cys(Mmt)-OH and its application in solid-phase peptide synthesis.

    PubMed

    Barlos, K; Gatos, D; Hatzi, O; Koch, N; Koutsogianni, S

    1996-03-01

    S-4-methoxytrityl cysteine was synthesized and converted into the corresponding Fmoc-Cys(Mmt)-OH by its reaction with Fmoc-OSu. As compared to the corresponding Fmoc-Cys(Trt)-OH, the S-Mmt-function was found to be considerably more acid labile. Quantitative S-Mmt-removal occurs selectively in the presence of groups of the tert butyl type and S-Trt by treatment with 0.5-1.0% TFA. The new derivative was successfully utilized in the SPPS of Tyr1-somatostatin on 2-chlorotrityl resin. In this synthesis groups of the Trt-type were exclusively used for amino acid side-chain protection. Quantitative cleavage from the resin and complete deprotection was performed by treatment with 3% TFA in DCM-TES (95:5) for 30 min at RT. We observed no reduction of tryptophan under these conditions.

  10. Microwave spectrum, molecular structure, and quadrupole coupling of vinyl chloroformate

    NASA Astrophysics Data System (ADS)

    Bimler, Jonathan; Broadbent, Stacy; Utzat, Karissa A.; Bohn, Robert K.; Restrepo, Albeiro; Harvey Michels, H.; True, Nancy S.

    2012-09-01

    Vinyl chloroformate is confirmed to have the planar structure reported in an earlier study [1]. Our study uses much higher resolution microwave rotational spectra and ab initio calculations have been extended to a higher level. Naturally abundant isotopologs with single substitution of 37Cl, 13C, and 18O isotopes have also been measured and a substitution structure obtained. The quadrupole coupling constants of the 35Cl and 37Cl isotopologs have been determined. The potential energy profiles of internal rotation about the Odbnd Csbnd Osbnd C and Csbnd Osbnd Cdbnd C dihedral angles have been calculated.

  11. Grunwald-Winstein Analysis - Isopropyl Chloroformate Solvolysis Revisited

    PubMed Central

    D’Souza, Malcolm J.; Reed, Darneisha N.; Erdman, Kevin J.; Kyong, Jin Burm; Kevill, Dennis N.

    2009-01-01

    Specific rates of solvolysis at 25 °C for isopropyl chloroformate (1) in 24 solvents of widely varying nucleophilicity and ionizing power, plus literature values for studies in water and formic acid, are reported. Previously published solvolytic rate constants at 40.0 °C are supplemented with two additional values in the highly ionizing fluoroalcohols. These rates are now are analyzed using the one and two-term Grunwald-Winstein Equations. In the more ionizing solvents including ten fluoroalcohols negligible sensitivities towards changes in solvent nucleophilicity (l) and very low sensitivities towards changes in solvent ionizing power (m) values are obtained, evocative to those previously observed for 1-adamantyl and 2-adamantyl chloroformates 2 and 3. These observations are rationalized in terms of a dominant solvolysis-decomposition with loss of the CO2 molecule. In nine of the more nucleophilic pure alchohols and aqueous solutions an association-dissociation mechanism is believed to be operative. Deficiencies in the acid production indicate 2-33% isopropyl chloride formation, with the higher values in less nucleophilic solvents. PMID:19399225

  12. Reliable test methods for the determination of a natural production of chloroform in soils.

    PubMed

    Grøn, Christian; Laturnus, Frank; Jacobsen, Ole Stig

    2012-03-01

    Chloroform is one of the most frequently found anthropogenic groundwater contaminants. Recent investigations, however, suggested that chloroform in groundwater may also originate from a natural production in soils. As societies response to the occurrence of chloroform in groundwater may depend upon its origin as anthropogenic or naturally produced, test methods are needed to measure the potential of natural soil chloroform production. Field measurements of ambient air and soil air, and field and laboratory incubation studies were evaluated for measurement of relative soil chloroform production at a site with four different vegetation types (spruce forest, beech forest, grassland, and grain field) on comparable geological soil. All test methods showed varying soil production of chloroform with spruce forest soil being most productive and grain field soil being least productive. Field measurements of the ratio of soil air to ambient air chloroform concentrations exhibited the smallest difference between high production and low production areas, whereas laboratory incubation studies showed the largest difference. Thus, laboratory incubation studies are suggested as most efficient for estimating relative chloroform production in soil. The study indicated that soil samples should be tested not more than 14 days after sampling. Furthermore, it was found that potentially limiting compounds, such as chloride or nitrate, are not needed to be added in spike experiments to obtain reliable production results. However, it should be recognized that the processes of soil chloroform production are not known yet in all details. Other factors than those studied here may affect the test methods for soil chloroform production too.

  13. Novel N omega-xanthenyl-protecting groups for asparagine and glutamine, and applications to N alpha-9-fluorenylmethyloxycarbonyl (Fmoc) solid-phase peptide synthesis.

    PubMed

    Han, Y; Solé, N A; Tejbrant, J; Barany, G

    1996-01-01

    The N alpha-9-fluorenylmethyloxycarbonyl (Fmoc), N omega-9H-xanthen-9-yl (Xan), N omega-2-methoxy-9H-xanthen-9-yl (2-Moxan) or N omega-3-methoxy-9H-xanthen-9-yl (3-Moxan) derivatives of asparagine and glutamine were prepared conveniently by acid-catalyzed reactions of appropriate xanthydrols with Fmoc-Asn-OH and Fmoc-Gln-OH. The Xan and 2-Moxan protected derivatives have been used in Fmoc solid-phase syntheses of several challenging peptides: a modified Riniker's peptide to probe tryptophanalkylation side reactions, Briand's peptide to assess deblocking, at the N-terminus and Marshall's ACP (65-74) to test difficult couplings. Removal of the Asn and Gln side-chain protection occurred concomitantly with release of peptide from the support, under the conditions for acidolytic cleavage of the tris(alkoxy)benzylamide (PAL) anchoring linkage by use of trifluoroacetic acid/scavenger mixtures. For each of the model peptides, the products obtained by the new protection schemes were purer than those obtained with N omega-2,4,6-trimethoxybenzyl (Tmob) or N omega-triphenylmethyl (Trt) protection for Asn and Gln.

  14. Phase transition in conjugated oligomers suspended in chloroform

    NASA Astrophysics Data System (ADS)

    Dwivedi, Shikha; Kumar, Anupam; Yadav, S. N. S.; Mishra, Pankaj

    2015-08-01

    Density functional theory (DFT) has been used to investigate the isotropic-nematic (I-N) phase transition in a system of high aspect ratio conjugated oligomers suspended in chloroform. The interaction between the oligomers is modeled using Gay-Berne potential in which effect of solvent is implicit. Percus-Yevick integral equation theory has been used to evaluate the pair correlation functions of the fluid phase at several temperatures and densities. These pair correlation function has been used in the DFT to evaluate the I-N freezing parameters. Highly oriented nematic is found to stabilize at low density. The results obtained are in qualitative agreement with the simulation and are verifiable.

  15. Atmospheric halocarbons - A discussion with emphasis on chloroform

    NASA Technical Reports Server (NTRS)

    Yung, Y. L.; Mcelroy, M. B.; Wofsy, S. C.

    1975-01-01

    Bleaching of paper pulp represents a major industrial use of chlorine and could provide an environmentally significant source of atmospheric halocarbons. The related global production of chloroform is estimated at 300,000 ton per year and there could be additional production associated with atmospheric decomposition of perchloroethylene. Estimates are given for the production of methyl chloride, methyl bromide and methyl iodide, 5.2 million, 77 thousand, and 740 thousand ton per year respectively. The relative yields of CH3Cl, CH3Br and CH3I are consistent with the hypothesis of a marine biological source for these compounds. Concentrations of other halocarbons observed in the atmosphere appear to indicate industrial sources.

  16. Chloroform in Indoor Air and Wastewater: The Role of Residential Washing Machines.

    PubMed

    Shepherd, Jennifer L; Corsi, Richard L; Kemp, Jeff

    1996-07-01

    A residential washing machine was studied in order to determine the extent of chloroform formation following the application of a laundry bleach containing sodium hypochlorite. A dynamic model was also developed to estimate chloroform formation, mass transfer, and gaseous emissions during a typical wash cycle. A series of 22 experiments was completed to determine model parameters, including chemical reaction and mass transfer rate coefficients, as well as headspace air exchange rates. Three additional experiments were completed to evaluate model performance. Experimental and model results suggest that washing machine environments are very conducive to chloroform formation, with chloroform levels frequently exceeding 1 mg/L in washwater. Chloroform stripping efficiencies were observed to be greater than those previously reported for ethanol, but less than those reported for radon. Mass emissions of chloroform to indoor air during a ten-minute wash cycle were predicted to be between 5.3 and 9.8 mg. On a unit activity basis, chloroform emissions associated with hypochlorite-containing bleach addition to washing machines far exceeded emissions from showers. Each source was estimated to emit similar quantities of chloroform on an annual basis. Finally, it was estimated that the use of hypochlorite-containing laundry bleaches may contribute a significant fraction of chloroform mass loadings to municipal wastewater.

  17. Cysteine racemization during the Fmoc solid phase peptide synthesis of the Nav1.7-selective peptide--protoxin II.

    PubMed

    Park, Jae H; Carlin, Kevin P; Wu, Gang; Ilyin, Victor I; Kyle, Donald J

    2012-07-01

    Protoxin II is biologically active peptide containing the inhibitory cystine knot motif. A synthetic version of the toxin was generated with standard Fmoc solid phase peptide synthesis. If N-methylmorpholine was used as a base during synthesis of the linear protoxin II, it was found that a significant amount of racemization (approximately 50%) was observed during the process of cysteine residue coupling. This racemization could be suppressed by substituting N-methylmorpholine with 2,4,6-collidine. The crude linear toxin was then air oxidized and purified. Electrophysiological assessment of the synthesized protoxin II confirmed its previously described interactions with voltage-gated sodium channels. Eight other naturally occurring inhibitory knot peptides were also synthesized using this same methodology. The inhibitory potencies of these synthesized toxins on Nav1.7 and Nav1.2 channels are summarized.

  18. Potential of vancomycin for the enantiomeric resolution of FMOC-amino acids by capillary electrophoresis-ion-trap-mass spectrometry.

    PubMed

    Sánchez-Hernández, Laura; Domínguez-Vega, Elena; Montealegre, Cristina; Castro-Puyana, María; Marina, María Luisa; Crego, Antonio L

    2014-05-01

    The potential of the antibiotic vancomycin (VC) as chiral selector for the enantiomeric separation of amino acids by CE-ESI-MS/MS² was investigated for the first time in this work. Derivatization of amino acids with FMOC-Cl was carried out to enable their interaction with VC as well as the formation of precursor ions with larger m/z which were employed in MS² experiments. The partial filling of a coated capillary was employed to avoid the loss in MS sensitivity originated by the introduction of VC in the ionization source. Under optimized conditions, the simultaneous enantiomeric separation and unequivocal identification of 17 amino acids (two of them being nonprotein amino acids) took place in about 20 min with LODs in the micromolar range.

  19. Synthesis of a-factor peptide from Saccharomyces cerevisiae and photoactive analogues via Fmoc solid phase methodology.

    PubMed

    Mullen, Daniel G; Kyro, Kelly; Hauser, Melinda; Gustavsson, Martin; Veglia, Gianluigi; Becker, Jeffery M; Naider, Fred; Distefano, Mark D

    2011-01-01

    a-Factor from Saccharomyces cerevisiae is a farnesylated dodecapeptide involved in mating. The molecule binds to a G-protein coupled receptor and hence serves as a simple system for studying the interactions between prenylated molecules and their cognate receptors. Here, we describe the preparation of a-factor and two photoactive analogues via Fmoc solid-phase peptide synthesis using hydrazinobenzoyl AM NovaGel™ resin; the structure of the synthetic a-factor was confirmed by MS-MS analysis and NMR; the structures of the analogues were confirmed by MS-MS analysis. Using a yeast growth arrest assay, the analogues were found to have activity comparable to a-factor itself.

  20. Synthesis of a-factor peptide from Saccharomyces cerevisiae and photoactive analogues via Fmoc solid phase methodology

    PubMed Central

    Mullen, Daniel G.; Kyro, Kelly; Hauser, Melinda; Gustavsson, Martin; Veglia, Gianluigi; Becker, Jeffery M.; Naider, Fred; Distefano, Mark D.

    2010-01-01

    a-Factor from S. cerevisiae is a farnesylated dodecapeptide involved in mating. The molecule binds to a G-protein coupled receptor and hence serves as a simple system for studying the interactions between prenylated molecules and their cognate receptors. Here, we describe the preparation of a-factor and two photoactive analogues via Fmoc solid-phase peptide synthesis using hydrazinobenzoyl AM NovaGel™ resin; the structure of the synthetic a-factor was confirmed by MS-MS analysis and NMR; the structures of the analogues were confirmed by MS-MS analysis. Using a yeast growth arrest assay, the analogues were found to have activity comparable to a-factor itself. PMID:21134758

  1. 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...

  2. 21 CFR 700.18 - Use of chloroform as an ingredient in cosmetic products.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 7 2011-04-01 2010-04-01 true Use of chloroform as an ingredient in cosmetic products. 700.18 Section 700.18 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN... products. Recent information has become available associating chloroform with carcinogenic effects...

  3. Transformation of chloroform in model treatment wetlands: from mass balance to microbial analysis.

    PubMed

    Chen, Yi; Wen, Yue; Zhou, Junwei; Zhou, Qi; Vymazal, Jan; Kuschk, Peter

    2015-05-19

    Chloroform is one of the common disinfection byproducts, which is not susceptible to degradation and poses great health concern. In this study, the chloroform removal efficiencies and contributions of sorption, microbial degradation, plant uptake, and volatilization were evaluated in six model constructed wetlands (CWs). The highest chloroform removal efficiency was achieved in litter-added CWs (99%), followed by planted (46-54%) and unplanted CWs (39%). Mass balance study revealed that sorption (73.5-81.2%) and microbial degradation (17.6-26.2%) were the main chloroform removal processes in litter-added CWs, and that sorption (53.6-66.1%) and plant uptake (25.3-36.2%) were the primary contributors to chloroform removal in planted CWs. Around 60% of chloroform got accumulated in the roots after plant uptake, and both transpiration and gas-phase transport were expected to be the drivers for the plant uptake. Sulfate-reducing bacteria and methanogens were found to be the key microorganisms for chloroform biodegradation through cometabolic dechlorination, and positive correlations were observed between functional genes (dsrA, mcrA) and biodegradation rates. Overall, this study suggests that wetland is an efficient ecosystem for sustainable chloroform removal, and that plant and litter can enhance the removal performance through root uptake and microbial degradation stimulation, respectively.

  4. Kinetics of the reaction of diethylene glycol bis-chloroformate with allyl alcohol

    SciTech Connect

    Alekseev, N.N.; Shtoda, N.F.; Dzumedzei, N.V.

    1988-10-01

    The kinetics of diethylene glycol bis-chloroformate solvolysis by excess allyl alcohol in toluene and carbon tetrachloride has been studied. Under conditions of a pseudofirst order reaction with respect to diethylene glycol bis-chloroformate the activation parameters confirm an addition-detachment mechanism.

  5. Sequential release of antigens from chloroform-treated Staphylococcus epidermidis: application towards a possible vaccine.

    PubMed

    Ahmad, A; Clarke, S; Buchan, A; Skinner, G R

    1990-11-01

    This study describes the properties of two potential Staphylococcus epidermidis vaccines prepared by chloroform treatment of bacteria and release of antigen from these chloroform-treated organisms. Both vaccines were antigenic on testing with homologous hyperimmune serum and induced immune reactivity in immunized rabbits. There was protective efficacy in mice against intraperitoneal challenge by Staph. epidermidis.

  6. Formation of chloroform during chlorination of alanine in drinking water.

    PubMed

    Chu, Wen-Hai; Gao, Nai-Yun; Deng, Yang; Dong, Bing-Zhi

    2009-11-01

    Currently, dissolved nitrogenous organic matters in water, important precursors of disinfection by-products (DBPs), are of significant concern. This study was to explore the formation of chloroform (CF) during chlorination of alanine (Ala), an important nitrogenous organic compound commonly present in water sources. Our results indicated that the CF yield reached a maximum value of 0.143% at the molar ratio of chlorine atom to nitrogen atom (Cl/N)=1.0 over a Cl/N range of 0.2-5.0 (pH=7.0, reaction time=5d, and initial Ala=0.1mM). At an acidic-neutral condition (pH 4-7), the formation of CF was suppressed. However, the highest CF yield (0.227%) occurred at weakly alkaline condition (pH 8.0) (initial Ala=0.1mM, and Cl/N=1.0). The increase of Br(-) in water can increase total trihalomethanes (THMs) and bromo-THMs. However, the bromo-THMs level reached a plateau at Br(-)/Cl>0.04. Finally, based on the computation of frontier electron density and identification and measurement of key intermediates during Ala chlorination, we proposed a formation pathway of CF from Ala chlorination: Ala-->monochloro-N-alanine (MC-N-Ala)-->acetaldehyde (AAld)-->monochloroacetaldehyde acetaldehyde (MCAld)-->dichloroacetaldehyde (DCAld)-->trichloroacetaldehyde (TCAld)-->CF.

  7. Alternative solvents to chloroform for gutta-percha removal.

    PubMed

    Wourms, D J; Campbell, A D; Hicks, M L; Pelleu, G B

    1990-05-01

    The purpose of this study was to evaluate new or previously identified solvents for effectiveness in dissolving gutta-percha. The solubility of a standardized weight of gutta-percha was tested in 5-ml quantities of 32 solvents. Ten samples of 30 solvents were tested at both 22 and 37 degrees C for 15 min. If the gutta-percha samples dissolved within 15 min, the times were recorded and statistically analyzed. There was no significant difference (p greater than 0.05, one-way analysis of variance and Scheffe's test) between dissolving times of chloroform and trichloroethylene at 22 and 37 degrees C. At 22 degrees C, three solvents dissolved the gutta-percha sample, whereas at 37 degrees C, nine solvents dissolved the gutta-percha. At 37 degrees C, halothane (Fluothane) was twice as effective as cineole and demonstrated many favorable working qualities. An important finding from the experimentation with halothane was the discovery of a potential obturating material, which was termed "halopercha".

  8. Carbonyl sulfide uptake and chloroform emissions from an Arctic site

    NASA Astrophysics Data System (ADS)

    Elkins, J. W.; Dutton, G. S.; Montzka, S. A.; Nance, J. D.; Hall, B. D.; Thoning, K. W.; Miller, J. B.; White, J.; Vaugh, B.; Manning, A.

    2008-12-01

    The Arctic Region is most sensitive to future climate change. Quantifying emissions and sinks of many important biogenic trace gases there may become important indicators of potential climate feedback. Once snowmelt at Pt. Barrow, Alaska (77o N) occurs, ground cover is exposed by sunlight and higher temperatures, then photosynthesis starts up. Peaks of chloroform (CHCl3) appear throughout the summer from southerly-based air masses based over northern Alaska and northwest Canada. Carbonyl sulfide (COS) undergoes uptake throughout the summer season through the same enzymes that uptake carbon dioxide (CO2). We will calculate the footprint of emissions of CHCl3 and uptake of COS using high frequency in situ observations, and the NAME and FLEXPART models. Previous studies show a large source of CHCl3 (8% of the total budget) may be coming from soils in high latitude pine forests. We will examine emissions of CHCl3 to see whether or not they are coming from the tundra just south of Pt. Barrow. We will identify the regions for uptake of COS and CO2 from the footprint generated from the models.

  9. QSPR for predicting chloroform formation in drinking water disinfection.

    PubMed

    Luilo, G B; Cabaniss, S E

    2011-01-01

    Chlorination is the most widely used technique for water disinfection, but may lead to the formation of chloroform (trichloromethane; TCM) and other by-products. This article reports the first quantitative structure-property relationship (QSPR) for predicting the formation of TCM in chlorinated drinking water. Model compounds (n = 117) drawn from 10 literature sources were divided into training data (n = 90, analysed by five-way leave-many-out internal cross-validation) and external validation data (n = 27). QSPR internal cross-validation had Q² = 0.94 and root mean square error (RMSE) of 0.09 moles TCM per mole compound, consistent with external validation Q2 of 0.94 and RMSE of 0.08 moles TCM per mole compound, and met criteria for high predictive power and robustness. In contrast, log TCM QSPR performed poorly and did not meet the criteria for predictive power. The QSPR predictions were consistent with experimental values for TCM formation from tannic acid and for model fulvic acid structures. The descriptors used are consistent with a relatively small number of important TCM precursor structures based upon 1,3-dicarbonyls or 1,3-diphenols.

  10. The antimicrobial effect of chloroform on Enterococcus faecalis after gutta-percha removal.

    PubMed

    Edgar, Scott W; Marshall, J Gordon; Baumgartner, J Craig

    2006-12-01

    The purpose of this in vitro study was to evaluate the antimicrobial effectiveness of chloroform on Enterococcus faecalis when used as a gutta-percha solvent during endodontic retreatment. Bilaterally matched human teeth were instrumented, infected with E. faecalis, and obturated. The gutta-percha was then removed using either chloroform or saline. Bacterial samples were collected after gutta-percha removal and following additional apical enlargement. A significant difference was seen (p<0.05) between the number of colony forming units (CFU) of E. faecalis for teeth retreated using chloroform (mean 21+56 CFU/ml) versus saline (mean 280+480 CFU/ml). Negative cultures were obtained in 11 of 17 chloroform samples and none of the saline samples. Samples taken after apical enlargement two sizes larger than the original master apical file showed a significant difference (p<0.05) between teeth retreated using chloroform versus saline. Negative cultures were seen in 9 of 17 chloroform samples and 1 of 17 saline samples. This study demonstrated that the use of chloroform during endodontic retreatment significantly reduced intracanal levels of cultivatable E. faecalis.

  11. Effects of combined UV and chlorine treatment on chloroform formation from triclosan.

    PubMed

    Ben, Weiwei; Sun, Peizhe; Huang, Ching-Hua

    2016-05-01

    The co-exposure to UV irradiation and free chlorine may occur in certain drinking water and wastewater treatment systems. This study investigated the effects of simultaneous low pressure ultraviolet (LPUV) irradiation and free chlorination on the formation of chloroform from triclosan which is a commonly used antibacterial agent. Different treatment systems (i.e., combined UV/chlorine, UV alone, and chlorine alone) were applied to examine the degradation of triclosan and formation of chloroform. The fate of representative intermediates, including chlorinated triclosan, dechlorinated triclosan intermediates and 2,4-dichlorophenol, were tracked to deduce the effect of combined UV/chlorine on the transformation of chloroform formation precursors. The relation between intermediates degradation and chloroform formation was investigated in depth by conducting stepwise experiments with UV and chlorine in different sequences. Results indicate that the combined UV/chlorine notably enhanced the chloroform formation from triclosan. From the reaction mechanism perspective the combined UV/chlorine, where the direct photolysis may play an important role, could accelerate the decay of intermediates and facilitate the generation of productive chloroform precursors. The radicals had modest influence on the degradation of triclosan and intermediates and partly hindered the formation of chloroform. These results emphasize the necessity of considering disinfection by-products formation in the application of combined UV/chlorine technology during water treatment.

  12. Adaptive tolerance in mice upon subchronic exposure to chloroform: Increased exhalation and target tissue regeneration

    SciTech Connect

    Anand, Sathanandam S. . E-mail: sanand@rx.uga.edu; Philip, Binu K.; Palkar, Prajakta S.; Mumtaz, Moiz M.; Latendresse, John R.; Mehendale, Harihara M. . E-mail: mehendale@ulm.edu

    2006-06-15

    The aims of the present study were to characterize the subchronic toxicity of chloroform by measuring tissue injury, repair, and distribution of chloroform and to assess the reasons for the development of tolerance to subchronic chloroform toxicity. Male Swiss Webster (SW) mice were given three dose levels of chloroform (150, 225, and 300 mg/kg/day) by gavage in aqueous vehicle for 30 days. Liver and kidney injury were measured by plasma ALT and BUN, respectively, and by histopathology. Tissue regeneration was assessed by {sup 3}H-thymidine incorporation into hepato- and nephro-nuclear DNA and by proliferating cell nuclear antigen staining. In addition, GSH and CYP2E1 in liver and kidney were assessed at selected time points. The levels of chloroform were measured in blood, liver, and kidney during the dosing regimen (1, 7, 14, and 30 days). Kidney injury was evident after 1 day with all three doses and sustained until 7 days followed by complete recovery. Mild to moderate liver injury was observed from 1 to 14 days with all three dose levels followed by gradual decrease. Significantly higher regenerative response was evident in liver and kidney at 7 days, but the response was robust in kidney, preventing progression of injury beyond first week of exposure. While the kidney regeneration reached basal levels by 21 days, moderate liver regeneration with two higher doses sustained through the end of the dosing regimen and 3 days after that. Following repeated exposure for 7, 14, and 30 days, the blood and tissue levels of chloroform were substantially lower with all three dose levels compared to the levels observed with single exposure. Increased exhalation of {sup 14}C-chloroform after repeated exposures explains the decreased chloroform levels in circulation and tissues. These results suggest that toxicokinetics and toxicodynamics (tissue regeneration) contribute to the tolerance observed in SW mice to subchronic chloroform toxicity. Neither bioactivation nor

  13. Systemic inflammatory response due to chloroform intoxication--an uncommon complication.

    PubMed

    Dettling, A; Stadler, K; Eisenbach, C; Skopp, G; Haffner, H T

    2016-03-01

    Well-known adverse effects of chloroform are drowsiness, nausea, and liver damage. Two cases with an uncommon complication due to chloroform intoxication are presented. In the first case, a general physician, because of nausea and dyspnea, admitted a 34-year-old woman to hospital. She developed a toxic pulmonary edema requiring mechanical ventilation for a few days, and a systemic inflammatory response syndrome (SIRS) with elevated white blood cell counts, a moderate increase of C-reactive protein, and slightly elevated procalcitonin levels. There were inflammatory altered skin areas progressing to necrosis later on. However, bacteria could be detected neither in blood culture nor in urine. Traces of chloroform were determined from a blood sample, which was taken 8 h after admission. Later, the husband confessed to the police having injected her chloroform and put a kerchief soaked with chloroform over her nose and mouth. In the second case, a 50-year-old man ingested chloroform in a suicidal attempt. He was found unconscious in his house and referred to a hospital. In the following days, he developed SIRS without growth of bacteria in multiple blood cultures. He died several days after admission due to multi-organ failure. SIRS in response to chloroform is a rare but severe complication clinically mimicking bacterial-induced sepsis. The mechanisms leading to systemic inflammation after chloroform intoxication are currently unclear. Possibly, chloroform and/or its derivates may interact with pattern recognition receptors and activate the same pro-inflammatory mediators (cytokines, interleukins, prostaglandins, leukotrienes) that cause SIRS in bacterial sepsis.

  14. Chloroform formation in Arctic and Subarctic soils - mechanism and emissions to the atmosphere

    NASA Astrophysics Data System (ADS)

    Albers, Christian N.; Johnsen, Anders R.; Jacobsen, Ole S.

    2015-04-01

    It is well established that halogenated organic compounds are formed naturally in the terrestrial environment. These compounds include volatiles such as trihalomethanes that may escape to the atmosphere. In deed most of the atmospheric chloroform (and other trihalomethane species) is regarded to have a natural origin. This origin may be both marine and terrestrial. Chloroform formation in soil has been reported in a number of studies, mostly conducted in temperate and (sub-) tropical environments. We hereby report that also colder soils emit chloroform naturally. We measured in situ the fluxes of chloroform from soil to atmosphere in 6 Subarctic and 5 Arctic areas covering different dwarf heath, wetland and forest biotopes in Greenland and Northern Sweden. Emissions were largest from the forested areas, but all areas emitted measurable amounts of chloroform. Also the brominated analog bromodichloromethane was formed in Arctic and Subarctic soils but the fluxes to the atmosphere were much lower than the corresponding chloroform emissions. No other volatile poly-halogenated organic compounds were found to be emitted from the study areas. It has previously been proposed that chloroform is formed in temperate forest soils through trichloroacetyl intermediates formed by unspecific enzymatic chlorination of soil organic matter. We found positive relationships between chloroform emissions and the concentration of trichloroacetyl groups in soil within the various biotopes. The hydrolysis of trichloroacetyl compounds is, however, very pH dependent, excluding a simple relationship between trichloroacetyl concentration and chloroform emission in any given soil. However, our results show that at low pH, turnover time of soil trichloroacetyl compounds may be counted in decades while at pH above 6, turnover time may be just a few months. We found no relationship between trichloroacetyl concentration and total organic chlorine concentration in the soils indicating that more than

  15. 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

  16. Polyamine Homeostasis in Wild Type and Phenolamide Deficient Arabidopsis thaliana Stamens

    PubMed Central

    Fellenberg, Christin; Ziegler, Jörg; Handrick, Vinzenz; Vogt, Thomas

    2012-01-01

    Polyamines (PAs) like putrescine, spermidine, and spermine are ubiquitous polycationic molecules that occur in all living cells and have a role in a wide variety of biological processes. High amounts of spermidine conjugated to hydroxycinnamic acids are detected in the tryphine of Arabidopsis thaliana pollen grains. Tapetum localized spermidine hydroxycinnamic acid transferase (SHT) is essential for the biosynthesis of these anther specific tris-conjugated spermidine derivatives. Sht knockout lines show a strong reduction of hydroxycinnamic acid amides (HCAAs). The effect of HCAA-deficient anthers on the level of free PAs was measured by a new sensitive and reproducible method using 9-fluorenylmethyl chloroformate (FMOC) and fluorescence detection by HPLC. PA concentrations can be accurately determined even when very limited amounts of plant material, as in the case of A. thaliana stamens, are available. Analysis of free PAs in wild type stamens compared to sht deficient mutants and transcript levels of key PA biosynthetic genes revealed a highly controlled regulation of PA homeostasis in A. thaliana anthers. PMID:22912643

  17. [Determination of netilmicin in rat plasma by reversed-phase high performance liquid chromatography with fluorescence detection and pre-column derivatization].

    PubMed

    Chang, Xiaojuan; Peng, Jingdong; Liu, Shaopu; Liu, Limin; Dai, Yongkuang

    2009-11-01

    A new, simple and sensitive method based on pre-column derivatization by reversed-phase high performance liquid chromatography (HPLC) is described for the separation and quantification of netilmicin in plasma, using 9-fluorenylmethyl chloroformate (FMOC-Cl) as the derivatization reagent. Its pharmacokinetics is also presented. The derivatization modes and chromatographic conditions were optimized. The separation was performed on an Agilent ZORBAX Eclipse XDB-C8 column (150 mm x 4.6 mm, 5 microm) with a mixture of water-acetonitrile (15:85, v/v) as mobile phase and the flow rate was 1.0 mL/min. The excitation wavelength was 265 nm and the emission wavelength was 315 nm. The linear range was 0.045-8.88 mg/L and the correlation coefficient (r) was 0.9993. The limit of detection (LOD) (S/N = 3) was about 0.01 mg/L, and the limit of quantification was 0.03 mg/L (3LOD) for netilmicin. The relative standard deviation was less than 3% for intra-day assay (n = 5) and 3.5% for inter-day assay (n = 5) and the relative recovery was in the range of 96.62%-100.84% (n = 3). The plasma volume of 30 microL was sufficient for the determination of netilmicin. The method provides a reliable bioanalytical methodology to carry out netilmicin pharmacokinetics in rat plasma.

  18. Application of solid-phase microextraction combined with derivatization to the determination of amphetamines by liquid chromatography.

    PubMed

    Cháfer-Pericás, C; Campíns-Falcó, P; Herráez-Hernández, R

    2004-10-15

    This work evaluates the utility of solid-phase microextraction (SPME) in the analysis of amphetamines by liquid chromatography (LC) after chemical derivatization of the analytes. Two approaches have been tested and compared, SPME followed by on-fiber derivatization of the extracted amphetamines, and solution derivatization followed by SPME of the derivatives formed. Both methods have been applied to measure amphetamine (AP), methamphetamine (MA), and 3,4-methylenedioxymethamphetamine (MDMA), using the fluorogenic reagent 9-fluorenylmethyl chloroformate (FMOC) and carbowax-templated resin (CW-TR)-coated fibers. Data on the application of the proposed methods for the analysis of different kind of samples are presented. When analyzing aqueous solutions of the analytes, both approaches gave similar analytical performance, but the sensitivity attainable with the solution derivatization/SPME method was better. The efficiencies observed when processing spiked urine samples by the SPME/on-fiber derivatization approach were very low. This was because the extraction of matrix components into the fiber coating prevented the extraction of the reagent. In contrast, the efficiencies obtained for spiked urine samples by the solution derivatization/SPME approach were similar to those obtained for aqueous samples. Therefore, the later method would be the method of choice for the quantification of amphetamines in urine.

  19. Determination of amphetamines in hair by integrating sample disruption, clean-up and solid phase derivatization.

    PubMed

    Argente-García, A; Moliner-Martínez, Y; Campíns-Falcó, P; Verdú-Andrés, J; Herráez-Hernández, R

    2016-05-20

    The utility of matrix solid phase dispersion (MSPD) for the direct analysis of amphetamines in hair samples has been evaluated, using liquid chromatography (LC) with fluorescence detection and precolumn derivatization. The proposed approach is based on the employment of MSPD for matrix disruption and clean-up, followed by the derivatization of the analytes onto the dispersant-sample blend. The fluorogenic reagent 9-fluorenylmethyl chloroformate (FMOC) has been used for derivatization. Different conditions for MSPD, analyte purification and solid phase derivatization have been tested, using amphetamine (AMP), methamphetamine (MET), ephedrine (EPE) and 3,4-methylenedioxymethamphetamine (MDMA) as model compounds. The results have been compared with those achieved by using ultrasound-assisted alkaline digestion and by MSPD combined with conventional solution derivatization. On the basis of the results obtained, a methodology is proposed for the analysis of amphetamines in hair which integrates sample disruption, clean-up and derivatization using a C18 phase. Improved sensitivity is achieved with respect to that obtained by the alkaline digestion or by the MSPD followed by solution derivatization methods. The method can be used for the quantification of the tested amphetamines within the 2.0-20.0ng/mg concentration interval, with limits of detection (LODs) of 0.25-0.75ng/mg. The methodology is very simple and rapid (the preparation of the sample takes less than 15min).

  20. Residue determination of glufosinate in plant origin foods using modified Quick Polar Pesticides (QuPPe) method and liquid chromatography coupled with tandem mass spectrometry.

    PubMed

    Han, Yongtao; Song, Le; Zhao, Pengyue; Li, Yanjie; Zou, Nan; Qin, Yuhong; Li, Xuesheng; Pan, Canping

    2016-04-15

    A sensitive and specific method for the determination of glufosinate in plant origin foods was developed. The method involves extraction using modified QuPPe method, clean-up by multi-walled carbon nanotubes (MWCNTs), derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) and detection with liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The method was validated on twelve matrices spiked at 10 or 20, 100 and 500 μg/kg. The recovery ranged from 80% to 108% with intra-day RSDs (n=5) of 0.6-9.8% and inter-day RSDs (n=15) of 3.0-9.4%. Good linearities (R(2)⩾0.9991) were obtained for all matrices. The limit of detection (LOD) and limit of quantification (LOQ) for the selected matrices ranged from 0.3 to 3.3 μg kg(-1) and from 1 to 10 μg kg(-1), respectively. The method was demonstrated to be reliable and sensitive for the routine monitoring of glufosinate in plant origin foods.

  1. A Fmoc-compatible Method for the Solid-Phase Synthesis of Peptide C-Terminal (alpha)-Thioesters based on the Safety-Catch Hydrazine Linker

    SciTech Connect

    Camarero, J A; Hackel, B J; de Yoreo, J J; Mitchell, A R

    2003-11-22

    C-terminal peptide thioesters are key intermediates for the synthesis/semisynthesis of proteins and for the production of cyclic peptides by native chemical ligation. They can be synthetically prepared by solid-phase peptide synthesis (SPPS) methods or biosynthetically by protein splicing techniques. Until recently, the chemical synthesis of C-terminal a-thioester peptides by SPPS was largely restricted to the Boc/Benzyl methodology because of the poor stability of the thioester bond to the basic conditions employed for the deprotection of the N{sup {alpha}}-Fmoc group. In the present work, we describe a new method for the SPPS of C-terminal thioesters by Fmoc/t-Bu chemistry. This method is based on the use of an aryl hydrazide linker, which is totally stable to the Fmoc-SPPS conditions. Once the peptide synthesis has been completed, activation of the linker can be achieved by mild oxidation. This step transforms the hydrazide group into a highly reactive diazene intermediate which can react with different H-AA-SEt to yield the corresponding {alpha}-thioester peptide in good yields. This method has been successfully used for the generation of different thioester peptides, circular peptides and a fully functional SH3 protein domain.

  2. Abiotic degradation rates for carbon tetrachloride and chloroform: Final report.

    SciTech Connect

    Amonette, James E.; Jeffers, Peter M.; Qafoku, Odeta; Russell, Colleen K.; Humphrys, Daniel R.; Wietsma, Thomas W.; Truex, Michael J.

    2012-12-01

    This report documents the objectives, technical approach, and progress made through FY 2012 on a project initiated in FY 2006 to help address uncertainties related to the rates of hydrolysis in groundwater for carbon tetrachloride (CT) and chloroform (CF). The project also sought to explore the possible effects of contact with minerals and sediment (i.e., heterogeneous hydrolysis) on these rates. We conducted 114 hydrolysis rate experiments in sealed vessels across a temperature range of 20-93 °C for periods as long as 6 years, and used the Arrhenius equation to estimate activation energies and calculate half-lives for typical Hanford groundwater conditions (temperature of 16 °C and pH of 7.75). We calculated a half-life of 630 years for hydrolysis for CT under these conditions and found that CT hydrolysis was unaffected by contact with sterilized, oxidized minerals or Hanford sediment within the sensitivity of our experiments. In contrast to CT, hydrolysis of CF was generally slower and very sensitive to pH due to the presence of both neutral and base-catalyzed hydrolysis pathways. We calculated a half-life of 3400 years for hydrolysis of CF in homogeneous solution at 16 °C and pH 7.75. Experiments in suspensions of Hanford sediment or smectite, the dominant clay mineral in Hanford sediment, equilibrated to an initial pH of 7.2, yielded calculated half-lives of 1700 years and 190 years, respectively, at 16 °C. Experiments with three other mineral phases at the same pH (muscovite mica, albite feldspar, and kaolinite) showed no change from the homogeneous solution results (i.e., a half-life of 3400 years). The strong influence of Hanford sediment on CF hydrolysis was attributed to the presence of smectite and its ability to adsorb protons, thereby buffering the solution pH at a higher level than would otherwise occur. The project also determined liquid-vapor partition coefficients for CT under the temperatures and pressures encountered in the sealed vessels that

  3. Antigenicity of chloroform-methanol-treated Coxiella burnetii preparations.

    PubMed

    Kazár, J; Schramek, S; Lisák, V; Brezina, R

    1987-03-01

    Phase I Coxiella burnetii (C.b.) cells untreated (Cb I) or treated with chloroform-methanol (CM) mixture (Cb I-CM) were compared as to their capacity to induce antibodies in laboratory animals and cattle, their ability to elicit delayed type hypersensitivity (DTH) reaction in mice and rabbits and protective effect in mice. In all animal species (mice, guinea pigs, rabbits, cattle) tested, the same doses of Cb I-CM cells induced lower levels of both phase I and phase II microagglutinating (MA) antibodies than Cb I cells at different intervals post-immunization (p.i.). Though for elicitation of DTH reaction in rabbits immunized with different C.b. preparations lower doses of Cb I than of Cb I-C M cells were necessary, C.b. cells caused inflammatory reaction at lower doses also in control rabbits. In mice immunized with Cb I and Cb I-CM cells, but not with trichloracetic acid extract (TCAE) from intact Cb I cells, DTH reaction was elicited by the same doses of Cb I and Cb I-CM cells. Higher immunizing doses of Cb I-CM than of Cb I cells were required, however, to induce DTH reaction (as tested by TCAE) as well as protection to phase I virulent challenge. TCAE from intact Cb I cells was protective in mice also at lower doses than TCAE from Cb I-CM cells (TCAE-CM). In humans who suffered from Q fever one year ago, higher proportion of positive skin test (ST) reactions and antibody recalls with higher mean geometric titres (MGT) of phase II MA antibodies was noticed following intradermal administration of TCAE than of TCAE-CM. When humans with no evidence of Q fever in past were vaccinated with TCAE or TCAE-CM, the former preparation not only caused higher proportion of both local and general post-vaccination reactions, but also of phase II MA antibody response and positive ST reactions as tested by TCAE 3 months post-vaccination in addition to higher proportion of phase II MA antibody recalls.

  4. PHYSIOLOGICALLY BASED PHARMACOKINEITC (PBPK) MODELING OF METABOLIC INHIBITION FOR INTERACTION BETWEEN TRICHLOROETHYLENE AND CHLOROFORM

    EPA Science Inventory

    Trichloroethylene (TCE) and chloroform (CHCl3) are two of the most common environmental contaminants found in water. PBPK models have been increasingly used to predict target dose in internal tissues from available environmental exposure concentrations. A closed inhalation (or g...

  5. THE RESISTANCE OF PUPS TO LATE CHLOROFORM POISONING IN ITS RELATION TO LIVER GLYCOGEN

    PubMed Central

    Graham, Evarts A.

    1915-01-01

    The relative difficulty with which the characteristic central lobular liver necrosis can be produced in young pups after chloroform administration is in some way referable to the high glycogen contents of their livers. Evidence for this conclusion lies in the following facts: 1. Pups can readily be made to show the central liver necrosis which is found in chloroform poisoning in adults, if, prior to the administration of chloroform, they have been starved or starved and made diabetic by phlorhizin. 2. A single quantitative experiment showed that the liver of a normal, well nourished pup, twenty-four hours old, contained as much as 9.07 per cent. of glycogen. 3. The feeding of carbohydrates to adult animals lessens their susceptibility to the production of liver necrosis by chloroform. PMID:19867861

  6. Fmoc-based peptide thioester synthesis with self-purifying effect: heading to native chemical ligation in parallel formats.

    PubMed

    Thomas, Franziska

    2013-03-01

    The chemical synthesis of proteins has facilitated functional studies of proteins due to the site-specific incorporation of post-translational modifications, labels, and non-proteinogenic amino acids. Moreover, native chemical ligation provides facile access to proteins by chemical means. However, the application of the native chemical ligation reaction in the synthesis of parallel formats such as protein arrays has been complicated because of the often cumbersome and time-consuming synthesis of the required peptide thioesters. An Fmoc-based peptide thioester synthesis with self-purification on the sulfonamide 'safety-catch' linker widens this bottleneck because HPLC purification can be avoided. The method is based on an on-resin cyclization-thiolysis reaction sequence. A macrocyclization via the N-terminus of the full-length peptide followed by a thiolytic C-terminal ring opening allows selective detachment of the truncation products and the full-length peptide. A brief overview of the chemical aspects of this method is provided including the optimization steps and the automation process. Furthermore, the application of the cyclization-thiolysis approach combined with the native chemical ligation reaction in the parallel synthesis of a library of 16 SH3-domain variants of SHO1 in yeast is described, demonstrating the value of this new technique for the chemical synthesis of protein arrays.

  7. Mechanism of chloroform formation by chlorine and its inhibition by chlorine dioxide.

    PubMed

    Suh, D H; Abdel-Rahman, M S

    1985-04-01

    Chlorination of drinking waters leads to the formation of trihalomethanes arising from the reaction of chlorine and organic substances. Therefore, chlorine dioxide (ClO2) which does not produce trihalomethanes is being considered as an alternative disinfectant. It has been reported that rat blood chloroform levels were significantly decreased after treatment with ClO2. Studies were conducted to investigate the mechanisms of chloroform formation by chlorine (HOCl) and its inhibition by ClO2 (5 mg/liter) in the presence of HOCl (5, 10, 20 mg/liter) using sodium citrate (1 mM) as an organic substance. When citrate was reacted with HOCl, beta-ketoglutaric acid, monochloroacetone, dichloroacetone, and trichloroacetone were produced as reaction intermediates and chloroform as a final product. There was a linear relationship between the concentrations of HOCl and the formation of chloroform. When ClO2 was substituted for HOCl, neither chloroform was formed nor citrate concentration was changed. Further, chloroform formation was inhibited by ClO2 in the presence of HOCl and citrate and the degree of inhibition depends on the ratio of ClO2/HOCl. Gas chromatograph/mass spectrometer analysis indicates that this inhibition is related to the reaction of ClO2 with beta-ketoglutaric acid to form malonic acid. Chlorine dioxide also oxidizes other intermediates such as monochloroacetone and dichloroacetone to acetic acid. These studies indicate that ClO2 inhibits chloroform formation from citrate and HOCl by the oxidation of the intermediates which were involved in the reaction of chloroform formation.

  8. Hot spot formation of chloroform in forest soils caused pollution of groundwater

    NASA Astrophysics Data System (ADS)

    Jacobsen, Ole S.; Albers, Christian N.; Laier, Troels; Hunkeler, Daniel

    2015-04-01

    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 from 1 - 10 µg/L. Groundwater containing chloroform that exceeds 1 µg/L could not be used for drinking water according to Danish regulations. The strict demands on groundwater quality may have to be taken into account when decisions are made regarding the change of land use in order to protect major recharge areas from pollution with nitrate and pesticides resulting from high-yield agriculture production. The terrestrial environment and especially hot spots in forest soils seem to be important contributors to apparent pollution of groundwater with chloroform. We performed a field study to investigate concentration and fluxes of chloroform to the groundwater from in four coniferous forests in order to increase knowledge on the hot spot formation and fate of natural chloroform. We investigated four stations over a period of several years in order to measure the net-formation of chloroform. Field measurements soil air concentrations of chloroform were monitored in five soil profiles down to the groundwater table. Meteorological data were recorded at all stations In the hotspots up to 120 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 with a maximum in August-September. The chloroform concentration decreased with depth in all profiles during the summer half-year to about 20 % of concentration in the production layer. However, the concentration is still high enough to give an equilibrium concentration in the upper groundwater of 1-10 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

  9. 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.

  10. 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

  11. Redox control of GPx catalytic activity through mediating self-assembly of Fmoc-phenylalanine selenide into switchable supramolecular architectures.

    PubMed

    Huang, Zupeng; Luo, Quan; Guan, Shuwen; Gao, Jianxiong; Wang, Yongguo; Zhang, Bo; Wang, Liang; Xu, Jiayun; Dong, Zeyuan; Liu, Junqiu

    2014-12-28

    Artificial enzymes capable of achieving tunable catalytic activity through stimuli control of enzymatic structure transition are of significance in biosensor and biomedicine research. Herein we report a novel smart glutathione peroxidise (GPx) mimic with modulatory catalytic activity based on redox-induced supramolecular self-assembly. First, an amphiphilic Fmoc-phenylalanine-based selenide was designed and synthesized, which can self-assemble into nanospheres (NSs) in aqueous solution. The NSs demonstrate extremely low GPx activity. Upon the oxidation of hydroperoxides (ROOH), the selenide can be quickly transformed into the selenoxide form. The change of the molecular structure induces complete morphology transition of the self-assemblies from NSs to nanotubes (NTs), resulting in great enhancement in the GPx catalytic activity. Under the reduction of GSH, the selenoxide can be further reversibly reduced back into the selenide; therefore the reversible switch between the NSs and NTs can be successfully accomplished. The relationship between the catalytic activity and enzymatic structure was also investigated. The dual response nature makes this mimic play roles of both a sensor and a GPx enzyme at the same time, which can auto-detect the signal of ROOH and then auto-change its activity to achieve quick or slow/no scavenging of ROOH. The dynamic balance of ROOH is vital in organisms, in which an appropriate amount of ROOH does benefit to the metabolism, whereas surplus ROOH can cause oxidative damage of the cell instead and this smart mimic is of remarkable significance. We expect that such a mimic can be developed into an effective antioxidant drug and provide a new platform for the construction of intelligent artificial enzymes with multiple desirable properties.

  12. High Performance Liquid Chromatography Coupled with Pre-column Derivatization for Determination of Oxidized Glutathione Level in Rats Exposed to Paraquat.

    PubMed

    Hami, Zahra; Amini, Mohsen; Kiani, Amir; Ghazi-Khansari, Mahmoud

    2013-01-01

    Glutathione (GSH) is one of the most important antioxidants that plays an essential role in detoxification of reactive oxygen species (ROS) which oxidizes to glutathione disulfide (GSSG). Paraquat (PQ), awidely used herbicide, causes pulmonary injury with the productionof ROS. Excessive ROS accumulation as a consequence of PQ exposure are frequently targeted by GSH thereby oxidative stress leads to depletion of cellular GSH by transforming of GSH to glutathione disulfide (GSSG). A precise method of measuring of GSSG concentration in plasma as indicator of oxidative stress is needed. Some analytical techniques such as high-performance liquid chromatography (HPLC), gas chromatography and capillary electrophoresis have been used for determination of GSSG concentration. In the present study, a new HPLC method with fluorescence detection based on derivatization of the amine group of glutathione with 9-fluorenylmethyl chloroformate (FMOC-Cl) was developed. Male Wistar albino rats exposed to different doses of PQ (20-60 mg/kg) and control group were used and after protein precipitation, their plasma was subjected to derivatization with FMOC in the presence of borate buffer. The derivatized samples were injected to HPLC system with C18 column, mobile phase consisting of methanol and phosphate buffer, λem= 315 nm, λex= 260 nm. Among all experimental groups, the rats which received 60 mg/kg PQ, showed a significant increase in the amount of oxidized glutathione (GSSG) compared to the control group. In this study, the applied derivatization and HPLC method made it possible to measure small amounts of glutathione in plasma using a precise and sensitive technique.

  13. High Performance Liquid Chromatography Coupled with Pre-column Derivatization for Determination of Oxidized Glutathione Level in Rats Exposed to Paraquat

    PubMed Central

    Hami, Zahra; Amini, Mohsen; Kiani, Amir; Ghazi-Khansari, Mahmoud

    2013-01-01

    Glutathione (GSH) is one of the most important antioxidants that plays an essential role in detoxification of reactive oxygen species (ROS) which oxidizes to glutathione disulfide (GSSG). Paraquat (PQ), awidely used herbicide, causes pulmonary injury with the productionof ROS. Excessive ROS accumulation as a consequence of PQ exposure are frequently targeted by GSH thereby oxidative stress leads to depletion of cellular GSH by transforming of GSH to glutathione disulfide (GSSG). A precise method of measuring of GSSG concentration in plasma as indicator of oxidative stress is needed. Some analytical techniques such as high-performance liquid chromatography (HPLC), gas chromatography and capillary electrophoresis have been used for determination of GSSG concentration. In the present study, a new HPLC method with fluorescence detection based on derivatization of the amine group of glutathione with 9-fluorenylmethyl chloroformate (FMOC-Cl) was developed. Male Wistar albino rats exposed to different doses of PQ (20-60 mg/kg) and control group were used and after protein precipitation, their plasma was subjected to derivatization with FMOC in the presence of borate buffer. The derivatized samples were injected to HPLC system with C18 column, mobile phase consisting of methanol and phosphate buffer, λem= 315 nm, λex= 260 nm. Among all experimental groups, the rats which received 60 mg/kg PQ, showed a significant increase in the amount of oxidized glutathione (GSSG) compared to the control group. In this study, the applied derivatization and HPLC method made it possible to measure small amounts of glutathione in plasma using a precise and sensitive technique. PMID:24523771

  14. Residue determination of glyphosate in environmental water samples with high-performance liquid chromatography and UV detection after derivatization with 4-chloro-3,5-dinitrobenzotrifluoride.

    PubMed

    Qian, Kun; Tang, Tao; Shi, Tianyu; Wang, Fang; Li, Jianqiang; Cao, Yongsong

    2009-03-09

    A pre-column derivatization high-performance liquid chromatographic method for glyphosate analysis has been developed. Derivatization of glyphosate was performed with 4-chloro-3,5-dinitrobenzotrifluoride (CNBF). In pH 9.5 H(3)BO(3)-Na(2)B(4)O(7) media, the reaction of glyphosate with CNBF completed at 60 degrees C for 30min. The labeled glyphosate was separated on a Kromasil C18 column (250mmx4.6mm, 5microm) at room temperature and UV detection was applied at 360nm. The separation of labeled glyphosate was achieved within 15min by gradient elution mode. Compared to other pre-column derivatization, this derivatization was performed more mildly, the derivative was more stable, and the detection limits of a few reagents were higher than CNBF, except 9-fluorenylmethyl chloroformate (FMOC-Cl) using fluorescence and mass spectrometry, however, this reagent avoid to be removed after derivatization like FMOC-Cl. The detection limit of glyphosate was 0.009mgL(-1) (S/N=3) without preconcentration and reach MRL, which is set at the level of 0.1mgL(-1) in China. The method linearity correlation coefficient was 0.9999, in concentrations ranging from 0.3 to 48.5mgL(-1). The proposed method has been applied to the quantitative determination of glyphosate in environmental water with recoveries of 91.80-100.20% and R.S.D. of 2.27-6.80, depending on the sample investigated.

  15. 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

  16. Fmoc-conjugated PEG-vitamin E2 micelles for tumor-targeted delivery of paclitaxel: enhanced drug-carrier interaction and loading capacity.

    PubMed

    Zhang, Yifei; Huang, Yixian; Zhao, Wenchen; Lu, Jianqin; Zhang, Peng; Zhang, Xiaolan; Li, Jiang; Gao, Xiang; Venkataramanan, Raman; Li, Song

    2014-11-01

    The purpose of this study is to develop an improved drug delivery system for enhanced paclitaxel (PTX) loading capacity and formulation stability based on PEG5K-(vitamin E)2 (PEG5K-VE2) system. PEG5K-(fluorenylmethoxycarbonyl)-(vitamin E)2 (PEG5K-FVE2) was synthesized using lysine as the scaffold. PTX-loaded PEG5K-FVE2 micelles were prepared and characterized. Fluorescence intensity of Fmoc in the micelles was measured as an indicator of drug-carrier interaction. Cytotoxicity of the micelle formulations was tested on various tumor cell lines. The therapeutic efficacy and toxicity of PTX-loaded micelles were investigated using a syngeneic mouse model of breast cancer (4T1.2). Our data suggest that the PEG5K-FVE2 micelles have a low CMC value of 4 μg/mL and small sizes (~60 nm). The PTX loading capacity of PEG5K-FVE2 micelles was much higher than that of PEG5K-VE2 micelles. The Fmoc/PTX physical interaction was clearly demonstrated by a fluorescence quenching assay. PTX-loaded PEG5K-FVE2 micelles exerted more potent cytotoxicity than free PTX or Taxol formulation in vitro. Finally, intravenous injection of PTX-loaded PEG5K-FVE2 micelles showed superior anticancer activity compared with PEG5K-VE2 formulation with minimal toxicity in a mouse model of breast cancer. In summary, incorporation of a drug-interactive motif (Fmoc) into PEG5K-VE2 micelles represents an effective strategy to improve the micelle formulation for the delivery of PTX.

  17. Temperature Dependence of Densities and Excess Molar Volumes of the Ternary Mixture (1-Butanol + Chloroform + Benzene) and its Binary Constituents (1-Butanol + Chloroform and 1-Butanol + Benzene)

    NASA Astrophysics Data System (ADS)

    Smiljanić, Jelena D.; Kijevčanin, Mirjana Lj.; Djordjević, Bojan D.; Grozdanić, Dušan K.; Šerbanović, Slobodan P.

    2008-04-01

    Densities ρ of the 1-butanol + chloroform + benzene ternary mixture and the 1-butanol + chloroform and 1-butanol + benzene binaries have been measured at six temperatures (288.15, 293.15, 298.15, 303.15, 308.15, and 313.15) K and atmospheric pressure, using an oscillating U-tube densimeter. From these densities, excess molar volumes ( V E) were calculated and fitted to the Redlich Kister equation for all binary mixtures and to the Nagata and Tamura equation for the ternary system. The Radojković et al. equation has been used to predict excess molar volumes of the ternary mixtures. Also, V E data of the binary systems were correlated by the van der Waals (vdW1) and Twu Coon Bluck Tilton (TCBT) mixing rules coupled with the Peng Robinson Stryjek Vera (PRSV) equation of state. The prediction and correlation of V E data for the ternary system were performed by the same models.

  18. Native Chemical Ligation Strategy to Overcome Side Reactions during Fmoc-Based Synthesis of C-Terminal Cysteine-Containing Peptides.

    PubMed

    Lelièvre, Dominique; Terrier, Victor P; Delmas, Agnès F; Aucagne, Vincent

    2016-03-04

    The Fmoc-based solid phase synthesis of C-terminal cysteine-containing peptides is problematic, due to side reactions provoked by the pronounced acidity of the Cα proton of cysteine esters. We herein describe a general strategy consisting of the postsynthetic introduction of the C-terminal Cys through a key chemoselective native chemical ligation reaction with N-Hnb-Cys peptide crypto-thioesters. This method was successfully applied to the demanding peptide sequences of two natural products of biological interest, giving remarkably high overall yields compared to that of a state of the art strategy.

  19. [Determination of chloroform and tetrachloro-methane in water with capillary column-microECD].

    PubMed

    Xu, Yinghua; Du, Daan

    2002-04-01

    The method of determination of chloroform and tetrachloro-methane in water with capillary column-microECD had better and wider linear range and lower detection limit. At the range of 0-10 micrograms/L chloroform and 0-1 microgram/L tetrachloro-methane, the regression equation were calculated as Y = 1.959X + 4.816 (r = 0.9910) and Y = 3.947X + 42.58 (r = 0.9970). The lowest detection limit were 0.1032 microgram/L chloroform and 0.01219 microgram/L tetrachloro-methane. This method not only simplified the operating processes, but also increase the sensitivity, and reavled better precision and accuracy.

  20. Self-aggregation and optical absorption of stilbazolium merocyanine in chloroform.

    PubMed

    Silva, Daniel L; Murugan, N Arul; Kongsted, Jacob; Ågren, Hans; Canuto, Sylvio

    2014-02-20

    Dipolar aggregation is in many cases detrimental for the functioning of optical materials. In this study we investigate self-aggregation and optical absorption of stilbazolium merocyanine (SM) in chloroform solution by performing classical Molecular Dynamics (MD) simulations under ambient conditions. The reversal solvatochromic shift, the large bathochromic shift, and the structured absorption band presented by SM in chloroform solution are all aspects of its optical absorption behavior for which the existence of self-aggregation is yet not completely understood. Moreover, the spectroscopic properties of SM oligomers and their occurrence in solvent of low polarity remain a relevant topic that deserves to be investigated. Our analysis of the aggregation behavior of SM in chloroform verified that the majority of the chromophores are involved in the formation of oligomers in solution, where the whole dimer and part of the trimer populations present a stable π-stacking structure. The optical properties of the monomers and oligomers in solution were evaluated by means of a discrete polarizable embedding quantum mechanical/molecular mechanical (PE-QM/MM) response scheme where the quantum part is described at the level of density functional theory. The visible absorption spectrum of SM in chloroform is simulated using time average values obtained for the monomeric and oligomeric forms of SM from the PE-QM/MM calculations performed on uncorrelated configurations extracted from the classical MD simulations. This study shows that the self-aggregation of SM in chloroform may exist, but it is not essential for reproducing the reversal solvatochromic shift in chloroform and that the process does not contribute to enhance the bathochromic shift nor explain the structure observed in its absorption band. Moreover, it is verified that since the electronic transitions of the monomer and oligomers are close together, changes in the interplane separation between the monomeric units

  1. Modeling the interaction of ozone with chloroform and bromoform under conditions close to stratospheric

    NASA Astrophysics Data System (ADS)

    Strokova, N. E.; Yagodovskaya, T. V.; Savilov, S. V.; Lukhovitskaya, E. E.; Vasil'ev, E. S.; Morozov, I. I.; Lunin, V. V.

    2013-02-01

    The reactions of ozone with chloroform and bromoform are studied using a flow gas discharge vacuum unit under conditions close to stratospheric (temperature range, 77-250 K; pressure, 10-3-0.1 Torr in the presence of nitrate ice). It is shown that the reaction with bromoform begins at 160 K; the reaction with chloroform, at 190 K. The reaction products are chlorine and bromine oxides of different composition, identified by low-temperature FTIR spectroscopy. The presence of nitrate ice raises the temperature of reaction onset to 210 K.

  2. Analysis of amino acids by HPLC/electrospray negative ion tandem mass spectrometry using 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl) derivatization.

    PubMed

    Ziegler, Jörg; Abel, Steffen

    2014-12-01

    A new method for the determination of amino acids is presented. It combines established methods for the derivatization of primary and secondary amino groups with 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl) with the subsequent amino acid specific detection of the derivatives by LC-ESI-MS/MS using multiple reaction monitoring (MRM). The derivatization proceeds within 5 min, and the resulting amino acid derivatives can be rapidly purified from matrix by solid-phase extraction (SPE) on HR-X resin and separated by reversed-phase HPLC. The Fmoc derivatives yield several amino acid specific fragment ions which opened the possibility to select amino acid specific MRM transitions. The method was applied to all 20 proteinogenic amino acids, and the quantification was performed using L-norvaline as standard. A limit of detection as low as 1 fmol/µl with a linear range of up to 125 pmol/µl could be obtained. Intraday and interday precisions were lower than 10 % relative standard deviations for most of the amino acids. Quantification using L-norvaline as internal standard gave very similar results compared to the quantification using deuterated amino acid as internal standards. Using this protocol, it was possible to record the amino acid profiles of only a single root from Arabidopsis thaliana seedlings and to compare it with the amino acid profiles of 20 dissected root meristems (200 μm).

  3. The Pore Loop Domain of TRPV1 Is Required for Its Activation by the Volatile Anesthetics Chloroform and Isoflurane.

    PubMed

    Kimball, Corinna; Luo, Jialie; Yin, Shijin; Hu, Hongzhen; Dhaka, Ajay

    2015-07-01

    The environmental irritant chloroform, a naturally occurring small volatile organohalogen, briefly became the world's most popular volatile general anesthetic (VGA) before being abandoned because of its low therapeutic index. When chloroform comes in contact with skin or is ingested, it causes a painful burning sensation. The molecular basis for the pain associated with chloroform remains unknown. In this study, we assessed the role of transient receptor potential (TRP) channel family members in mediating chloroform activation and the molecular determinants of VGA activation of TRPV1. We identified the subpopulation of dorsal root ganglion (DRG) neurons that are activated by chloroform. Additionally, we transiently expressed wild-type or specifically mutated TRP channels in human embryonic kidney cells and used calcium imaging or whole-cell patch-clamp electrophysiology to assess the effects of chloroform or the VGA isoflurane on TRP channel activation. The results revealed that chloroform activates DRG neurons via TRPV1 activation. Furthermore, chloroform activates TRPV1, and it also activates TRPM8 and functions as a potent inhibitor of the noxious chemical receptor TRPA1. The results also indicate that residues in the outer pore region of TRPV1 previously thought to be required for either proton or heat activation of the channel are also required for activation by chloroform and isoflurane. In addition to identifying the molecular basis of DRG neuron activation by chloroform and the opposing effects chloroform has on different TRP channel family members, the findings of this study provide novel insights into the structural basis for the activation of TRPV1 by VGAs.

  4. 21 CFR 700.18 - Use of chloroform as an ingredient in cosmetic products.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... animals. Studies conducted by the National Cancer Institute have demonstrated that the oral administration of chloroform to mice and rats induced hepatocellular carcinomas (liver cancer) in mice and renal... gastrointestinal tract, through the respiratory system, and through the skin. The Commissioner concludes that,...

  5. 21 CFR 700.18 - Use of chloroform as an ingredient in cosmetic products.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... animals. Studies conducted by the National Cancer Institute have demonstrated that the oral administration of chloroform to mice and rats induced hepatocellular carcinomas (liver cancer) in mice and renal... gastrointestinal tract, through the respiratory system, and through the skin. The Commissioner concludes that,...

  6. 21 CFR 700.18 - Use of chloroform as an ingredient in cosmetic products.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... animals. Studies conducted by the National Cancer Institute have demonstrated that the oral administration of chloroform to mice and rats induced hepatocellular carcinomas (liver cancer) in mice and renal... gastrointestinal tract, through the respiratory system, and through the skin. The Commissioner concludes that,...

  7. 21 CFR 700.18 - Use of chloroform as an ingredient in cosmetic products.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... animals. Studies conducted by the National Cancer Institute have demonstrated that the oral administration of chloroform to mice and rats induced hepatocellular carcinomas (liver cancer) in mice and renal... gastrointestinal tract, through the respiratory system, and through the skin. The Commissioner concludes that,...

  8. The Role of Atomic Polarization in the Thermodynamics of Chloroform Partitioning to Lipid Bilayers.

    PubMed

    Vorobyov, Igor; Bennett, W F Drew; Tieleman, D Peter; Allen, Toby W; Noskov, Sergei

    2012-02-14

    In spite of extensive research and use in medical practice, the precise molecular mechanism of volatile anesthetic action remains unknown. The distribution of anesthetics within lipid bilayers and potential targeting to membrane proteins is thought to be central to therapeutic function. Therefore, obtaining a molecular level understanding of volatile anesthetic partitioning into lipid bilayers is of vital importance to modern pharmacology. In this study we investigate the partitioning of the prototypical anesthetic, chloroform, into lipid bilayers and different organic solvents using molecular dynamics simulations with potential models ranging from simplified coarse-grained MARTINI to additive and polarizable CHARMM all-atom force fields. Many volatile anesthetics display significant inducible dipole moments, which correlate with their potency, yet the exact role of molecular polarizability in their stabilization within lipid bilayers remains unknown. We observe that explicit treatment of atomic polarizability makes it possible to accurately reproduce solvation free energies in solvents with different polarities, allowing for quantitative studies in heterogeneous molecular distributions, such as lipid bilayers. We calculate the free energy profiles for chloroform crossing lipid bilayers to reveal a role of polarizability in modulating chloroform partitioning thermodynamics via the chloroform-induced dipole moment and highlight competitive binding to the membrane core and toward the glycerol backbone that may have significant implications for understanding anesthetic action.

  9. THE INFLUENCE OF ADVANCED AGE ON THE HEPATIC AND RENAL TOXICITY OF CHLOROFORM

    EPA Science Inventory

    THE INFLUENCE OF ADVANCED AGE ON THE HEPATIC AND RENAL TOXICITY OF CHLOROFORM (CHC13). A McDonald, Y M Sey and J E Simmons. NHEERL, ORD, U.S. EPA, RTP, NC.
    Disinfection, by chlorination or by ozonation followed by treatment with either chlorine or chloramine, of water containi...

  10. Halothane and eucalyptol as alternatives to chloroform for softening gutta-percha.

    PubMed

    Hunter, K R; Doblecki, W; Pelleu, G B

    1991-07-01

    Because chloroform was identified as a potential carcinogen by the Food and Drug Administration, interest has been revived to identify an alternative solvent to soften gutta-percha for removal from obturated root canals. This study compared the effectiveness of halothane, eucalyptol, and chloroform in softening gutta-percha in simulated root canals. One milliliter of a solvent was placed into a small glass funnel whose stem was obturated with a 30-mm column of gutta-percha. After 30 s, softening was evaluated for each solvent by recording the time required to reach a depth of 10 mm by hand filing with a #100 Hedstrom file. The depth of penetration of a #40 finger plugger under constant weight for 15 min was also determined for each solvent. By using a one-way analysis of variance and Scheffe's test, all comparisons were not significant except for the depth of penetration with constant weight between chloroform and halothane (p less than 0.05). The results indicate that halothane and eucalyptol are suitable alternatives to chloroform as gutta-percha softening solvents.

  11. A Comparison of the Effectiveness of Chloroform in Dissolving Resilon and Gutta-Percha

    PubMed Central

    Azar, MR.; Khojastehpour, L.; Iranpour, N.

    2011-01-01

    Objective: Application of chemical solvents especially in problematic canals is usually a part of the retreatment process. This study was performed to compare the solubility of Gutta-Percha and Resilon in chloroform and to find the effect of sample thickness as well as the time of shaking on their solubility. Materials and Methods: Specific weight of Resilon and gutta-percha was placed in a sample tube and after adding 1.0 ml of chloroform at 37ºC, the tubes were capped and shaked for 1, 3 and 5 minutes. The amount of non dissolved material was determined by reweighting of each sample and the percent of solubility was assessed according to the exact weight loss of the samples. The procedure was repeated three times for a given thickness and time of shaking. The difference in the solubility of Gutta-Percha and Resilon as well as the effect of sample thickness and time of shaking on solubility were assessed by repeated measurement ANOVA (p<0.05). Results: Resilon has significantly higher solubility than Gutta-Percha in chloroform (p<0.05). Resilon as well as Gutta-Percha Solubility are increased significantly over the time .The amount of solubility is not affected by sample thickness. Conclusion: Comparison of Resilon and Gutta-Percha solubility in chloroform shows that one of the advantages for Resilon could be the chance for using possible safer organic solvents during retreatment. PMID:21998803

  12. Chloroform-free permeabilization for improved detection of β-galactosidase activity in Vibrio cholerae.

    PubMed

    Toulouse, Charlotte; Häse, Claudia C; Steuber, Julia

    2017-03-21

    LacZ (β-galactosidase) is used to monitor the transcription of genes in reporter strains carrying the lacZ gene under the control of a promotor of interest. This protocol for LacZ activity determinations in Vibrio cholerae following detergent lysis results in 2.5-fold increase of LacZ activities compared to lysis with chloroform.

  13. Persulfate Oxidation of MTBE- and Chloroform-Spent Granular Activated Carbon

    EPA Science Inventory

    Activated persulfate (Na2S2O8) regeneration of methyl tert-butyl ether (MTBE) and chloroform-spent GAC was evaluated in this study. Thermal-activation of persulfate was effective and resulted in greater MTBE removal than either alkaline-activation or H2O2–persulfate binary mixtur...

  14. ESTIMATING CHLOROFORM BIOTRANSFORMATION IN F-344 RAT LIVER USING IN VITRO TECHNIQUES AND PHARMACOKINETIC MODELING

    EPA Science Inventory

    ESTIMATING CHLOROFORM BIOTRANSFORMATION IN F-344 RAT LIVER USING IN VITRO TECHNIQUES AND PHARMACOKINETIC MODELING

    Linskey, C.F.1, Harrison, R.A.2., Zhao, G.3., Barton, H.A., Lipscomb, J.C4., and Evans, M.V2., 1UNC, ESE, Chapel Hill, NC ; 2USEPA, ORD, NHEERL, RTP, NC; 3 UN...

  15. 'My beloved chloroform'. Attitudes to childbearing in colonial Queensland: a case study.

    PubMed

    Woolcock, H R; Thearle, M J; Saunders, K

    1997-12-01

    In 1847 the anaesthetic and analgesic properties of chloroform were discovered. This technology generated a new era for midwifery: mothers could be relieved of pain in childbirth. The introduction of chloroform for childbirth saw increasing medical dominance in obstetrics, traditionally in the hands of the midwife. At the same time the use of chloroform sparked a medical and moral controversy which lasted for several decades. On the one hand women were destined by the 'curse of Eve' to experience pain during childbirth; on the other, medical humanitarians and practitioners believed that there were technical and moral reasons for alleviating pain in childbirth. In concentrating on the debate historians have largely ignored the reactions of mothers to the introduction of the technology. This paper explores changing attitudes to childbearing within the context of colonial Queensland society, 1860-90, by examining the correspondence of an upper-class mother. Her education and liberal outlook, and a certain ambivalence towards motherhood, all influenced her attitude to the use of chloroform and the process of childbirth.

  16. Treatment of natural ovine malignant theileriosis with a chloroform extract of the plant Peganum harmala.

    PubMed

    Mirzaiedehaghi, M

    2006-06-01

    One hundred sheep naturally infected with Theileria lestoquardi were treated with a chloroform extract of the plant Peganum harmala. The treatment was continued for 5 days, the dose of extract being 5 mg/kg per day. Sixty-five of the sheep responded to treatment and recovered but 35 did not and died. The cure rate was 65%.

  17. 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 ...

  18. Detection of chloroform in water using an azo dye-modified β-cyclodextrin - Epichlorohydrin copolymer as a fluorescent probe

    NASA Astrophysics Data System (ADS)

    Ncube, Phendukani; Krause, Rui W. M.; Mamba, Bhekie B.

    Chlorination disinfection by-products (DBPs) in water pose a health threat to humans and the aquatic environment. Their detection in water sources is therefore vital. Herein we present the detection of chloroform, a DBP, using a molecular fluorescent probe. The detection was based on the quenching of fluorescence of the probe by chloroform due to host-guest complex formation between β-cyclodextrin in the probe and the chloroform molecule. The stability constant for the host-guest complex was high at 3.825 × 104 M-1. Chloroform quenched the fluorescence of the copolymer the most compared to the other small chlorinated compounds studied, suggesting that the probe was more sensitive to chloroform, with a sensing factor of 0.35 compared to as low as 0.0073 for dichloromethane. There was no interference from other chloroalkanes on the quenching efficiency of chloroform. The probe was used to detect chloroform in dam water as well as in bottled water. Detection of chloroform in both water samples using the probe was possible without chemically treating the water samples which may introduce other pollutants.

  19. 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...

  20. Heterogeneous catalysis by solid superacids--14. Perfluorinated resinsulfonic acid (nafion-h) catalyzed Friedel-crafts alkylation of toluene and phenol with alkyl chloroformates and oxalates

    SciTech Connect

    Olah, G.A.; Meidar, D.; Malhotra, R.; Olah, J.A.; Narang, S.C.

    1980-01-01

    Product distributions obtained in the alkylation of toluene with methyl, ethyl, and isopropyl chloroformate and dimethyl oxalate and of phenol with methyl and ethyl chloroformate and dimethyl- and diethyloxalate.

  1. A non-phenol-chloroform extraction of double-stranded RNA from plant and fungal tissues.

    PubMed

    Balijja, Alitukiriza; Kvarnheden, Anders; Turchetti, Tullio

    2008-09-01

    Double-stranded RNA (dsRNA) molecules of viruses are found in nature at a very high frequency. Their detection in plants and fungi has been carried out with difficulty due to the complicated dsRNA extraction techniques used commonly which includes phenol-chloroform extractions. In this study, an extraction method for isolation of dsRNA is described that is free of phenol and chloroform. A lysis buffer, containing beta-mercaptoethanol and polyvinylpolypyrrolidone (PVPP-40), was added to homogenised tissues and the subsequent supernatant was filtered through a cellulose CF-11 mini-column. DsRNA molecules were separated based on the differing affinity of nucleic acids for the cellulose CF-11 resin in 20% ethanol buffer. This easy, rapid and cheap technique has been successfully tested on fungi and plants containing different dsRNA virus molecules, indicating the possibility of a wide use of the method.

  2. A case of drug-facilitated sexual assault leading to death by chloroform poisoning.

    PubMed

    Gaillard, Yvan; Masson-Seyer, Marie Françoise; Giroud, Michel; Roussot, Jean François; Prevosto, Jean Michel

    2006-07-01

    The purpose of this investigation was to determine the cause of death of a 13-year-old girl, where none was immediately evident. Our analysis showed it to be a very unusual case of a drug-facilitated sexual assault (DFSA), which led to the tragic death of the young rape victim and then to the suicide of the rapist. The incapacitating agent used was chloroform. The post-mortem analysis revealed a blood concentration of 833.9 mg/l for the girl, whereas the quantitation of chloroform in various fluids and viscera of the rapist proved that he had recently been handling the solvent (with concentrations in fat tissues 20 times higher than in his blood). This case draws attention to the need for broad searches for volatile substances in such investigations.

  3. Determination of biogenic amines in infusions of tea (Camellia sinensis) by HPLC after derivatization with 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl).

    PubMed

    Brückner, H; Flassig, S; Kirschbaum, J

    2012-02-01

    The reagent 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl) was used for the pre-column derivatization of the biogenic amines (BAs) cadaverine (Cad), histamine (Him), octopamine (Ocp), phenylethylamine (Pea), putrescine (Put), spermidine (Spd), spermine (Spm), tyramine (Tym) and the internal standard 1,6-diaminohexane (Dhx). The resulting Fmoc-derivatives were resolved by high-performance liquid chromatography on a Superspher(©) C(18) column using a binary gradient generated from sodium acetate and acetonitrile. For quantification, the fluorescence of derivatives was used at 263 nm excitation and 313 nm emission wavelength. This approach was applied to free BAs extractable with boiling water from 14 black, 5 green, 1 Oolong, and 1 instant tea. Infusions were prepared by adding 35 ml boiling water to one gram of tea and extracted for 20 min. In the Oolong tea and two black teas, no BAs could be detected. Limits of detection were 0.07-1.0 pmol for BAs at signal-to-noise ratio 3:1. Besides most abundant Tym and Spm lower quantities of Pea, Put, and Spd were detected, albeit not in all teas. Quantities of Tym ranged from 16 to 431 μg Tym/L infusion (1.1-25.3 μg Tym/g tea) and 31 to 319 μg Spm/L infusion (1.5-16.9 μg Spm/g tea). In none of the teas, Him was detected. Owing to the low amounts of free BAs in tea infusions, no health risks are to be expected even on consumption of large quantities of tea as beverage.

  4. A reactive nitrone-based organogel that self-assembles from its constituents in chloroform.

    PubMed

    Richards, Josh E; Philp, Douglas

    2016-04-11

    The reversible reaction of an aldehyde with a hydroxylamine affords a nitrone which is capable of forming a stiff gel with chloroform at concentrations as low as 0.20 wt% (6 mM). The gelator forms dynamically from its constituents and the gel assembly can be degraded in a controlled manner through a recognition-mediated reaction that targets the nitrone component of the gel network.

  5. Comparison of Gutta Percha Filling Techniques. Part 2. Three Chloroform-Gutta Percha Filling Techniques.

    DTIC Science & Technology

    1980-06-01

    Inlo Di-5r7W~f~ IU* Report) DTIC illE LECTE f_ IS. KEY WORDS (Conthue on revere* side It necessay and Identify by block numbei) -. , endodontics ...TECHNIQUES. PART 2: THREE CHLOROFORM-GUTTA PERCHA FILLING TECHNIQUES M. Wong, DDS Resident, Endodontics Dept. of Dentistry - PO Box 59 Madigan Army Medical...Center Tacoma, Washington 98431 D. 0. Peters, BA, DDS, MS Mentor, Endodontics US Army Institute of Dental Research Walter Reed Army Medical Center

  6. Effect of Chloroform, Eucalyptol and Orange Oil Solvents on the Microhardness of Human Root Dentin

    PubMed Central

    Khedmat, Sedigheh; Hashemi, Alaleh; Dibaji, Fatemeh; Kharrazifard, Mohammad Javad

    2015-01-01

    Objectives: This study aimed to assess the effect of chloroform, eucalyptol and orange oil solvents on the microhardness of human root dentin. Materials and Methods: Sixty-eight single-rooted single-canal extracted human premolar teeth were used. Tooth crowns were separated from the roots at the cementoenamel junction (CEJ). Roots were buccolingually sectioned into mesial and distal halves. Specimens were randomly divided into 5 groups, with 20 teeth in each solvent group and 4 teeth in each control group. Primary microhardness of specimens was measured using Vickers microhardness tester. Specimens were exposed to solvents for 15 minutes and were subjected to microhardness testing again. Data were recorded and analyzed using repeated measure ANOVA. Results: No significant difference was found in dentin microhardness before and after exposure to solvents in any of the orange oil, eucalyptol, chloroform or saline groups (P=0.727). None of the experimental groups showed any significant difference in terms of dentin microhardness reduction (P=0.99) and had no significant difference with the negative control group. Conclusion: This study showed that chloroform, eucalyptol and orange oil as gutta percha solvents did not decrease the microhardness of root dentin. Thus, none of the mentioned solvents has any superiority over the others in terms of affecting dentin properties. PMID:26005451

  7. Comprehensive study of the chemical reactions resulting from the decomposition of chloroform in alkaline aqueous solution

    NASA Astrophysics Data System (ADS)

    Estevez Mews, Jorge

    Chloroform (CHCl3) is a volatile liquid, which has a rather slow rate of decomposition in ground water. It is a known carcinogen and one of the most common contaminants found at toxic waste sites. The dominant degradation process for chloroform in both the atmosphere and the groundwater is the reaction with the hydroxyl radical or hydroxide ion. This process triggers a sequence of reactions which ultimately yield carbon monoxide, hydrogen chloride, and formic acid. The rate of chloroform degradation is considerably larger in solution than that in the gas phase and it increases dramatically with increasing pH. However, only one of the viable reactions had been studied previously at a high level of theory in solution. It is of great interest to gain a deeper understanding of the decomposition reaction mechanism. Quantum mechanical methods are well suited for studying the mechanism of organic reactions. However, a full quantum mechanical treatment of the entire fluid system is not computationally feasible. In this work, combined quantum mechanical and molecular mechanical (QM/MM) methods are used for studying chemical reactions in condensed phases. In these calculations, the solute molecules are treated quantum mechanically (QM), whereas the solvent molecules are approximated by empirical (MM) potential energy functions. The use of quantum mechanics and statistical sampling simulation is necessary to determine the reaction free energy profile. In the present study, the ab initio Hartree-Fock theory along with the 3-21G basis set was used in the quantum mechanical calculations to elucidate the reaction pathways of chloroform decomposition, with a focus on basic reaction conditions. Statistical mechanical Monte Carlo approach was then applied in molecular mechanical simulations, employing the empirical TIP3P model for water. We employed state-of-the-art electronic structure methods to determine the gas-phase inter-nuclear potential energy profile for all the relevant

  8. Quantitative extraction and concentration of synthetic water-soluble acid dyes from aqueous media using a quinine-chloroform solution

    SciTech Connect

    Kobayashi, F.; Ozawa, N.; Hanai, J.; Isobe, M.; Watabe, T.

    1986-12-01

    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 diacidic 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.

  9. Resonance Raman intensity analysis of chlorine dioxide dissolved in chloroform: The role of nonpolar solvation

    NASA Astrophysics Data System (ADS)

    Foster, Catherine E.; Barham, Bethany P.; Reid, Philip J.

    2001-05-01

    Absolute resonance Raman cross sections for chlorine dioxide (OClO) dissolved in chloroform are obtained at several excitation wavelengths spanning the photochemically relevant 2B1-2A2 optical transition. The absolute scattering cross sections of OClO are determined by reference to the 666 cm-1 transition of chloroform whose absolute scattering cross sections are reported here. The time-dependent theory for Raman and absorption are used to develop a mode-specific description of the 2A2 excited state surface. This description demonstrates that photoexcitation of OClO leads to significant structural evolution along the symmetric stretch and bend coordinates, with only limited evolution occurring along the asymmetric stretch. This description is similar to that determined for OClO dissolved in cyclohexane and water demonstrating that the excited-state structural evolution of OClO is similar in these solvents. Analysis of the OClO absolute scattering cross sections establishes that the homogeneous linewidth is 95±15 cm-1 in chloroform, essentially identical to the linewidths in cyclohexane and water. To establish the origin of this linewidth, the fluorescence cross section for OClO dissolved in cyclohexane is measured and found to be consistent with an excited-state lifetime of ˜200 fs. Comparison of this lifetime to the homogeneous linewidth establishes that the homogeneous broadening is dominated by solvent-induced pure dephasing. It is proposed that the apparent solvent independence of the homogeneous linewidth reflects the mechanical response of the solvent to the photoinitiated change in solute geometry. In support of this hypothesis, the homogeneous linewidth is reproduced using the viscoelastic continuum model of nonpolar solvation. Finally, it is argued that the restricted evolution along the asymmetric-stretch coordinate is due to dielectric solvent-solute interactions consistent with the increase in inhomogeneous linewidth with an increase in solvent

  10. Slow motion, trapping, and sorting of water- and chloroform-soluble porphyrins in nanowells.

    PubMed

    Bhosale, Sheshanath; Bhosale, Sidhanath; Wang, Tianyu; Li, Guangtao; Siggel, Ulrich; Fuhrhop, Jürgen-Hinrich

    2004-10-13

    A two-step self-assembly procedure on smooth, aminated silica particles established holey monolayers. At first, single, flat-lying porphyrin tetraamides (A) were bound covalently, followed by the build-up of a rigid monolayer made of diamido bolaamphiphiles (bolas) around the porphyrin islands. "Nanowells" around porphyrin (A) bottoms with a uniform diameter of 2.2 nm and varying depths of 0.6, 1.0, or 1.5 nm depending on the length of the applied bolas were thus obtained. Oligoethylene headgroups solubilized the particles in water, ethanol, and chloroform/ethanol, and two hydrogen bond chains between the secondary amide groups prevented swelling of the monolayer. Manganese(III) porphyrinates (B) migrated from the bulk solution to the bottom of the form-stable nanowells with a speed of about 1 pm/s and were trapped there above porphyrin (A). After isolation of the (A,B) particles by centrifugation or ultrafiltration, the particles were suspended in a chloroform solution of a chlorin (C), which was also fixated irreversibly on the bottom of the nanowells. The nanowells thus contained three different porphyrins A,B,C in a noncovalent stack. The reverse sequence A,C,B was built-up correspondingly, first in chloroform/ethanol, and then in water. The "sorting" of A,B,C and A,C,B systems was characterized by visible spectra, sequence-dependent fluorescence quenching, and cyclic voltammetry of the top component. The molecular sorting method is the first of its kind and should be generally useful for the production of noncovalent reaction systems on any smooth surface.

  11. Comparing the Apical Microleakage of Lateral Condensation and Chloroform Dip Techniques with a New Obturation Method

    PubMed Central

    Saatchi, Masoud; Barekatain, Behnaz; Behzadian, Masoumeh

    2011-01-01

    Background: The final objective of root canal therapy is to create a hermetic seal along the length of the root canal system. For this purpose, many methods and materials have been introduced. The purpose of this study was to compare the apical microleakage in a new obturation technique (true-tug-back) with two other obturation techniques (lateral condensation and chloroform dip technique). Methods: In this in vitro study 102 single canal teeth were selected. The crowns were removed, and the canals were prepared using step-back technique. The master apical file was K-file #40. The teeth were divided into 3 experimental groups of 32 teeth. First group were obturated with lateral condensation technique and second group with chloroform dip technique and the third group with true-tug-back technique. Six teeth were used as control group. The teeth were placed in incubator at 100% humidity and 37°c for three days. The roots of the teeth were coated with two layers of nail varnish except for the apical 2 millimeter. Teeth were placed in Methylene blue 2% for one week. The teeth were sectioned vertically and the depth of maximum dye penetration for each tooth was recorded by stereomicroscope. Data were analyzed using ANOVA and Dunkan test. Results: The mean liner dye penetration differences between lateral condensation group (6.88 ± 4.06 mm) and chloroform dip technique group (7.16 ± 3.37 mm) were not statistically significant (P = 0.719). The differences between true-tug-back group (3.15 ± 0.52 mm) and two other groups were statistically significant (P < 0.001). Conclusion: The results of this study showed that the true-tug-back technique can improve apical seal. Further studies are needed for this purpose. PMID:22132011

  12. Fast and Facile Synthesis of 4-Nitrophenyl 2-Azidoethylcarbamate Derivatives from N-Fmoc-Protected α-Amino Acids as Activated Building Blocks for Urea Moiety-Containing Compound Library.

    PubMed

    Chen, Ying-Ying; Chang, Li-Te; Chen, Hung-Wei; Yang, Chia-Ying; Hsin, Ling-Wei

    2017-03-13

    A fast and facile synthesis of a series of 4-nitrophenyl 2-azidoethylcarbamate derivatives as activated urea building blocks was developed. The N-Fmoc-protected 2-aminoethyl mesylates derived from various commercially available N-Fmoc-protected α-amino acids, including those having functionalized side chains with acid-labile protective groups, were directly transformed into 4-nitrophenyl 2-azidoethylcarbamate derivatives in 1 h via a one-pot two-step reaction. These urea building blocks were utilized for the preparation of a series of urea moiety-containing mitoxantrone-amino acid conjugates in 75-92% yields and parallel solution-phase synthesis of a urea compound library consisted of 30 members in 38-70% total yields.

  13. Anti-Inflammatory Effects of Hyptis albida Chloroform Extract on Lipopolysaccharide-Stimulated Peritoneal Macrophages

    PubMed Central

    Sánchez Miranda, Elizabeth; Pérez Ramos, Julia; Fresán Orozco, Cristina; Zavala Sánchez, Miguel Angel; Pérez Gutiérrez, Salud

    2013-01-01

    We examined the effects of a chloroform extract of Hyptis albida (CHA) on inflammatory responses in mouse lipopolysaccharide (LPS) induced peritoneal macrophages. Our findings indicate that CHA inhibits LPS-induced production of tumor necrosis factor (TNF-α) and interleukin-6 (IL-6). During the process, levels of cyclooxygenase-2 (COX-2), nitric oxide synthase (iNOS), and nitric oxide (NO) increased in the mouse peritoneal macrophages; however, the extract suppressed them significantly. These results provide novel insights into the anti-inflammatory actions of CHA and support its potential use in the treatment of inflammatory diseases. PMID:23970974

  14. Influence of Sulfur for Oxygen Substitution in the Solvolytic Reactions of Chloroformate Esters and Related Compounds

    PubMed Central

    D’Souza, Malcolm J.; Kevill, Dennis N.

    2014-01-01

    The replacement of oxygen within a chloroformate ester (ROCOCl) by sulfur can lead to a chlorothioformate (RSCOCl), a chlorothionoformate (ROCSCl), or a chlorodithioformate (RSCSCl). Phenyl chloroformate (PhOCOCl) reacts over the full range of solvents usually included in Grunwald-Winstein equation studies of solvolysis by an addition-elimination (A-E) pathway. At the other extreme, phenyl chlorodithioformate (PhSCSCl) reacts across the range by an ionization pathway. The phenyl chlorothioformate (PhSCOCl) and phenyl chlorothionoformate (PhOCSCl) react at remarkably similar rates in a given solvent and there is a dichotomy of behavior with the A-E pathway favored in solvents such as ethanol-water and the ionization mechanism favored in aqueous solvents rich in fluoroalcohol. Alkyl esters behave similarly but with increased tendency to ionization as the alkyl group goes from 1° to 2° to 3°. N,N-Disubstituted carbamoyl halides favor the ionization pathway as do also the considerably faster reacting thiocarbamoyl chlorides. The tendency towards ionization increases as, within the three contributing structures of the resonance hybrid for the formed cation, the atoms carrying positive charge (other than the central carbon) change from oxygen to sulfur to nitrogen, consistent with the relative stabilities of species with positive charge on these atoms. PMID:25310653

  15. Project Work Plan Carbon Tetrachloride and Chloroform Attenuation Parameter Studies: Heterogeneous Hydrolytic Reactions

    SciTech Connect

    Amonette, James E.; Truex, Michael J.; Fruchter, Jonathan S.

    2006-06-01

    Between 1955 and 1973, an estimated 750,000 kg of carbon tetrachloride were discharged to the soil in the 200 West Area of the Hanford Site as part of the plutonium production process. Of this amount, some carbon tetrachloride reached the groundwater more than 70 m below the ground surface and formed a plume of 10 km2. Recent information has shown that the carbon tetrachloride plume extends to a depth of at least 60 m below the water table. Some carbon tetrachloride has been degraded either by the original process or subsequent transformations in the subsurface to form a co-existing chloroform plume. Although current characterization efforts are improving the conceptual model of the source area, more information is needed to effectively assess the fate and transport of carbon tetrachloride and chloroform to support upcoming remediation decisions for the plume. As noted in a simulation study by Truex et al. (2001), parameters describing porosity, sorption, and abiotic degradation have the largest influence on predicted plume behavior. The work proposed herein will improve the ability to predict future plume movement by better quantifying abiotic degradation mechanisms and rates. This effort will help define how much active remediation may be needed and estimate where the plume will eventually stabilize – key factors in determining the most appropriate remedy for the plume.

  16. Liquid chloroform structure from computer simulation with a full ab initio intermolecular interaction potential

    NASA Astrophysics Data System (ADS)

    Yin, Chih-Chien; Li, Arvin Huang-Te; Chao, Sheng D.

    2013-11-01

    We have calculated the intermolecular interaction energies of the chloroform dimer in 12 orientations using the second-order Møller-Plesset perturbation theory. Single point energies of important geometries were calibrated by the coupled cluster with single and double and perturbative triple excitation method. Dunning's correlation consistent basis sets up to aug-cc-pVQZ have been employed in extrapolating the interaction energies to the complete basis set limit values. With the ab initio potential data we constructed a 5-site force field model for molecular dynamics simulations. We compared the simulation results with recent experiments and obtained quantitative agreements for the detailed atomwise radial distribution functions. Our results were also consistent with previous results using empirical force fields with polarization effects. Moreover, the calculated diffusion coefficients reproduced the experimental data over a wide range of thermodynamic conditions. To the best of our knowledge, this is the first ab initio force field which is capable of competing with existing empirical force fields for liquid chloroform.

  17. Abiotic Degradation Rates for Carbon Tetrachloride and Chloroform: Progress in FY 2010

    SciTech Connect

    Amonette, James E.; Jeffers, Peter M.; Qafoku, Odeta; Russell, Colleen K.; Humphrys, Daniel R.; Wietsma, Thomas W.; Truex, Michael J.

    2010-12-08

    This report documents the progress made through FY 2010 on a project initiated in FY 2006 to help address uncertainties related to the rates of hydrolysis in groundwater at the Hanford Site for carbon tetrachloride (CT) and chloroform (CF). The study also explores the possible effects of contact with minerals and sediment (i.e., heterogeneous hydrolysis) on these rates. The research was initiated to decrease the uncertainties in abiotic degradation rates of CT and chloroform CF associated with temperature and possible heterogeneous effects. After 2 years of data collection, the first evidence for heterogeneous effects was identified for hydrolysis of CT, and preliminary evidence for the effects of different mineral types on CF hydrolysis rates also was reported. The CT data showed no difference among mineral types, whereas significant differences were seen in the CF results, perhaps due to the fact that CF hydrolyzes by both neutral and base-catalyzed mechanisms whereas CT follows only the neutral hydrolysis path. In this report, we review the project objectives, organization, and technical approaches taken, update the status and results of the hydrolysis-rate experiments after 4 years of experimentation (i.e., through FY 2010), and provide a brief discussion of how these results add to scientific understanding of the behavior of the CT/CF plume at the Hanford Site.

  18. Development and validation of a high-performance liquid chromatography-fluorescence detection method for the accurate quantification of colistin in human plasma.

    PubMed

    Chepyala, Divyabharathi; Tsai, I-Lin; Sun, Hsin-Yun; Lin, Shu-Wen; Kuo, Ching-Hua

    2015-02-01

    Recently, colistin has become one of the most important drugs for treating infections caused by multidrug-resistant Gram-negative bacteria. Therapeutic drug monitoring is recommended to ensure the safety and efficacy of colistin and to improve clinical outcomes. This study developed an accurate and sensitive high-performance liquid chromatography-fluorescence detection (HPLC-FLD) method for the quantification of colistin in human plasma. The sample preparation included protein precipitation using trichloroacetic acid (TCA) and methanol, followed by in-solid phase extraction (In-SPE) derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl). A Poroshell 120 EC-C18 2.1×100mm (2.7μm) column was used in the HPLC method with a mobile phase composed of acetonitrile (ACN), tetrahydrofuran (THF), and deionized (DI) water (82%, 2%, 16% (v/v), respectively). Polymyxin B1 was used as the internal standard. The total analysis time was 22min under optimal separation conditions. The HPLC-FLD method was validated over a therapeutic range of 0.3-6.0μgmL(-1). The intra-day and inter-day precisions for colistin A and colistin B were below 9.9% and 4.5% relative standard deviations, respectively. The accuracy test results were between 100.2 and 118.4%. The extraction recoveries were between 81.6 and 94.1%. The method was linear over the test range, with a 0.9991 coefficient of determination. The limit of detection was 0.1μgmL(-1). The validated HPLC-FLD method was successfully applied to quantify the colistin concentrations in 2 patient samples for therapeutic drug monitoring.

  19. The herbicide glyphosate and its metabolite AMPA in the Lavaux vineyard area, Western Switzerland: proof of widespread export to surface waters. Part I: method validation in different water matrices.

    PubMed

    Daouk, Silwan; Grandjean, Dominique; Chevre, Nathalie; De Alencastro, Luiz F; Pfeifer, Hans-Rudolf

    2013-01-01

    An analytical method for the quantification of the widely used herbicide, glyphosate, its main by-product, aminomethylphosphonic acid (AMPA) and the herbicide glufosinate at trace level was developed and tested in different aqueous matrices. Their derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) was done prior to their concentration and purification by solid phase extraction. The concentrated derivates were then analyzed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Spiking tests at three different concentrations were realized in several water matrices: ultrapure water, Evian(©) mineral water, river water, soil solution and runoff water of a vineyard. Except for AMPA in runoff water, obtained regression curves for all matrices of interest showed no statistical differences of their slopes and intercepts, validating the method for the matrix effect correction in relevant environmental samples. The limits of detection and quantification of the method were as low as 5 and 10 ng/l respectively for the three compounds. Spiked Evian(©) and river water samples at two different concentrations (30 and 130 ng/l) showed mean recoveries between 86 and 109%, and between 90 and 133% respectively. Calibration curves established in spiked Evian(©) water samples between 10 and 1000 ng/l showed r(2) values above 0.989. Monitoring of a typical vineyard river showed peaks of pollution by glyphosate and AMPA during main rain events, sometimes above the legal threshold of 100 ng/l, suggesting the diffuse export of these compounds by surface runoff. The depth profile sampled in the adjacent lake near a waste water treatment plant outlet showed a concentration peak of AMPA at 25m depth, indicating its release with treated urban wastewater.

  20. Influence of Ultrasonic Irrigation and Chloroform on Cleanliness of Dentinal Tubules During Endodontic Retreatment-An Invitro SEM Study

    PubMed Central

    Singhal, Anurag; Gurtu, Anuraag; Vinayak, Vineet

    2015-01-01

    Background Ultrasonic irrigation has been proved for its remarkable cleaning efficiency in the field of endodontics. But its role in endodontic re-treatment has been understated. There is not much data available to understand the effect of ultrasonic irrigation for the evaluation of cleanliness of dentinal tubules when it is used with or without chloroform, a gutta percha solvent during endodontic retreatment. Aim To compare the influence of ultrasonic irrigation with syringe irrigation on cleanliness of dentinal tubules after gutta perch removal for endodontic retreatment with or without the use of chloroform a gutta percha solvent using scanning electron microscope (SEM). Materials and Methods Freshly extracted 45 human mandibular premolar teeth for periodontal and orthodontic reasons were taken and were occlusally adjusted to a working length of 19 mm. The root canals of all teeth were prepared chemo mechanically to a master apical file size 40 and were divided in various groups. In Group 1 (n = 5; control group), the canals remained unfilled. In Groups 2 and 3 (n = 20 each), the canals were filled using lateral compaction with gutta-percha and AH plus sealer, removal of root fillings was undertaken after 2 weeks using Gates Glidden drills and H files without chloroform in Group 2 and with chloroform in group 3. The specimen of Group 2 and 3 were further divided into two subgroups I and II (n=10). In subgroup I, irrigation was done using side vented needles and sodium hypochlorite. In subgroup II irrigation was done using passive ultrasonic irrigation with sodium hypochlorite. Thereafter, the roots were split and the sections were observed under SEM. The number of occluded dentinal tubules /total number of dentinal tubules were calculated for the coronal, middle and apical third of each root half. Statistical analysis was performed using one-way ANOVA followed by Tukey’s test using standardized technique. Result Results indicated that the cleanest dentinal

  1. Expanding the scope of PNA-encoded synthesis (PES): Mtt-protected PNA fully orthogonal to fmoc chemistry and a broad array of robust diversity-generating reactions.

    PubMed

    Chouikhi, Dalila; Ciobanu, Mihai; Zambaldo, Claudio; Duplan, Vincent; Barluenga, Sofia; Winssinger, Nicolas

    2012-10-01

    Nucleic acid-encoded libraries are emerging as an attractive and highly miniaturized format for the rapid identification of protein ligands. An important criterion in the synthesis of nucleic acid encoded libraries is the scope of reactions that can be used to introduce molecular diversity and devise divergent pathways for diversity-oriented synthesis (DOS). To date, the protecting group strategies that have been used in peptide nucleic acid (PNA) encoded synthesis (PES) have limited the choice of reactions used in the library synthesis to just a few prototypes. Herein, we describe the preparation of PNA monomers with a protecting group combination (Mtt/Boc) that is orthogonal to Fmoc-based synthesis and compatible with a large palette of reactions that have been productively used in DOS (palladium cross-couplings, metathesis, reductive amination, amidation, heterocycle formation, nucleophilic addition, conjugate additions, Pictet-Spengler cyclization). We incorporate γ-modifications in the PNA backbone that are known to enhance hybridization and solubility. We demonstrate the robustness of this strategy with a library synthesis that is characterized by MALDI MS analysis at every step.

  2. [Kenzo Yoshida of the Hiroshima Domain and Chloroform Anesthesia at the End of the Edo Era in Japan].

    PubMed

    Matsuiki, Akitomo

    2016-02-01

    Japanese physicians encountered difficulties in obtaining information from The Netherlands on general anesthetics since 1848. It was a turning point when Chinese medical textbooks were imported in the latter half of the 1850 s. They were written by Benjamin Hobson (1816-1873), a British missionary and physician who lived in China for many years. Among them, Siyi Luelun (Sei-i Ryakuron) published in 1857, was widely welcome by Japanese surgeons and medical students. Kenzo Yoshida (1848-1924), a physician from the Hiroshima Domain, was impressed by a two-page description of chloroform anesthesia in it He observed William Willis administrating chloroform anesthesia on several injured Satsuma domain soldiers in January 1868 at the Shokokuji Temple in Kyoto. A few days later, Yoshida had a chance to administer chloroform to a man who committed suicide by cutting the abdomen. The intestine, which was 90 cm long, protruded from the wound, was put back into the abdominal cavity under chloroform anesthesia given by Yoshida. This is the second case of chloroform anesthesia administered by a Japanese physician.

  3. Acute Toxicity Investigation and Anti-diarrhoeal Effect of the Chloroform-Methanol Extract of the Leaves of Persea americana.

    PubMed

    Christian E, Odo; Okwesili Fc, Nwodo; Parker E, Joshua; Okechukwu Pc, Ugwu

    2014-01-01

    Persea americana is a plant used by traditional medicine practitioners to treat ailments including diarrhoea and diabetes mellitus in Nigeria. Hence, the chloroform and the methanol fractions of the chloroform-methanol extract of the leaves of P. americana were evaluated for their acute toxicity as well as anti-diarrhoeal effects in Wistar rats to substantiate this claim. The chloroform and methanol fractions [at graded doses of 100 and 200 mg/Kg body weight (b.w) of each] were studied for their anti-diarrhoeal effects in terms of the reductions in the wetness of faeces and the frequency of defaecation of castor oil-induced diarrhoea. To understand the mechanism of their anti-diarrhoeal effects, their actions were further evaluated on castor oil-induced enteropooling (intestinal fluid accumulation). The median lethal dose (LD50) of the methanol fraction was found to be less than 5000 mg/Kg b.w. At the two doses, the chloroform and the methanol fractions showed dose-dependent significant (p < 0.05) reductions in the wetness of faeces and the frequency of defaecation with the 200 mg/Kg b.w of the chloroform fraction being the most effective. Results of the fractions were comparable with those of the standard anti-diarrhoeal drug, hyoscine butylbromide (3 mg/Kg b.w). Both fractions produced remarkable (p < 0.05) dose-related inhibition of castor oil-induced enteropooling as shown by the significant (p < 0.05) decreases in the weight and volume of the intestinal contents. Experimental findings show that the chloroform-methanol extract of the leaves of P. americana possesses significant anti-diarrhoeal effect and may be a potent source of anti-diarrhoeal drug(s) in future.

  4. Capillary gas chromatography with cryogenic oven temperature for headspace samples: analysis of chloroform or methylene chloride in whole blood.

    PubMed

    Watanabe, K; Seno, H; Ishii, A; Suzuki, O; Kumazawa, T

    1997-12-15

    A new and sensitive gas chromatography (GC) method for measurement of chloroform or methylene chloride in whole blood is presented. Trace levels of these analytes present in the headspace of samples were cryogenically trapped prior to on-line GC analysis. After heating of a blood sample containing chloroform and methylene chloride (internal standard, and vice versa) in a 7.0-mL vial at 55 degrees C for 20 min, 5 mL of the headspace vapor was drawn into a glass syringe. All vapor was introduced into an Rtx-Volatiles middle-bore capillary column in the splitless mode at -30 degrees C oven temperature to trap the entire analytes, and the oven temperature was programmed up to 280 degrees C for detection of the compounds and for cleaning of the column. The present conditions gave sharp peaks for both chloroform and methylene chloride and very low background noises for whole blood samples. As much as 11.5 and 20.0% of chloroform and methylene chloride, respectively, which had been added to whole blood in a vial, could be introduced into the GC column. The calibration curves showed linearity in the range of 0.05-5.0 micrograms/0.5 mL of whole blood. The detection limit was estimated to be about 2 ng/0.5 mL. The coefficients of intraday and interday variations were 1.31 and 8.90% for chloroform and 1.37 and 9.03% for methylene chloride, respectively. The data on chloroform or methylene chloride in rat blood after inhalation of each compound were also presented.

  5. Antinociceptive Activity of the Chloroform Fraction of Dioclea virgata (Rich.) Amshoff (Fabaceae) in Mice

    PubMed Central

    Mota, Vanine Gomes; de Carvalho, Fabíola Lélis; de Morais, Liana Clébia Soares Lima; Bhattacharyya, Jnanabrata; de Almeida, Reinaldo Nóbrega; de Alencar, Jacicarlos Lima

    2011-01-01

    Acute treatment with the chloroform fraction of Dioclea virgata (Rich.) Amshoff (CFDv) in mice produced decreased ambulation and sedation in the behavioral pharmacological screening. Doses of 125 and 250 mg/kg CFDv decreased latency of sleep onset in the test of sleeping time potentiation. In the open field, animals treated with CFDv reduced ambulation and rearing (250 mg/kg), as well as defecation (125; 250 mg/kg). Regarding the antinociceptive activity, CFDv (125, 250, 500 mg/kg) increased latency to first writhing and decreased the number of writhings induced by acetic acid. In the formalin test, CFDv (250 mg/kg) decreased paw licking time in the first and second phases indicating antinociceptive activity that can be mediated both peripherally and at the central level. CFDv did not affect motor coordination until 120 minutes after treatment. CFDv shows psychopharmacological effects suggestive of CNS-depressant drugs with promising antinociceptive activity. PMID:21776190

  6. [Ether and chloroform narcosis in the pages of Zagreb newspapers from 1847 to 1849].

    PubMed

    Belicza, B

    2000-01-01

    Overviews of the development of anesthesiology in Croatia always emphasize that the first report on the employment of ether anesthesia was published on March 13, 1847, in the Zadar newspaper Gazzeta di Zara. In the world as well, daily newspapers played an important part in spreading the news about the discovery and first use of ether anesthesia. This fact stimulated us to do a systematic search through Zagreb press of that time. Agramer Zeitung (Zagrebacke novine), with its supplement Luna--Beiblatt zur Agramer politischen Zeitung, then Narodne novine and Sudslavische Zeitung (Slavenski jug), were examined for the period from 1846 to the beginning of 1849. Research was directed to give the following answers: when the first writings about discovery and surgery in ether and chloroform anesthesia appeared, which primary and secondary sources of information were used; who wrote them; what kind of attitudes and opinions were promoted, and whose; and finally, did Zagreb newspapers communicate or record the news about first employment of ether and chloroform anesthesia in surgery in Croatia. Research results reveal that Zagreb press had an important role in informing Croatian public about breakthroughs and first experiences in the use of ether and chloroform anesthesia in Europe, and that it contributed to their popularization and employment in our settings. From January 26, 1847, Zagreb papers began publishing first articles about surgeries in ether anesthesia, referring to successes achieved in England and Scotland, and later in Vienna, Berlin, Prague and other big European towns. However, there were no reviews on first successes in America. Sources of information were mainly foreign daily papers, mostly Austrian, and much less often European professional medical journals. Zagreb papers did not report on first surgeries performed in Zadar, Dubrovnik and Split during 1847. The only news from the country referred to the procedure performed in August 1847 in Sisak. This

  7. Effect of chloroform, orange solvent and eucalyptol on the accuracy of four electronic apex locators.

    PubMed

    Al-Hadlaq, Solaiman M

    2013-12-01

    The aim of this study was to evaluate the effect of three retreatment solutions on the accuracy of four electronic apex locators, the Root ZX mini, the Mini Apex Locator, the Root ZX and the Elements Diagnostic Unit and Apex Locator. Forty extracted single-rooted human teeth were used in this study. The four electronic apex locators were operated according to the manufacturer's instructions to locate the 'apical constriction' in the presence of chloroform, orange solvent or eucalyptol in the canal. The accuracy of each apex locator was not affected by the type of retreatment solution present in the root canal. In addition, the accuracy of the four apex locators was similar in the presence of each of the tested solutions.

  8. Abiotic Degradation Rates for Carbon Tetrachloride and Chloroform: Progress in FY2009

    SciTech Connect

    Amonette, James E.; Jeffers, Peter M.; Qafoku, Odeta; Russell, Colleen K.; Wietsma, Thomas W.; Truex, Michael J.

    2010-03-31

    This report documents the progress made through FY 2009 on a project initiated in FY 2006 to help address uncertainties related to the rates of hydrolysis in groundwater for carbon tetrachloride (CT) and chloroform (CF). The study seeks also to explore the possible effects of contact with minerals and sediment (i.e., heterogeneous hydrolysis) on these rates. In previous years the work was funded as two separate projects by various sponsors, all of whom received their funding from the U.S. Department of Energy (DOE). In FY2009, the projects were combined and funded by CH2MHill Plateau Remediation Corporation (CHPRC). Work in FY2009 was performed by staff at the Pacific Northwest National Laboratory (PNNL). Staff from the State University of New York at Cortland (SUNY–Cortland) contributed in previous years.

  9. Antinociceptive activity of the chloroform fraction of Dioclea virgata (Rich.) Amshoff (Fabaceae) in mice.

    PubMed

    Mota, Vanine Gomes; de Carvalho, Fabíola Lélis; de Morais, Liana Clébia Soares Lima; Bhattacharyya, Jnanabrata; de Almeida, Reinaldo Nóbrega; de Alencar, Jacicarlos Lima

    2011-01-01

    Acute treatment with the chloroform fraction of Dioclea virgata (Rich.) Amshoff (CFDv) in mice produced decreased ambulation and sedation in the behavioral pharmacological screening. Doses of 125 and 250 mg/kg CFDv decreased latency of sleep onset in the test of sleeping time potentiation. In the open field, animals treated with CFDv reduced ambulation and rearing (250 mg/kg), as well as defecation (125; 250 mg/kg). Regarding the antinociceptive activity, CFDv (125, 250, 500 mg/kg) increased latency to first writhing and decreased the number of writhings induced by acetic acid. In the formalin test, CFDv (250 mg/kg) decreased paw licking time in the first and second phases indicating antinociceptive activity that can be mediated both peripherally and at the central level. CFDv did not affect motor coordination until 120 minutes after treatment. CFDv shows psychopharmacological effects suggestive of CNS-depressant drugs with promising antinociceptive activity.

  10. Carbon Tetrachloride and Chloroform Attenuation Parameter Studies: Heterogeneous Hydrolytic Reactions -- Status Report

    SciTech Connect

    Amonette, James E.; Qafoku, Odeta; Wietsma, Thomas W.; Jeffers, Peter M.; Russell, Colleen K.; Truex, Michael J.

    2009-09-18

    This report documents a project initiated in FY 2006 to help address uncertainties related to the rates of hydrolysis in groundwater for carbon tetrachloride (CT) and chloroform (CF). The study sought also to explore the possible effects of contact with minerals and sediment (i.e., heterogeneous hydrolysis) on these rates. It was funded by the U.S. Department of Energy (DOE) through the Office of Environmental Management Columbia River Protection Supplemental Technologies (CRPST) project and complements work initiated by the Hanford Groundwater Project in FY 2006 that focused primarily on CT in homogenous solution. Work was performed by staff at the Pacific Northwest National Laboratory (PNNL) and the State University of New York at Cortland (SUNY-Cortland).

  11. [Solvents for the removal of gutta-percha from root canals. 2. Side effects of chloroform, halothane and xylene].

    PubMed

    Schuur, A H B; Moorer, W R; Wesselink, P R

    2004-08-01

    High concentrations of organic solvents have adverse effects on the health of professional workers. In endodontics, halothane, chloroform and xylene are used to dissolve gutta-percha from root canals. It is therefore questioned whether the use of these solvents could affect the health of patients or of the dental team. It seems warranted to conclude that the amounts and concentrations of chloroform, xylene and halothane such as used in endodontic retreatment, are low and therefore are safe. However, the dentist should be aware of possible hypersensitivity induced by previous application of halothane as a narcotic.

  12. Spectral and electroluminescent properties of coordination compounds of terbium (III) with ibuprofen (in solid form, chloroform solutions, and polyvinylcarbazole films)

    NASA Astrophysics Data System (ADS)

    Samsonova, L. G.; Kopylova, T. N.; Degtyarenko, K. M.; Ponarin, N. V.; Meshkova, S. B.; Zheltvai, I. I.

    2015-08-01

    Spectral properties of terbium (III) complexes with composition of TbL3DL, where L is an anion of d,l-2-(4-isobutylphenyl)propanoic acid (ibuprofen) and DL is 2,2'-dipyridyl (Dipy), 1,10-phenanthroline (Phen), or triphenylphosphine oxide (TPPO), have been studied in a solid form, chloroform solutions, and polyvinylcarbazole (PVC) films. It has been demonstrated that, in PVC films, occupation of the emitting level of terbium (III) involves the participation of polymer. The emission decay lifetimes of terbium in the chloroform solutions and PVC films have been measured. The possibility of the appearance of electroluminescence of complexes in PVC films has been studied.

  13. Construction of recombinant Nitrosomonas europaea expressing green fluorescent protein in response to co-oxidation of chloroform.

    PubMed

    Gvakharia, Barbara O; Bottomley, Peter J; Arp, Daniel J; Sayavedra-Soto, Luis A

    2009-04-01

    Transcriptional fusions with gfp driven by the promoter region of mbla (NE2571) in pPRO/mbla4 and clpB (NE2402) in pPRO/clpb7 were used to transform the ammonia-oxidizing bacterium Nitrosomonas europaea (ATCC 19718). The two genes were chosen because their transcript levels were found at much higher levels in N. europaea in response to oxidation of chloroform and chloromethane. In N. europaea transformed with pPRO/mbla4, green fluorescent protein (GFP)-dependent fluorescence increased from 3- to 18-fold above control levels in response to increasing chloroform concentrations (7 to 28 microM), and from 8- to 10-fold in response to increasing hydrogen peroxide concentrations (2.5-7.5 mM). The GFP-dependent fluorescence of N. europaea transformed with pPRO/clpb7 also showed an increase of 6- to 10-fold in response to chloroform (28-100 microM) but did not respond to H(2)O(2). Our data provide proof of concept that biosensors can be fabricated in ammonia-oxidizing bacteria using "sentinel" genes that up-regulate in response to stress caused either by co-oxidation of chlorinated solvents or by the presence of H(2)O(2). The fabricated biosensors had a consistent concentration-dependent response to chloroform; however, these did not respond to other chlorinated compounds that cause similar cellular stress.

  14. Factors associated with sources, transport, and fate of chloroform and three other trihalomethanes in untreated groundwater used for drinking water

    USGS Publications Warehouse

    Carter, Janet M.; Moran, Michael J.; Zogorski, John S.; Price, Curtis V.

    2012-01-01

    Multiple lines of evidence for indicating factors associated with the sources, transport, and fate of chloroform and three other trihalomethanes (THMs) in untreated groundwater were revealed by evaluating low-level analytical results and logistic regression results for THMs. Samples of untreated groundwater from wells used for drinking water were collected from 1996-2007 from 2492 wells across the United States and analyzed for chloroform, bromodichloromethane, dibromochloromethane, and bromoform by a low-level analytical method implemented in April 1996. Using an assessment level of 0.02 μg/L, chloroform was detected in 36.5% of public-well samples and 17.6% of domestic-well samples, with most concentrations less than 1 μg/L. Brominated THMs occurred less frequently than chloroform but more frequently in public-well samples than domestic-well samples. For both public and domestic wells, THMs occurred most frequently in urban areas. Logistic regression analyses showed that the occurrence of THMs was related to nonpoint sources such as urban land use and to point sources like septic systems. The frequent occurrence and concentration distribution pattern of THMs, as well as their frequent co-occurrence with other organic compounds and nitrate, all known to have anthropogenic sources, and the positive associations between THM occurrence and dissolved oxygen and recharge indicate the recycling of water that contains THMs and other anthropogenic contaminants.

  15. Attenuation of nonenzymatic glycation, hyperglycemia, and hyperlipidemia in streptozotocin-induced diabetic rats by chloroform leaf extract of Azadirachta indica

    PubMed Central

    Gutierrez, Rosa Martha Pérez; Gómez, Yolanda Gómez Y.; Guzman, Mónica Damián

    2011-01-01

    Background: The hypoglycemic effects of hexane, chloroform and methanol extracts of leaves of Azadirachta indica (AI) were evaluated by oral administration in streptozotocin-induced severe diabetic rats (SD). Materials and Methods: The effect of chronic oral administration of the extract for 28 days was evaluated in streptozotozin diabetic rats. Lipid peroxidation, glycogen content of liver and skeletal muscles, insulin, superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), oxidized glutathione (GSSG) levels were determined. In addition, advanced glycation end product formation (AGEs) was evaluated. Results: The most active extracts were obtained with chloroform. Chloroform extract from AI shows increased levels of SOD, GSH, GSSG and CAT, hepatic glycogen content, glucose-6-phosphatase and insulin plasma levels, which also decreased the glucokinase (GK), lipid peroxidation and insulin resistance. The chloroform extract exhibited significant inhibitory activity against advanced glycation end product formation with an IC50 average range of 79.1 mg/ml. Conclusion: Azadirachta indica can improve hyperlipidemia and hyperinsulinema in streptozocin-induced diabetic rats and, therefore, AI can be potentially considered to be an antidiabetic-safe agent. PMID:21969798

  16. Chloroform in the hydrologic system--sources, transport, fate, occurrence, and effects on human health and aquatic organisms

    USGS Publications Warehouse

    Ivahnenko, Tamara; Barbash, Jack E.

    2004-01-01

    Chloroform is one of the volatile organic compounds (VOCs) detected most frequently in both ground and surface water. Because it is also one of the four trihalomethanes (THMs) produced in the highest concentrations during the chlorination of drinking water and wastewater, the frequent detection of this compound in ground and surface water of the United States is presumed to be caused primarily by the input of chlorinated water to the hydrologic system. Although anthropogenic sources of the compound are substantial, they are currently estimated to constitute only 10 percent of the total global input to the hydrologic system. Natural sources of the compound include volcanic gases, biomass burning, marine algae, and soil microorganisms. Under most conditions (except in the presence of unusually high bromide concentrations), chloroform is the THM produced in the highest concentrations during chlorination. Furthermore, in most cases where more than one THM is produced from chlorination, the relative concentrations among the different compounds usually decrease with increasing bromination (chloroform > dichlorobromomethane > chlorodibromomethane > bromoform). This phenomenon is presumed to be responsible for the common observation that when more than one THM is detected during investigations of the occurrence of these compounds in the hydrologic system, this same trend is typically observed among their relative concentrations or, for a uniform reporting limit, their relative frequencies of detection. This pattern could provide a valuable means for distinguishing between chlorinated water and other potential sources of chloroform in the environment. Chloroform has been widely detected in national, regional, and local studies of VOCs in ground, surface, source, and drinking waters. Total THM (TTHM) concentrations of the compound, however, were typically less than the Maximum Contaminant Level (MCL) of 80 ?g/L (micrograms per liter) established by the U.S. Environmental

  17. The natural chlorine cycle - Formation of the carcinogenic and greenhouse gas compound chloroform in drinking water reservoirs.

    PubMed

    Forczek, Sándor T; Pavlík, Milan; Holík, Josef; Rederer, Luděk; Ferenčík, Martin

    2016-08-01

    Chlorine cycle in natural ecosystems involves formation of low and high molecular weight organic compounds of living organisms, soil organic matter and atmospherically deposited chloride. Chloroform (CHCl3) and adsorbable organohalogens (AOX) are part of the chlorine cycle. We attempted to characterize the dynamical changes in the levels of total organic carbon (TOC), AOX, chlorine and CHCl3 in a drinking water reservoir and in its tributaries, mainly at its spring, and attempt to relate the presence of AOX and CHCl3 with meteorological, chemical or biological factors. Water temperature and pH influence the formation and accumulation of CHCl3 and affect the conditions for biological processes, which are demonstrated by the correlation between CHCl3 and ΣAOX/Cl(-) ratio, and also by CHCl3/ΣAOX, CHCl3/AOXLMW, CHCl3/ΣTOC, CHCl3/TOCLMW and CHCl3/Cl(-) ratios in different microecosystems (e.g. old spruce forest, stagnant acidic water, humid and warm conditions with high biological activity). These processes start with the biotransformation of AOX from TOC, continue via degradation of AOX to smaller molecules and further chlorination, and finish with the formation of small chlorinated molecules, and their subsequent volatilization and mineralization. The determined concentrations of chloroform result from a dynamic equilibrium between its formation and degradation in the water; in the Hamry water reservoir, this results in a total amount of 0.1-0.7 kg chloroform and 5.2-15.4 t chloride. The formation of chloroform is affected by Cl(-) concentration, by concentrations and ratios of biogenic substrates (TOC and AOX), and by the ratios of the substrates and the product (feedback control by chloroform itself).

  18. 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...

  19. 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...

  20. Low-Dose Priming Before Vaccination with the Phase I Chloroform-Methanol Residue Vaccine Against Q Fever Enhances Humoral and Cellular Immune Responses to Coxiella Burnetii

    DTIC Science & Technology

    2008-10-01

    Vaccination with the Phase I Chloroform-Methanol Residue Vaccine against Q Fever Enhances Humoral and Cellular Immune Responses to Coxiella burnetii David... I Coxiella burnetii cellular vaccine is completely efficacious in humans, adverse local and systemic reactions may develop if immune individuals are...inadvertently vaccinated. The phase I chloroform- methanol residue (CMRI) vaccine was developed as a potentially safer alternative. Human volunteers

  1. Site-specific PEGylation for high-yield preparation of Lys(21)-amine PEGylated growth hormone-releasing factor (GRF) (1-29) using a GRF(1-29) derivative FMOC-protected at Tyr(1) and Lys(12).

    PubMed

    Youn, Yu Seok; Lee, Kang Choon

    2007-01-01

    PEGylation has been viewed as an effective means of overcoming the therapeutic restriction of growth hormone-releasing factor (1-29) (GRF(1-29)) due to its short biological lifetime caused by severe proteolysis and rapid glomerular filtration. Of three isomers according to the PEGylation sites (Tyr1, Lys12, or Lys21), PEGylated GRF(1-29) at Lys21-amine (Lys21-PEG-GRF(1-29)) was shown to have the highest bioactivity. In this report, we propose a unique two-step site-specific PEGylation method capable of producing only Lys21-PEG-GRF(1-29) with a single composition in high yield using a GRF(1-29) derivative protected at Tyr1 and Lys12 and remained available at Lys21 (FMOC1,12-GRF(1-29)). The first step of this reaction involved the PEG attachment to FMOC1,12-GRF(1-29), and the second step involved the removal of FMOC moieties. This PEGylation process was optimized at the following conditions: 0.2-0.3% (v/v) triethylamine concentration, 5.0-6.0-fold molar amount of PEG, reaction temperature of 25-45 degrees C, and reaction time of 30 min. Under these conditions, the maximum yield of Lys21-PEG-GRF(1-29) produced was ca. approximately 95%, 6.3-fold higher than that by nonspecific PEGylation at pH 8.5. Significantly, this site-specific Lys21-PEG-GRF(1-29) was found to have greatly increased resistance to rat plasma, liver, and kidney homogenates, with 7.0-, 25.4-, and 16.4-fold longer half-lives vs GRF(1-29), respectively. Furthermore, 125I-Lys21-PEG-GRF(1-29) displayed significantly reduced liver and kidney distributions and extended blood presence vs 125I-GRF(1-29) in rats. Due to these benefits, Lys21-PEG-GRF(1-29) displayed an enhanced initial growth hormone release in vivo despite having 15% remaining activity in vitro. This devised PEGylation method using an FMOC-protection/deprotection strategy would provide great usefulness for PEGylating bioactive peptides in terms of improved biological potency, elevated production yield, and a uniform composition.

  2. Probing the excited state relaxation dynamics of pyrimidine nucleosides in chloroform solution.

    PubMed

    Röttger, Katharina; Marroux, Hugo J B; Böhnke, Hendrik; Morris, David T J; Voice, Angus T; Temps, Friedrich; Roberts, Gareth M; Orr-Ewing, Andrew J

    2016-12-16

    Ultrafast transient electronic and vibrational absorption spectroscopy (TEAS and TVAS) of 2'-deoxy-cytidine (dC) and 2'-deoxy-thymidine (dT) dissolved in chloroform examines their excited-state dynamics and the recovery of ground electronic state molecules following absorption of ultraviolet light. The chloroform serves as a weakly interacting solvent, allowing comparisons to be drawn with prior experimental studies of the photodynamics of these nucleosides in the gas phase and in polar solvents such as water. The pyrimidine base nucleosides have some propensity to dimerize in aprotic solvents, but the monomer photochemistry can be resolved clearly and is the focus of this study. UV absorption at a wavelength of 260 nm excites a (1)ππ* ← S0 transition, but prompt crossing of a significant fraction (50% in dC, 17% in dT) of the (1)ππ* population into a nearby (1)nπ* state is too fast for the experiments to resolve. The remaining flux on the (1)ππ* state leaves the vertical Franck-Condon region and encounters a conical intersection with the ground electronic state of ethylenic twist character. In dC, the (1)ππ* state decays to the ground state with a time constant of 1.1 ± 0.1 ps. The lifetime of the (1)nπ* state is much longer in the canonical forms of both molecules: recovery of the ground state population from these states occurs with time constants of 18.6 ± 1.1 ps in amino-oxo dC and ∼114 ps in dT, indicating potential energy barriers to the (1)nπ*/S0 conical intersections. The small fraction of the imino-oxo tautomer of dC present in solution has a longer-lived (1)nπ* state with a lifetime for ground state recovery of 193 ± 55 ps. No evidence is found for photo-induced tautomerization of amino-oxo dC to the imino-oxo form, or for population of low lying triplet states of this nucleoside. In contrast, ∼8% of the UV-excited dT molecules access the long-lived T1 ((3)ππ*) state through the (1)nπ* state. The primary influence of the solvent

  3. Characterization of N-methylated amino acids by GC-MS after ethyl chloroformate derivatization.

    PubMed

    Reddy, B Sudarshana; Chary, V Naresh; Pavankumar, P; Prabhakar, S

    2016-08-01

    Methylation is an essential metabolic process in the biological systems, and it is significant for several biological reactions in living organisms. Methylated compounds are known to be involved in most of the bodily functions, and some of them serve as biomarkers. Theoretically, all α-amino acids can be methylated, and it is possible to encounter them in most animal/plant samples. But the analytical data, especially the mass spectral data, are available only for a few of the methylated amino acids. Thus, it is essential to generate mass spectral data and to develop mass spectrometry methods for the identification of all possible methylated amino acids for future metabolomic studies. In this study, all N-methyl and N,N-dimethyl amino acids were synthesized by the methylation of α-amino acids and characterized by a GC-MS method. The methylated amino acids were derivatized with ethyl chloroformate and analyzed by GC-MS under EI and methane/CI conditions. The EI mass spectra of ethyl chloroformate derivatives of N-methyl (1-18) and N,N-dimethyl amino acids (19-35) showed abundant [M-COOC2 H5 ](+) ions. The fragment ions due to loss of C2 H4 , CO2 , (CO2  + C2 H4 ) from [M-COOC2 H5 ](+) were of structure indicative for 1-18. The EI spectra of 19-35 showed less number of fragment ions when compared with those of 1-18. The side chain group (R) caused specific fragment ions characteristic to its structure. The methane/CI spectra of the studied compounds showed [M + H](+) ions to substantiate their molecular weights. The detected EI fragment ions were characteristic of the structure that made easy identification of the studied compounds, including isomeric/isobaric compounds. Fragmentation patterns of the studied compounds (1-35) were confirmed by high-resolution mass spectra data and further substantiated by the data obtained from (13) C2 -labeled glycines and N-ethoxycarbonyl methoxy esters. The method was applied to human plasma samples for the identification

  4. Antiproliferative activity of chloroformic extract of Persian Shallot, Allium hirtifolium, on tumor cell lines.

    PubMed

    Ghodrati Azadi, Hamideh; Ghaffari, Seyed Mahmood; Riazi, Gholam Hossein; Ahmadian, Shahin; Vahedi, Fatemeh

    2008-03-01

    Allium hirtifolim (Persian Shallot) belongs to Allium genus (Alliaceae family). We investigated the in vitro effects of chloroformic extract of A. hirtifolium and its Allicin on the proliferation of HeLa (cervical cancer), MCF7 (human, caucasion, breast, adenocarcinoma) and L929 (mouse, C3H/An, connective) cell lines. Our results showed that components of A. hirtifolium might inhibit proliferation of tumor cell lines. This inhibition in HeLa and MCF-7 cells was dose-dependent. The presence of Allicin was evaluated by TLC method in bulbs and the extract of A. hirtifolium was analyzed by HPLC. MTT test was performed 24, 48 and 72 h after cell culture. A significant decrease in cell lines was observed in HeLa and MCF-7 as compared to L929 cell lines. DNA fragmentation analysis revealed a large number of apoptotic cells in treated HeLa and MCF-7 cell groups, but no effects in L929 cells. Therefore A. hirtifolium might be a candidate for tumor suppression.

  5. Antimicrobial activity of essential oils and chloroform alone and combinated with cetrimide against Enterococcus faecalis biofilm

    PubMed Central

    Ferrer Luque, Carmen Maria; González-Rodríguez, Maria Paloma; Arias-Moliz, Maria Teresa; Baca, Pilar

    2013-01-01

    Abstract The Enterococcus faecalis bacteria have been identified as the most commonly recovered species from teeth with persistent endodontic infections. The antimicrobial activity of essential oils and chloroform (CHL), alone and in association with various concentrations of cetrimide (CTR), against biofilm of Enterococcus faecalis was investigated. Solutions of CHL, eucalyptus oil (EO) and orange oil (OO) associated with CTR at 0.3%, 0.2%, 0.1%, and 0.05% were used to determine antimicrobial activity by exposing treated bovine dentine blocks to E. faecalis. Biofilms grown in the dentine blocks for 7 days were exposed to solutions for 2 and 5 min. Biofilm reduction between OO and EO at 2 min did not show any significant differences; however, OO had a higher kill percentage of biofilms than did the eucalyptus oil at 5 min (p < 0.01). Combinations with CTR at all concentrations achieved a 100% kill rate at 2 and 5 min. The association of CTR with solvent agents achieved the maximum antimicrobial activity against E. faecalis biofilms in dentine. PMID:24265917

  6. Gastroprotective activity of the chloroform extract of the roots from Arctium lappa L.

    PubMed

    Dos Santos, Ana C; Baggio, Cristiane H; Freitas, Cristina S; Lepieszynski, Juliana; Mayer, Bárbara; Twardowschy, André; Missau, Fabiana C; dos Santos, Elide P; Pizzolatti, Moacir G; Marques, Maria C A

    2008-06-01

    Arctium lappa L. is used in folk medicine as a diuretic, depurative and digestive stimulant and in dermatological conditions. The objective of this study was to evaluate the effect and the possible mechanisms involved in the gastroprotective effects of a chloroform extract (CE) of the roots from A. lappa and its fractions. Oral pretreatment with CE (10, 30 and 100 mg kg(-1)) significantly reduced gastric lesions induced by ethanol by 61%, 70% and 76%, respectively. Oral administration of CE (100 mg kg(-1) per day for 7 days) reduced the chronic gastric ulceration induced by acetic acid by 52%. Intraduodenal CE (100, 300 and 600 mg kg(-1)) reduced the total acidity of gastric secretion by 22%, 22% and 33%, respectively, while i.p. administration (10, 30 and 100 mg kg(-1)) inhibited total acidity by 50%, 60% and 67%, respectively. In-vitro, CE inhibited H+, K+ -ATPase activity with an EC50 of 53 microgmL(-1) and fraction A (30 and 100 microgmL(-1)) reduced this by 48% and 89%, respectively. CE had no effect on gastrointestinal motility. CE (250 microgmL(-1)) and fraction B (100 and 250 microgmL(-1)) had free-radical scavenging ability, inhibiting 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical activity by 50%, 20% and 55%, respectively. Collectively, the results show that the CE protects animals from gastric lesions by reducing gastric acid secretion via inhibition of gastric H+, K+ -ATPase.

  7. Replacing methyl chloroform for cleaning turbine generator components and NDE applications

    SciTech Connect

    Bailey, K.P.; O'Shanka, J.J.; Corley, T.J. . Power Generation Business Unit); Sadhir, R.K. )

    1993-08-01

    Industrial applications of methyl chloroform (1,1,1-trichloroethane) have proven to be a significant concern to the environment. As a chlorofluorocarbon (CFC), the chemical is classified by the Environmental Protection Agency as an ozone-layer-depleting substance (OLDS). CFCs are effective cleaners of organic-based materials (oils, greases, cutting fluids, etc.). The Westinghouse Power Generation Business Unit (PGBU) has taken a proactive approach to this problem and instituted two programs in 1991 and 1992 to eliminate their consumption of CFCs. The scope of the first program was to establish an alternate cleaner for the removal of oil on generator stator windings. The second program built on the work of the first program, extending the scope to include general purpose cleaning of various contaminants prior to and at the completion of nondestructive examinations (NDE). The article that follows details the methodology, results, discussions, and conclusions of the second program and the data extrapolated from the first program. The specific NDE qualification requirements are highlighted in the methodology section.

  8. Sensitive chemi-sensor for environmental applications as marker of chloroform in aqueous solution

    NASA Astrophysics Data System (ADS)

    Khan, Sher Bahadar; Faisal, M.; Rahman, Mohammed M.; Akhtar, Kalsoom; Asiri, Abdullah M.; Alamry, Khalid A.

    2013-04-01

    Sensor technology is a talented progression in the fundamental science and has vital role in the environmental and health monitoring. A lot of sensor materials with different sensitivities have been exposed but the prospective for industrial development as sensors and environmental supplement are still limited. Chloroform chemi-sensor has been fabricated by proficient exploitation of CuO microsphere as redox mediator. CuO microsphere showed good sensitivity (1.497 μM cm-2 mM-1) and lower limit of detection (LOD = 0.08 mM) with linear dynamic range (LDR) of 0.13 mM to 0.66 M in short response time (10.0 s). The hydrothermally synthesized CuO microsphere were characterized by using field emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and UV-vis spectra. XRD and FTIR approved that synthesized product is well crystalline, single phase monoclinic CuO. Morphological investigations revealed that CuO composed of nano-block which arranged and structured themselves in a proper manner to give sphere like micro-structures.

  9. Cooperative motion in liquids: On librational dynamics of chloroform throughout its normal liquid-phase range

    NASA Astrophysics Data System (ADS)

    Rothschild, Walter G.; Cavagnat, Raymond M.

    1994-03-01

    We have extended the Raman spectral accumulations of the ν3 mode (A1, 367 cm-1) of liquid CHCl3-Cl-35 and its simulation in terms of an orientational equilibrium renewal process [W. G. Rothschild, R. M. Cavagnat, and P. Maraval, J. Chem. Phys. 99, 8922 (1993)] to a temperature of 338 K, about the normal boiling point of the system (335 K). The values of the best-fit parameters predict that the orientational motion of liquid chloroform, even at such a relatively high kinetic energy, is described predominantly by libratory states; their lifetime (˜1 ps) is four times longer than that of the free-rotational steps. The character of the orientational motion of the system, when traversing the range of 213 to 338 K from just above its melting to near its boiling point at about atmospheric pressure, reflects the softening of the liquid-cage structure in terms of an increasing dispersion and/or a decreasing value of the mean libration frequency, a lowering of the depth of its potential well, but near-invariance of its lifetime. Simultaneously, there is an approximately twofold increase in the lifetime of the much shorter stages of free-rotational motion. In essence, the system dynamics remain that of an assembly of librators.

  10. Unexpected Temperature Behavior of Polyethylene Glycol Spacers in Copolymer Dendrimers in Chloroform

    PubMed Central

    Markelov, Denis A.; Matveev, Vladimir V.; Ingman, Petri; Nikolaeva, Marianna N.; Penkova, Anastasia V.; Lahderanta, Erkki; Boiko, Natalia I.; Chizhik, Vladimir I.

    2016-01-01

    We have studied copolymer dendrimer structure: carbosilane dendrimers with terminal phenylbenzoate mesogenic groups attached by poly(ethylene) glycol (PEG) spacers. In this system PEG spacers are additional tuning to usual copolymer structure: dendrimer with terminal mesogenic groups. The dendrimer macromolecules were investigated in a dilute chloroform solution by 1H NMR methods (spectra and relaxations). It was found that the PEG layer in G = 5 generations dendrimer is “frozen” at high temperatures (above 260 K), but it unexpectedly becomes “unfrozen” at temperatures below 250 K (i.e., melting when cooling). The transition between these two states occurs within a small temperature range (~10 K). Such a behavior is not observed for smaller dendrimer generations (G = 1 and 3). This effect is likely related to the low critical solution temperature (LCST) of PEG and is caused by dendrimer conformations, in which the PEG group concentration in the layer increases with growing G. We suppose that the unusual behavior of PEG fragments in dendrimers will be interesting for practical applications such as nanocontainers or nanoreactors. PMID:27052599

  11. Cation mobility and the sorption of chloroform in zeolite NaY: molecular dynamics study.

    PubMed

    Ramsahye, Naseem A; Bell, Robert G

    2005-03-17

    Molecular dynamics simulations at temperatures of 270, 330, and 390 K have been carried out to address the question of cation migration upon chloroform sorption in sodium zeolite Y. The results show that sodium cations located in different sites exhibit different types of mobility. These may be summarized as follows: (1) SII cations migrate toward the center of the supercage upon sorption, due to interactions with the polar sorbate molecules. (2) SI' cations hop from the sodalite cage into the supercage to fill vacant SII sites. (3) SI' cations migrate to other SI' sites within the same sodalite cage. (4) SI cations hop out of the double six-rings into SI' sites. In some instances, concerted motion of cations is observed. Furthermore, former SI' and SI cations, having crossed to SII sites, may then further migrate within the supercage, as in (1). The cation motion is dependent on the level of sorbate loading, with 10 molecules per unit cell not being enough to induce significant cation displacements, whereas the sorption of 40 molecules per unit cell results in a number of cations being displaced from their original positions. Further rearrangement of the cation positions is observed upon evacuation of the simulation cell, with some cations reverting back to sites normally occupied in bare NaY.

  12. Chloroform induces outstanding crystallization of poly(hydroxybutyrate) (PHB) vesicles within bacteria.

    PubMed

    Rebois, Rolando; Onidas, Delphine; Marcott, Curtis; Noda, Isao; Dazzi, Alexandre

    2017-03-01

    Poly[(R)-3-hydroxyalkanoate]s or PHAs are aliphatic polyesters produced by numerous microorganisms. They are accumulated as energy and carbon reserve in the form of small intracellular vesicles. Poly[(R)-3-hydroxybutyrate] (PHB) is the most ubiquitous and simplest PHA. An atomic force microscope coupled with a tunable infrared laser (AFM-IR) was used to record highly spatially resolved infrared spectra of commercial purified PHB and native PHB within bacteria. For the first time, the crystallinity degree of native PHB within vesicle has been directly evaluated in situ without alteration due to the measure or extraction and purification steps of the polymer: native PHB is in crystalline state at 15% whereas crystallinity degree reaches 57% in commercial PHB. Chloroform addition on native PHB induces crystallization of the polymer within bacteria up to 60%. This possibility of probing and changing the physical state of polymer in situ could open alternative ways of production for PHB and others biopolymers. Graphical abstract An atomic force microscope coupled with a tunable infrared laser (AFM-IR) has been used to record local infrared spectra of biopolymer PHB within bacteria. Deconvolution of those spectra has allowed to determine in situ the crystallinity degree of native PHB.

  13. LATE POISONING WITH CHLOROFORM AND OTHER ALKYL HALIDES IN RELATIONSHIP TO THE HALOGEN ACIDS FORMED BY THEIR CHEMICAL DISSOCIATION

    PubMed Central

    Graham, Evarts A.

    1915-01-01

    The central lobular necrosis in the liver, which has been regarded by some writers as characteristic of late chloroform poisoning, has been produced experimentally with a number of other drugs. It is, therefore, in no sense peculiar to chloroform poisoning. Substances which have been shown to produce a morphological picture indistinguishable from that of late chloroform poisoning are: (a) dichlor- and tetrachlormethane, (b) tribrom- and triiodomethane, (c) monochlor-, monobrom-, and monoiodoethane, also the dibromethane; that is, in general, the halogen substituted aliphatic hydrocarbons containing one or two carbon atoms. Presumably similar results might be obtained with the higher members of the same series. The mechanism by which chloroform produces its characteristic tissue changes must accordingly be considered as a group reaction. Outside the body the similarities between the chemical behavior of different members of this group have been correlated by Nef on the basis of the type of dissociation which these substances undergo and the differences in their behavior on the basis of the differences of the degree to which such dissociations occur. According to the work of Nef, the group of substances under discussion has the property of dissociating to yield a halogen acid and an unsaturated alkylidene rest. Thus with chloroform the type of dissociation may be expressed thus: See PDF for Equation In this paper the view is developed that the changes characteristic of late poisonings with the above named group, namely edema, multiple hemorrhages, fat infiltration, and necrosis are ascribable (1) to acids and (2) to the fact that the amount of acid formed parallels the chemical dissociability of the drug outside of the body. Favoring the view that acid is responsible for the changes are the following observations. 1. All the characteristic features of late chloroform poisoning have been produced merely by the administration of hydrochloric acid, except, however, for

  14. Single Laboratory Validation of a Method for Determination of Glucosamine in Raw Materials and Dietary Supplements Containing Glucosamine Sulfate and/or Glucosamine Hydrochloride by High-Performance Liquid Chromatography with FMOC-Su Derivatization

    PubMed Central

    Zhou, Joseph ZiQi; Waszkuc, Ted; Mohammed, Felicia

    2008-01-01

    Single laboratory validation of a method for determination of glucosamine in raw materials and dietary supplements containing glucosamine sulfate and/or glucosamine hydrochloride by with high-performance liquid Chromatography FMOC-Su derivatization. Tests with 2 blank matrixes containing SAMe, vitamin C, citric acid, chondroitin sulfates, methylsulfonylmethane, lemon juice concentrate, and other potential interferents showed the method to be selective and specific. Eight calibration curves prepared over 7 working days indicated excellent reproducibility with the linear range at least over 2.0–150 μg/mL, and determination coefficients >0.9999. Average spike recovery from the blank matrix (n = 8 over 2 days) was 93.5, 99.4, and 100.4% at respective spike levels of 15,100, and 150%, and from the sample matrix containing glucosamine (n = 3) was 99.9 and 102.8% at respective levels of 10 and 40%, with relative standard deviations <0.9%. The method was also applied to 12 various glucosamine finished products and raw materials. The stability tests confirmed that glucosamine–FMOC-Su derivative once formed is stable at room temperature for at least 5 days. Limit of quantitation was 1 μg/mL and limit of detection was 0.3 μg/mL. The method is ready to proceed for the collaborative study. PMID:15493664

  15. Comparative Dynamics of Leucine Methyl Groups in FMOC-Leucine and in a Protein Hydrophobic Core Probed by Solid-State Deuteron Nuclear Magnetic Resonance over 7-324 K Temperature Range

    SciTech Connect

    Vugmeyster, Liliya; Ostrovsky, Dmitry; Moses, Mark; Ford, Joseph J.; Lipton, Andrew S.; Hoatson, Gina; Vold, Robert L.

    2010-12-09

    Quantitative dynamics of methyl groups in 9-fluorenylmethyloxycarbonyl-leucine (FMOC-leu) have been analyzed and compared with earlier studies of methyl dynamics in chicken villin headpiece subdomain protein (HP36) labeled at L69, a key hydrophobic core position. A combination of deuteron solid-state nuclear magnetic resonance experiments over the temperature range of 7-324 K and computational modeling indicated that while the two compounds show the same modes of motions, there are marked differences in the best-fit parameters of these motions. One of the main results is that the crossover observed in the dynamics of the methyl groups in the HP36 sample at 170 K is absent in FMOC-leu. A second crossover at around 95-88 K is present in both samples. The differences in the behavior of the two compounds suggest that some of the features of methyl dynamics reflect the complexity of the protein hydrophobic core and are not determined solely by local interactions.

  16. Oxidation of trichloroethylene, 1,1-dichloroethylene, and chloroform by toluene/o-xylene monooxygenase from Pseudomonas stutzeri OX1

    SciTech Connect

    Chauhan, S.; Wood, T.K.; Barbieri, P.

    1998-08-01

    Toluene/o-xylene monooxygenase (ToMO) from Pseudomonas stutzeri OX1, which oxidizes toluene and o-xylene, was examined for its ability to degrade the environmental pollutants trichloroethylene (TCE), 1,1-dichloroethylene (1,1-DCE), cis-1,2-DCE, trans-1,2-DCE, chloroform, dichloromethane, phenol, 2,4-dichlorophenol, 2,4,5-trichlorophenol, 2,4,6-trichlorophenol, 2,3,5,6-tetrachlorophenol, and 2,3,4,5,6-pentachlorophenol. Escherichia coli JM109 that expressed ToMO from genes on plasmid pBZ1260 under control of the lac promoter degraded TCE, 1,1-DCE, and chloroform at initial rates of 3.1, 3.6, and 1.6 nmol, respectively. Stoichiometric amounts of chloride release were seen, indicating mineralization. Thus, the substrate range of ToMO is extended to include aliphatic chlorinated compounds.

  17. Spectrophotometric determination of tungsten in ores and steel by chloroform extraction of the tungsten-thiocyanate-diantipyrylmethane complex.

    PubMed

    Donaldson, E M

    A method for determining up to about 6% of tungsten in ores and mill products is described. It is based on the extraction of the yellow tungsten(V)-thiocyanate-diantipyrylmethane ion-association complex into chloroform from a 2.4M sulphuric acid-7.8M hydrochloric acid medium containing ammonium hydrogen fluoride as masking agent for niobium. The molar absorptivity of the complex is 1510 1. mole(-1).mm(-1) at 404 nm, the wavelength of maximum absorption. Moderate amounts of molybdenum and selenium may be present in the sample solution without causing appreciable error in the result. Interference from large amounts is avoided by separating these elements from tungsten by chloroform extraction of their xanthate complexes. Large amounts of copper interfere during the extraction of tungsten because of the precipitation of cuprous thiocyanate. Common ions, including uranium, vanadium, cobalt, titanium, arsenic and tellurium, do not interfere. The proposed method is also applicable to steel.

  18. Estimates of European emissions of methyl chloroform using a Bayesian inversion method

    NASA Astrophysics Data System (ADS)

    Maione, M.; Graziosi, F.; Arduini, J.; Furlani, F.; Giostra, U.; Blake, D. R.; Bonasoni, P.; Fang, X.; Montzka, S. A.; O'Doherty, S. J.; Reimann, S.; Stohl, A.; Vollmer, M. K.

    2014-03-01

    Methyl chloroform (MCF) is a man-made chlorinated solvent contributing to the destruction of stratospheric ozone and is controlled under the Montreal Protocol on Substances that Deplete the Ozone Layer. Long-term, high-frequency observations of MCF carried out at three European sites show a constant decline of the background mixing ratios of MCF. However, we observe persistent non-negligible mixing ratio enhancements of MCF in pollution episodes suggesting unexpectedly high ongoing emissions in Europe. In order to identify the source regions and to give an estimate of the magnitude of such emissions, we have used a Bayesian inversion method and a point source analysis, based on high-frequency long-term observations at the three European sites. The inversion identified south-eastern France (SEF) as a region with enhanced MCF emissions. This estimate was confirmed by the point source analysis. We performed this analysis using an eleven-year data set, from January 2002 to December 2012. Overall emissions estimated for the European study domain decreased nearly exponentially from 1.1 Gg yr-1 in 2002 to 0.32 Gg yr-1 in 2012, of which the estimated emissions from the SEF region accounted for 0.49 Gg yr-1 in 2002 and 0.20 Gg yr-1 in 2012. The European estimates are a significant fraction of the total semi-hemisphere (30-90° N) emissions, contributing a minimum of 9.8% in 2004 and a maximum of 33.7% in 2011, of which on average 50% are from the SEF region. On the global scale, the SEF region is thus responsible from a minimum of 2.6% (in 2003) to a maximum of 10.3% (in 2009) of the global MCF emissions.

  19. Estimates of European emissions of methyl chloroform using a Bayesian inversion method

    NASA Astrophysics Data System (ADS)

    Maione, M.; Graziosi, F.; Arduini, J.; Furlani, F.; Giostra, U.; Blake, D. R.; Bonasoni, P.; Fang, X.; Montzka, S. A.; O'Doherty, S. J.; Reimann, S.; Stohl, A.; Vollmer, M. K.

    2014-09-01

    Methyl chloroform (MCF) is a man-made chlorinated solvent contributing to the destruction of stratospheric ozone and is controlled under the "Montreal Protocol on Substances that Deplete the Ozone Layer" and its amendments, which called for its phase-out in 1996 in developed countries and 2015 in developing countries. Long-term, high-frequency observations of MCF carried out at three European sites show a constant decline in the background mixing ratios of MCF. However, we observe persistent non-negligible mixing ratio enhancements of MCF in pollution episodes, suggesting unexpectedly high ongoing emissions in Europe. In order to identify the source regions and to give an estimate of the magnitude of such emissions, we have used a Bayesian inversion method and a point source analysis, based on high-frequency long-term observations at the three European sites. The inversion identified southeastern France (SEF) as a region with enhanced MCF emissions. This estimate was confirmed by the point source analysis. We performed this analysis using an 11-year data set, from January 2002 to December 2012. Overall, emissions estimated for the European study domain decreased nearly exponentially from 1.1 Gg yr-1 in 2002 to 0.32 Gg yr-1 in 2012, of which the estimated emissions from the SEF region accounted for 0.49 Gg yr-1 in 2002 and 0.20 Gg yr-1 in 2012. The European estimates are a significant fraction of the total semi-hemisphere (30-90° N) emissions, contributing a minimum of 9.8% in 2004 and a maximum of 33.7% in 2011, of which on average 50% are from the SEF region. On the global scale, the SEF region is thus responsible for a minimum of 2.6% (in 2003) and a maximum of 10.3% (in 2009) of the global MCF emissions.

  20. Mechanisms of chloroform and carbon tetrachloride toxicity in primary cultured mouse hepatocytes.

    PubMed Central

    Ruch, R J; Klaunig, J E; Schultz, N E; Askari, A B; Lacher, D A; Pereira, M A; Goldblatt, P J

    1986-01-01

    Mechanisms of chloroform (CHCl3) and carbon tetrachloride (CCl4) toxicity to primary cultured male B6C3F1 mouse hepatocytes were investigated. The cytotoxicity of both CHCl3 and CCl4 was dose- and duration-dependent. Maximal hepatocyte toxicity, as determined by lactate dehydrogenase leakage into the culture medium, occurred with the highest concentrations of CHCl3 (5 mM) and CCl4 (2.5 mM) used and with the longest duration of treatment (20 hr). CCl4 was approximately 16 times more toxic than CHCl3 to the hepatocytes. The toxicity of these compounds was decreased by adding the mixed function oxidase system (MFOS) inhibitor, SKF-525A (25 microM) to the cultures. The addition of diethyl maleate (0.25 mM), which depletes intracellular glutathione (GSH)-potentiated CHCl3 and CCl4 toxicity. The toxicity of CHCl3 and CCl4 could also be decreased by adding the antioxidants N,N'-diphenyl-p-phenylenediamine (DPPD) (25 microM), alpha-tocopherol acetate (Vitamin E) (0.1 mM), or superoxide dismutase (SOD) (100 U/mL) to the cultures. These results suggest that: in mouse hepatocytes, both CHCl3 and CCl4 are metabolized to toxic components by the MFOS; GSH plays a role in detoxifying those metabolites; free radicals are produced during the metabolism of CHCl3 and CCl4; and free radicals may be important mediators of the toxicity of these two halomethanes. PMID:3816733

  1. Desorption behavior of carbon tetrachloride and chloroform in contaminated low organic carbon aquifer sediments.

    PubMed

    Riley, Robert G; Szecsody, James E; Sklarew, Debbie S; Mitroshkov, Alex V; Gent, Philip M; Brown, Christopher F; Thompson, Christopher J

    2010-05-01

    Slow release behavior of carbon tetrachloride (CCl(4)) and chloroform (CHCl(3)) in low organic carbon (<0.1%) deep aquifer sediments was quantified by 1-D column desorption studies with intact cores. The compounds had been in contact with the sediments for 30years. Comparison of the CCl(4) distribution coefficient (K(d)) from this study with those from short contact time experiments suggested that CCl(4)K(d)'s calculated from site contaminated sediments of long contact time are likely a factor of 10 or more higher than those calculated from short contact-time lab experiments. A significant portion of the CHCl(3) mass (55% to more than 90%) was resistant to aqueous desorption in sediments with clay contents ranging from 2.0% to 36.7% and organic carbon content ranging from 0.017% to 0.088%. In contrast, CCl(4) showed greatest mass retention (31% or more) only in the highest clay and organic carbon content sediment. Relatively easy solvent extraction of the residual masses of CCl(4) and CHCl(3) from the sediments indicated the compounds were not permanently sequestered. Tracer breakthrough in columns was well behaved, indicating interparticle diffusion was not causing the slow release behavior. Diffusion out of intraparticle pores is suggested to be the main process governing the observed behavior although, diffusion out of natural organic matter cannot be ruled out as a potential contributing factor. The half-life for release of the slow fraction of CHCl(3) mass from sediments was estimated to be in the range of weeks (100h) to months (1100h). Neither CCl(4) or CHCl(3) were detected at measurable levels in the column effluent of one of the sediments even though a significant mass fraction of CHCl(3) was found present on the sediment following desorption suggesting that our estimate of hundreds to thousands of hours for complete release of CHCl(3) masses from such sediment is conservative.

  2. Extracellular lipids of Camelina sativa: characterization of chloroform-extractable waxes from aerial and subterranean surfaces.

    PubMed

    Razeq, Fakhria M; Kosma, Dylan K; Rowland, Owen; Molina, Isabel

    2014-10-01

    Camelina sativa (L.) Crantz is an emerging low input, stress tolerant crop with seed oil composition suitable for biofuel and bioproduct production. The chemical compositions and ultrastructural features of surface waxes from C. sativa aerial cuticles, seeds, and roots were analyzed using gas chromatography and microscopy. Alkanes, primary fatty alcohols, and free fatty acids were common components of all analyzed organs. A particular feature of leaf waxes was the presence of alkyl esters of long-chain fatty acids and very long-chain fatty alcohols, ranging from C38 to C50 and dominated by C42, C44 and C46 homologues. Stem waxes were mainly composed of non-sterol pentacyclic triterpenes. Flowers accumulated significant amounts of methyl-branched iso-alkanes (C29 and C31 total carbon number) in addition to straight-chain alkanes. Seed waxes were mostly primary fatty alcohols of up to 32 carbons in length and unbranched C29 and C31 alkanes. The total amount of identified wax components extracted by rapid chloroform dipping of roots was 280μgg(-1) (fresh weight), and included alkyl hydroxycinnamates, predominantly alkyl coumarates and alkyl caffeates. This study provides qualitative and quantitative information on the waxes of C. sativa root, shoot, and seed boundary tissues, allowing the relative activities of wax biosynthetic pathways in each respective plant organ to be assessed. This detailed description of the protective surface waxes of C. sativa may provide insights into its drought-tolerant and pathogen-resistant properties, and also identifies C. sativa as a potential source of renewable high-value natural products.

  3. Mechanisms of chloroform and carbon tetrachloride toxicity in primary cultured mouse hepatocytes

    SciTech Connect

    Ruch, R.J.; Klaunig, J.E.; Schultz, N.E.; Askari, A.B.; Lacher, D.A.; Pereira, M.A.; Goldblatt, P.J.

    1986-11-01

    Mechanisms of chloroform (CHCl/sub 3/) and carbon tetrachloride (CCl/sub 4/) toxicity to primary cultured male B6C3F1 mouse hepatocytes were investigated. The cytotoxicity of both CHCl/sub 3/ and CCl/sub 4/ was dose- and duration-dependent. Maximal hepatocyte toxicity, as determined by lactate dehydrogenase leakage into the culture medium, occurred with the highest concentrations of CHCl/sub 3/ (5 mM) and CCl/sub 4/ (2.5 mM) used and with the longest duration of treatment (20 hr). CCl/sub 4/ was approximately 16 times more toxic than CHCl/sub 3/ to the hepatocytes. The toxicity of these compounds was decreased by adding the mixed function oxidase system (MFOS) inhibitor, SKF-525A (25..mu..M) to the cultures. The addition of diethyl maleate (0.25 mM), which depletes intracellular glutathione (GSH)-potentiated CHCl/sub 3/ and CCl/sub 4/ toxicity. The toxicity of CHCl/sub 3/ and CCl/sub 4/ could also be decreased by adding the antioxidants N,N'-diphenyl-p-phenylenediamine (DPPD) (25..mu..M), ..cap alpha..-tocopherol acetate (Vitamin E) (0.1 mM), or superoxide dismutase (SOD) (100 U/mL) to the cultures. These results suggest that: in mouse hepatocytes, both CHCl/sub 3/ and CCl/sub 4/ are metabolized to toxic components by the MFOS; GSH plays a role in detoxifying those metabolites; free radicals are produced during the metabolism of CHCl/sub 3/ and CCl/sub 4/; and free radicals may be important mediators of the toxicity of these two halomethanes.

  4. A practical route to tertiary diarylmethylamides or -carbamates from imines, organozinc reagents and acyl chlorides or chloroformates

    PubMed Central

    Pignon, Antoine; Martens, Thierry

    2011-01-01

    Summary A practical route to tertiary diarylmethylamides or -carbamates from imines, organozinc reagents and acyl chlorides or chloroformates is described. This route involves the formation of an imine, which is used without isolation, followed by its activation by the carbonyl-containing electrophile and the trapping of the acyliminium by an organozinc reagent. Most steps are conducted concomitantly to render the procedure as practical and straightforward as possible. Therefore, the whole experimental protocol takes less than two hours. PMID:21915199

  5. In Vitro Comparison of Gutta-Percha Removal with H-File and ProTaper with or without Chloroform

    PubMed Central

    Khalilak, Zohreh; Vatanpour, Mehdi; Dadresanfar, Bahareh; Moshkelgosha, Pouneh; Nourbakhsh, HamidReza

    2013-01-01

    Introduction Removal of root filling materials is one of the key steps in success of root canal retreatment. The purpose of this study was to evaluate the efficacy of H-File and ProTaper with or without chloroform in the removal of gutta-percha during retreatment of mandibular premolars. Materials and Methods Sixty mandibular premolars with one canal, and curvatures less than 30 degrees were used in this experimental study. They were instrumented with K-files and laterally obturated with condensed gutta-percha using AH26 as the sealer and were stored in 100% humidity at 37°C for 2 weeks. The teeth were randomly divided into four groups of 15 teeth each. Removal of gutta-percha was performed with H-File and ProTaper. All techniques were used with or without chloroform. The teeth were split longitudinally and the area of remaining gutta-percha/sealer on the root canal wall was explored under stereomicroscope. Retreatment time duration was also recorded for each sample. Data were analyzed statistically by Two-way ANOVA, t-test and Tukey’s. Results In all groups, no significant difference was found in remaining gutta-percha and sealer with or without using chloroform, but chloroform shortened the time of retreatment. ProTaper left significantly less remaining filling materials than H-File (P<0.05). Retreatment time was significantly different between the studied groups (P<0.001). Conclusion ProTaper Ni-Ti instruments proved to be more efficient and time-saving devices for removal of gutta-percha compared to H-File in canals with no or slight curvature. PMID:23413203

  6. A practical route to tertiary diarylmethylamides or -carbamates from imines, organozinc reagents and acyl chlorides or chloroformates.

    PubMed

    Le Gall, Erwan; Pignon, Antoine; Martens, Thierry

    2011-01-01

    A practical route to tertiary diarylmethylamides or -carbamates from imines, organozinc reagents and acyl chlorides or chloroformates is described. This route involves the formation of an imine, which is used without isolation, followed by its activation by the carbonyl-containing electrophile and the trapping of the acyliminium by an organozinc reagent. Most steps are conducted concomitantly to render the procedure as practical and straightforward as possible. Therefore, the whole experimental protocol takes less than two hours.

  7. Determination of a Kinetic Rate Expression for the Oxidation of Chloroform Over a 2% Platinum/Alpha-Alumina Catalyst

    DTIC Science & Technology

    1992-04-01

    equations in Table 2 are presented in Appendix B. 3. EXPERIMENTAL METHODS 3.1 Materials. Chloroform ( HPLC grade, 99.9 %) was obtained from Aldrich...schematic representation of the fixed bed catalytic reactor system. Compressed air from an oil -free compressor was purified using a commer- cial PSA drier...to 77 % during the following three hour period, and remained essentially constant for the sample collected overnight. The same trend was observed for

  8. Survey on efficacy of chloroformic extract of Artemisia annua against Giardia lamblia trophozoite and cyst in vitro.

    PubMed

    Golami, Shirzad; Rahimi-Esboei, Bahman; Mousavi, Parisa; Marhaba, Zahra; Youssefi, Mohammad Reza; Rahimi, Mohammad Taghi

    2016-03-01

    Giardiasis is a parasitic cosmopolitan disease that the rate of infection in developing countries is considerable. This infection directly is associated with poor hygienic conditions, poor water quality control, and overcrowding. Reinfection and drug resistance are two major problems in endemic areas. Recently, researchers are concentrating on herbal drugs as a proper solution. Therefore, the objective of the present study was to survey on efficacy of chloroformic extract of Artemisia annua against Giardia lamblia trophozoite and cyst in vitro. G. lamblia cysts were prepared from faces of giardiasis patients from different hospitals of Mazandaran Medical University. Four concentrations (1, 10, 50 and 100 mg/ml) of chloroformic extract of A. annua were utilized for 1, 5, 30, 60 and 180 min. Viability of G. lamblia cysts was confirmed by 0.1 % Eosin staining. Cyst and trophozoite contact (intermix) of G. lamblia with extract of A. annua with variant concentrations (1, 10, 50 and 100 mg/ml) after 1 and 180 min caused following cyst and trophozoite elimination rates: (67, 69, 71 and 73 %), (65, 67, 67 and 72 %), (94, 96, 97 and 99 %) and (100, 100, 100 and 100 %), respectively. Authors from the current investigation draw a conclusion that chloroformic extract of A. annua has the ability to eliminate G. lamblia cysts and trophozoites in vitro.

  9. Development of an improved method to extract pesticide residues in foods using acetonitrile with magnesium sulfate and chloroform.

    PubMed

    Liu, Guozhu; Rong, Lei; Guo, Bin; Zhang, Mingshan; Li, Shengjun; Wu, Qing; Chen, Jitao; Chen, Bo; Yao, Shouzhuo

    2011-03-18

    A multiresidue method was developed based on extraction of 10 g sample with 10 mL acetonitrile and subsequent liquid-liquid partitioning formed by adding 4 g MgSO₄ plus 1 mL chloroform. During the partitioning process, the extraction recoveries of polar analytes were found to be essentially determined by the acetonitrile content in the aqueous phase. The use of MgSO₄ gave the least acetonitrile left in the aqueous phase (lower than 5%) and thus promoting complete partitioning of analytes into the organic phase. At the same time, removal of water from the acetonitrile phase was achieved by adding only a small amount of chloroform with no influence on the acetonitrile content in the aqueous phase, thus leading to decreasing the co-extraction of polar matrix components. The most complete mutual separation of acetonitrile and water was achieved by the joint use of MgSO₄ and chloroform and thus the optimal extraction recovery and analytical selectivity were obtained simultaneously. The new method, with higher recoveries of polar analytes, better analytical selectivity and simpler manipulation, is a claimed improvement to the original QuEChERS method. The proposed method was finally validated by the determination of 20 pesticides in a mixed food matrix by using liquid chromatography tandem mass spectrum (LC-MS/MS). Acceptable linearity, sensitivity, recovery, precision and selectivity results were obtained.

  10. The Utilization of Chloroform Post-Treatment to Improve the Adhesion of Au Thin Films onto PMMA

    NASA Astrophysics Data System (ADS)

    Krist, Kathleen; Hughes, Chris; Hu, Xiaofeng; Augustine, Brian

    2015-03-01

    The metallization of Au onto plastics is an important processing step in the fabrication of microfluidic devices. While its corrosion resistance and excellent electrical and thermal conductivity make Au a good choice, its inertness results in poor adhesion to polymer surfaces. Previous studies have indicated that exposing commercially available Poly(methyl methacrylate) (PMMA) sheets to chloroform vapor following Au deposition significantly improves adhesion. In this study, we deposited 6 nm of Au onto 1.50 mm thick PMMA and exposed the samples to vapor released from chloroform heated on a hot plate set at 70 °C. The force required to remove the Au thin films was determined by placing samples on a polisher spinning at 150 rpm and utilizing UV-VIS spectroscopy to measure the transmittance of 700 nm light through the films to quantify their removal as a function of applied polishing force. The Au thin films were also characterized using AFM. AFM images demonstrated a progressive roughening of the surface corresponding to an increase in applied force. Additionally, these images support a model in which the chloroform treatment softens the PMMA surface, producing a softened layer that the polisher removes simultaneously with the Au thin film. Undergraduate.

  11. Adsorption characteristics of acetone, chloroform and acetonitrile on sludge-derived adsorbent, commercial granular activated carbon and activated carbon fibers.

    PubMed

    Tsai, Jiun-Horng; Chiang, Hsiu-Mei; Huang, Guan-Yinag; Chiang, Hung-Lung

    2008-06-15

    The adsorption characteristics of chloroform, acetone, and acetonitrile on commercial activated carbon (C1), two types of activated carbon fibers (F1 and F2), and sludge adsorbent (S1) was investigated. The chloroform influent concentration ranged from 90 to 7800 ppm and the acetone concentration from 80 to 6900 ppm; the sequence of the adsorption capacity of chloroform and acetone on adsorbents was F2>F1 approximately C1 approximately S1. The adsorption capacity of acetonitrile ranged from 4 to 100 mg/g, corresponding to the influent range from 43 to 2700 ppm for C1, S1, and F1. The acetonitrile adsorption capacity of F2 was approximately 20% higher than that of the other adsorbents at temperatures<30 degrees C. The Freundlich equation fit the data better than the Langmuir and Dubinin-Radushkevich (D-R) equations. The adsorption rate of carbon fibers is higher than that of the other adsorbents due to their smaller fiber diameter and higher surface area. The micropore diffusion coefficient of VOC on activated carbon and sludge adsorbent was approximately 10(-4) cm2 s(-1). The diffusion coefficient of VOC on carbon fibers ranged from 10(-8) to 10(-7) cm2 s(-1). The small carbon fiber pore size corresponds to a smaller diffusion coefficient.

  12. Polarographic behaviour of uranium beta-diketonates in chloroform Application of ac polarography to the analysis of uranium minerals.

    PubMed

    Pournaghi-Azar, M H; Ordoukhanian, J

    1991-12-01

    The polarographic behaviour of U(VI) beta-diketonates has been studied in chloroform. The conditions for reversible electrochemical reduction of U(VI) acetylacetonate and benzoylacetonate at a dropping mercury electrode was optimized by using a suitable ratio of piperidinium perchlorate and piperidine mixture as supporting electrolyte in chloroform. The one-electron nature of the reduction wave of U(VI) complexes was confirmed by controlled potential coulometry. The ac method preceded by a solvent extraction of U(VI) benzoylacetonate in chloroform was used for the determination of uranium. The calibration curve was linear over the range 0.5-20 mu/ml. The correlation coefficient was 0.9998 and the detection limit was about 0.2 mu/ml. The interference of some concomitant ions were examined and EDTA was used as an effective masking agent to separate uranium from other metals. The proposed method has been applied to the determination of uranium in uranium minerals.

  13. Application of the Grunwald-Winstein Equations to Studies of Solvolytic Reactions of Chloroformate and Fluoroformate Esters

    PubMed Central

    D’Souza, Malcolm J.; Kevill, Dennis N.

    2014-01-01

    Chloroformates are important laboratory and industrial chemicals with almost one hundred listed in the catalogs of leading suppliers. They are, for example, of prime importance as protecting groups in peptide synthesis. In some instances, the more stable fluoroformate is preferred. In recent years, the specific rates of solvolysis (k) for chloroformates and fluoroformates in solvents of widely ranging nucleophilicity and ionizing power have been studied. Analysis of these rates using the extended (two-term) Grunwald-Winstein equation has led to important information concerning reaction mechanism. Also assisting in this effort have been studies of kinetic solvent isotope effects (KSIE), of leaving group effects (especially kF/kCl ratios), and of entropies of activation from studies of specific rate variations with temperature. For solvolyses of chloroformate esters, two mechanisms (addition-elimination and ionization) are commonly encountered. For solvolyses of fluoroformates, mainly because of a strong C–F bond, the ionization pathway is rare and the addition-elimination pathway is in most situations the one encountered. PMID:25364780

  14. Non-linear optics and local-field factors in liquid chloroform: A time-dependent density-functional theory study

    NASA Astrophysics Data System (ADS)

    Strubbe, David A.; Andrade, Xavier; Rubio, Angel; Louie, Steve G.

    2009-03-01

    Chloroform is often used as a solvent and reference when measuring non-linear optical properties of organic molecules. We calculate directly the non-linear susceptibilities of liquid chloroform at optical frequencies, using molecular dynamics and the Sternheimer equation in time-dependent density-functional theory [X. Andrade et al., J. Chem. Phys. 126, 184106 (2007)]. We compare the results to those of chloroform in the gas and solid phases, and experimental values, and make an ab initio calculation of the local-field factors which are needed to extract molecular properties from liquid calculations and experimental measurements.

  15. Hepatoprotection with a chloroform extract of Launaea procumbens against CCl4-induced injuries in rats

    PubMed Central

    2012-01-01

    Background Launaea procumbens (Asteraceae) is used as a folk medicine to treat hepatic disorders in Pakistan. The effect of a chloroform extract of Launaea procumbens (LPCE) was evaluated against carbon-tetrachloride (CCl4)-induced liver damage in rats. Methods To evaluate the hepatoprotective effects of LPCE, 36 male Sprague–Dawley rats were equally divided into six groups. Animals of group 1 (control) had free access to food and water. Group II received 3 ml/kg of CCl4 (30% in olive oil v/v) via the intraperitoneal route twice a week for 4 weeks. Group III received 1 ml of silymarin via gavage (100 mg/kg b.w.) after 48 h of CCl4 treatment whereas groups IV and V were given 1 ml of LPCE (100 and 200 mg/kg b.w., respectively) after 48 h of CCl4 treatment. Group VI received 1 ml of LPCE (200 mg/kg b.w.) twice a week for 4 weeks. The activities of the antioxidant enzymes catalase, peroxidase (POD), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), glutathione reductase (GSR), glutathione (GSH) and lipid peroxidation (thiobarbituric acid reactive substances (TBARS)) were measured in liver homogenates. DNA damage, argyrophilic nucleolar organizer regions (AgNORs) counts and histopathology were studied in liver samples. Serum was analyzed for various biochemical parameters. Phytochemical composition in LPCE was determined through high-performance liquid chromatography (HPLC). Results LPCE inhibited lipid peroxidation, and reduced the activities of aspartate transaminase, alanine transaminase, alkaline phosphatase, and lactate dehydrogenase in serum induced by CCl4. GSH contents were increased as were the activities of antioxidant enzymes (catalase, SOD, GST, GSR, GSH-Px) when altered due to CCl4 hepatotoxicity. Similarly, absolute liver weight, relative liver weight and the number of hepatic lesions were reduced with co-administration of LPCE. Phyochemical analyses of LPCE indicated that it contained catechin, kaempferol

  16. Suppression of Pulmonary Host Defenses and Enhanced Susceptibility to Respiratory bacterial Infection in mice Following Inhalation Exposure to Trichloroethylene and Chloroform

    EPA Science Inventory

    Numerous epidemiologic studies have associated episodes of increased air pollution with increased incidence of respiratory disease, including pneumonia, croup, and bronchitis. Trichloroethylene (TCE) and chloroform are among 33 hazardous air pollutants identified by the U.S. Env...

  17. Chloroform inhibits the development of diethylnitrosamine-initiated, phenobarbital-promoted gamma-glutamyltranspeptidase and placental form glutathione S-transferase-positive foci in rat liver.

    PubMed

    Reddy, T V; Daniel, F B; Lin, E L; Stober, J A; Olson, G R

    1992-08-01

    In this study we demonstrate that chloroform, a widely used industrial solvent, a medicinal chemical and a common drinking water contaminant, reduces the number of detectable preneoplastic enzyme-altered foci [gamma-glutamyltranspeptidase-positive (GGT+) and placental form glutathione S-transferase-positive (GST-P+)] in the liver of male Fischer 344 rats. The animals were given a partial hepatectomy and 18 h later received a single oral dose of either 0.5 mmol/kg diethylnitrosamine (DENA) or saline. Two weeks later, groups of 12 animals were started on drinking water containing phenobarbital with varying concentrations (200-1800 mg/l) of chloroform fro 12 weeks. Treated and control animals were killed and the number and the volume of GGT+ and GST-P+ expressing hepatic foci were tabulated. The numbers of foci per unit volume (and per unit area), the percent focal volume and the focal liver were reduced by chloroform in a dose-dependent manner. The mean focal volume was not influenced by chloroform. A plausible explanation for these results could be that chloroform exerts its focal inhibitory effect either by selectively killing the putative initiated cells, by retarding the inherent growth rate of enzyme-altered cells or by reducing the effectiveness of the promoter, phenobarbital. The available evidence suggests that the first hypothesis is the most likely explanation for these observations. These results are consistent with earlier studies showing that chloroform inhibits tumorigenesis in rodents.

  18. Preparation of the very acid-sensitive Fmoc-Lys(Mtt)-OH. Application in the synthesis of side-chain to side-chain cyclic peptides and oligolysine cores suitable for the solid-phase assembly of MAPs and TASPs.

    PubMed

    Aletras, A; Barlos, K; Gatos, D; Koutsogianni, S; Mamos, P

    1995-05-01

    N alpha-9-Fluorenylmethoxycarbonyl-N epsilon-4=methyltrityl-lysine, [Fmoc-Lys(Mtt)-OH], was prepared in two steps from lysine, in 42% overall yield. The N epsilon-Mtt function can be quantitatively removed upon treatment with 1% TFA in dichloromethane or with a 1:2:7 mixture of acetic acid/trifluoroethanol/dichloromethane for 30 min and 1 h at room temperature, respectively. Under these conditions, groups of the tert-butyl type and peptide ester bonds to TFA-labile resins, such as the 2-chlorodiphenylmethyl- and the Wang-resin, remained intact. The utility of the new derivative in peptide synthesis has been exemplified with the synthesis of a cyclic cholecystokinin analog. As an example of further application, five types of lysine cores suitable for the solid-phase synthesis of one, two or three epitopes containing antigenic peptides or template-assembled synthetic proteins have been synthesized on Merrifield, Wang and 2-chlorodiphenylmethyl resin.

  19. Gelation behaviour of a bent-core dihydrazide derivative: effect of incubation temperature in chloroform and toluene.

    PubMed

    Zhang, Chunxue; Zhang, Tianren; Ji, Nan; Zhang, Yan; Bai, Binglian; Wang, Haitao; Li, Min

    2016-02-07

    In this work, a new kind of gelator, 1,3-bis[(3,4-dioctyloxy phenyl) hydrazide]phenylene (BP8-C), containing two dihydrazide units as the rigid bent-core, has been synthesized and investigated. It was demonstrated that BP8-C is an efficient gelator which can gel various organic solvents, such as ethanol, benzene, toluene, chloroform, etc. Both an opaque gel (O-gel) and a transparent gel (T-gel), which is more stable, were obtained with BP8-C in chloroform at different incubation temperatures. Kinetic data based on fluorescence spectra revealed that the T-gels showed a larger Avrami parameter (n = 1.44 at 20 °C) than that of the O-gels (n = 1.21 for gelation at temperatures below 0 °C). While BP8-C did form the opaque gel in toluene, gelation took longer at lower incubation temperatures and even precipitated out below 0 °C. The kinetic Avrami analysis on sols of BP8-C with different concentrations shows a two-phrase mechanism, i.e. the n values are between 0.88 and 1.74 followed by 1.69 and 3.01 throughout the temperature range of 5 °C and 35 °C for 5.34 mg mL(-1) BP8-C in toluene, indicating that the fibers formed first and then bundled to produce compact networks. We propose that supersaturation governs the formation of gel in chloroform and that the diffusion process denominates gelation in toluene. XRD and FT-IR measurements confirmed that the xerogels prepared at different temperatures in different solvents exhibited a Col(h) structure and that there are three molecules in one columnar slice. Our results indicate that the gelation process, morphology of the gels and thus the final properties of the gels depend strongly on the preparation conditions such as temperature, solvent, concentration, etc.

  20. Effectiveness of rotary and hand files in gutta-percha and sealer removal using chloroform or chlorhexidine gel.

    PubMed

    Bueno, Carlos Eduardo da Silveira; Delboni, Maraisa Greggio; de Araújo, Roberta Aranha; Carrara, Hilton José; Cunha, Rodrigo Sanches

    2006-01-01

    The purpose of this study was to assess in vitro the efficacy of nickel-titanium K3 rotary files and hand files for removal of gutta-percha and sealer from obturated root canals using either chloroform or chlorhexidine as solvents. Sixty extracted single-rooted bovine teeth with straight, large canals were prepared, obtured and randomly assigned to 3 groups (n=20). The teeth were stored at 37 degrees C for 1 month and then the gutta-percha and sealer were removed using different techniques, as follows. Group I: size 3 Gates-Glidden drills plus size 30 hand K-files and Hedström files and chloroform; Group II: K3 NiTi rotary files and chloroform; and Group III: K3 NiTi rotary files and 2% chlorhexidine gel. Radiographs were taken and scanned and the images were digitized. The total area of the canal and the area with remaining obturation material were measured in millimeters using a computed image analysis system (ImageLab). Data were analyzed statistically by one-way ANOVA and Tukey test at 5% significance level. The groups differed statistically (p<0.05) with respect to the average percentage of remaining gutta-percha and sealer, presenting the following sequence of effectiveness (from most to least effective): Group I (15.48%), Group II (28.42%) and Group III (35.96%). The findings of this study showed that, despite the technique used for removal of filling material, none of the retreated canals were completely free of gutta-percha and sealer remnants. The use of stainless steel hand files resulted in a lesser amount of filling debris than the use of nickel-titanium rotary instruments.

  1. Fasting increases the concentrations of carbon tetrachloride and of its metabolite chloroform in the liver of mice.

    PubMed

    Pentz, R; Strubelt, O

    1983-05-01

    Fasting mice for 24 h strongly enhanced hepatic triglyceride concentrations as well as the hepatic levels of carbon tetrachloride (CCl4) and chloroform (CHCl3) after i.p. injection of 0.1 ml/kg CCl4. The ratio CHCl3:CCl4 was lower in the livers of the fasted than in those of the fed mice. Fasting-induced steatosis leading to an increased affinity of the liver to a lipophilic compound like CCl4 is considered to be the cause for the increase in CCl4 hepatotoxicity induced by fasting in mice.

  2. The role of renal proximal tubule P450 enzymes in chloroform-induced nephrotoxicity: Utility of renal specific P450 reductase knockout mouse models

    SciTech Connect

    Liu, Senyan; Yao, Yunyi; Lu, Shijun; Aldous, Kenneth; Ding, Xinxin; Mei, Changlin; Gu, Jun

    2013-10-01

    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 with 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.

  3. The use of alkaline hydrolysis as a novel strategy for chloroform remediation: the feasibility of using construction wastes and evaluation of carbon isotopic fractionation.

    PubMed

    Torrentó, Clara; Audí-Miró, Carme; Bordeleau, Geneviève; Marchesi, Massimo; Rosell, Mònica; Otero, Neus; Soler, Albert

    2014-01-01

    Laboratory and field-scale pilot experiments were performed to evaluate the feasibility of chloroform degradation by alkaline hydrolysis and the potential of δ(13)C values to assess this induced reaction process at contaminated sites. In batch experiments, alkaline conditions were induced by adding crushed concrete (pH 12.33 ± 0.07), a filtered concrete solution (pH 12.27 ± 0.04), a filtered cement solution (pH 12.66 ± 0.02) and a pH 12 buffer solution (pH 11.92 ± 0.11). The resulting chloroform degradation after 28 days was 94, 96, 99, and 72%, respectively. The experimental data were described using a pseudo-first-order kinetic model, resulting in pseudo-first-order rate constant values of 0.10, 0.12, 0.20, and 0.05 d(-1), respectively. Furthermore, the significant chloroform carbon isotopic fractionation associated with alkaline hydrolysis of chloroform (-53 ± 3‰) and its independence from pH in the admittedly limited tested pH range imply a great potential for the use of δ(13)C values for in situ monitoring of the efficacy of remediation approaches based on alkaline hydrolysis. The carbon isotopic fractionation obtained at the lab scale allowed the calculation of the percentage of chloroform degradation in field-scale pilot experiments where alkaline conditions were induced in two recharge water interception trenches filled with concrete-based construction wastes. A maximum of approximately 30-40% of chloroform degradation was achieved during the two studied recharge periods. Although further research is required, the treatment of chloroform in groundwater through the use of concrete-based construction wastes is proposed. This strategy would also imply the recycling of construction and demolition wastes for use in value-added applications to increase economic and environmental benefits.

  4. An inelastic neutron scattering and NIR-FT Raman spectroscopy study of chloroform and trichloroethylene in faujasites

    SciTech Connect

    Davidson, A.M.; Mellot, C.F.; Eckert, J.; Cheetham, A.K.

    2000-01-27

    Molecular information about the nature and the strength of the interactions between chloroform and trichloroethylene (TCE) sorbates and siliceous FAU, NaY, and NaX zeolites was obtained by inelastic neutron scattering (INS) and Raman spectroscopies. The spectral features of the two sorbates differ in terms of their frequencies, splittings, and line widths from the ones of chloroform and TCE molecules in the gas phase. In conjunction with the authors simulation results, these differences are rationalized by assuming that, in siliceous FAU, the two sorbates undergo a nondissociative adsorption involving the formation of an H{sub sorbate}{hor{underscore}ellipsis}O{sub framework} hydrogen bond and Cl{sub sorbate}{hor{underscore}ellipsis}O{sub framework} van der Waals interactions, whereas in NaY and NaX, additional Cl{sub sorbate}{hor{underscore}ellipsis}Na{sup +} electrostatic interactions are involved. Interestingly, no {pi}/Na{sup +} interaction takes place for TCE. These findings, which are in agreement with previous calorimetric and simulation results, confirm that the strength of the sorbate/zeolite interactions is correlated to the basicity of the zeolite and therefore increases in the sequence siliceous FAU < NaY < NaX, following the sequence of the heats of adsorption.

  5. A pillar-layered metal-organic framework as luminescent sensor for selective and reversible response of chloroform

    NASA Astrophysics Data System (ADS)

    Wang, Kun; Li, Shuni; Jiang, Yucheng; Hu, Mancheng; Zhai, Quan-Guo

    2017-03-01

    A new 3D metal-organic framework, namely, {Zn4(H2BPTC)2(HCOO)4}n (SNNU-1, H4BPTC=biphenyl-3,3',5,5'-tetracarboxylic acid, SNNU=Shaanxi Normal University) has been solvothermal synthesized. Four independent tetrahedral Zn atoms are connected by organic ligands to form a 2D Zn-H2BPTC layer, which is further bridged by in-situ generated HCOO- to give the 3D pillar-layered framework of SNNU-1. Unique Zn and H2BPTC all act as 4-connected nodes leading to a new 4,4,4-connected topological net with point symbol of {4·5·62·82}{4·52·62·8}{52·63·7}. Notably, intense blue emission band is observed for SNNU-1, which exhibits solvent-dependent effect. Compared to other common organic solvents, chloroform can specially improve the photoluminescent intensity of SNNU-1. Further repeated response and release experiments clearly showed that SNNU-1 can act as luminescent sensor for selective and reversible detection of chloroform.

  6. Morphological Evolution of Noble Metal Nanoparticles in Chloroform: Mechanism of Switching on/off by Protic Species

    PubMed Central

    Douglas-Gallardo, O. A.; Gomez, C. G.; Macchione, M. A.; Cometto, F. P.; Coronado, E. A.; Macagno, V. A.; Pérez, M. A.

    2015-01-01

    The morphological stability/morphological reshaping of noble metal nanoparticles are studied experimentally in order to unravel the chemical mechanisms lying beneath. Gold and silver nanoparticles (AuNPs and AgNPs, respectively) formed in chloroformic environment are used, as model synthetic systems, to study phenomena of morphological change. The morphological evolution of NPs that follows their formation, is characterized by spectroscopy (UV-Visible, Raman and FTIR) and TEM (Transmission Electron Microscopy). The change of NP morphology involves the increase of the average NP size and the broadening of size distribution, in a close resemblance with the effect characteristically obtained from the Ostwald ripening. The effect of the poor solvating properties of chloroform in stabilizing small charged species (H+, Ag+, Au+) as well as the principle of electroneutrality of matter are analyzed in order to formulate a feasible reaction scheme consisting of a three-step processes: the generation of soluble intermediary species by corrosion of nanoparticles, the diffusion of intermediary species from one nanoparticle to another, and the re-deposition process involving the reduction of intermediary species. This basic reaction scheme is used as hypothesis to plan and perform experiments, which reveal that molecular oxygen dissolved in the dispersive medium can drive NP corrosion, however, protic species are also required as co-reactant. The polarity of the hydrogen bond and the ligand properties of the anions produced by deprotonation are feature of the protic species that enable/disable the corrosion and, in turn, the NP morphological evolution. PMID:26889378

  7. Spectroscopic analysis of porphyrin compounds irradiated with visible light in chloroform with addition of β-myrcene

    NASA Astrophysics Data System (ADS)

    Makarska-Bialokoz, Magdalena; Gladysz-Plaska, Agnieszka

    2016-12-01

    The behaviour of two porphyrins, 5,10,15,20-tetraphenyl-21H,23H-porphine (H2TPP) and 5,10,15,20-tetra(4-pyridyl)-21H, 23H-porphine (H2TPyP), as well as their Zn(II) complexes (ZnTPP and ZnTPyP), have been studied analysing their absorption and steady-state fluorescence spectra in chloroform with addition of β-myrcene. After irradiation with visible light the free-base porphyrins have been converted to the form of dication on account of hydrochloric acid generated as a result of chloroform decomposition induced by β-myrcene. Whereas in case of their Zn(II) complexes the mechanism of action is more complicated, leading presumably to the formation of the aggregated metalloporphyrin species with chloride ions playing the bridging role. The pseudo-first-order rate constants of the absorption quenching process were calculated for all the systems examined, with respect to the porphyrin concentration. The most effective irradiation was observed in case of H2TPP porphyrin.

  8. Inductively coupled plasma torch efficiency at atmospheric pressure for organo-chlorine liquid waste removal: chloroform destruction in oxidative conditions.

    PubMed

    Kamgang-Youbi, Georges; Poizot, Karine; Lemont, Florent

    2013-01-15

    The performance of a plasma reactor for the degradation of chlorinated hydrocarbon waste is reported. Chloroform was used as a target for a recently patented destruction process based using an inductive plasma torch. Liquid waste was directly injected axially into the argon plasma with a supplied power of ~4kW in the presence of oxygen as oxidant and carrier gas. Decomposition was performed at CHCl(3) feed rates up to 400 g h(-1) with different oxygen/waste molar ratios, chloroform destruction was obtained with at least 99% efficiency and the energy efficiency reached 100 g kWh(-1). The conversion end products were identified and assayed by online FTIR spectroscopy (CO(2), HCl and H(2)O) and redox titration (Cl(2)). Considering phosgene as representative of toxic compounds, only very small quantities of toxics were released (<1 g h(-1)) even with high waste feed rates. The experimental results were very close to the equilibrium composition predicted by thermodynamic calculations. At the bottom of the reactor, the chlorinated acids were successfully trapped in a scrubber and transformed into mineral salts, hence, only CO(2) and H(2)O have been found in the final off-gases composition.

  9. HPLC and anti-inflammatory studies of the flavonoid rich chloroform extract fraction of Orthosiphon stamineus leaves.

    PubMed

    Yam, Mun Fei; Lim, Vuanghao; Salman, Ibrahim Muhammad; Ameer, Omar Ziad; Ang, Lee Fung; Rosidah, Noersal; Abdulkarim, Muthanna Fawzy; Abdullah, Ghassan Zuhair; Basir, Rusliza; Sadikun, Amirin; Asmawi, Mohd Zaini

    2010-06-21

    The aim of the present study was to verify the anti-inflammatory activity of Orthosiphon stamineus leaf extracts and to identify the active compound(s) contributing to its anti-inflammatory activity using a developed HPLC method. Active chloroform extract of O. stamineus was fractionated into three fractions using a dry flash column chromatography method. These three fractions were investigated for anti-peritoneal capillary permeability, in vitro nitric oxide scavenging activity, anti-inflammatory and nitric oxide (NO) inhibition using carrageenan-induced hind paw edema method. The flavonoid rich chloroform extract fraction (CF2) [containing sinensetin (2.86% w/w), eupatorin (5.05% w/w) and 3'-hydroxy-5,6,7,4'-tetramethoxyflavone (1.101% w/w)], significantly reduced rat hind paw edema, NO and decreased dye leakage to peritoneal cavity at p < 0.05. IC(50) of in vitro NO scavenging of CF2 was 0.3 mg/mL. These results suggest that the anti-inflammatory properties of these CF2 may possibly be due to the presence of flavonoid compounds capable of affecting the NO pathway.

  10. Genotoxicity of drinking water disinfection by-products (bromoform and chloroform) by using both Allium anaphase-telophase and comet tests.

    PubMed

    Khallef, Messaouda; Liman, Recep; Konuk, Muhsin; Ciğerci, İbrahim Hakkı; Benouareth, Djameleddine; Tabet, Mouna; Abda, Ahlem

    2015-03-01

    Genotoxic effects of bromoform and chloroform, disinfection by-products of the chlorination of drinking water, were examined by using mitotic index (MI), mitotic phase, chromosome aberrations (CAs) and comet assay on root meristematic cells of Allium cepa. Different concentrations of bromoform (25, 50, 75 and 100 μg/mL) and chloroform (25, 50, 100 and 200 μg/mL) were introduced to onion tuber roots. Distilled water was used as a negative control and methyl methansulfonate (MMS-10 μg/mL) as positive control. All obtained data were subjected to statistical analyses by using SPSS 15.0 for Windows software. For comparison purposes, Duncan multiple range tests by using one-way analysis of variance were employed and p < 0.05 was accepted as significant value. Exposure of both chemicals (except 25 μg/mL applications of bromoform) significantly decreased MI. Bromoform and chloroform (except 25 μg/mL applications) increased total CAs in Allium anaphase-telophase test. A significant increase in DNA damage was also observed at all concentrations of both bromoform and chloroform examined by comet assay. The damages were higher than that of positive control especially at 75-100 μg/mL for bromoform and 100-200 μg/mL for chloroform.

  11. Formation of Carbonized Polystyrene Sphere/hemisphere Shell Arrays by Ion Beam Irradiation and Subsequent Annealing or Chloroform Treatment

    PubMed Central

    Song, Xianyin; Dai, Zhigao; Xiao, Xiangheng; Li, Wenqing; Zheng, Xudong; Shang, Xunzhong; Zhang, Xiaolei; Cai, Guangxu; Wu, Wei; Meng, Fanli; Jiang, Changzhong

    2015-01-01

    Heat-resistant two-dimensional (2D) sphere/hemisphere shell array is significant for the fabrication of novel nanostructures. Here large-area, well-ordered arrays of carbonized polystyrene (PS) hollow sphere/hemisphere with controlled size and morphology are prepared by combining the nanosphere self-assembly, kV Ag ion beam modification, and subsequent annealing or chloroform treatment. Potential mechanisms for the formation and evolution of the heat-resistant carbonized PS spherical shell with increasing ion fluence and energy are discussed. Combined with noble metal or semiconductor, these modified PS sphere arrays should open up new possibilities for high-performance nanoscale optical sensors or photoelectric devices. PMID:26640125

  12. Amelioration of oxygen and glucose deprivation-induced neuronal death by chloroform fraction of bay leaves (Laurus nobilis).

    PubMed

    Cho, Eun-Young; Lee, Sung-Jin; Nam, Kung-Woo; Shin, Jongheon; Oh, Ki-bong; Kim, Kyeong Ho; Mar, Woongchon

    2010-01-01

    The purpose of this study was to determine whether the Laurus nobilis chloroform fraction (LNCF) protects against cerebral ischemia neuronal damage. Human neuroblastoma SH-SY5Y cells and brain slices from rats were subjected to oxygen and glucose deprivation (OGD), followed by reoxgenation with and without LNCF. The viabilities of SH-SY5Y cells and brain slices from the rats were 58.5±4.9% and 79.7±5.9% in the group subjected to OGD, and 80.4±0.4% and 97.2±1.9% at 4 µg/ml of LNCF, respectively. LNCF also significantly inhibited death-associated protein kinase (DAPK) dephosphorylation. Pretreatment with LNCF at 4 mg/kg significantly decreased infarct size by 79% of vehicle control in the middle cerebral artery occlusion (MCAO) in vivo model. LNCF is a neuroprotective drug candidate against cerebral ischemia neuronal damage.

  13. Determination of buoyant density and sensitivity to chloroform and freon for the etiological agent of infectious salmonid anaemia

    USGS Publications Warehouse

    Christie, K.E.; Hjeltnes, B.; Uglenes , I.; Winton, J.R.

    1993-01-01

    Plasma was collected from Atlantic salmon Salrno salar with acute infectious salmon anaemia (ISA) and used to challenge Atlantic salmon parr by intraperitoneal injection. Treatment of plasma with the lipid solvent, chloroform, showed that the etiological agent of ISA contained essential lipids, probably as a viral envelope. Some infectivity remained following treatment with freon. Injection challenges using fractions from equilibrium density gradient centrifugation of plasma from fish with acute ISA revealed a band of infectivity in the range 1.184 to 1.262 g cm-3. The band was believed to conta~n both complete ISA-virus particles and infectious particles lacking a complete envelope, nucleocapsid or genome. Density gradient centrifugation of infectious plasma for enrichment of the putative ISA virus appeared to offer a suitable method for obtaining virus-specific nucleic acid for use in the construction of cDNA libraries. 

  14. Effects of chloroformic extracts from washed and unwashed papaya seeds (Carica papaya) on the sperm concentration of dogs.

    PubMed

    Ortega-Pacheco, A; Jiménez-Coello, M; Gutiérrez-Blanco, E; Acosta-Viana, K Y; Guzmán-Marín, E; Zavala-Sánchez, M A; Montalvo-Beltrán, N E; Pérez-Gutiérrez, M S

    2010-12-01

    Papaya seeds (Carica papaya Linn) have been found to have a significant effect on sperm characteristics in some mammals, including humans, but no studies have investigated the effects on dogs. In the present study, a significant decrease in sperm concentration was observed in a group of dogs treated with extracts from washed papaya seeds, but no decrease was observed in the group of dogs treated with non-washed seeds. An important effect of extract components from washed seeds such as fatty acids is probably involved in the reduction of sperm production because of Sertoli cell damage, as has been suggested for langur monkeys. Dilution of the active components in the non-washed papaya seeds or interference with some of the components may reduce the expected effect on spermatogenesis. This first report on the effects of a chloroformic extract of papaya seeds in dogs suggests that an increased dose is necessary to achieve azoospermia.

  15. Induced regenerative cell proliferation in livers and kidneys of male F-344 rats given chloroform in corn oil by gavage or ad libitum in drinking water.

    PubMed

    Larson, J L; Wolf, D C; Butterworth, B E

    1995-01-06

    These studies were designed to establish the dose response relationships for the induction of cytolethality and regenerative cell proliferation in the liver and kidneys of male F-344 rats given chloroform by gavage or in drinking water. Rats were administered oral doses of 0, 10, 34, 90 or 180 mg/kg/day chloroform dissolved in corn oil by gavage for 4 days or for 5 days/week for 3 weeks. A second group of rats was given chloroform ad libitum in the drinking water at concentrations of 0, 60, 200, 400, 900 or 1800 ppm for 4 days or 3 weeks. Bromodeoxyuridine (BrdU) was administered via an implanted osmotic pump 3.5 days prior to necropsy to label cells in S-phase. Cells having incorporated BrdU were visualized in tissue sections immunohistochemically and the labelling index (LI) evaluated as the percentage of S-phase cells. Rats treated with 90 or 180 mg/kg/day by gavage for 4 days had mild to moderate degeneration of renal proximal tubules and centrilobular hepatocytes. These alterations were absent or slight after 3 weeks of treatment. LI were increased in the kidney cortex only in the rats treated with 180 mg/kg/day for 4 days. A dose-dependent increase in LI was seen in rat liver after 4 days of treatment with 90 and 180 mg/kg/day by gavage, but the LI remained elevated after 3 weeks of treatment only at the 180 mg/kg/day dose. When chloroform was administered in the drinking water, no microscopic alterations were seen in the kidneys after 4 days of treatment. As a general observation, rats treated for 3 weeks with 200 ppm chloroform and greater had slightly increased numbers of focal areas of regenerating renal proximal tubular epithelium and cell proliferation than were noted in the controls, but no clear dose response relationship was evident. However, the overall renal LI was not increased at any dose or time point. Similarly, only mild hepatocyte vacuolation was observed in rats given 1800 ppm chloroform in the water for 3 weeks with no increase in the

  16. 1H NMR spectra of alcohols and diols in chloroform: DFT/GIAO calculation of chemical shifts.

    PubMed

    Lomas, John S

    2014-12-01

    Proton nuclear magnetic resonance (NMR) shifts of aliphatic alcohols in chloroform have been computed on the basis of density functional theory, the solvent being included by the integral-equation-formalism polarisable continuum model of Gaussian 09. Relative energies of all conformers are calculated at the Perdew, Burke and Ernzerhof (PBE)0/6-311+G(d,p) level, and NMR shifts by the gauge-including atomic orbital method with the PBE0/6-311+G(d,p) geometry and the cc-pVTZ basis set. The 208 computed CH proton NMR shifts for 34 alcohols correlate very well with the experimental values, with a gradient of 1.00 ± 0.01 and intercept close to zero; the overall root mean square difference (RMSD) is 0.08 ppm. Shifts for CH protons of diols in chloroform are well correlated with the theoretical values for (isotropic) benzene, with similar gradient and intercept (1.02 ± 0.01, -0.13 ppm), but the overall RMSD is slightly higher, 0.12 ppm. This approach generally gives slightly better results than the CHARGE model of Abraham et al. The shifts of unsaturated alcohols in benzene have been re-examined with Gaussian 09, but the overall fit for CH protons is not improved, and OH proton shifts are worse. Shifts of vinyl protons in alkenols are systematically overestimated, and the correlation of computed shifts against the experimental data for unsaturated alcohols follows a quadratic equation. Splitting the 20 compounds studied into two sets, and applying empirical scaling based on the quadratic for the first set to the second set, gives an RMSD of 0.10 ppm. A multi-standard approach gives a similar result.

  17. Chloroform extract of hog barn dust modulates skeletal muscle ryanodine receptor calcium-release channel (RyR1).

    PubMed

    Tian, Chengju; Shao, Chun Hong; Fenster, Danielle S; Mixan, Mark; Romberger, Debra J; Toews, Myron L; Bidasee, Keshore R

    2010-09-01

    Skeletal muscle weakness is a reported ailment in individuals working in commercial hog confinement facilities. To date, specific mechanisms responsible for this symptom remain undefined. The purpose of this study was to assess whether hog barn dust (HBD) contains components that are capable of binding to and modulating the activity of type 1 ryanodine receptor Ca2+-release channel (RyR1), a key regulator of skeletal muscle function. HBD collected from confinement facilities in Nebraska were extracted with chloroform, filtered, and rotary evaporated to dryness. Residues were resuspended in hexane-chloroform (20:1) and precipitates, referred to as HBDorg, were air-dried and studied further. In competition assays, HBDorg dose-dependently displaced [3H]ryanodine from binding sites on RyR1 with an IC50 of 1.5±0.1 microg/ml (Ki=0.4±0.0 microg/ml). In single-channel assays using RyR1 reconstituted into a lipid bilayer, HBDorg exhibited three distinct dose-dependent effects: first it increased the open probability of RyR1 by increasing its gating frequency and dwell time in the open state, then it induced a state of reduced conductance (55% of maximum) that was more likely to occur and persist at positive holding potentials, and finally it irreversibly closed RyR1. In differentiated C2C12 myotubes, addition of HBD triggered a rise in intracellular Ca2+ that was blocked by pretreatment with ryanodine. Since persistent activation and/or closure of RyR1 results in skeletal muscle weakness, these new data suggest that HBD is responsible, at least in part, for the muscle ailment reported by hog confinement workers.

  18. Chloroform extract of hog barn dust modulates skeletal muscle ryanodine receptor calcium-release channel (RyR1)

    PubMed Central

    Tian, Chengju; Shao, Chun Hong; Fenster, Danielle S.; Mixan, Mark; Romberger, Debra J.; Toews, Myron L.

    2010-01-01

    Skeletal muscle weakness is a reported ailment in individuals working in commercial hog confinement facilities. To date, specific mechanisms responsible for this symptom remain undefined. The purpose of this study was to assess whether hog barn dust (HBD) contains components that are capable of binding to and modulating the activity of type 1 ryanodine receptor Ca2+-release channel (RyR1), a key regulator of skeletal muscle function. HBD collected from confinement facilities in Nebraska were extracted with chloroform, filtered, and rotary evaporated to dryness. Residues were resuspended in hexane-chloroform (20:1) and precipitates, referred to as HBDorg, were air-dried and studied further. In competition assays, HBDorg dose-dependently displaced [3H]ryanodine from binding sites on RyR1 with an IC50 of 1.5 ± 0.1 μg/ml (Ki = 0.4 ± 0.0 μg/ml). In single-channel assays using RyR1 reconstituted into a lipid bilayer, HBDorg exhibited three distinct dose-dependent effects: first it increased the open probability of RyR1 by increasing its gating frequency and dwell time in the open state, then it induced a state of reduced conductance (55% of maximum) that was more likely to occur and persist at positive holding potentials, and finally it irreversibly closed RyR1. In differentiated C2C12 myotubes, addition of HBD triggered a rise in intracellular Ca2+ that was blocked by pretreatment with ryanodine. Since persistent activation and/or closure of RyR1 results in skeletal muscle weakness, these new data suggest that HBD is responsible, at least in part, for the muscle ailment reported by hog confinement workers. PMID:20576841

  19. A comparative VCD study of methyl mandelate in methanol, dimethyl sulfoxide, and chloroform: explicit and implicit solvation models.

    PubMed

    Poopari, Mohammad Reza; Dezhahang, Zahra; Xu, Yunjie

    2013-02-07

    Vibrational absorption (VA) and vibrational circular dichroism (VCD) spectra of methyl mandelate, a prototype chiral molecule, in a series of organic solvents, namely methanol (MeOH-d(4)), dimethyl sulfoxide (DMSO-d(6)), and chloroform (CDCl(3)), have been measured in the finger print region from 1800 to 1150 cm(-1). Implicit solvation models in the form of polarizable continuum model and explicit solvation models have been employed independently and simultaneously. The goal is to evaluate their efficiencies in dealing with solvent effects in each solution and to establish a general strategy to adequately account for effects of solvents. Molecular dynamics (MD) simulation and radial distribution function analysis have been performed to aid the construction of the explicit solvation models. Initial geometry searches have been carried out at the B3LYP/6-31G(d) level for the methyl mandelate monomer and its explicit 1 : 1 and 1 : 2 solute-solvent hydrogen-bonded complexes. B3LYP/cc-pVTZ has been used for all the final geometry optimizations, the vibrational frequency, VA and VCD intensity, and optical rotation dispersion (ORD) calculations. The results show that inclusion of solvent explicitly and implicitly at the same time has significant impacts on the appearance of the VA and VCD spectra, and is crucial for reliable spectral assignments when solvents are capable of hydrogen-bonding interactions with solutes. When no strong solvent-solute hydrogen-bonding interactions in the case of chloroform are expected, the gas phase monomer model is adequate for spectral interpretation, while inclusion of implicit solvation improves the frequency agreement with experiment. ORD spectra of methyl mandelate in the aforementioned solvents at different concentrations under 5 excitation wavelengths have also been measured. The comparison between the calculated and the experimental ORD spectra supports the conclusions drawn from the VA and VCD investigations.

  20. Physiologically-based pharmacokinetic (PBPK) modeling of two binary mixtures: metabolic activation of carbon tetrachloride by trichloroethylene and metabolic inhibition of chloroform by trichloroethylene.

    EPA Science Inventory

    The interaction between trichloroethylene (TCE) and chloroform (CHCI3) has been described as less than additive, with co-exposure to TCE and CHC13 resulting in less hepatic and renal toxicity than observed with CHCl3 alone. In contrast, the nonadditive interaction between TCE and...

  1. In Vivo Antiprotozoal Activity of the Chloroform Extract from Carica papaya Seeds against Amastigote Stage of Trypanosoma cruzi during Indeterminate and Chronic Phase of Infection

    PubMed Central

    Ortega-Pacheco, Antonio; Perez-Gutierrez, Salud; Guzman-Marin, Eugenia

    2014-01-01

    In order to evaluate the antiprotozoal activity of the chloroform extract of Carica papaya seeds during the subacute and chronic phase of infection of Trypanosoma cruzi, doses of 50 and 75 mg/kg were evaluated during the subacute phase, including a mixture of their main components (oleic, palmitic, and stearic acids). Subsequently, doses of 50 and 75 mg/kg in mice during the chronic phase of infection (100 dpi) were also evaluated. It was found that chloroform extract was able to reduce the amastigote nests numbers during the subacute phase in 55.5 and 69.7% (P > 0.05) as well as in 56.45% in animals treated with the mixture of fatty acids. Moreover, the experimental groups treated with 50 and 75 mg/kg during the chronic phase of the infection showed a significant reduction of 46.8 and 53.13% respectively (P < 0.05). It is recommended to carry out more studies to determine if higher doses of chloroformic extract or its administration in combination with other antichagasic drugs allows a better response over the intracellular stage of T. cruzi in infected animal models and determine if the chloroform extract of C. papaya could be considered as an alternative for treatment during the indeterminate and chronic phase of the infection. PMID:25276216

  2. Ameliorative Effects of Chloroform Fraction of Cocos nucifera L. Husk Fiber Against Cisplatin-induced Toxicity in Rats

    PubMed Central

    Adaramoye, Oluwatosin Adekunle; Azeez, Adesola Fausat; Ola-Davies, Olufunke Elizabeth

    2016-01-01

    Background: Cisplatin (Cis) is used in the treatment of solid tumors and is known to elicit serious side effects. Objective: The present study investigated the protective effects of chloroform fraction of Cocos nucifera husk fiber (CFCN) against Cis-induced organs’ damage and chromosomal defect in rats. Quercetin (QUE), standard antioxidant, served as positive control. Materials and Methods: Thirty male Wistar rats were assigned into six groups and treated with corn oil (control), Cis alone, Cis + CFCN, CFCN alone, Cis + QUE, and QUE alone. QUE and CFCN were given at 50 and 200 mg/kg/day, respectively, by oral gavage for 7 days before the rats were exposed to a single dose of Cis (10 mg/kg, intraperitoneal) at the last 36 h of study. Results: Administration of Cis alone caused a significant (P < 0.05) increase in the levels of serum creatinine and urea by 72% and 70%, respectively, when compared with the control. The activity of serum aspartate aminotransferase was significantly (P < 0.05) increased while alanine aminotransferase and alkaline phosphatase were insignificantly (P > 0.05) affected in Cis-treated rats. Furthermore, the activities of hepatic and renal catalase, superoxide dismutase, glutathione S-transferase, glutathione peroxidase, and levels of reduced glutathione were significantly (P < 0.05) decreased in Cis-treated rats with concomitant elevation of malondialdehyde. Cis exposure increased the frequency of micro nucleated polychromatic erythrocytes (mPCE) by 92%. Pretreatment with CFCN inhibited lipid peroxidation, enhanced the activities of some antioxidative enzymes and reduced the frequency of mPCE. Conclusions: Chloroform fraction of CFCN may protect against organs damage by Cis. Further studies are required to determine the component of the plant responsible for this activity. SUMMARY Cisplatin (Cis) is used in the treatment of solid tumors and is known to elicit serious side effects. This study investigated the protective effects of

  3. Carcinogenicity of chloroform in drinking water to male Osborne-Mendel rats and female B6C3F1 mice.

    PubMed

    Jorgenson, T A; Meierhenry, E F; Rushbrook, C J; Bull, R J; Robinson, M

    1985-08-01

    The carcinogenic activity of chloroform administered at 0, 200, 400, 900, and 1800 mg/liter in drinking water was studied in male Osborne-Mendel rats and female B6C3F1 mice. A second control group was included in the study and was restricted to the water consumption of the high-dose group. Animals were maintained on study for 104 weeks. Group sizes were adjusted at low doses such that a detectable tumor response would result at the lowest dose if there was a linear relationship with dose, and the higher doses produced responses similar to previous carcinogenesis bioassays of chloroform. The primary finding was that chloroform increased the yield of renal tubular adenomas and adenocarcinomas in male rats in a dose-related manner. For the high-dose group, which corresponded to a time-weighted average dose of 160 mg/kg per day for 104 weeks, there was a 14% incidence of renal tubular adenomas and adenocarcinomas, vs 1% in the control group. This compares to a 24% incidence observed when 180 mg/kg per day of chloroform was administered for 78 weeks in earlier studies. In contrast, chloroform in the drinking water of mice failed to increase the incidence of hepatocellular carcinomas in female B6C3F1 mice. The highest dose group received a time-weighted average dose of 263 mg/kg for 104 weeks, resulting in a 5% combined incidence of hepatocellular adenoma and carcinoma relative to a 6% incidence in the control groups.(ABSTRACT TRUNCATED AT 250 WORDS)

  4. A rapid method for profiling of volatile and semi-volatile phytohormones using methyl chloroformate derivatisation and GC-MS.

    PubMed

    Rawlinson, Catherine; Kamphuis, Lars G; Gummer, Joel P A; Singh, Karam B; Trengove, Robert D

    Phytohormones are central components of complex signalling networks in plants. The interplay between these metabolites, which include abscisic acid (ABA), auxin (IAA), ethylene, jasmonic acid (JA) and salicylic acid (SA), regulate plant growth and development and modulate responses to biotic and abiotic stress. Few methods of phytohormone profiling can adequately quantify a large range of plant hormones simultaneously and without the requirement for laborious or highly specialised extraction protocols. Here we describe the development and validation of a phytohormone profiling protocol, based on methyl-chloroformate derivatisation of the plant metabolites and analysis by gas chromatography/mass spectrometry (GC-MS). We describe the analysis of 11 metabolites, either plant phytohormones or intermediates of phytohormone metabolism; ABA, azelaic acid, IAA, JA and SA, and the phytohormone precursors 1-aminocyclopropane 1-carboxylic acid, benzoic acid, cinnamic acid, 13-epi-12-oxophytodienoic acid (13-epi-OPDA), linoleic acid and linolenic acid, and validate the isolation from foliar tissue of the model legume Medicago truncatula. The preparation is insensitive to the presence of water, facilitating measurement of the volatile metabolites. Quantitation was linear over four orders of magnitude, and the limits of detection between two and 10 ng/mL for all measured metabolites using a single quadrupole GC-MS.

  5. 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).

  6. The room temperature preservation of filtered environmental DNA samples and assimilation into a phenol-chloroform-isoamyl alcohol DNA extraction.

    PubMed

    Renshaw, Mark A; Olds, Brett P; Jerde, Christopher L; McVeigh, Margaret M; Lodge, David M

    2015-01-01

    Current research targeting filtered macrobial environmental DNA (eDNA) often relies upon cold ambient temperatures at various stages, including the transport of water samples from the field to the laboratory and the storage of water and/or filtered samples in the laboratory. This poses practical limitations for field collections in locations where refrigeration and frozen storage is difficult or where samples must be transported long distances for further processing and screening. This study demonstrates the successful preservation of eDNA at room temperature (20 °C) in two lysis buffers, CTAB and Longmire's, over a 2-week period of time. Moreover, the preserved eDNA samples were seamlessly integrated into a phenol-chloroform-isoamyl alcohol (PCI) DNA extraction protocol. The successful application of the eDNA extraction to multiple filter membrane types suggests the methods evaluated here may be broadly applied in future eDNA research. Our results also suggest that for many kinds of studies recently reported on macrobial eDNA, detection probabilities could have been increased, and at a lower cost, by utilizing the Longmire's preservation buffer with a PCI DNA extraction.

  7. Atmospheric histories of halocarbons from analysis of Antarctic firn air: Methyl bromide, methyl chloride, chloroform, and dichloromethane

    NASA Astrophysics Data System (ADS)

    Trudinger, C. M.; Etheridge, D. M.; Sturrock, G. A.; Fraser, P. J.; Krummel, P. B.; McCulloch, A.

    2004-11-01

    We reconstruct atmospheric levels of methyl bromide (CH3Br), methyl chloride (CH3Cl), chloroform (CHCl3), and dichloromethane (CH2Cl2) back to before 1940 using measurements of air extracted from firn on Law Dome in Antarctica. The firn air at this site has a relatively narrow age spread, giving high time resolution reconstructions. The CH3Br reconstructions confirm previously measured firn records but with more temporal structure. Our CH3Cl reconstruction is slightly different from previous reconstructions, raising some questions about CH3Cl in the firn. Our reconstructions for CHCl3 and CH2Cl2 are the first published records of concentration prior to direct atmospheric measurements. A two-box atmospheric model is used to investigate the budgets of these gases. Much of the variation in CH3Cl can be explained by biomass burning emissions that increase up to 1980 and then are relatively stable apart from some high burning years such as 1997-1998. The CHCl3 firn reconstruction suggests that the anthropogenic source for CHCl3 is greater than previously thought, with human influence on the soil source a possible important contributor here. The CH2Cl2 firn reconstruction is consistent with industrial emission estimates based on audited sales data but suggests that the ocean source of CH2Cl2 is less than previously estimated.

  8. Anaerobic Biotransformation of High Concentrations of Chloroform by an Enrichment Culture and Two Bacterial Isolates ▿ †

    PubMed Central

    Shan, Huifeng; Kurtz, Harry D.; Mykytczuk, Nadia; Trevors, Jack T.; Freedman, David L.

    2010-01-01

    A fermentative enrichment culture (designated DHM-1) was developed that is capable of cometabolically biotransforming high concentrations of chloroform (CF) to nontoxic end products. Two Pantoea spp. were isolated from DHM-1 that also possess this dechlorination capability. Following acclimation to increasing levels of CF, corn syrup-grown DHM-1 was able to transform over 500 mg/liter CF in the presence of vitamin B12 (approximately 3% of CF on a molar basis) at a rate as high as 22 mg/liter/day in a mineral salts medium. CO, CO2, and organic acids were the predominant biodegradation products, suggesting that hydrolytic reactions predominate during CF transformation. DHM-1 was capable of growing on corn syrup in the presence of high concentrations of CF (as may be present near contaminant source zones in groundwater), which makes it a promising culture for bioaugmentation. Strains DHM-1B and DHM-1T transform CF at rates similar to that of the DHM-1 enrichment culture. The ability of these strains to grow in the presence of high concentrations of CF appears to be related to alteration of membrane fluidity or homeoviscous and homeophasic adaptation. PMID:20693443

  9. Vasorelaxant Action of the Chloroform Fraction of Orthosiphon stamineus via NO/cGMP Pathway, Potassium and Calcium Channels.

    PubMed

    Yam, Mun Fei; Tan, Chu Shan; Ahmad, Mariam; Ruan, Shibao

    2016-01-01

    Orthosiphon stamineus Benth. (Lamiaceae) is an important plant in traditional folk medicine that is used to treat hypertension and kidney stones. In humans, this plant has been tested as an addition regiment for antihypertensive treatment. Among the treatments for hypertension, O. stamineus had been to have diuretic and vasorelaxant effects in animal models. There is still very little information regarding the vasorelaxant effect of O. stamineus. Therefore, the present study was designed to investigate the vasorelaxant activity and mechanism of action of the fractions of O. stamineus. The vasorelaxant activity and the underlying mechanisms of the chloroform fraction of the 50% methanolic extract of O. stamineus (CF) was evaluated on thoracic aortic rings isolated from Sprague Dawley rats. CF caused relaxation of the aortic ring pre-contracted with phenylephrine in the presence and absence of endothelium, and pre-contracted with potassium chloride in endothelium-intact aortic ring. In the presence of endothelium, both indomethacin (a nonselective cyclooxygenase inhibitor) and [Formula: see text]-[1,2,4]Oxadiazolo[4,3-[Formula: see text

  10. DNA extraction and quantitation of forensic samples using the phenol-chloroform method and real-time PCR.

    PubMed

    Köchl, Silvano; Niederstätter, Harald; Parson, Walther

    2005-01-01

    Forensic laboratories are increasingly confronted with problematic samples from the scene of crime, containing only minute amounts of deoxyribonucleic acid (DNA), which may include polymerase chain reaction (PCR)-inhibiting substances. Efficient DNA extraction procedures, as well as accurate DNA quantification methods, are critical steps involved in the process of successful DNA analysis of such samples. The phenol-chloroform method is a sensitive method for the extraction of DNA from a wide variety of forensic samples, although it is known to be laborious compared with single-tube extraction methods. The relatively high DNA recovery and the quality of the extracted DNA speak for itself. For reliable and sensitive DNA quantitation, the application of real-time PCR is described. We modified a published real-time PCR assay, which allows for the combined analysis of nuclear and mitochondrial DNA, by introducing 1) improved hybridization probes with the use of minor groove binders; 2) an internal positive control (for both nuclear and mitochondrial DNA) for the detection of PCR inhibitors; and 3) different amplicon lengths for the determination of the degradation state of the DNA. The internal positive controls were constructed by site directed mutagenesis by overlap extension of the wild-type mitochondrial and nuclear DNA target with the advantage that no additional probes, which are cost-intensive, are required. The quantitation system is accomplished as a modular concept, which allows for the combined determination of the above-mentioned features (quantity/inhibition or quantity/degradation) depending on the situation.

  11. Evaluation of chloroform/methanol extraction to facilitate the study of membrane proteins of non-model plants.

    PubMed

    Vertommen, Annelies; Panis, Bart; Swennen, Rony; Carpentier, Sebastien Christian

    2010-04-01

    Membrane proteins are of great interest to plant physiologists because of their important function in many physiological processes. However, their study is hampered by their low abundance and poor solubility in aqueous buffers. Proteomics studies of non-model plants are generally restricted to gel-based methods. Unfortunately, all gel-based techniques for membrane proteomics lack resolving power. Therefore, a very stringent enrichment method is needed before protein separation. In this study, protein extraction in a mixture of chloroform and methanol in combination with gel electrophoresis is evaluated as a method to study membrane proteins in non-model plants. Benefits as well as disadvantages of the method are discussed. To demonstrate the pitfalls of working with non-model plants and to give a proof of principle, the method was first applied to whole leaves of the model plant Arabidopsis. Subsequently, a comparison with proteins extracted from leaves of the non-model plant, banana, was made. To estimate the tissue and organelle specificity of the method, it was also applied on banana meristems. Abundant membrane or lipid-associated proteins could be identified in both tissues, with the leaf extract yielding a higher number of membrane proteins.

  12. Calculated Third Order Rate Constants for Interpreting the Mechanisms of Hydrolyses of Chloroformates, Carboxylic Acid Halides, Sulfonyl Chlorides and Phosphorochloridates

    PubMed Central

    Bentley, T. William

    2015-01-01

    Hydrolyses of acid derivatives (e.g., carboxylic acid chlorides and fluorides, fluoro- and chloroformates, sulfonyl chlorides, phosphorochloridates, anhydrides) exhibit pseudo-first order kinetics. Reaction mechanisms vary from those involving a cationic intermediate (SN1) to concerted SN2 processes, and further to third order reactions, in which one solvent molecule acts as the attacking nucleophile and a second molecule acts as a general base catalyst. A unified framework is discussed, in which there are two reaction channels—an SN1-SN2 spectrum and an SN2-SN3 spectrum. Third order rate constants (k3) are calculated for solvolytic reactions in a wide range of compositions of acetone-water mixtures, and are shown to be either approximately constant or correlated with the Grunwald-Winstein Y parameter. These data and kinetic solvent isotope effects, provide the experimental evidence for the SN2-SN3 spectrum (e.g., for chloro- and fluoroformates, chloroacetyl chloride, p-nitrobenzoyl p-toluenesulfonate, sulfonyl chlorides). Deviations from linearity lead to U- or V-shaped plots, which assist in the identification of the point at which the reaction channel changes from SN2-SN3 to SN1-SN2 (e.g., for benzoyl chloride). PMID:26006228

  13. Induced cytotoxicity and cell proliferation in the hepatocarcinogenicity of chloroform in female B6C3F1 mice: comparison of administration by gavage in corn oil vs ad libitum in drinking water.

    PubMed

    Larson, J L; Wolf, D C; Butterworth, B E

    1994-01-01

    Chloroform increases the incidence of liver tumors in B6C3F1 mice when administered in by gavage in corn oil, but not when given in the drinking water at similar daily doses. Since cytotoxicity and regenerative cell proliferation have been implicated in the tumorigenic process for this nongenotoxic agent, these effects of chloroform in corn oil and drinking water were evaluated under conditions similar to the two bioassays. Female B6C3F1 mice were administered oral doses of 0, 3, 10, 34, 90, 238, or 477 mg/kg chloroform dissolved in corn oil 5 days/week for periods of 4 days or 3 weeks, or were continually exposed to chloroform in the drinking water at concentrations of 0, 60, 200, 400, 900, or 1800 ppm for 4 days or 3 weeks, at which time they were necropsied. 5-Bromo-2'-deoxyuridine (BrdU) was delivered via osmotic pumps implanted 3.5 days prior to necropsy. Cell proliferation was evaluated as the percentage of hepatocytes that entered S-phase over 3.5 days (labeling index, LI), measured by immunohistochemical detection of BrdU incorporated into the DNA. Dose-dependent changes included centrilobular necrosis and markedly elevated LI in mice given 238 or 477 mg/kg chloroform in corn oil (the average daily doses that produced tumors in the cancer bioassay). The no-observed-effect level for histopathological changes was 10 mg/kg/day and for induced cell proliferation was 34 mg/kg/day for chloroform given in corn oil. Chloroform given in the drinking water did not increase the hepatic LI after either 4 days or 3 weeks in any of the dose groups, nor were any microscopic alterations observed in the livers, even though the cumulative daily amount of chloroform ingested in the 1800-ppm exposure group was 329 mg/kg/day. The sustained increase in LI in the livers of mice administered hepatocarcinogenic doses of chloroform in corn oil, but not for chloroform in drinking water, is evidence that chloroform-induced mouse liver cancer is secondary to events associated with

  14. Stereoselective alkoxycarbonylation of unactivated C(sp3)–H bonds with alkyl chloroformates via Pd(II)/Pd(IV) catalysis

    PubMed Central

    Liao, Gang; Yin, Xue-Song; Chen, Kai; Zhang, Qi; Zhang, Shuo-Qing; Shi, Bing-Feng

    2016-01-01

    Several examples on Pd-catalysed carbonylation of methyl C(sp3)–H bonds with gaseous CO via Pd(II)/Pd(0) catalysis have been reported. However, methylene C(sp3)–H carbonylation remains a great challenge, largely due to the lack of reactivity of C–H bonds and the difficulty in CO migratory insertion. Herein, we report the stereoselective alkoxycarbonylation of both methyl and methylene C(sp3)–H bonds with alkyl chloroformates through a Pd(II)/Pd(IV) catalytic cycle. A broad range of aliphatic carboxamides and alkyl chloroformates are compatible with this protocol. In addition, this process is scalable and the directing group could be easily removed under mild conditions with complete retention of configuration. PMID:27678161

  15. Assessment of the in vivo genotoxicity of cadmium chloride, chloroform, and D,L-menthol as coded test chemicals using the alkaline comet assay.

    PubMed

    Wada, Kunio; Fukuyama, Tomoki; Nakashima, Nobuaki; Matsumoto, Kyomu

    2015-07-01

    As part of the Japanese Center for the Validation of Alternative Methods (JaCVAM) international validation study of in vivo rat alkaline comet assays, we examined cadmium chloride, chloroform, and D,L-menthol under blind conditions as coded chemicals in the liver and stomach of Sprague-Dawley rats after 3 days of administration. Cadmium chloride showed equivocal responses in the liver and stomach, supporting previous reports of its poor mutagenic potential and non-carcinogenic effects in these organs. Treatment with chloroform, which is a non-genotoxic carcinogen, did not induce DNA damage in the liver or stomach. Some histopathological changes, such as necrosis and degeneration, were observed in the liver; however, they did not affect the comet assay results. D,L-Menthol, a non-genotoxic non-carcinogen, did not induce liver or stomach DNA damage. These results indicate that the comet assay can reflect genotoxic properties under blind conditions.

  16. Investigating Migration Inhibition and Apoptotic Effects of Fomitopsis pinicola Chloroform Extract on Human Colorectal Cancer SW-480 Cells

    PubMed Central

    Wang, Pan; Wang, Lu; Fan, Jianping; Wang, Xiaobing; Liu, Quanhong

    2014-01-01

    Background Fomitopsis pinicola (Sw. Ex Fr.m) Karst (FPK) which belongs to the Basidiomycota fungal class is one of the most popular medical fungi in China. It has been used for many diseases: cancer, heart diseases, diabetes and so on. However, little study on the pro-apoptotic effect and migration inhibition of FPK chloroform extract (FPKc) has been reported and the possible involved mechanism has not been illuminated. Methodology/Principal Findings Chemical analysis was performed by HPLC which showed ergosterol (ES) concentration was 105 µg/mg. MTT assay revealed that FPKc could selectively inhibit SW-480 cells viability with the IC50 of 190.28 µg/ml. Wound healing and transwell assay indicated that FPKc could inhibit the migration of SW-480 cells obviously, FPKc could also dramatically decreased the matrix metalloproteinases-2, 9 (MMP-2 and MMP-9) expression. Annexin V–FITC/PI staining, nuclear Hoechst 33342 staining and DNA fragmentation analysis revealed that FPKc and ES could induce SW-480 cells apoptosis. The apoptosis process closely involved in ROS accumulation and depletion of GSH, activation of caspase 3, poly (ADP-ribose) polymerase (PARP) degradation. FPKc could also up-regulate P53 expression and thus lead to G1 phase arrest. When SW-480 cells were pretreated with N-acetylcysteine (NAC), the ROS generation, cell viability and apoptotic ratio were partially declined, which indicated that ROS was vertical in the pro-apoptosis process induced by FPKc. Moreover, in the whole process, ES which has been previously found in FPKc had the similar effect to FPKc. Thus we could conclude that ES, as one of the highest abundant components in FPKc, might also be one of the active constituents. Conclusion/Significance FPKc could inhibit the migration of SW-480 cells, induce SW-480 cells G1 phase arrest and cause ROS-mediated apoptosis effect. And ES might be one of the effective constituents in the whole process. PMID:24992193

  17. NOAEL and LOAEL determinations of acute hepatotoxicity for chloroform and bromodichloromethane delivered in an aqueous vehicle to F344 rats.

    PubMed

    Keegan, T E; Simmons, J E; Pegram, R A

    1998-09-11

    Chloroform (CHCl3) and bromodichloromethane (BDCM) are generally the two most prevalent disinfection by-products formed during chlorination of drinking water, and both have been shown to be hepatotoxic, nephrotoxic, and carcinogenic in rodents. As the toxicity of these trihalomethanes (THMs) has most often been studied with corn oil as the vehicle of administration, the objectives of this study were to assess hepatotoxicity after exposure to single, low dosages of CHCl3 and BDCM given orally in an aqueous vehicle to estimate a lowest-observed-adverse-effect level (LOAEL) and a no-observed-adverse-effect level (NOAEL) and to compare toxic potency. Ninety-day-old male Fischer 344 rats were gavaged with either 0.125, 0.1875, 0.25, 0.5, 0.75, 1.0, or 1.5 mmol CHCl3 or BDCM/kg body weight in 10% Alkamuls EL-620 (5 ml/kg body weight). At 24 h postgavage, serum was collected for analysis of clinical chemistry indicators of liver damage. Both CHCl3 and BDCM induced dose-dependent hepatotoxicity; serum alanine aminotransferase, aspartate aminotransferase, and sorbitol dehydrogenase were elevated significantly over control at 1.5, 1.0, and 0.5 mmol/kg. At these dose levels after 24 h, the two THMs appeared to be equipotent hepatotoxicants. Additional assessments at later time points demonstrated that BDCM causes more persistent liver damage than CHCl3 (Lilly et al., 1997). At 0.25, 0. 1875, and 0. 125 mmol of either THM/kg, significant increases over control were not detected for any measured endpoint. Therefore, these data indicate that the acute, oral NOAELs and LOAELs for liver toxicity are 0.25 and 0.5 mmol/kg, respectively, for both CHCl3 and BDCM. These determinations should provide a basis to establish new exposure limits for One-Day Health Advisories for these prevalent THMs.

  18. Cytotoxic effects of chloroform and hydroalcoholic extracts of aerial parts of Cuscuta chinensis and Cuscuta epithymum on Hela, HT29 and MDA-MB-468 tumor cells

    PubMed Central

    Jafarian, A.; Ghannadi, A.; Mohebi, B.

    2014-01-01

    Previous studies have indicated that some species of Cuscuta possess anticancer activity on various cell lines. Due to the lack of detailed researches on the cytotoxic effects of Cuscuta chinensis and Cuscuta epithymum, the aim of the present study was to evaluate cytotoxic effects of chloroform and hydroalcoholic extracts of these plants on the human breast carcinoma cell line (MDA-MB-468), human colorectal adenocarcinoma cell line (HT29) and human uterine cervical carcinoma (Hela). Using maceration method, different extracts of aerial parts of C. chinensis and C. epithymum were prepared. Extraction was performed using chloroform and ethanol/water (70/30). Total phenolic contents of the extracts were determined according to the Folin-Ciocalteu method. Using MTT assay, the cytotoxic activity of the extracts against HT29, Hela and MDA-MB-468 tumor cells was evaluated. Extracts were considered cytotoxic when more than 50% reduction on cell survival was observed. The poly-phenolic content of the hydroalcoholic and chloroform extracts of C. chinensis and C. epithymum were 56.08 ± 4.11, 21.49 ± 2.00, 10.64 ± 0.86 and 4.81 ± 0.38, respectively. Our findings showed that the chloroform extracts of C. chinensis and C. epithyum significantly reduced the viability of Hela, HT-29 and MDA-MB-468 cells. Also, hydroalcoholic extracts of C. chinensis significantly decreased the viability of HT29, Hela and MDA-MB-468 cells. However, in the case of hydroalcoholic extracts of C. epithymum only significant decrease in the viability of MDA-MB-468 cells was observed (IC50 = 340 μg/ml). From these findings it can be concluded that C. chinensis and C. epithymum are good candidates for further study to find new possible cytotoxic agents. PMID:25657780

  19. Cytotoxic effects of chloroform and hydroalcoholic extracts of aerial parts of Cuscuta chinensis and Cuscuta epithymum on Hela, HT29 and MDA-MB-468 tumor cells.

    PubMed

    Jafarian, A; Ghannadi, A; Mohebi, B

    2014-01-01

    Previous studies have indicated that some species of Cuscuta possess anticancer activity on various cell lines. Due to the lack of detailed researches on the cytotoxic effects of Cuscuta chinensis and Cuscuta epithymum, the aim of the present study was to evaluate cytotoxic effects of chloroform and hydroalcoholic extracts of these plants on the human breast carcinoma cell line (MDA-MB-468), human colorectal adenocarcinoma cell line (HT29) and human uterine cervical carcinoma (Hela). Using maceration method, different extracts of aerial parts of C. chinensis and C. epithymum were prepared. Extraction was performed using chloroform and ethanol/water (70/30). Total phenolic contents of the extracts were determined according to the Folin-Ciocalteu method. Using MTT assay, the cytotoxic activity of the extracts against HT29, Hela and MDA-MB-468 tumor cells was evaluated. Extracts were considered cytotoxic when more than 50% reduction on cell survival was observed. The poly-phenolic content of the hydroalcoholic and chloroform extracts of C. chinensis and C. epithymum were 56.08 ± 4.11, 21.49 ± 2.00, 10.64 ± 0.86 and 4.81 ± 0.38, respectively. Our findings showed that the chloroform extracts of C. chinensis and C. epithyum significantly reduced the viability of Hela, HT-29 and MDA-MB-468 cells. Also, hydroalcoholic extracts of C. chinensis significantly decreased the viability of HT29, Hela and MDA-MB-468 cells. However, in the case of hydroalcoholic extracts of C. epithymum only significant decrease in the viability of MDA-MB-468 cells was observed (IC50 = 340 μg/ml). From these findings it can be concluded that C. chinensis and C. epithymum are good candidates for further study to find new possible cytotoxic agents.

  20. RNA extraction from various recalcitrant plant tissues with a cethyltrimethylammonium bromide-containing buffer followed by an acid guanidium thiocyanate-phenol-chloroform treatment.

    PubMed

    Suzuki, Yuji; Mae, Tadahiko; Makino, Amane

    2008-07-01

    High-quality total RNA was extracted using a cethyltrimethylammonium bromide-containing buffer followed by an acid guanidium thiocyanate-phenol-chloroform treatment from recalcitrant plant tissues such as tree leaves (pine, Norway spruce, ginkgo, Japanese cedar, rose), flowers (rose, Lotus japonicus) and storage tissues (seeds of Lotus japonicus and rice, sweet potato tuber, banana fruit). This protocol greatly reduced the time required for RNA extraction.

  1. Survey of several methods deproteinizing human plasma before and within the chloroformate-mediated treatment of amino/carboxylic acids quantitated by gas chromatography.

    PubMed

    Hušek, Petr; Svagera, Zdeněk; Hanzlíková, Dagmar; Simek, Petr

    2012-01-01

    Trichloroacetic acid, perchloric acid, phosphotungstic acid, acetonitrile, methanol and some other organic solvents were evaluated for their ability to provide protein and lipid-free plasma supernatants. The residual proteins, total cholesterol and triacylglycerols were assayed in the supernatant on a Beckman Analyzer instrument. The free cholesterol and the neutral lipids were further analyzed by means of high-temperature GC analysis. The conditions for the deproteinizing step were optimized for minimal lipoprotein disruption. A substantial difference regarding contamination by the lipids was found if the plasma supernatant or the whole serum were treated with an alkyl chloroformate reagent. Three plasma sulfur amino acids and the aromatic ones were chosen as model compounds to evaluate compatibility of the precipitation methods with a subsequent methyl chloroformate-mediated derivatization and GC-MS analysis. The results of the total homocysteine assay matched well with that obtained using a commercial immunoassay. Precipitation with trichloroacetic acid has proven to be a method of choice for the analysis of the acido-basic analytes by GC-MS via chloroformates.

  2. Theoretical calculations of infrared absorption, vibrational circular dichroism, and two-dimensional vibrational spectra of acetylproline in liquids water and chloroform.

    PubMed

    Hahn, Seungsoo; Lee, Hochan; Cho, Minhaeng

    2004-07-22

    Infrared absorption, vibrational circular dichroism, and two-dimensional infrared pump-probe and photon echo spectra of acetylproline solutions are theoretically calculated and directly compared with experiments. In order to quantitatively determine interpeptide interaction-induced amide I mode frequency shifts, high-level quantum chemistry calculations were performed. The solvatochromic amide I mode frequency shift and fluctuation were taken into account by carrying out molecular dynamics simulations of acetylproline dissolved in liquids water and chloroform and by using the extrapolation method developed recently. We first studied correlation time scales of the two amide I vibrational frequency fluctuations, cross correlation between the two fluctuating local mode frequencies, ensemble averaged conformations of the acetylproline molecule in liquids water and chloroform. The corresponding conformations of the acetylproline in liquids water and chloroform are close to the ideal 3(10) helix and the C(7) structure, respectively. A few methods proposed to determine the angle between the two transition dipoles associated with the amide I vibrations were tested and their limitations are discussed.

  3. Reliable solution processed planar perovskite hybrid solar cells with large-area uniformity by chloroform soaking and spin rinsing induced surface precipitation

    SciTech Connect

    Chern, Yann-Cherng; Wu, Hung-Ruei; Chen, Yen-Chu; Horng, Sheng-Fu; Zan, Hsiao-Wen; Meng, Hsin-Fei

    2015-08-15

    A solvent soaking and rinsing method, in which the solvent was allowed to soak all over the surface followed by a spinning for solvent draining, was found to produce perovskite layers with high uniformity on a centimeter scale and with much improved reliability. Besides the enhanced crystallinity and surface morphology due to the rinsing induced surface precipitation that constrains the grain growth underneath in the precursor films, large-area uniformity with film thickness determined exclusively by the rotational speed of rinsing spinning for solvent draining was observed. With chloroform as rinsing solvent, highly uniform and mirror-like perovskite layers of area as large as 8 cm × 8 cm were produced and highly uniform planar perovskite solar cells with power conversion efficiency of 10.6 ± 0.2% as well as much prolonged lifetime were obtained. The high uniformity and reliability observed with this solvent soaking and rinsing method were ascribed to the low viscosity of chloroform as well as its feasibility of mixing with the solvent used in the precursor solution. Moreover, since the surface precipitation forms before the solvent draining, this solvent soaking and rinsing method may be adapted to spinless process and be compatible with large-area and continuous production. With the large-area uniformity and reliability for the resultant perovskite layers, this chloroform soaking and rinsing approach may thus be promising for the mass production and commercialization of large-area perovskite solar cells.

  4. Relative source allocation of TDI to drinking water for derivation of a criterion for chloroform: a Monte-Carlo and multi-exposure assessment.

    PubMed

    Niizuma, Shun; Matsui, Yoshihiko; Ohno, Koichi; Itoh, Sadahiko; Matsushita, Taku; Shirasaki, Nobutaka

    2013-10-01

    Drinking water quality standard (DWQS) criteria for chemicals for which there is a threshold for toxicity are derived by allocating a fraction of tolerable daily intake (TDI) to exposure from drinking water. We conducted physiologically based pharmacokinetic model simulations for chloroform and have proposed an equation for total oral-equivalent potential intake via three routes (oral ingestion, inhalation, and dermal exposures), the biologically effective doses of which were converted to oral-equivalent potential intakes. The probability distributions of total oral-equivalent potential intake in Japanese people were estimated by Monte Carlo simulations. Even when the chloroform concentration in drinking water equaled the current DWQS criterion, there was sufficient margin between the intake and the TDI: the probability that the intake exceeded TDI was below 0.1%. If a criterion that the 95th percentile estimate equals the TDI is regarded as both providing protection to highly exposed persons and leaving a reasonable margin of exposure relative to the TDI, then the chloroform drinking water criterion could be a concentration of 0.11mg/L. This implies a daily intake equal to 34% of the TDI allocated to the oral intake (2L/d) of drinking water for typical adults. For the highly exposed persons, inhalation exposure via evaporation from water contributed 53% of the total intake, whereas dermal absorption contributed only 3%.

  5. Apoptotic effects of Physalis minima L. chloroform extract in human breast carcinoma T-47D cells mediated by c-myc-, p53-, and caspase-3-dependent pathways.

    PubMed

    Ooi, Kheng Leong; Tengku Muhammad, Tengku Sifzizul; Lim, Chui Hun; Sulaiman, Shaida Fariza

    2010-03-01

    The chloroform extract of Physalis minima produced a significant growth inhibition against human T-47D breast carcinoma cells as compared with other extracts with an EC(50) value of 3.8 microg/mL. An analysis of cell death mechanisms indicated that the extract elicited an apoptotic cell death. mRNA expression analysis revealed the coregulation of apoptotic genes, that is, c-myc , p53, and caspase-3. The c-myc was significantly induced by the chloroform extract at the earlier phase of treatment, followed by p53 and caspase-3. Biochemical assay and ultrastructural observation displayed typical apoptotic features in the treated cells, including DNA fragmentation, blebbing and convolution of cell membrane, clumping and margination of chromatin, and production of membrane-bound apoptotic bodies. The presence of different stages of apoptotic cell death and phosphatidylserine externalization were further reconfirmed by annexin V and propidium iodide staining. Thus, the results from this study strongly suggest that the chloroform extract of P. minima induced apoptotic cell death via p53-, caspase-3-, and c-myc-dependent pathways.

  6. Anticancer Activity of Chloroform Extract and Sub-fractions of Nepeta deflersiana on Human Breast and Lung Cancer Cells: An In vitro Cytotoxicity Assessment

    PubMed Central

    Al-Oqail, Mai M.; Al-Sheddi, Ebtesam S.; Siddiqui, Maqsood A.; Musarrat, Javed; Al-Khedhairy, Abdulaziz A.; Farshori, Nida N.

    2015-01-01

    Background: Cancer is one of the major causes of death worldwide. The plant-derived natural products have received considerable attention in recent years due to their diverse pharmacological properties including anticancer effects. Nepeta deflersiana (ND) is used in the folk medicine as antiseptic, carminative, antimicrobial, antioxidant, and for treating rheumatic disorders. However, the anticancer activity of ND chloroform extract has not been explored so far. Objectives: The present study was aimed to investigate the anticancer activities of chloroform Nepeta deflersiana extract and various sub-fractions (ND-1–ND-15) of ND against human breast cancer cells (MCF-7) and human lung cancer cells (A-549). Materials and Methods: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and neutral red uptake assays, and cellular morphological alterations using phase contrast light microscope were studied. Cells were exposed with 10–1000 μg/ml of sub-fractions of ND for 24 h. Results: Results showed that selected sub-fractions of the chloroform extract significantly reduced the cell viability of MCF-7 and A-549 cells, and altered the cellular morphology in a concentration-dependent manner. Among the sub-fractions, ND-10 fraction showed relatively higher cytotoxicity compared to other fractions whereas, ND-1 did not cause any cytotoxicity even at higher concentrations. The A-549 cells were found to be more sensitive to growth inhibition by all the extracts as compared to the MCF-7 cells. Conclusion: The present study provides preliminary screening of anticancer activities of chloroform extract and sub-fractions of ND, which can be further used for the development of a potential therapeutic anticancer agent. SUMMARY Nepeta deflersiana extract exhibit cytotoxicity and altered the cellular morphology. Sub-fractions of the chloroform extract of Nepeta deflersiana reduced the cell viability of MCF-7 and A-549 cells. Among the sub-fractions, ND-10 fraction showed

  7. Sources and occurrence of chloroform and other trihalomethanes in drinking-water supply wells in the United States, 1986-2001

    USGS Publications Warehouse

    Ivahnenko, Tamara; Zogorski, J.S.

    2006-01-01

    Chloroform and three other trihalomethanes (THMs)--bromodichloromethane, dibromochloromethane, and bromoform--are disinfection by-products commonly produced during the chlorination of water and wastewater. Samples of untreated ground water from drinking-water supply wells (1,096 public and 2,400 domestic wells) were analyzed for THMs and other volatile organic compounds (VOCs) during 1986-2001, or compiled, as part of the U.S. Geological Survey's National Water-Quality Assessment Program. This report provides a summary of potential sources of THMs and of the occurrence and geographical distribution of THMs in samples from public and domestic wells. Evidence for an anthropogenic source of THMs and implications for future research also are presented. Potential sources of THMs to both public and domestic wells include the discharge of chlorinated drinking water and wastewater that may be intentional or inadvertent. Intentional discharge includes the use of municipally supplied chlorinated water to irrigate lawns, golf courses, parks, gardens, and other areas; the use of septic systems; or the regulated discharge of chlorinated wastewater to surface waters or ground-water recharge facilities. Inadvertent discharge includes leakage of chlorinated water from swimming pools, spas, or distribution systems for drinking water or wastewater sewers. Statistical analyses indicate that population density, the percentage of urban land, and the number of Resource Conservation and Recovery Act hazardous-waste facilities near sampled wells are significantly associated with the probability of detection of chloroform, especially for public wells. Domestic wells may have several other sources of THMs, including the practice of well disinfection through shock chlorination, laundry wastewater containing bleach, and septic system effluent. Chloroform was the most frequently detected VOC in samples from drinking-water supply wells (public and domestic wells) in the United States. Although

  8. Measurement and deduction of emissions of trichloroethene, tetrachloroethene, and trichloromethane (chloroform) in the northeastern United States and southeastern Canada

    NASA Astrophysics Data System (ADS)

    Kleiman, G.; Prinn, R. G.

    2000-12-01

    We present optimal estimates of the emission patterns for trichloroethene (TCE, CHCl = CCl2), tetrachloroethene (perchloroethylene, CCl2 = CCl2), and trichloromethane (chloroform, CHCl3) utilizing hourly gas chromatographie measurements at Nahant, Massachusetts (approximately 10 km northeast of Boston). Our analysis combines the measurements with back trajectory information obtained from the Hybrid Single-Particle Lagrangian Integrated Trajectory (HYSPLIT-4) model (National Oceanic and Atmospheric Administration Air Resources Laboratory, Silver Spring, Maryland). Using a Kaiman filter inverse method and an analytical solution of the continuity equation to estimate the effects of eddy diffusion, we calculate the surface emissions on a 1°×1° grid for the selected species necessary to optimally match the observations. These emissions are compared with the estimates determined by the Reactive Chlorine Emissions Inventory (RCEI) working group of the International Global Atmospheric Chemistry Program Global Emissions Inventory Activity (GEIA). The new emissions scenarios computed here provide an observation-based assessment for comparison with the RCEI emissions inventories for the northeastern United States and southeastern Canada. Results indicate that the RCEI estimates of the anthropogenic emissions of these chemicals in the geographical domain studied differ from our optimal estimates but generally lie within the estimated error of these optimal estimates. Results are sensitive to the assumed vertical distributions and hence to the assumed vertical mixing rates. The current accuracy achievable through this observation-based technique (±45%) is limited in large part by the uncertainty in the vertical distribution of these compounds in the troposphere over highly emitting regions. Within this accuracy the optimal estimates of emissions presented here indicate that the emissions for many grid cells do not need to be corrected significantly from the 1990 estimates

  9. Solvent effects on the absorption and emission spectra of novel (E)-4-((4-(heptyloxy)phenyl)diazenyl)benzyl (((9H-fluoren-9-yl)methoxy)carbonyl)-D-alaninate (Fmoc-al-az): Determination of dipole moment by experimental and theoretical study

    NASA Astrophysics Data System (ADS)

    Y, Tej Varma; Agarwal, Devesh S.; Sarmah, Amrit; Joshi, Lata; Sakhuja, Rajeev; Pant, Debi D.

    2017-02-01

    Amino acid appended azobenzene hybrid has been synthesized (Fmoc-al-az) and its electronic absorbance and fluorescence spectra were recorded at room temperature in a series of polar and non-polar solvents. The ground state and excited state dipole moments were calculated using solvatochromic shift method. A DFT based study was also performed using- Gaussian09 program package. We have observed that the absorption spectra don't show sensitive behavior to the change in the polarity of the solvent, whereas a bathochromic shift was observed in the fluorescence spectra as we moved from non-polar to polar solvents indicating a π→π* transition. It was observed that the dipole moment in the excited state is much higher than the ground state, due to the amino acid containing hydrogen bond acceptor (Osbnd Cdbnd O) and hydrogen bond donor (sbnd NH) in Fmoc-al-az increasing the reorientation tendency of the solvent molecule around the dye therefore enhancing the stabilization of the excited state which is attributed to the high polarity of the excited states. DFT level electronic structure calculations are also performed for a better molecular level understanding of the experimental observations. We obtained a good correlation between the theoretical studies and experimental results.

  10. Chloroform extract of alfalfa (Medicago sativa) inhibits lipopolysaccharide-induced inflammation by downregulating ERK/NF-κB signaling and cytokine production.

    PubMed

    Choi, Ki-Choon; Hwang, Jung-Min; Bang, Sung-Jun; Kim, Beom-Tae; Kim, Dong-Hern; Chae, Minseon; Lee, Seung-Ah; Choi, Gi Jun; Kim, Da Hye; Lee, Jeong-Chae

    2013-05-01

    Alfalfa (Medicago sativa L.) is commonly used as a traditional medicine and functional food. This study investigated the anti-inflammatory potential of alfalfa and the mechanisms involved. The chloroform extract of alfalfa aerial parts inhibited lipopolysaccharide (LPS)-stimulated immune responses more than ether, butanol, or water soluble extracts. Treatment with 1 μg/mL LPS increased nitrite concentrations to 44.3 μM in RAW267.4 macrophages, but it was reduced to 10.6 μM by adding 100 μg/mL chloroform extract. LPS treatment also increased the concentrations of tumor necrosis factor-α, interleukin (IL)-6, and IL-1β to 41.3, 11.6, and 0.78 ng/mL in culture supernatants of the cells, but these cytokine levels decreased to 12.5, 3.1, and 0.19 ng/mL, respectively, by pretreating with 100 μg/mL of the extract. ICR mice injected with LPS (30 mg/kg body weight) alone showed a 0% survival rate after 48 h of the injection, but 48-h survival of the mice increased to 60% after oral administration of the extract. Subfractions of the chloroform extract markedly suppressed LPS-mediated activation of the extracellular signal-regulated kinase and nuclear factor kappa-B. Cinnamic acid derivatives and fatty acids were found to be active constituents of the extract. This research demonstrated that alfalfa aerial parts exert anti-inflammatory activity and may be useful as a functional food for the prevention of inflammatory disorders.

  11. Screening of Ethanol, Petroleum Ether and Chloroform Extracts of Medicinal Plants, Lawsonia inermis L. and Mimosa pudica L. for Antibacterial Activity

    PubMed Central

    Akter, A.; Neela, F. A.; Khan, M. S. I.; Islam, M. S.; Alam, M. F.

    2010-01-01

    Organic extracts (ethanol, petroleum ether and chloroform) of two medicinal plants Lawsonia inermis L. and Mimosa pudica L. were proven for antibacterial properties against 15 Gram-positive and Gram-negative human pathogenic bacteria. Among the three types of extracts tested, ethanol extract was found to possess maximum antibacterial activity. The diameter of the zone of inhibition of bacterial growth showed that Gram-negative bacteria are more sensitive than Gram-positive bacteria to plant extracts. Between the two plants species studied, Lawsonia inermis extract showed more antibacterial activity compared to Mimosa pudica extract. PMID:21188055

  12. Studies on a complex mechanism for the activation of plasminogen by kaolin and by chloroform: the participation of Hageman factor and additional cofactors

    PubMed Central

    Ogston, Derek; Ogston, C. Marie; Ratnoff, Oscar D.; Forbes, Charles D.

    1969-01-01

    As demonstrated by others, fibrinolytic activity was generated in diluted, acidified normal plasma exposed to kaolin, a process requiring Hageman factor (Factor XII). Generation was impaired by adsorbing plasma with glass or similar agents under conditions which did not deplete its content of Hageman factor or plasminogen. The defect could be repaired by addition of a noneuglobulin fraction of plasma or an agent or agents eluted from diatomaceous earth which had been exposed to normal plasma. The restorative agent, tentatively called Hageman factor-cofactor, was partially purified by chromatography and had an apparent molecular weight of approximately 165,000. It could be distinguished from plasma thromboplastin antecedent (Factor XI) and plasma kallikrein, other substrates of Hageman factor, and from the streptokinase-activated pro-activator of plasminogen. Evidence is presented that an additional component may be needed for the generation of fibrinolytic activity in mixtures containing Hageman factor, HF-cofactor, and plasminogen. The long-recognized generation of plasmin activity in chloroform-treated euglobulin fractions of plasma was found to be dependent upon the presence of Hageman factor. Whether chloroform activation of plasminogen requires Hageman factor-cofactor was not determined, but glass-adsorbed plasma, containing Hageman factor and plasminogen, did not generate appreciable fibrinolytic or caseinolytic activity. These studies emphasize the complex nature of the mechanisms which lead to the generation of plasmin in human plasma. PMID:4241814

  13. A study of the sources and sinks of methane and methyl chloroform using a global three-dimensional Lagrangian tropospheric tracer transport model

    NASA Technical Reports Server (NTRS)

    Taylor, John A.; Brasseur, G. P.; Zimmerman, P. R.; Cicerone, R. J.

    1991-01-01

    Sources and sinks of methane and methyl chloroform are investigated using a global three-dimensional Lagrangian tropospheric tracer transport model with parameterized hydroxyl and temperature fields. Using the hydroxyl radical field calibrated to the methyl chloroform observations, the globally averaged release of methane and its spatial and temporal distribution were investigated. Two source function models of the spatial and temporal distribution of the flux of methane to the atmosphere were developed. The first model was based on the assumption that methane is emitted as a proportion of net primary productivity (NPP). The second model identified source regions for methane from rice paddies, wetlands, enteric fermentation, termites, and biomass burning based on high-resolution land use data. The most significant difference between the two models were predictions of methane fluxes over China and South East Asia, the location of most of the world's rice paddies, indicating that either the assumption that a uniform fraction of NPP is converted to methane is not valid for rice paddies, or that NPP is underestimated for rice paddies, or that present methane emission estimates from rice paddies are too high.

  14. 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.

  15. Determination of CD- and CH-bond energies in d-chloroform and fluoroform from vibrational spectra using the Morse potential

    SciTech Connect

    Apostolova, E.S.; Tulub, A.V.

    1995-04-01

    Based on the six-parametric model energy operator containing Morse potentials, the spectrum of valence vibrations of d-chloroform and fluoroform is calculated to the vibrational quantum number v= 2 with an accuracy of 5 and 6.3 cm{sup -1}, respectively. Upon HCCl{sub 3}{r_arrow} DCC1{sub 3} isotopic substitution, the CH-bond dissociation energy increases by 25.2 kJ/mol, which agrees with experimental estimates. The CH-bond dissociation energy 444.2 kJ/mol in the HCF{sub 3} molecule is quite close to its experimental value of 446.4 {+-} 4.2 kJ/mol. The influence of the high-frequency laser field on the vibrational spectrum of HCC{sub 3} is considered using averaging over the vibration period.

  16. Probabilistic human health risk assessment for quarterly exposure to high chloroform concentrations in drinking-water distribution network of the Province of Quebec, Canada.

    PubMed

    Buteau, Stephane; Valcke, Mathieu

    2010-01-01

    Because quarterly concentrations of total trihalomethanes (THM) exceeding the 80 μg/L guideline are often tolerated by the public health authorities of the Province of Quebec (Canada), this study examined whether quarterly episodes of high concentrations of THM may pose a risk to the health of its population. Using Monte Carlo simulations, a probabilistic risk assessment was performed for infants (0-<6 mo), toddlers (6 mo-<5 yr) and adults (≥20 yr). Multiroute exposure including ingestion of drinking water as well as inhalation and dermal exposure while showering or bathing was considered. The resulting absorbed doses were compared to short-term reference values for chloroform, used as surrogate for THM, by calculating risk quotients (RQ). On the basis of THM concentrations values in Quebec's drinking water distribution systems during the months of July to October and exceeding the guideline value (>80 μg/L), the 95th percentile value of RQ were 0.65, 0.46, and 0.24 for infants, toddlers, and adults, respectively. Back-calculation allowed determining that a chloroform concentration of 330 μg/L would result in RQ ≤ 1 for 99% of infants, the subgroup considered the most susceptible among the general population. Overall, this study showed that episodes of high THM concentration encountered in Quebec drinking-water distribution network need not be considered as an immediate health concern for the general population. However, these results should not be interpreted as an authorization to exceed the 80 μg/L standard but rather as a risk management tool for public health authorities.

  17. 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.

  18. Application of the Novel 5-chloro-2,2,3,3,4,4,5,5-octafluoro-1-pentyl Chloroformate Derivatizing Agent for the Direct Determination of Highly Polar Water Disinfection Byproducts

    EPA Science Inventory

    A novel derivatizing agent, 5-chloro-2,2,3,3,4,4,5,5-octafluoropentyl chloroformate (ClOFPCF), was synthesized and tested as a reagent for direct water derivatization of highly polar and hydrophilic analytes. Its analytical performance satisfactorily compared to a perfluorinated ...

  19. Effect of chloroform and aqueous basic fraction of ethanolic extract from Apium graveolens L. in experimentally-induced hyperlipidemia in rats.

    PubMed

    Iyer, Deepa; Patil, U K

    2011-09-27

    The present study was undertaken to explore the antihyperlipidemic effect of ethanolic extract of seeds of Apium graveolens L. and its chloroform and aqueous basic fraction in olive oil induced hyperlipidemic rats. The antihyperlipidemic activity of Apium graveolens was compared with a standard drug Atrovastatin (50mg/kg). The study involved phytochemical screening and chromatographic studies of extract and fractions. The ethanolic extract and its fractions were administered orally at doses of 200 and 400 mg/kg body weight in rats. Olive oil (5ml/kg oral dose) was administered 30 min after treatment. Blood was collected by ocular puncture 2 and 4 h after olive oil treatment and centrifuged at 3000 rpm for 15-20 min. Serum samples were further subjected to biochemical analysis. The study dose dependently inhibited the total cholesterol (TC) triglycerides (TG), low density lipoproteins (LDL) level, and significantly increased high density lipoprotein (HDL) level. Phytochemical screening revealed the presence of terpenoid, tannin, alkaloid, glycoside, flavanoid and sterols. UV λmax was found to be 206 nm with a melting point of 137-138°C for the isolated component. The antihyperlipidemic effect was evaluated in olive oil loaded rats. Acute treatment caused stimulatory effect on HDL level and inhibition in TC and TG elevation induced by olive oil.

  20. A spectrophotometric and thermodynamic study of the sitting-atop complex formation from reaction between free base meso-tetraarylporphyrins and zirconyl nitrate in chloroform solution

    NASA Astrophysics Data System (ADS)

    Dehghani, Hossein; Mansournia, Mohammad Reza

    2009-10-01

    The sitting-atop complexation of meso-tetraarylporphyrins and its para-substituted derivatives (H 2t(4-X)pp, X:H, Br, Cl, CH(CH 3) 2, OCH 3, CH 3), as electron donors, with zirconyl, as an electron acceptor, have been investigated spectrophotometrically in chloroform. The mole ratio studies based on physicochemical techniques were employed clearly and revealed the formation of 1:1 sitting-atop complexes which was confirmed by UV-vis, 1H NMR and IR spectroscopic data. The value of the formation constant was estimated for each complex using a nonlinear optimization of the complex absorbance vs. mole ratio data by package KINFIT. The results showed that the stability of these complexes decreases with the temperature enhancement. Thermodynamic parameters, Δ G°, Δ H° and Δ S°, of the SAT complexes have been determined from the temperature dependence of formation constants by Van't Hoff equation. Also, the influence of the substituents of the aryl rings in H 2t(4-X)pp on the stability of the SAT complexes is discussed.

  1. Molecular mechanisms involved in the protective effect of the chloroform extract of Selaginella lepidophylla (Hook. et Grev.) Spring in a lithiasic rat model.

    PubMed

    Mirian, Estévez-Carmona María; Juanita, Narvaéz-Morales; Christophe, Barbier Olivier; Estela, Meléndez-Camargo María

    2013-06-01

    Urolithiasis is a multifaceted process, progressing from urine supersaturation to the formation of mature renal calculi. Calcium oxalate, the main component of kidney stones, has toxicological effects on renal epithelial cells. Some medicinal plants have shown pharmacological effects against renal lithiasis, such as Selaginella lepidophylla (Hook. et Grev) Spring, a plant empirically used in Mexico for its diuretic and antilithiasic activity. The plant was identified and ground, and a chloroform extract (CE) was obtained. Urolithiasis was induced in Wistar female rats by administration of ethylene glycol and ammonium chloride for 21 days. Urolithiasis rats were treated with the CE (50 mg/kg) for 21 days. Osmolality, creatinine, sodium and potassium concentrations were measured in blood and urine. Glomerular filtration rate (GFR), and electrolytic and water balances were calculated. Urinary oxalic acid concentration was measured. Apoptosis, lipoperoxidation, ROS and p-amino hippuric acid were determined in cortical tissue. Urolithiasis rats showed a decrease of urinary flow, GFR, electrolytic balance, renal tubular secretion and ATP concentration and increase of urinary oxalic acid, lipoperoxidation, oxidative stress and apoptosis in cortical tissue. After treatment with the CE, urinary flow rate, GFR and renal tubular secretion levels were recovered; on the other hand, serum creatinine and urinary oxalic acid decreased on day 21. CE of Selaginella lepidophylla prevented the damage caused by lithiasic process by improving the active secretion in the proximal tubules, counteracting the ROS and lipoperoxidation effects by oxalate and decreased the OAT3 expression on kidney.

  2. Optimization of a gas chromatography-mass spectrometry method with methyl chloroformate derivatization for quantification of amino acids in plant tissue.

    PubMed

    Vancompernolle, Bram; Croes, Kim; Angenon, Geert

    2016-04-01

    Rapid, easy and reliable quantification of amino acids is crucial in research on plant amino acid metabolism and nutritional improvement of crops via enrichment of essential amino acids. A recently reported analysis method, based on solid phase extraction (SPE), derivatization with methyl chloroformate and gas chromatography-mass spectrometry was optimized and tested on three-week-old Arabidopsis thaliana leaf tissues. Optimization of the SPE cleanup yielded recovery rates of minimum 95% for all amino acids (except arginine). Variations in accuracy and precision did not exceed 12.5%, except for cysteine, histidine and tryptophane, which were excluded from analysis. Quantification of overlapping peaks for isoleucine/threonine and proline/asparagine was possible by selection of two specific fragment ions for each amino acid. Of the 16 selected amino acids, 14 were quantified successfully in at least 75% of the samples, while methionine and tyrosine were only quantifiable in 6% and 42%, respectively. A case study on the aspartate super pathway confirmed the applicability of the optimized method on wild type and genetically modified plants: external supplementation of methionine or lysine yielded a 146-fold or 27-fold increase in the respective absolute amino acid levels compared with the control treatment. Induced expression of dhdps-r1 (a mutated lysine biosynthesis gene encoding a feedback insensitive enzyme) caused an 83-fold increase in absolute lysine levels.

  3. Continuous water infusion enhances atmospheric pressure chemical ionization of methyl chloroformate derivatives in gas chromatography coupled to time-of-flight mass spectrometry-based metabolomics.

    PubMed

    Wachsmuth, Christian J; Dettmer, Katja; Lang, Sven A; Mycielska, Maria E; Oefner, Peter J

    2014-09-16

    The effects of continuous water infusion on efficiency and repeatability of atmospheric pressure chemical ionization of both methyl chloroformate (MCF) and methoxime-trimethylsilyl (MO-TMS) derivatives of metabolites were evaluated using gas chromatography-time-of-flight mass spectrometry. Water infusion at a flow-rate of 0.4 mL/h yielded not only an average 16.6-fold increase in intensity of the quasimolecular ion for 20 MCF-derivatized metabolite standards through suppression of in-source fragmentation but also the most repeatable peak area integrals. The impact of water infusion was the greatest for dicarboxylic acids and the least for (hetero-) aromatic compounds. Water infusion also improved the ability to detect reliably fold changes as small as 1.33-fold for the same 20 MCF-derivatized metabolite standards spiked into a human serum extract. On the other hand, MO-TMS derivatives were not significantly affected by water infusion, neither in their fragmentation patterns nor with regard to the detection of differentially regulated compounds. As a proof of principle, we applied MCF derivatization and GC-APCI-TOFMS to the detection of changes in abundance of metabolites in pancreatic cancer cells upon treatment with 17-DMAG. Water infusion increased not only the number of metabolites identified via their quasimolecular ion but also the reproducibility of peak areas, thereby almost doubling the number of significantly regulated metabolites (false discovery rate < 0.05) to a total of 23.

  4. Cytotoxic Activities of Physalis minima L. Chloroform Extract on Human Lung Adenocarcinoma NCI-H23 Cell Lines by Induction of Apoptosis

    PubMed Central

    Leong, Ooi Kheng; Muhammad, Tengku Sifzizul Tengku; Sulaiman, Shaida Fariza

    2011-01-01

    Physalis minima L. is reputed for having anticancer property. In this study, the chloroform extract of this plant exhibited remarkable cytotoxic activities on NCI-H23 (human lung adenocarcinoma) cell line at dose- and time-dependent manners (after 24, 48 and 72 h of incubation). Analysis of cell-death mechanism demonstrated that the extract exerted apoptotic programed cell death in NCI-H23 cells with typical DNA fragmentation, which is a biochemical hallmark of apoptosis. Morphological observation using transmission electron microscope (TEM) also displayed apoptotic characteristics in the treated cells, including clumping and margination of chromatins, followed by convolution of the nuclear and budding of the cells to produce membrane-bound apoptotic bodies. Different stages of apoptotic programed cell death as well as phosphatidylserine externalization were confirmed using annexin V and propidium iodide staining. Furthermore, acute exposure to the extract produced a significant regulation of c-myc, caspase-3 and p53 mRNA expression in this cell line. Due to its apoptotic effect on NCI-H23 cells, it is strongly suggested that the extract could be further developed as an anticancer drug. PMID:19541726

  5. Suppressive activity of the chloroform extract of Tripterygium wilfordii Hook f on effector T cell activation during Hymenolepis nana infection in mice.

    PubMed

    Asano, K; Matsuishi, J; Yu, Y; Nemoto, K; Nakazawa, M; Kasahara, T; Hisamitsu, T

    1998-01-01

    The chloroform extract of Tripterygium wilfordii Hook f (TWH extract) administered into mice daily at doses of 80.0 to 200.0 micrograms/kg (but not 40.0 micrograms/kg) caused suppression of protective immunity to Hymenolepis nana when the extract was injected subcutaneously during the induction phase of protective immunity. Daily administration of 200.0 micrograms/kg TWH extract, during the course of larval development from challenge, also suppressed protective immunity. Inhibition of protective immunity was only observed in mice that received TWH extract for 6 days at a daily dose of 200.0 micrograms/kg and were challenged 24 h after the final injection. TWH extract did not inhibit formation of effector cells that mediate delayed type hypersensitivity (DTH) to H. nana egg antigen when the extract was administered subcutaneously at a dose of 200.0 micrograms/kg/day for 5 days before cell preparation. However, TWH extract did inhibit DTH effector cell activation when cells prepared from infected, PBS-injected mice were transferred into 200.0 micrograms/kg TWH extract-treated recipient mice. These results strongly indicate that TWH extract cannot inhibit the generation of effector cells but will suppress their function in vivo.

  6. C-H⋯O Hydrogen Bonded Complexes Between Chloroform and Cyclic Ketones: Correlation of Spectral Shifts and Complex Stability with Ring Size

    NASA Astrophysics Data System (ADS)

    Mukhopadhyay, Anamika; Chakraborty, Tapas

    2010-06-01

    Stable C-H⋯O hydrogen bonded complexes between choloroform and three small cyclic ketones (cyclohexanone, cyclopentanone, cyclobutanone) are identified by use of FTIR spectroscopy in CCl4 solution at room temperature. The C-H stretching fundamental of chloroform (νC-H) in the said three complexes exhibits blue shifting with enhancement in νC-H transition intensity. However, the red shifts of the νC=O bands of the cyclic ketones in the complexes show no apparent correlation with the corresponding blue shifts of νC-H. The spectral analysis reveals that the stability of the complexes decreases with the ring size of the cyclic ketones. Electronic structure calculation at DFT/B3LYP/6-311++G(d,p) and MP2/6-31+G(d) levels predict that the complex binding energies are correlated with the dipole moment, proton affinity, and n(O)→σ∗(C-H) hyperconjugative charge transfer ability of the cyclic ketones.

  7. C-H⋯O Hydrogen Bonded Complexes Between Chloroform and Cyclic Ketones: Correlation of Spectral Shifts and Complex Stability with Ring Size

    NASA Astrophysics Data System (ADS)

    Mukhopadhyay, Anamika; Chakraborty, Tapas

    Stable C-H⋯O hydrogen bonded complexes between choloroform and three small cyclic ketones (cyclohexanone, cyclopentanone, cyclobutanone) are identified by use of FTIR spectroscopy in CCl4 solution at room temperature. The C-H stretching fundamental of chloroform (νC-H) in the said three complexes exhibits blue shifting with enhancement in νC-H transition intensity. However, the red shifts of the νC=O bands of the cyclic ketones in the complexes show no apparent correlation with the corresponding blue shifts of νC-H. The spectral analysis reveals that the stability of the complexes decreases with the ring size of the cyclic ketones. Electronic structure calculation at DFT/B3LYP/6-311++G(d,p) and MP2/6-31+G(d) levels predict that the complex binding energies are correlated with the dipole moment, proton affinity, and n(O)→σ∗(C-H) hyperconjugative charge transfer ability of the cyclic ketones.

  8. A study of the sources and sinks of methane and methyl chloroform using a global three-dimensional Lagrangian tropospheric tracer transport model

    SciTech Connect

    Taylor, J.A. National Center for Atmospheric Research, Boulder, CO ); Brasseur, G.P.; Zimmerman, P.R.; Cicerone, R.J. )

    1991-02-20

    Using the hydroxyl radical field calibrated to the methyl chloroform observations, the globally averaged release of methane and its spatial and temporal distribution were investigated. Two source function models of the spatial and temporal distribution of the flux of methane to the atmosphere were developed. The first model was based on the assumption that methane is emitted as a proportion of net primary productivity (NPP). With the average hydroxyl radical concentration fixed, the methane source term was computed as {approximately}623 Tg CH{sub 4}, giving an atmospheric lifetime for methane {approximately}8.3 years. The second model identified source regions for methane from rice paddies, wetlands, enteric fermentation, termites, and biomass burning based on high-resolution land use data. This methane source distribution resulted in an estimate of the global total methane source of {approximately}611 Tg CH{sub 4}, giving an atmospheric lifetime for methane {approximately}8.5 years. The most significant difference between the two models were predictions of methane fluxes over China and South East Asia, the location of most of the world's rice paddies. Using a recent measurement of the reaction rate of hydroxyl radical and methane leads to estimates of the global total methane source for SF1 of {approximately}524 Tg CH{sub 4} giving an atmospheric lifetime of {approximately}10.0 years and for SF2{approximately}514 Tg CH{sub 4} yielding a lifetime of {approximately}10.2 years.

  9. Chloroform Extract of Rasagenthi Mezhugu, a Siddha Formulation, as an Evidence-Based Complementary and Alternative Medicine for HPV-Positive Cervical Cancers

    PubMed Central

    Riyasdeen, Anvarbatcha; Periasamy, Vaiyapuri S.; Paul, Preethy; Alshatwi, Ali A.; Akbarsha, Mohammad A.

    2012-01-01

    Rasagenthi Mezhugu (RGM) is a herbomineral formulation in the Siddha system of traditional medicine and is prescribed in the southern parts of India as a remedy for all kinds of cancers. However, scientific evidence for its therapeutic efficacy in cervical cancer is lacking, and it contains heavy metals. To overcome these limitations, RGM was extracted, and the fractions were tested on HPV-positive cervical cancer cells, ME-180 and SiHa. The extracts, free from the toxic heavy metals, affected the viability of both the cells. The chloroform fraction (cRGM) induced DNA damage and apoptosis. Mitochondria-mediated apoptosis was indicated. Though both the cells responded to the treatment, ME-180 was more responsive. Thus, this study brings up scientific evidence for the efficacy of RGM against the HPV-mediated cervical cancer cells and, if the toxic heavy metals are the limitation in its use, cRGM would be a suitable candidate as evidence-based complementary and alternative medicine for HPV-positive cervical cancers. PMID:22114617

  10. C-H···Y hydrogen bonds in the complexes of p-cresol and p-cyanophenol with fluoroform and chloroform.

    PubMed

    Shirhatti, Pranav R; Maity, Dilip K; Wategaonkar, Sanjay

    2013-03-21

    In this work, we present spectroscopic investigations of hydrogen bonded complexes of CHF3 and CHCl3 with p-cresol and p-cyanophenol. The systems were chosen as the potential candidates bound by C-H···Y type hydrogen bonds that are known to exhibit unconventional blue shifts in the C-H stretching frequency. The two phenol derivatives chosen offer multiple hydrogen bonding acceptor sites. They also differ from each other in regard to the electron-donating/withdrawing ability of the para substituents which could dictate the global minimum structure in each case. The complexes were formed using the supersonic jet expansion method and were investigated using a variant of the IR-UV double resonance technique, namely fluorescence depletion IR (FDIR) spectroscopy. It was found that in the case of p-cresol the complexes were C-H···π bound in which the C-H stretch was blue-shifted. In the case of p-cyanophenol the complexes were C-H···N bound. In its fluoroform complex the C-H frequency was blue-shifted by 27 cm(-1), whereas the chloroform complex gave an example of zero-shifted hydrogen bond. The ab initio computational studies indicated that for the CHCl3 complexes it is necessary to optimize the structures on the BSSE-corrected PES using the counterpoise method to correctly predict the magnitudes of the C-H frequency shift.

  11. Integrative analysis of genomics and proteomics data on clinical breast cancer tissue specimens extracted with acid guanidinium thiocyanate-phenol-chloroform.

    PubMed

    Braakman, René B H; Bezstarosti, Karel; Sieuwerts, Anieta M; de Weerd, Vanja; van Galen, Anne M; Stingl, Christoph; Luider, Theo M; Timmermans, Mieke A M; Smid, Marcel; Martens, John W M; Foekens, John A; Demmers, Jeroen A A; Umar, Arzu

    2015-03-06

    Acid guanidinium thiocyanate, phenol, and chloroform extraction (AGPC) is a commonly used procedure to extract RNA from fresh frozen tissues and cell lines. In addition, DNA and proteins can be recovered, which makes AGPC an attractive source for integrative analysis on tissues of which little material is available, such as clinical specimens. Despite this potential, AGPC has only scarcely been used for proteomic analysis, mainly due to difficulties in extracting proteins. We have used a quantitative mass spectrometry method to show that proteins can readily be recovered from AGPC extracted tissues with high recovery and repeatability, which allows this method to be used for global proteomic profiling. Protein expression data for a selected number of clinically relevant markers, of which transcript and protein levels are known to be correlated, were in agreement with genomic and transcriptomic data obtained from the same AGPC lysate. Furthermore, global proteomic profiling successfully discriminated breast tumor tissues according to their clinical subtype. Lastly, a reference gene set of differentially expressed transcripts was strongly enriched in the differentially abundant proteins in our cohort. AGPC lysates are therefore well suited for comparative protein and integrative analyses.

  12. Spectrophotometric determination of boron in iron and steel with curcumin after separation by 2-ethyl-1,3-hexanediol-chloroform extraction.

    PubMed

    Donaldson, E M

    1981-11-01

    A simple and reliable method for determining approximately 0.0001% or more of total boron in iron and low- and high-alloy steels is described. After the sample is decomposed at <70 degrees in the presence of hydrogen peroxide and potassium hydrogen fluoride, the insoluble material is filtered off and ultimately fused with sodium carbonate. The cooled melt is dissolved in dilute hydrochloric acid and the solution is combined with the main solution. Fluoride is subsequently complexed with zirconium and boron is separated from iron and other elements by extraction as borate from 1M sulphuric acid medium into chloroform containing 2-ethyl-1,3-hexanediol. Boron, in a 1-ml portion of the extract, is ultimately determined spectrophotometrically at 550 nm in an ethanol medium, after formation of the curcumin rosocyanin complex in a glacial acetic acid-concentrated sulphuric acid medium. Acid-soluble and acid-insoluble boron can also be determined. Common ions, including large amounts of manganese, chromium, vanadium, titanium, molybdenum, tungsten, niobium and tantalum do not interfere.

  13. Determination of chromium in ores, rocks and related materials, iron, steel and non-ferrous alloys by atomic-absorption spectrophotometry after separation by tribenzylamine-chloroform extraction.

    PubMed

    Donaldson, E M

    1980-10-01

    A method for determining trace and moderate amounts of chromium in ores, concentrates, rocks, soils and clays is described. After fusion of the sample with sodium peroxide, the melt is dissolved in dilute sulphuric acid. The chromium(III) produced by the hydrogen peroxide formed is co-precipitated with hydrous ferric oxide. The precipitate is dissolved in 0.7M sulphuric acid and chromium oxidized to chromium(VI) with ceric ammonium sulphate. The chromium(VI) is extracted as an ion-association complex into chloroform containing tribenzylamine and stripped with ammoniacal hydrogen peroxide. This solution is acidified with perchloric acid and chromium determined by atomic-absorption spectrophotometry in an air-acetylene flame, at 357.9 nm. Barium and strontium do not interfere. The procedure is also applicable to iron and steel, and nickel-copper, aluminium and zirconium alloys. Up to 5 mg of manganese and 10 mg each of molybdenum and vanadium will not interfere. In the absence of vanadium, up to 10 mg of tungsten will not interfere. In the presence of 1 mg of vanadium, up to 1 mg of tungsten will not interfere.

  14. Dependence of solvent quality on the composition of copolymers: experiment and theory for solutions of P(MMA-ran-t-BMA) in toluene and in chloroform.

    PubMed

    Bercea, Maria; Wolf, Bernhard A

    2015-01-21

    The interaction of toluene with P(MMA-ran-t-BMA) and with the corresponding homopolymers was determined via vapor pressure measurements at 30, 50 and 70 °C. A unified thermodynamic approach served for the modeling of the results. It is capable of describing the behavior of the different solutions by means of two adjustable parameters, one representing the effective number of solvent segments and the other accounting for the interactions between the components. The solvent quality of toluene passes a maximum, a minimum and another maximum upon an increase of the t-BMA content of the copolymer at all temperatures. A similar behavior is discernable from vapor pressure data of chloroform published for the same copolymers. The heats of mixing for toluene depend strongly on temperature; at 50 °C they are all endothermal with the exception of PMMA, for which the value obtained from vapor pressures at 30 °C agrees very well with published caloric data.

  15. Micronuclei in Bone Marrow and Liver in relation to Hepatic Metabolism and Antioxidant Response due to Coexposure to Chloroform, Dichloromethane, and Toluene in the Rat Model

    PubMed Central

    Belmont-Díaz, Javier; López-Gordillo, Ana Paulina; Molina Garduño, Eunice; Serrano-García, Luis; Coballase-Urrutia, Elvia; Cárdenas-Rodríguez, Noemí; Arellano-Aguilar, Omar; Montero-Montoya, Regina D.

    2014-01-01

    Genotoxicity in cells may occur in different ways, direct interaction, production of electrophilic metabolites, and secondary genotoxicity via oxidative stress. Chloroform, dichloromethane, and toluene are primarily metabolized in liver by CYP2E1, producing reactive electrophilic metabolites, and may also produce oxidative stress via the uncoupled CYP2E1 catalytic cycle. Additionally, GSTT1 also participates in dichloromethane activation. Despite the oxidative metabolism of these compounds and the production of oxidative adducts, their genotoxicity in the bone marrow micronucleus test is unclear. The objective of this work was to analyze whether the oxidative metabolism induced by the coexposure to these compounds would account for increased micronucleus frequency. We used an approach including the analysis of phase I, phase II, and antioxidant enzymes, oxidative stress biomarkers, and micronuclei in bone marrow (MNPCE) and hepatocytes (MNHEP). Rats were administered different doses of an artificial mixture of CLF/DCM/TOL, under two regimes. After one administration MNPCE frequency increased in correlation with induced GSTT1 activity and no oxidative stress occurred. Conversely, after three-day treatments oxidative stress was observed, without genotoxicity. The effects observed indicate that MNPCE by the coexposure to these VOCs could be increased via inducing the activity of metabolism enzymes. PMID:24949447

  16. Spectrophotometric study of interaction of iodine with 4'-aminobenzo-15-crown-5 in chloroform, dichloromethane and 1,2-dichloroethane solutions.

    PubMed

    Hasani, Masoumeh; Akbari, Sakineh

    2007-11-01

    The formation of charge transfer complex between 4'-aminobenzo-15-crown-5 (AB15C5) and iodine is investigated spectrophotometrically in chloroform, dichloromethane (DCM) and 1,2-dichloroethane (DCE) solutions at 25 degrees C. The continuous variation method clearly revealed the formation of 1:1 charge transfer complex in solution. The observed time dependence of the charge transfer band and subsequent formation of I 3- in solution were related to the slow transformation of the initially formed 1:1 AB15C5.I2 outer complex to an inner electron donor-acceptor (EDA) complex, followed by fast reaction of the inner complex with iodine to form a triiodide ion. The pseudo-first-order rate constants for the transformation process were evaluated from the absorbance-time data and found to vary in the order of 1,2-DCE>DCM>CHCl3. The values of the formation constant, KDA, for each complex are evaluated from Benesi-Hilebrand equation. Stability of the resulting complex in three solvents was also found to vary in the order of 1,2-DCE>DCM>CHCl3.

  17. A spectrophotometric and thermodynamic study of the charge-transfer complexes of iodine with 2-aminomethyl-15-crown-5 in chloroform and 1,2-dichloroethane solutions.

    PubMed

    Hasani, Masoumeh; Akbari, Sakineh

    2007-05-01

    Interaction of 2-aminomethyl-15-crown-5 (AM15C5) with iodine has been investigated spectrophotometrically in chloroform and 1,2-dichloroethane (1,2-DCE) solutions. The observed time dependence of the charge-transfer band and subsequent formation of I(3)(-) in solution were related to the slow transformation of the initially formed 1:1 AM15C5.I(2) outer complex to an inner electron donor-acceptor (EDA) complex, followed by fast reaction of the inner complex with iodine to form a triiodide ion. The pseudo-first-order rate constants were evaluated from the absorbance- and conductivity-time data. The stoichiometry and formation constants of the resulting EDA complexes have also been determined. Thermodynamic parameters, Delta H degrees and Delta S degrees , of the complexes have been determined from the temperature dependence of stability constants by Van't Hoff equation. The results indicate that iodine complexes of AM15C5 in both solvents are enthalpy stabilized but entropy destabilized. The influence of solvent properties on the kinetics and stability of the resulting charge-transfer complexes are discussed.

  18. Cytotoxic Activities of Physalis minima L. Chloroform Extract on Human Lung Adenocarcinoma NCI-H23 Cell Lines by Induction of Apoptosis.

    PubMed

    Leong, Ooi Kheng; Muhammad, Tengku Sifzizul Tengku; Sulaiman, Shaida Fariza

    2011-01-01

    Physalis minima L. is reputed for having anticancer property. In this study, the chloroform extract of this plant exhibited remarkable cytotoxic activities on NCI-H23 (human lung adenocarcinoma) cell line at dose- and time-dependent manners (after 24, 48 and 72 h of incubation). Analysis of cell-death mechanism demonstrated that the extract exerted apoptotic programed cell death in NCI-H23 cells with typical DNA fragmentation, which is a biochemical hallmark of apoptosis. Morphological observation using transmission electron microscope (TEM) also displayed apoptotic characteristics in the treated cells, including clumping and margination of chromatins, followed by convolution of the nuclear and budding of the cells to produce membrane-bound apoptotic bodies. Different stages of apoptotic programed cell death as well as phosphatidylserine externalization were confirmed using annexin V and propidium iodide staining. Furthermore, acute exposure to the extract produced a significant regulation of c-myc, caspase-3 and p53 mRNA expression in this cell line. Due to its apoptotic effect on NCI-H23 cells, it is strongly suggested that the extract could be further developed as an anticancer drug.

  19. 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.

  20. Fabrication and optical nonlinearities of composite films derived from the water-soluble Keplerate-type polyoxometalate and chloroform-soluble porphyrin.

    PubMed

    Shi, Zonghai; Zhou, Yunshan; Zhang, Lijuan; Yang, Di; Mu, Cuncun; Ren, Haizhou; Shehzad, Farooq Khurum; Li, Jiaqi

    2015-03-07

    Composite films derived from the water-soluble Keplerate-type polyoxometalate (NH4)42[Mo132O372(CH3COO)30(H2O)72]·ca. 300H2O·ca. 10CH3COONH4 (denoted (NH4)42{Mo132}) and chloroform-soluble tetraphenylporphyrin perchlorate [H2TPP](ClO4)2 are successfully fabricated by a layer-by-layer self-assembly method and characterized by UV-vis spectroscopy and X-ray photoelectron spectroscopy (XPS). The structure of the {Mo132} and [H2TPP](2+) in the films remain intact in light of the results of UV-vis spectroscopy and XPS. UV-vis spectra measurements reveal that the amounts of deposition of {Mo132} and [H2TPP](2+) remain constant in every adsorption cycle in the composite films assembly process. Nonlinear optical properties of the composite films have been investigated by using the Z-scan technique at a wavelength of 532 nm and pulse width of 7 ns. The results show that the composite films have notable nonlinear saturated absorption and self-defocusing effects. The combination of {Mo132} with [H2TPP](2+) can result in composite films with remarkably enhanced optical nonlinearities. The interfacial charge transfer induced by laser from porphyrin to POM in the films is thought to play a key role in the enhancement of NLO response. The third-order NLO susceptibility χ((3)) of the composite films increases with the increase of film thickness.

  1. Potent Anti-inflammatory and Analgesic Actions of the Chloroform Extract of Dendropanax morbifera Mediated by the Nrf2/HO-1 Pathway.

    PubMed

    Akram, Muhammad; Kim, Kyeong-A; Kim, Eun-Sun; Syed, Ahmed Shah; Kim, Chul Young; Lee, Jong Soo; Bae, Ok-Nam

    2016-01-01

    Dendropanax morbifera LEVEILLE (DP) has been used in traditional Korean medicines to treat a variety of inflammatory diseases. Although the in vitro anti-inflammatory potential of this plant is understood, its in vivo efficacy and underlying molecular mechanism of anti-inflammatory effects are largely unknown. We elucidated the anti-inflammatory and analgesic activities and the underlying molecular mechanisms of DP using in vitro and in vivo models. Lipopolysaccharide (LPS)-stimulated murine macrophages were used to analyze the in vitro anti-inflammatory potential of DP extract and to elucidate the underlying mechanisms. In vivo animal models of phorbol 12-myristate 13-acetate (TPA)-induced ear edema and acetic acid-induced writhing response tests were used to analyze the in vivo anti-inflammatory effects and anti-nociceptive effects of DP extract, respectively. Methanolic extract of DP (DPME) significantly inhibited the release of nitric oxide (NO) and prostaglandin E2 (PGE2) in LPS-activated macrophages. Among the five sub-fractions, the chloroform fraction (DP-C) showed the most potent suppressive effects against pro-inflammatory mediators and cytokines in LPS-stimulated macrophages. These effects were attributed to inhibition of nuclear factor-κB (NF-κB) nuclear translocation and c-Jun N terminal kinase (JNK) 1/2 phosphorylation and to activation of NF-E2-related factor 2/heme oxygenase-1 (Nrf2/HO-1) signaling. DP-C exhibited strong protective in vivo effects in TPA-induced ear edema mouse model and acetic acid-induced writhing response test. Our data suggest that DP-C has potent anti-inflammatory and analgesic activities and may be a promising treatment against a variety of inflammatory diseases.

  2. Reduction of nitro-BT by some components of the chloroform-methanol extract of the grey matter of the rat brain.

    PubMed

    Kaluza, J; Szydlowska, H

    1977-01-01

    An attempt was made to extract the substance present in the nerve fibres of the cerebral cortex and basal ganglia which reduced Nitro-BT in alkaline medium pH 9-5 [4,5]. The investigations were performed on 40 white Wistar rats of both sexes and ca. 200 g of body weight. The brains taken for studies were fixed in Baker's formalin for 1,5 hr, thereafter grinded and homogenized in chloroform-methanol solution supplemented up to 100 ml of final volume after homogenisation. Thus prepared solution was left for 24 hr at 4 degrees C temperature and than centrifuged 2000 g/min. The extract was evaporated and the sediment further investigated. The following fraction were received: 1. fraction soluble in aceton, 2. fraction soluble in ethylalcohol, 3. fraction soluble in ether, 4. fraction soluble in water and 5. the sediment. Each fraction was incubated with a standard medium containing Nitro-BT. After incubation the amount of reduced Nitro-BT in each of the incubation medium was spectrophotometrically measured. The most intense reduction was found in the incubation medium with the water fraction. The spectrophotometrically determined reduction of Nitro-BT of the water fraction compared with that of the gangliosides solution were similar. The addition into the incubation media biogenic amines and N,N-diethyl-p-phenylenediamine enhances the reduction of Nitro-BT significantly. According to the results obtained an interference of gangliosides and of the investigated amines in the reduction of Nitro-BT is suggested.

  3. Positive and Negative Contributions in the Solvation Enthalpy due to Specific Interactions in Binary Mixtures of C1-C4 n-Alkanols and Chloroform with Butan-2-one.

    PubMed

    Varfolomeev, Mikhail A; Rakipov, Ilnaz T; Solomonov, Boris N; Lodowski, Piotr; Marczak, Wojciech

    2015-06-25

    In the paper, results of calorimetric measurements, IR spectra, and calculated ab initio stabilization energies of dimers are reported for binary systems butan-2-one + (methanol, ethanol, propan-1-ol, butan-1-ol, and chloroform). Changes in the total enthalpy of specific interactions due to dissolution of butan-2-one in the alcohols, calculated using equations derived in previous works, are positive. That results from the endothermic breaking of the O-H···O-H bonds not completely compensated by the exothermic effects of formation of the O-H···O═C ones. Moreover, the concentration of nonbonded molecules of butan-2-one is significant even in dilute solutions, as is evidenced by the shape of the C═O stretching vibrations band in the IR spectra. Apart from that, the spectra do not confirm 1:2 complexes in spite of two lone electron pairs in the carbonyl group of butan-2-one capable of forming the hydrogen bonds. The changes in enthalpy of specific interactions are negative for dilute solutions of alcohols and chloroform in butan-2-one and of butan-2-one in chloroform, because no hydrogen bonds occur in pure butan-2-one. The experimental results are positively correlated with the enthalpies estimated from the ab initio energies using a simple "chemical reaction" approach.

  4. Profiling of urinary amino-carboxylic metabolites by in-situ heptafluorobutyl chloroformate mediated sample preparation and gas chromatography-mass spectrometry.

    PubMed

    Hušek, Petr; Švagera, Zdeněk; Hanzlíková, Dagmar; Řimnáčová, Lucie; Zahradníčková, Helena; Opekarová, Iva; Šimek, Petr

    2016-04-22

    A novel 1,1,1,2,2,3,3-heptafluorobutyl chloroformate reagent (HFBCF) was examined for in-situ derivatization of amino-carboxylic metabolites in human urine. The arising reaction products exhibit greatly reduced polarity which facilitates combining the derivatization and liquid-liquid microextraction (LLME) from an aqueous urine into an isooctane phase and immediate gas chromatographic-mas spectrometric analysis (GC-MS). The sample preparation protocol is simple, proceeds without an alcohol excess and provides cleaner extracts than other urinary GC-MS based methods. Moreover, thiol metabolites bound in disulfide bonds can be released by reduction with tris(3-hydroxypropyl)phosphine (THP) prior to the developed derivatization and LLME step. In order to evaluate potential of the novel method for GC-MS metabolomics, reaction products of 153 urinary metabolites with HFBCF, particularly those possessing amino and carboxyl groups (56 amino acids and their conjugates, 84 organic acids, 9 biogenic amines, 4 other polar analytes) and two internal standards were investigated in detail by GC-MS and liquid chromatography-mass spectrometry (LC-MS). One hundred and twenty metabolites (78%) yielded a single product, 25 (16%) and 2 metabolites (2-methylcitrate, citrate) generated two and more derivatives. From the examined set, analytically applicable products of 5 metabolites were not detected; the derivatives of 3 metabolites were only suitable for LC-MS analysis. Electron ionization (EI) of the examined analytes contained characteristic, diagnostic ions enabling to distinguish related and isomeric structures. The new method was validated for 132 metabolites using two internal standards in artificial urine and with special attention to potential disease biomarker candidates. The developed sample preparation protocol was finally evaluated by means of a certified organic acid standard mixture in urine and by GC-MS analysis of 100 morning urines obtained from healthy patients (50

  5. Classification and analysis of corn steep liquor by UPLC/Q-TOF MS and HPLC.

    PubMed

    Xiao, Xue; Hou, Yuanyuan; Liu, Yang; Liu, Yanjie; Zhao, Hongzhi; Dong, Linyi; Du, Jun; Wang, Yiming; Bai, Gang; Luo, Guoan

    2013-03-30

    Corn steep liquor (CSL), an important raw material with high nutritional value, serves as a nitrogen source in the fermentation industry. The CSL quality directly affects the yield and quality of fermentation products. In this work, a fingerprinting technique was used to identify the potential markers of CSL. Forty-two CSL samples from different manufacturers were profiled by ultra-performance liquid chromatography with tandem quadrupole time-of-flight mass spectrometry. Sixteen compounds, almost all of which were amino acids and their derivatives, were considered as the potential markers. Then, o-phthalaldehyde-9-fluorenylmethyl chloroformate precolumn derivatization by high-performance liquid chromatography was performed to identify the free amino acids in CSL. Principal component analysis (PCA) was used to distinguish among the samples from different manufacturers. The results demonstrated that the fingerprinting technique combined with PCA analysis was a powerful tool for determining the CSL quality.

  6. A new method for immediate derivatization of hydroxyl groups by fluoroalkyl chloroformates and its application for the determination of sterols and tocopherols in human serum and amniotic fluid by gas chromatography-mass spectrometry.

    PubMed

    Řimnáčová, L; Hušek, P; Šimek, P

    2014-04-25

    A new method has been described for efficient derivatization of secondary alicyclic hydroxyl groups in steroids by alkyl chloroformates (RCFs). Cholesterol, an essential human sterol and a steroid precursor in eukaryotic cells, was used as a model for treatment with various RCFs in an aqueous and non-aqueous environment. While the cholesterol hydroxyl group did not react completely with any of the tested RCFs reagents in the former case, trifluoroethyl chloroformate (TFECF) or heptafluorobutyl chloroformate (HFBCF) fully converts cholesterol and related metabolites into the corresponding mixed carbonates under anhydrous conditions in seconds. The acylation reaction was combined with liquid-liquid microextraction (LLME) between isooctane and acetonitrile phase. The sample preparation requires just a stepwise addition of 50μl isooctane with 5μl of a pyridine catalyst, 100μl acetonitrile and 100μl isooctane with dissolved 5μl of the fluoroalkyl chloroformate reagent to a dried sample. The protocol developed in this study was successfully tested for GC-MS analysis of 12 important model steroids and four main tocopherols. Each analyte provided a single peak with excellent GC separation properties, well defined EI spectra containing diagnostic fragment ions suitable for their identification and quantitation. The new method was further validated for the determination of six diagnostic non-cholesterol sterols and four main tocopherols in human serum and in amniotic fluid. Satisfactory data were obtained in terms of calibration, quantitation limits (for sterols and tocopherols, 0.05 and 0.15μg/ml, respectively), within-run precision (0.9-19.5%) and between-run precision (0.2-19.0%), accuracy (82-115%) and recovery (90-110%). The validated method was successfully applied to GC-MS analysis of the analytes in woman sera and amniotic fluids and the results are well-comparable with those reported by other authors. The presented work demonstrates for the first time

  7. A mixed quantum-classical molecular dynamics study of anti-tetrol and syn-tetrol dissolved in liquid chloroform: hydrogen-bond structure and its signature on the infrared absorption spectrum.

    PubMed

    Kwac, Kijeong; Geva, Eitan

    2013-12-27

    The intramolecular hydrogen-bond structure of stereoselectively synthesized syn-tetrol and anti-tetrol dissolved in deuterated chloroform is investigated via a mixed quantum-classical molecular dynamics simulation. An extensive conformational analysis is performed in order to determine the dominant conformations, the distributions among them, and their sensitivity to the method for assigning partial charges (RESP vs AM1-BCC). The signature of the conformational distribution and method of assigning partial charges on the infrared absorption spectra is analyzed in detail. The relationship between the spectra and the underlying hydrogen-bond structure is elucidated.

  8. The n-hexane and chloroform fractions of Piper betle L. trigger different arms of immune responses in BALB/c mice and exhibit antifilarial activity against human lymphatic filarid Brugia malayi.

    PubMed

    Singh, Meghna; Shakya, Shilpy; Soni, Vishal Kumar; Dangi, Anil; Kumar, Nikhil; Bhattacharya, Shailja-Misra

    2009-06-01

    Modulation of immune functions by using herbal plants and their products has become fundamental regime of therapeutic approach. Piper betle Linn. (Piperaceae) is a widely distributed plant in the tropical and subtropical regions of the world and has been attributed as traditional herbal remedy for many diseases. We have recently reported the antifilarial and antileishmanial efficacy in the leaf extract of Bangla Mahoba landrace of P. betle which is a female plant. The present report describes the in vivo immunomodulatory efficacy of the crude methanolic extract and its n-hexane, chloroform, n-butanol fractions of the female plant at various dose levels ranging between 0.3 and 500 mg/kg in BALB/c. Attempts were also made to observe antifilarial activity of the active extracts and correlate it with the antigen specific immune responses in another rodent Mastomys coucha infected with human lymphatic filarial parasite Brugia malayi. The crude methanol extract and n-hexane fraction were found to potentiate significant (p<0.001) enhancement of both humoral (plaque forming cells, hemagglutination titre) as well as cell-mediated (lymphoproliferation, macrophage activation, delayed type hypersensitivity) immune responses in mice. The flow cytometric analysis of splenocytes of treated mice indicated enhanced population of T-cells (CD4(+), CD8(+)) and B-cells (CD19(+)). The n-hexane fraction (3 mg/kg) was found to induce biased type 2 cytokine response as revealed by increased IL-4(+) and decreased IFN-gamma(+) T-cell population while the chloroform fraction (10 mg/kg) produced a predominant type 1 cytokines. Crude methanolic extract (100 mg/kg) demonstrated a mixed type 1 and type 2 cytokine responses thus suggesting a remarkable immunomodulatory property in this plant. The induction of differential T-helper cell immune response appears ideal to overcome immunosuppression as observed in case of lymphatic, filarial Brugia malayi infection which may also be extended to other

  9. 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.

  10. Quantitative determination of the macrolide antibiotics erythromycin, roxithromycin, azithromycin and clarithromycin in human serum by high-performance liquid chromatography using pre-column derivatization with 9-fluorenylmethyloxycarbonyl chloride and fluorescence detection.

    PubMed

    Sastre Toraño, J; Guchelaar, H J

    1998-12-11

    A validated, highly sensitive and precise high-performance liquid chromatographic (HPLC) method for the determination of the macrolides erythromycin, azithromycin, clarithromycin and roxithromycin in human serum is described. A diethyl ether extract, obtained from serum using a saturated sodium carbonate solution, was treated with 9-fluorenylmethyl-oxycarbonyl chloride (FMOC-Cl) for 40 min at 40 degrees C and chromatographed on a base-deactivated octadecyl column, maintained at 50 degrees C during elution, using an eluent composed of acetonitrile-hydrogenphosphate buffer, pH 7.5, with 0.125% triethylamine (3:2, v/v). Fluorescence detection was used at an excitation wavelength of 255 nm and an emission wavelength of 315 nm. Erythromycin, clarithromycin, roxithromycin and azithromycin were found to have retention times of 8.8, 15.7, 17.1 and 20.7 min, respectively. Recoveries ranging from 93 to 104% were found with reproducibility coefficients of variation of 1.1-5%. Mean correlation coefficients of 0.9997, 0.9998, 0.9996 and 0.9994 were found for the linear calibration curves (n = 2) of erythromycin (0.320-16.1 mg/l), roxithromycin (3.24-19.4 mg/l), clarithromycin (0.190-19.4 mg/l) and azithromycin (0.0988-4.94 mg/l), respectively.

  11. Crystal structure of bis-[μ-(4-meth-oxy-phen-yl)methane-thiol-ato-κ(2) S:S]bis-[chlorido-(η(6)-1-isopropyl-4-methyl-benzene)-ruthenium(II)] chloro-form disolvate.

    PubMed

    Stíbal, David; Süss-Fink, Georg; Therrien, Bruno

    2015-10-01

    The mol-ecular structure of the title complex, [Ru2(C8H9OS)2Cl2(C10H14)2]·2CHCl3 or (p-MeC6H4Pr (i) )2Ru2(SCH2-p-C6H5-OCH3)2Cl2·2CHCl3, shows inversion symmetry. The two symmetry-related Ru(II) atoms are bridged by two 4-meth-oxy-α-toluene-thiol-ato [(4-meth-oxy-phen-yl)methane-thiol-ato] units. One chlorido ligand and the p-cymene ligand complete the typical piano-stool coordination environment of the Ru(II) atom. In the crystal, the CH moiety of the chloro-form mol-ecule inter-acts with the chlorido ligand of the dinuclear complex, while one Cl atom of the solvent inter-acts more weakly with the methyl group of the bridging 4-meth-oxy-α-toluene-thiol-ato unit. This assembly leads to the formation of supra-molecular chains extending parallel to [021].

  12. A mixed quantum-classical molecular dynamics study of anti-tetrol and syn-tetrol dissolved in liquid chloroform II: infrared emission spectra, vibrational excited-state lifetimes, and nonequilibrium hydrogen-bond dynamics.

    PubMed

    Kwac, Kijeong; Geva, Eitan

    2013-11-21

    The effect of vibrational excitation and relaxation of the hydroxyl stretch on the hydrogen-bond structure and dynamics of stereoselectively synthesized syn-tetrol and anti-tetrol dissolved in deuterated chloroform are investigated via a mixed quantum-classical molecular dynamics simulation. Emphasis is placed on the changes in hydrogen-bond structure upon photoexcitation and the nonequilibrium hydrogen-bond dynamics that follows the subsequent relaxation from the excited to the ground vibrational state. The propensity to form hydrogen bonds is shown to increase upon photoexcitation of the hydroxyl stretch, thereby leading to a sizable red-shift of the infrared emission spectra relative to the corresponding absorption spectra. The vibrational excited state lifetimes are calculated within the framework of Fermi's golden rule and the harmonic-Schofield quantum correction factor, and found to be sensitive reporters of the underlying hydrogen-bond structure. The energy released during the relaxation from the excited to the ground state is shown to break hydrogen bonds involving the relaxing hydroxyl. The spectral signature of this nonequilibrium relaxation process is analyzed in detail.

  13. A simple strategy for determining ethanol in all types of alcoholic beverages based on its on-line liquid-liquid extraction with chloroform, using a flow injection system and Fourier transform infrared spectrometric detection in the mid-IR.

    PubMed

    Gallignani, Máximo; Ayala, Carlos; Brunetto, Maria Del Rosario; Burguera, Jose Luis; Burguera, Marcela

    2005-12-15

    In this work, a simple strategy for the determination of ethanol in all types of alcoholic beverages using Fourier transform infrared spectrometric detection has been developed. The methodological proposal includes the quantitative on-line liquid-liquid extraction of ethanol with chloroform, through a sandwich type cell equipped with a PTFE membrane, using a two-channel manifold; and direct measurement of the analyte in the organic phase, by means of Fourier transform infrared spectrometry. The quantification was carried out measuring the ethanol absorbance at 877cm(-1)(,) corrected by means of a baseline established between 844 and 929cm(-1). The procedure, which does not require any sample pretreatment (except for the simple degassing of beer and gassy wine samples, and a simple dilution of spirits with water), was applied to determine ethanol in different alcoholic beverages such as beers, wines and spirits. The results obtained highly agree with those obtained by a derivative FTIR spectrometric procedure, and by head space-gas chromatography with FID detection. The proposed method is simple, fast, precise and accurate. Moreover, it can be easily adapted to any infrared spectrometer equipped with a standard transmission IR cell, and provides attractive analytical features, which are comparable to, or better than those offered by other published methods. In consequence, it represents a valid alternative for the determination of ethanol in alcoholic beverages, and could be suitable for the routine control analysis.

  14. Chloroform-Methanol Residue of Coxiella burnetii Markedly Potentiated the Specific Immunoprotection Elicited by a Recombinant Protein Fragment rOmpB-4 Derived from Outer Membrane Protein B of Rickettsia rickettsii in C3H/HeN Mice

    PubMed Central

    Gong, Wenping; Wang, Pengcheng; Xiong, Xiaolu; Jiao, Jun; Yang, Xiaomei; Wen, Bohai

    2015-01-01

    The obligate intracellular bacteria, Rickettsia rickettsii and Coxiella burnetii, are the potential agents of bio-warfare/bio-terrorism. Here C3H/HeN mice were immunized with a recombinant protein fragment rOmp-4 derived from outer membrane protein B, a major protective antigen of R. rickettsii, combined with chloroform-methanol residue (CMR) extracted from phase I C. burnetii organisms, a safer Q fever vaccine. These immunized mice had significantly higher levels of IgG1 and IgG2a to rOmpB-4 and interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α), two crucial cytokines in resisting intracellular bacterial infection, as well as significantly lower rickettsial loads and slighter pathological lesions in organs after challenge with R. rickettsii, compared with mice immunized with rOmpB-4 or CMR alone. Additionally, after challenge with C. burnetii, the coxiella loads in the organs of these mice were significantly lower than those of mice immunized with rOmpB-4 alone. Our results prove that CMR could markedly potentiate enhance the rOmpB-4-specific immunoprotection by promoting specific and non-specific immunoresponses and the immunization with the protective antigen of R. rickettsii combined with CMR of C. burnetii could confer effective protection against infection of R. rickettsii or C. burnetii. PMID:25909586

  15. Chloroform-Methanol Residue of Coxiella burnetii Markedly Potentiated the Specific Immunoprotection Elicited by a Recombinant Protein Fragment rOmpB-4 Derived from Outer Membrane Protein B of Rickettsia rickettsii in C3H/HeN Mice.

    PubMed

    Gong, Wenping; Wang, Pengcheng; Xiong, Xiaolu; Jiao, Jun; Yang, Xiaomei; Wen, Bohai

    2015-01-01

    The obligate intracellular bacteria, Rickettsia rickettsii and Coxiella burnetii, are the potential agents of bio-warfare/bio-terrorism. Here C3H/HeN mice were immunized with a recombinant protein fragment rOmp-4 derived from outer membrane protein B, a major protective antigen of R. rickettsii, combined with chloroform-methanol residue (CMR) extracted from phase I C. burnetii organisms, a safer Q fever vaccine. These immunized mice had significantly higher levels of IgG1 and IgG2a to rOmpB-4 and interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α), two crucial cytokines in resisting intracellular bacterial infection, as well as significantly lower rickettsial loads and slighter pathological lesions in organs after challenge with R. rickettsii, compared with mice immunized with rOmpB-4 or CMR alone. Additionally, after challenge with C. burnetii, the coxiella loads in the organs of these mice were significantly lower than those of mice immunized with rOmpB-4 alone. Our results prove that CMR could markedly potentiate enhance the rOmpB-4-specific immunoprotection by promoting specific and non-specific immunoresponses and the immunization with the protective antigen of R. rickettsii combined with CMR of C. burnetii could confer effective protection against infection of R. rickettsii or C. burnetii.

  16. 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.

  17. Development, validation and comparison of two microextraction techniques for the rapid and sensitive determination of pregabalin in urine and pharmaceutical formulations after ethyl chloroformate derivatization followed by gas chromatography-mass spectrometric analysis.

    PubMed

    Mudiam, Mohana Krishna Reddy; Chauhan, Abhishek; Jain, Rajeev; Ch, Ratnasekhar; Fatima, Ghizal; Malhotra, Ekta; Murthy, R C

    2012-11-01

    The present article reports first time the use of solid-phase microextraction (SPME) and dispersive liquid-liquid microextraction (DLLME) to extract pregabalin (PRG) from urine and pharmaceutical formulations followed by GC-MS analysis after ethyl chloroformate (ECF) derivatization. PRG is an antiepileptic and analgesic drug, which is a structural analogue of γ-amino-butyric acid (GABA). It is approved by Food and Drug Administration (FDA) for the treatment of central nervous system (CNS) disorders and neuropathic pain. Initially PRG was derivatized with ECF in the presence of pyridine at room temperature for 30s. Experimental parameters were investigated for derivatization, SPME and DLLME conditions. The limit of detection (LOD) and limit of quantitation (LOQ) were found to be 0.019 μg/ml and 0.063 μg/ml for SPME and 0.022 μg/ml and 0.075 μg/ml for DLLME respectively. The percentage recovery, in case of SPME was in the range of 83-98% while for DLLME it is in the range of 84-98%. The intra and inter-day precisions were found to be less than 6%. The developed methods after ECF derivatization were found to be simple, fast, efficient and inexpensive. DLLME has several advantages like lesser extraction time and cost effectiveness as compared to SPME. The developed methods may find wide application for the routine determination of PRG in biological as well as in quality control samples of pharmaceutical formulations.

  18. Silver(I)-directed growth of metal-organic complex nanocrystals with bidentate ligands of hydroquinine anthraquinone-1,4-diyl diethers as linkers at the water-chloroform interface

    PubMed Central

    2014-01-01

    Immiscible liquid-liquid interfaces provide unique double phase regions for the design and construction of nanoscale materials. Here, we reported Ag(I)-directed growth of metal-organic complex nanocrystals by using AgNO3 as a connector in the aqueous solution and bidentate ligand of 1,4-bis(9-O-dihydroquininyl)anthraquinone [(DHQ)2AQN] and its enantiomer of (DHQD)2AQN in the chloroform solutions as linkers. The Ag-(DHQ)2AQN and Ag-(DHQD)2AQN complex nanocrystals were formed at the liquid-liquid interfaces and characterized by using UV-vis absorption and fluorescence spectroscopy and X-ray photoelectron spectroscopy, as well as by using scanning electron microscopy. Screw-like nanocrystals were formed at the initial 30 min after the interfacial coordination reaction started, then they grew into nanorods after several days, and finally became cubic microcrystals after 2 weeks. The pure ligand showed two emission bands centered at about 363 and 522 nm in the methanol solution, the second one of which was quenched and shifted to about 470 nm in the Ag-complex nanocrystals. Two couples of reversible redox waves were recorded for the Ag-complex nanocrystals; one centered at about -0.25 V (vs. Ag/AgCl) was designated to one electron transfer process of Ag - (DHQ)2AQN and Ag - (DHQ)2AQN+, and the other one centered at about 0.2 V was designated to one electron transfer process of Ag - (DHQ)2AQN and Ag+ - (DHQ)2AQN. PMID:25246874

  19. Rapid and simultaneous determination of twenty amino acids in complex biological and food samples by solid-phase microextraction and gas chromatography-mass spectrometry with the aid of experimental design after ethyl chloroformate derivatization.

    PubMed

    Mudiam, Mohana Krishna Reddy; Ratnasekhar, Ch; Jain, Rajeev; Saxena, Prem Narain; Chauhan, Abhishek; Murthy, R C

    2012-10-15

    Amino acids play a vital role as intermediates in many important metabolic pathways such as the biosynthesis of nucleotides, vitamins and secondary metabolites. A sensitive and rapid analytical method has been proposed for the first time for the simultaneous determination of twenty amino acids using solid-phase microextraction (SPME). The protein samples were hydrolyzed by 6M HCl under microwave radiation for 120 min. Then the amino acids were derivatized by ethyl chloroformate (ECF) and the ethoxy carbonyl ethyl esters of amino acids formed were extracted using SPME by direct immersion. Finally the extracted analytes on the SPME fiber were desorbed at 260°C and analyzed by gas chromatography-mass spectrometer (GC-MS) in electron ionization mode. Factors which affect the SPME efficiency were screened by Plackett-Burmann design; most significant factors were optimized with response surface methodology. The optimum conditions for SPME are as follows: pH of 1.7, ionic strength of 733 mg, extraction time of 30 min and fiber of divinyl benzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS). The recovery of all the amino acids was found to be in the range of 89.17-100.98%. The limit of detection (LOD) of all derivatized amino acids in urine, hair and soybean was found to be in the range of 0.20-7.52 μg L(-1), 0.21-8.40 μg L(-1) and 0.18-5.62 μg L(-1), respectively. Finally, the proposed technique was successfully applied for the determination of amino acids in complex biological (hair, urine) and food samples (soybean). The method can find wide applications in the routine analysis of amino acids in any biological as well as food samples.

  20. Section i: Thermodynamic Properties of Hydrocarbon Radicals, Peroxy Hydrocarbon and Peroxy Chlorohydrocarbon Molecules and Radicals. Section II. Kinetics and Reaction Mechanisms For: (1) Chloroform Pyrolysis and Oxidation; (2) Benzene and Toluene Oxidation Under Atmospheric Conditions.

    NASA Astrophysics Data System (ADS)

    Lay, Tsan-Horng

    1995-01-01

    Alkyl radicals are important active intermediates in gas phase photochemistry and combustion reaction systems. With the exception of a limited number of the most elementary radicals, accurate thermodynamic properties of alkyl radicals are either not available or only rough estimations exist. An H atom Bond Increment approach is developed and a data base is derived, for accurately estimating thermodynamic properties (Delta H_{f }^circ298, S ^circ298 and Cp(T)) for generic classes of hydrocarbon radical species. Reactions of alkyl radicals with molecular oxygen are one of the major reaction paths for these radicals in atmospheric photochemistry, oxidation of hydrocarbon liquids and combustion process. Alkyl hydroperoxides are subsequently formed through the alkyl peroxy radicals reactions with varied chemical species present in the reaction system. Thermodynamic properties of the alkyl hydroperoxides and related radicals are therefore frequently required in gas phase modeling and kinetic studies on these systems. The thermodynamic properties of alkyl hydroperoxides, alkyl peroxy radicals and hydroperoxyl-1-ethyl radicals including the species with fluorine and chlorine substituents on the alpha-carbon are evaluated using molecular orbital calculations. Chloroform is used as a model chlorocarbon system with high Cl/H ratio to investigate thermal decomposition processes of chlorocarbons in oxidative and pyrolytic reaction environments. A detailed reaction mechanism is developed to describe the important features of products and reagent loss and is shown to predict the experimental data well. Reaction pathways and rate constants are developed for CCl _3, CCl_2 and rm C_2Cl_3 radical addition to O_2 and combination with O, OH HO_2 and ClO. The reversible addition reaction of OH radical with benzene to form the hydroxyl-2,4-cyclohexadienyl (benzene -OH) adduct and the subsequent reactions of this benzene -OH adduct with O_2 are important initial steps for the

  1. Vortex-assisted surfactant-enhanced-emulsification liquid-liquid microextraction of biogenic amines in fermented foods before their simultaneous analysis by high-performance liquid chromatography.

    PubMed

    Donthuan, Jaruwan; Yunchalard, Sirinda; Srijaranai, Supalax

    2014-11-01

    A simple, rapid, sensitive, and environmentally friendly method, based on modified dispersive liquid-liquid microextraction coupled with high-performance liquid chromatography was developed for the simultaneous determination of five biogenic amines in fermented food samples. Biogenic amines were derivatized with 9-fluorenylmethyl chloroformate, extracted by vortex-assisted surfactant-enhanced emulsification liquid-liquid microextraction, and then analyzed by high-performance liquid chromatography. Five biogenic amine compounds were separated within 30 min using a C18 column and gradient elution with acetonitrile and 1% acetic acid. Factors influencing the derivatization and extraction efficiency such as type and volume of extraction solvent, type, and concentration of surfactant, pH, salt addition, and vortex time were optimized. Under the optimum conditions, the method provided the enrichment factors in the range of 161-553. Good linearity was obtained from 0.002-0.5 mg/L for cadaverine and tyramine, 0.003-1 mg/L for tryptamine and histamine, and 0.005-1 mg/L for spermidine with coefficient of determination (R(2) ) > 0.992. The limits of detection ranged from 0.0010 to 0.0026 mg/L. The proposed method was successfully applied to analysis of biogenic amines in fermented foods such as fermented fish (plaa-som), wine and beer where good recoveries were obtained in the range of 83.2-112.5%

  2. A sensitive fluorescence reagent, 2-[2-(7H-dibenzo[a,g]carbazol-7-yl)-ethoxy]ethyl chloroformate, for amino acids determination in Saussurea involucrate and Artemisia capillaris Thunb using high-performance liquid chromatography with fluorescence detection and identification with mass spectroscopy/electrospray ionization source.

    PubMed

    Li, Guoliang; Sun, Zhiwei; Song, Cuihua; Xia, Lian; Zheng, Jie; Suo, Yourui; You, Jinmao

    2011-06-01

    Recent researches shows that amino acids (AA) are not only cell signaling molecules but are also regulators of gene expression and the protein phosphorylation cascade. More precise analysis of AA composition is reckoned to be one of the most important applications in the biomedical and pharmaceutical fields. In this paper, we develop a sample, sensitive and mild method using 2-[2-(7H-dibenzo[a,g]carbazol-7-yl)-ethoxy]ethyl chloroformate (DBCEC) as A labeling reagent for AA determination by high-performance liquid chromatography (HPLC) with fluorescence detection (FLD) and identification with mass spectroscopy. The maximum excitation and emission wavelengths for DBCEC-AA derivatives were 300 and 395 nm, respectively. This method, in conjunction with a gradient elution, offered a baseline resolution of 20 AA on a reversed-phase Hypersil BDS C(18) column. LC separation for the derivatized AA showed good reproducibility, and all AA were found to give excellent linear responses with correlation coefficients > 0.9993. The calculated detection limits with a 25.0 fmol injection of each AA (at a signal-to-noise ratio of 3:1) ranged from 2.62 to 22.6 fmol. This method was applied to determine the AA composition in Saussurea involucrate and Artemisia capillaris Thunb. Meanwhile, this method exhibits a powerful potential for trace analysis of AA from biomedicine, foodstuff and other complex samples.

  3. Chloroform extraction of iodine in seawater: method development

    NASA Astrophysics Data System (ADS)

    Seidler, H. B.; Glimme, A.; Tumey, S.; Guilderson, T. P.

    2012-12-01

    While 129I poses little to no radiological health hazard, the isotopic ratio of 129I to stable iodine is very useful as a nearly conservative tracer for ocean mixing processes. The unfortunate disaster at the Fukushima Daiichi nuclear power plant released many radioactive materials into the environment, including 129I. The release allows the studying of oceanic processes through the tracking of 129I. However, with such a low iodine (~0.5 micromolar) and 129I concentrations (<10-11) accelerator mass spectrometry (AMS) is needed for accurate measurements. In order to prepare the samples of ocean water for analysis by AMS, the iodine needs to be separated from the various other salts in the seawater. Solvent extraction is the preferred method for preparation of seawater for AMS analysis of 129I. However, given the relatively low background 129I concentrations in the Pacific Ocean, we sought to optimize recovery of thismethod, which would minimize both the sample size and the carrier addition required for analysis. We started from a base method described in other research and worked towards maximum efficiency of the process while boosting the recovery of iodine. During development, we assessed each methodological change qualitatively using a color scale (I2 in CHCl3) and quantitatively using Inductively Coupled Plasma Mass Spectrometry (ICP-MS). The "optimized method" yielded a 20-40% increase in recovery of the iodine compared to the base method (80-85% recovery vs. 60%). Lastly, the "optimized method" was tested by AMS for fractionation of the extracted iodine.

  4. On-line sample preconcentration by sweeping and poly(ethylene oxide)-mediated stacking for simultaneous analysis of nine pairs of amino acid enantiomers in capillary electrophoresis.

    PubMed

    Lin, En-Ping; Lin, Kai-Cheng; Chang, Chia-Wei; Hsieh, Ming-Mu

    2013-09-30

    This study proposes a sensitive method for the simultaneous separation and concentration of 9 pairs of amino acid enantiomers by combining poly(ethylene oxide) (PEO)-based stacking, β-cyclodextrin (β-CD)-mediated micellar electrokinetic chromatography (MEKC), and 9-fluoroenylmethyl chloroformate (FMOC) derivatization. The 9 pairs of FMOC-derivatized amino acid enantiomers were baseline separated using a discontinuous system, and the buffer vials contained a solution of 150 mM Tris-borate (TB), 12.5% (v/v) isopropanol (IPA), 0.5% (w/v) PEO, 35 mM sodium taurodeoxycholate (STDC), and 35 mM β-CD, and the capillary was filled with a solution of 1.5 M TB, 12.5% (v/v) IPA, 35 mM STDC, and 35 mM β-CD. Based on the difference in viscosity between the sample zone and PEO solution and because of the STDC sweeping, the discontinuous system effectively stacked 670 nL of the 9 pairs of FMOC-derivatized amino acid enantiomers without losing chiral resolution. Consequently, the limits of detection for the 9 pairs of FMOC-derivatized amino acid enantiomers were reduced to 40-60 nM. This method was successfully used to determine d-Tryptophan (Trp), l-Trp, d-Phenylalanine (Phe), l-Phe, d-Glutamic acid (Glu), and l-Glu in various types of beers.

  5. Solventless mechanosynthesis of N-protected amino esters.

    PubMed

    Konnert, Laure; Lamaty, Frédéric; Martinez, Jean; Colacino, Evelina

    2014-05-02

    Mechanochemical derivatizations of N- or C-protected amino acids were performed in a ball mill under solvent-free conditions. A vibrational ball mill was used for the preparation of N-protected α- and β-amino esters starting from the corresponding N-unmasked precursors via a carbamoylation reaction in the presence of di-tert-butyl dicarbonate (Boc2O), benzyl chloroformate (Z-Cl) or 9-fluorenylmethoxycarbonyl chloroformate (Fmoc-Cl). A planetary ball mill proved to be more suitable for the synthesis of amino esters from N-protected amino acids via a one-pot activation/esterification reaction in the presence of various dialkyl dicarbonates or chloroformates. The spot-to-spot reactions were straightforward, leading to the final products in reduced reaction times with improved yields and simplified work-up procedures.

  6. Liquid chromatographic determination of the cyanobacterial toxin beta-n-methylamino-L-alanine in algae food supplements, freshwater fish, and bottled water.

    PubMed

    Scott, Peter M; Niedzwiadek, Barbara; Rawn, Dorothea F K; Lau, Ben P-Y

    2009-08-01

    Beta-N-Methylamino-L-alanine (BMAA) is a neurotoxin originally found in cycad seeds and now known to be produced by many species of freshwater and marine cyanobacteria. We developed a method for its determination in blue-green algae (BGA) food supplements, freshwater fish, and bottled water by using a strong cation-exchange, solid-phase extraction column for cleanup after 0.3 M trichloroacetic acid extraction of BGA supplements and fish. Bottled water was applied directly onto the solid-phase extraction column. For analysis of carbonated water, sonication and pH adjustment to 1.5 were needed. To determine protein-bound BMAA, the protein pellet left after extraction of the BGA supplement and fish was hydrolyzed by boiling with 6 M hydrochloric acid; BMAA was cleaned up on a C18 column and a strong cation-exchange, solid-phase extraction column. Determination of BMAA was by liquid chromatography of the fluorescent derivative formed with 9-fluorenylmethyl chloroformate. The method was validated by recovery experiments using spiking levels of 1.0 to 10 microg/g for BGA supplements, 0.5 to 5.0 microg/g for fish, and 0.002 microg/g for bottled water; mean recoveries were in the range of 67 to 89% for BGA supplements and fish, and 59 to 92% for bottled water. Recoveries of BMAA from spiked extracts of hydrolyzed protein from BGA supplements and fish ranged from 66 to 83%. The cleanup developed provides a useful method for surveying foods and supplements for BMAA and protein-bound BMAA.

  7. 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.

  8. Characterization, thermal stability studies, and analytical method development of Paromomycin for formulation development.

    PubMed

    Khan, Wahid; Kumar, Neeraj

    2011-06-01

    Paromomycin (PM) is an aminoglycoside antibiotic, first isolated in the 1950s, and approved in 2006 for treatment of visceral leishmaniasis. Although isolated six decades back, sufficient information essential for development of pharmaceutical formulation is not available for PM. The purpose of this paper was to determine thermal stability and development of new analytical method for formulation development of PM. PM was characterized by thermoanalytical (DSC, TGA, and HSM) and by spectroscopic (FTIR) techniques and these techniques were used to establish thermal stability of PM after heating PM at 100, 110, 120, and 130 °C for 24 h. Biological activity of these heated samples was also determined by microbiological assay. Subsequently, a simple, rapid and sensitive RP-HPLC method for quantitative determination of PM was developed using pre-column derivatization with 9-fluorenylmethyl chloroformate. The developed method was applied to estimate PM quantitatively in two parenteral dosage forms. PM was successfully characterized by various stated techniques. These techniques indicated stability of PM for heating up to 120 °C for 24 h, but when heated at 130 °C, PM is liable to degradation. This degradation is also observed in microbiological assay where PM lost ∼30% of its biological activity when heated at 130 °C for 24 h. New analytical method was developed for PM in the concentration range of 25-200 ng/ml with intra-day and inter-day variability of < 2%RSD. Characterization techniques were established and stability of PM was determined successfully. Developed analytical method was found sensitive, accurate, and precise for quantification of PM. Copyright © 2010 John Wiley & Sons, Ltd.

  9. Novel one-step headspace dynamic in-syringe liquid phase derivatization-extraction technique for the determination of aqueous aliphatic amines by liquid chromatography with fluorescence detection.

    PubMed

    Muniraj, Sarangapani; Shih, Hou-Kung; Chen, Ying-Fang; Hsiech, Chunming; Ponnusamy, Vinoth Kumar; Jen, Jen-Fon

    2013-06-28

    A novel one-step headspace (HS) dynamic in-syringe (DIS) based liquid-phase derivatization-extraction (LPDE) technique has been developed for the selective determination of two short-chain aliphatic amines (SCAAs) in aqueous samples using high performance liquid chromatography (HPLC) with fluorescence detection (FLD). Methylamine (MA) and dimethylamine (DMA) were selected as model compounds of SCAAs. In this method, a micro-syringe pre-filled with derivatizing reagent solution (9-fluorenylmethyl chloroformate) in the barrel was applied to achieve the simultaneous derivatization and extraction of two methylamines evolved from alkalized aqueous samples through the automated reciprocated movements of syringe plunger. After the derivatization-extraction process, the derivatized phase was directly injected into HPLC-FLD for analysis. Parameters influencing the evolution of methylamines and the HS-DIS-LPDE efficiency, including sample pH and temperature, sampling time, as well as the composition of derivatization reagent, reaction temperature, and frequency of reciprocated plunger movements, were thoroughly examined and optimized. Under optimal conditions, detections were linear in the range of 25-500μgL(-1) for MA and DMA with correlation coefficients all above 0.995. The limits of detection (based on S/N=3) were 5 and 19ngmL(-1) for MA and DMA, respectively. The applicability of the developed method was demonstrated for the determination of MA and DMA in real water samples without any prior cleanup of the sample. The present method provides a simple, selective, automated, low cost and eco-friendly procedure to determine aliphatic amines in aqueous samples.

  10. Separation and Measurement of Plant Alkaloid Enantiomers by RP-HPLC Analysis of their Fmoc-Alanine Analogs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plants often synthesize secondary metabolites that are enantiomers. Enantiomers can cause very different physiological responses. Ammodendrine (1) and anabasine (2) are teratogens that can cause congenital malformations in livestock and enantiomeric forms of each have been found in Lupinus spp. an...

  11. Cysteine pseudoprolines for thiol protection and peptide macrocyclization enhancement in Fmoc-based solid-phase peptide synthesis.

    PubMed

    Postma, Tobias M; Albericio, Fernando

    2014-03-21

    Contrary to other studies, here we describe cysteine (Cys) pseudoproline-containing peptides with short deprotection times in TFA. The deprotection times fell in the same range as other protecting groups commonly used in SPPS (e.g., 1-3 h). Moreover, when using Cys pseudoprolines as peptide macrocyclization-enhancing moieties a considerable reduction in reaction time was observed compared to a peptide containing trityl protected Cys.

  12. Study of a new derivatizing reagent that improves the analysis of amino acids by HPLC with fluorescence detection: application to hydrolyzed rape bee pollen.

    PubMed

    You, Jinmao; Liu, Lingjun; Zhao, Wenchen; Zhao, Xianen; Suo, Yourui; Wang, Honglun; Li, Yulin

    2007-04-01

    A simple and sensitive method for evaluating the chemical compositions of protein amino acids, including cystine (Cys)(2) and tryptophane (Try) has been developed, based on the use of a sensitive labeling reagent 2-(11H-benzo[alpha]-carbazol-11-yl) ethyl chloroformate (BCEC-Cl) along with fluorescence detection. The chromophore of the 1,2-benzo-3,4-dihydrocarbazole-ethyl chloroformate (BCEOC-Cl) molecule was replaced with the 2-(11H-benzo[alpha]-carbazol-11-yl) ethyl functional group, yielding the sensitive fluorescence molecule BCEC-Cl. The new reagent BCEC-Cl could then be substituted for labeling reagents commonly used in amino acid derivatization. The BCEC-amino acid derivatives exhibited very high detection sensitivities, particularly in the cases of (Cys)(2) and Try, which cannot be determined using traditional labeling reagents such as 9-fluorenyl methylchloroformate (FMOC-Cl) and ortho-phthaldialdehyde (OPA). The fluorescence detection intensities for the BCEC derivatives were compared to those obtained when using FMOC-Cl and BCEOC-Cl as labeling reagents. The ratios I (BCEC)/I (BCEOC) = 1.17-3.57, I (BCEC)/I (FMOC) = 1.13-8.21, and UV(BCEC)/UV(BCEOC) = 1.67-4.90 (where I is the fluorescence intensity and UV is the ultraviolet absorbance). Derivative separation was optimized on a Hypersil BDS C(18) column. The detection limits calculated from 1.0 pmol injections, at a signal-to-noise ratio of 3, ranged from 7.2 fmol for Try to 8.4 fmol for (Cys)(2). Excellent linear responses were observed, with coefficients of >0.9994. When coupled with high-performance liquid chromatography, the method established here allowed the development of a highly sensitive and specific method for the quantitative analysis of trace levels of amino acids including (Cys)(2) and Try from bee-collected pollen (bee pollen) samples.

  13. Tris(naphthalen-1-yl)phosphane chloro-form hemisolvate.

    PubMed

    Davis, Wade L; Muller, Alfred

    2012-12-01

    The title compound, P(C10H7)3·0.5CHCl3, was isolated after the unsuccessful reaction of KSeCN and tris-(naphthalen-1-yl)phosphane. The solvent mol-ecule is disordered about an inversion center. The effective cone angle of the phosphine is 203°. In the crystal, weak C-H⋯Cl and C-H⋯π inter-actions are observed.

  14. Carbon Tetrachloride and Chloroform Partition Coefficients Derived from Aqueous Desorption of Contaminated Hanford Sediments

    SciTech Connect

    Riley, Robert G.; Sklarew, Debbie S.; Brown, Christopher F.; Gent, Philip M.; Szecsody, Jim E.; Mitroshkov, Alexandre V.; Thompson, Christopher J.

    2005-07-08

    Researchers at PNNL determined CCl4 and CHCl3 groundwater/sediment partition coefficients (Kd values) for contaminated aquifer sediments collected from borehole C3246 (299-W15-46) located in the 200 West Area adjacent to the Z-9 trench. Having realistic values for this parameter is critical to predict future movement of CCl4 in groundwater from the 200 West Area.

  15. EXPERIMENTAL MEASUREMENT AND MODELING OF THE VAPOR-LIQUID EQUILIBRIUM OF CARBON DIOXIDE + CHLOROFORM. (R826734)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  16. A VCD robust mode analysis of induced chirality: the case of pulegone in chloroform.

    PubMed

    Nicu, Valentin Paul; Debie, Elke; Herrebout, Wouter; Van der Veken, Benjamin; Bultinck, Patrick; Baerends, Evert Jan

    2009-01-01

    Vibrational modes in an achiral molecule may acquire rotational strength by complexation to a chiral molecule, as happens for achiral solvent molecules complexed to a chiral solute. We investigate this transfer of chirality in vibrational circular dichroism for the pulegone molecule in CDCl(3) solvent from the point of view of the robustness concept introduced recently. It turns out that the transfer of chirality yields nonrobust modes, which means that, although they are observed in vibrational circular dichroism (VCD) experiments, the sign of these modes cannot be predicted reliably with standard (Density Functional Theory) VCD calculations. This limits the usefulness of the induced chirality phenomenon for obtaining information on the intermolecular interactions that give rise to it.

  17. Cell proliferation and apoptosis during chloroform-induced hepatocarcinogenesis in male F-344/N rats.

    EPA Science Inventory

    The carcinogenic potential of chlorinated organics is of direct importance in human risk assessment. Most drinking water chlorinated organics are disinfection by products (DBPs) of water chlorination and many test positive in rodent bioassays. Trihalomethanes (THMs) are the most ...

  18. 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...

  19. Abiotic Degradation Rates for Carbon Tetrachloride: and Chloroform: Progress in FY 2008

    SciTech Connect

    Amonette, James E.; Jeffers, Peter M.; Qafoku, Odeta; Russell, Colleen K.; Wietsma, Thomas W.; Truex, Michael J.

    2008-10-31

    This is a letter report summarizing work performed in FY2008 to determine the rates of carbon tetrachloride hydrolysis at temperatures close to actual groundwater temperatures. The report describes the project, the methodology, and the results obtained since the project's inception in FY2006. Measurements of hydrolysis rates in homogeneous solution have been completed for temperaturs of 70 C through 40 C, with additional data available at 30 C and 20 C. These results show no difference between the rates in deionized H2O and in filter-sterilized Hanford-Site groundwater. Moreover, the rates measured are 2-3 times slower than predicted from the open literature. Measurements of rates involving sterile suspensions of Hanford-Site sediment in Hanford-Site groundwater, however, show faster hydrolysis at temperatures below 40 C. Extrapolation of the current data available suggests a six-fold increase in rate would be expected at groundwater temperature of 16 C due to the presence of the sediment. This result translates into a 78-year half-life, rather than the 470-680 year half-life that would be predicted from rate determinations in homogeneous solution. The hydrolysis rate data at 20 C, in contrast to those at higher temperatures, are preliminary and have low statistical power. While significant (p < 0.05) differences between the heterogeneous and homogeneous systems are seen at 30 C, the results at 20 C are not statistically significant at this level due to limited data and the very slow nature of the reaction. More time is needed to collect data at these low temperatures to improve the statistical power of our observation. Given the critical need for hydrolysis rate data at temperatures relevant to groundwater systems, we have three recommendations for future work. First, we recommend a continuation of the sampling and analysis of the remaining long-term sealed-ampule experiments described in this report. These are primarily 20 C and 30 C experiments. The data at 20°C, which are most germane to the groundwater environment, will likely take two more years before they are complete. Second, due to the importance of heterogeneous effects, we recommend the continuation of sampling and analysis for a similar set of experiments looking at hydrolysis rates of CT and CF in contact with individual minerals (montmorillonite, kaolinite, albite, and muscovite) or Hanford-Site sediment, and, for CF, in homogeneous solutions. These experiments were set up under a separate project funded by a congressional earmark (EM-22) for which funding expired at the end of FY08. Third, we strongly recommend development of a 13C/12C isotope ratio mass spectrometry (IRMS) approach for determination of hydrolysis rates at groundwater temperatures and relevant CT and CF concentrations. The technique is sensitive enough that we expect to be able to shorten the time required for a rate-constant determination by 20-fold. Thus, a 5-year experiment conducted by the sealed-ampule approach could be shortened to 3 months by the IRMS approach. This sensitivity also would allow the IRMS technique to be used to follow hydrolysis rates at concentrations found in groundwater plumes at Hanford.

  20. Use of a temporary "solubilizing" peptide tag for the Fmoc solid-phase synthesis of human insulin glargine via use of regioselective disulfide bond formation.

    PubMed

    Hossain, Mohammed Akhter; Belgi, Alessia; Lin, Feng; Zhang, Suode; Shabanpoor, Fazel; Chan, Linda; Belyea, Chris; Truong, Hue-Trung; Blair, Amy R; Andrikopoulos, Sof; Tregear, Geoffrey W; Wade, John D

    2009-07-01

    Solid-phase peptide synthesis has been refined to a stage where efficient preparation of long and complex peptides is now achievable. However, the postsynthesis handling of poorly soluble peptides often remains a significant hindrance to their purification and further use. Several synthetic schemes have been developed for the preparation of such peptides containing modifications to aid their solubility. However, these require the use of complex chemistry or yield non-native sequences. We describe a simple approach based on the use of penta-lysine "tags" that are linked to the C-terminus of the peptide of interest via a base-labile linker. After ready purification of the now freely solubilized peptide, the "tag" is removed by simple, brief base treatment giving the native sequence in much higher overall yield. The applicability of the method was demonstrated by the novel preparation of insulin glargine via solid-phase synthesis of each of the two chains--including the notoriously poorly soluble A-chain--followed by their combination in solution via regioselective disulfide bond formation. At the conclusion of the chain combination, the solubilizing peptide tag was removed from the A-chain to provide synthetic human glargine in nearly 10% overall yield. This approach should facilitate the development of new insulin analogues as well as be widely applicable to the improved purification and acquisition of otherwise poorly soluble synthetic peptides.

  1. Aqueous microwave-assisted solid-phase peptide synthesis using Fmoc strategy. III: racemization studies and water-based synthesis of histidine-containing peptides.

    PubMed

    Hojo, Keiko; Shinozaki, Natsuki; Hidaka, Koushi; Tsuda, Yuko; Fukumori, Yoshinobu; Ichikawa, Hideki; Wade, John D

    2014-10-01

    In this study, we describe the first aqueous microwave-assisted synthesis of histidine-containing peptides in high purity and with low racemization. We have previously shown the effectiveness of our synthesis methodology for peptides including difficult sequences using water-dispersible 9-fluorenylmethoxycarbonyl-amino acid nanoparticles. It is an organic solvent-free, environmentally friendly method for chemical peptide synthesis. Here, we studied the racemization of histidine during an aqueous-based coupling reaction with microwave irradiation. Under our microwave-assisted protocol using 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride, the coupling reaction can be efficiently performed with low levels of racemization of histidine. Application of this water-based microwave-assisted protocol with water-dispersible 9-fluorenylmethoxycarbonyl-amino acid nanoparticles led to the successful synthesis of the histidine-containing hexapeptide neuropeptide W-30 (10-15), Tyr-His-Thr-Val-Gly-Arg-NH₂, in high yield and with greatly reduced histidine racemization.

  2. A study of the biological effect of continuous inhalation exposure of 1, 1, 1-trichloroethene (methyl chloroform) on animals

    NASA Technical Reports Server (NTRS)

    Macewen, J. D.; Kinkead, E. R.; Haun, C. C.

    1974-01-01

    The effects of continuous exposure to 1,1,1-trichloroethane on hepatic morphology and function are evaluated and compared with those produced by methylene chloride (dichloromethane) to determine environmental concentrations of each compound that would produce a similar biological response, i.e., a comparable increase in liver triglycerides over control levels. Experimental findings on mice, rats, dogs, and monkeys indicate that the pathological alternations observed with 1,1,1-trichloroethane are similar to those observed with dichloromethane except for different time courses of the effects and different degrees of recovery. A ten fold greater atmospheric concentration of 1,1,1-trichloroethane is required to produce the minimal liver changes found at 100 ppm dichloromethane.

  3. Identification of Coordinating Ligand Atoms in Cu(calcimycin) 2 Complex from EPR Linewidths in Chloroform Solutions

    NASA Astrophysics Data System (ADS)

    Prabhananda, B. S.; Kombrabail, M. H.

    The X-band EPR spectra of Cu(cal) 2 complex in CHCl 3 solutions (cal = calcimycin) do not show resolved 14N hyperfine structure (J. S. Puskin and T. E. Gunter, Biochemistry14, 187, 1975) even though they are expected from the nitrogen coordination to divalent metal ions inferred by NMR acid by model building (C. M. Deber and D. R. Pfeiffer, Biochemistry15, 132, 1976). In the present work, unresolved hyperfine structure from two equivalent 14N has been inferred in the EPR spectra of Cu(cal) 2 from an analysis of linewidths and lineshapes of mI = {3}/{2} and {1}/{2} transitions at 298 K and mI = {3}/{2} transition at 253 K, in CHCl 3 solutions ( mI = nuclear magnetic quantum number associated with the Cu hyperfine transition); g(parallel to); g|| - g⊥(=0.24), | A|| - A⊥| (=131 G), and 14N hyperfine constant AN (˜11.3 G) determined from liquid solution linewidth studies in this work favor a solution structure of the complex in which two nitrogens and two oxygens are involved in the square-planar coordination at the metal ion similar to that suggested on the basis of the model for Ca(cal) 2 complex.

  4. 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.

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

    PubMed Central

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

    2016-01-01

    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. PMID:27282822

  6. UTILIZATION OF CATHODIC HYDROGEN AS ELECTRON DONOR FOR CHLOROFORM COMETABOLISM BY A MIXED, METHANOGENIC CULTURE. (R825549C044)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  7. High-performance liquid chromatographic assay for catecholamines and metanephrines using fluorimetric detection with pre-column 9-fluorenylmethyloxycarbonyl chloride derivatization.

    PubMed

    Chan, E C; Wee, P Y; Ho, P Y; Ho, P C

    2000-12-01

    A convenient HPLC-fluorescent assay of norepinephrine (NE), epinephrine (E), dopamine (DA) and their 3-O-methylated metabolites, normetanephrine (NM) and metanephrine (MN) was developed. These analytes were coupled to 9-fluorenylmethyloxycarbonyl chloride (FMOC-Cl) before assays. Results showed that using a linear gradient elution, peaks of FMOC-NE, FMOC-E, FMOC-DA, FMOC-NM, FMOC-MN and FMOC-DHBA (3,4-dihydroxybenzylamine, internal standard) were simultaneously resolved within 40 min. Optimization of the chromatographic and derivatization conditions, and validation of the assay were further discussed in the paper. The structures of these derivatives were confirmed by atmospheric pressure chemical ionization mass spectrometry (APCI-MS). The molecular ions [M+H]- of FMOC-NE, FMOC-E, FMOC-DA, FMOC-NM and FMOC-MN were m/z 836, 850, 820, 628 and 642, respectively. Based on these findings, the FMOC-derivatives of metanephrines and catecholamines were confirmed to be bi-substituted and tri-substituted respectively at the amino and catechol functional groups. Finally, the assay was successfully applied to the measurement of urinary E, DA, NM and MN after direct derivatization and simple cleaning.

  8. An Active Insect Kinin Analog with 4-Aminopyroglutamate, A Novel cis-Peptide Bond, Type VI beta-Turn Motif

    DTIC Science & Technology

    2004-01-01

    modified FastMoc0.25 procedure as well as manually by the solid-phase method, using the Fmoc - strategy and starting from Rink Amide resin (Novabiochem...0.53 mM/g). The Fmoc protecting group was removed by 20% piperidine in DMF. A fourfold excess of the respective Fmoc -amino acids was activated in...equivalents). Amino acid side-chain protect- ing groups were Pbf for Arg and Boc for Trp. The coupling of Fmoc -(S,S)-4-aminopyroglutamic acid ( Fmoc -APy

  9. Photophysics and biological applications of the environment-sensitive fluorophore 6-N,N-dimethylamino-2,3-naphthalimide.

    PubMed

    Vázquez, M Eugenio; Blanco, Juan B; Imperiali, B

    2005-02-02

    We have synthesized a new environment-sensitive fluorophore, 6-N,N-dimethylamino-2,3-naphthalimide (6DMN). This chromophore exhibits valuable fluorescent properties as a biological probe with emission in the 500-600 nm range and a marked response to changes in the environment polarity. The 6DMN fluorescence is red-shifted in polar protic environments, with the maximum emission intensity shifting more than 100 nm from 491 nm in toluene to 592 nm in water. Additionally, the fluorescence quantum yield decreases more than 100-fold from chloroform (Phi = 0.225) to water (Phi = 0.002). The scope and applications of the 6DMN probe are expanded with the synthesis of an Fmoc-protected amino acid derivative (5), which contains the fluorophore. This unnatural amino acid has been introduced into several peptides, demonstrating that it can be manipulated under standard solid-phase peptide synthesis conditions. Peptides incorporating the new residue can be implemented for monitoring protein-protein interactions as exemplified in studies with Src homology 2 (SH2) phosphotyrosine binding domains. The designed peptides exhibit a significant increase in the quantum yield of the long wavelength fluorescence emission band (596 nm) upon binding to selected SH2 domains (e.g., Crk SH2, Abl SH2, and PI3K SH2). The peptides can be used as ratiometric sensors, since the short wavelength band (460 nm) was found almost invariable throughout the titrations.

  10. Cell-penetrating TAT peptide in drug delivery systems: Proteolytic stability requirements

    PubMed Central

    Koren, Erez; Apte, Anjali; Sawant, Rupa R.; Grunwald, Jacob; Torchilin, Vladimir P.

    2012-01-01

    The stability and activity of the HIV cell-penetrating TAT peptide (TATp) on the surface of TATp-modified micelles and liposomes in relation to its proteolytic cleavage was investigated. TATp moieties were attached to the surface of these nanocarriers using TATp modified with a conjugate of phosphatidyl ethanolamine with a ‘short’ PEG (PEG-PE). Following pre-incubation with trypsin, elastase, or collagenase, the proteolytic stability of TATp on the surface of these modified carriers was studied by HPLC with fluorescence detection using fluorenylmethyl chloroformate (FMOC) labeling. All tested enzymes produced a dose-dependent cleavage of TATp as shown by the presence of TATp Arg-Arg fragments. Inhibition of TATp cleavage occurred when these TATp-micelles were modified by the addition of longer PEG-PE blocks, indicating an effective shielding of TATp from proteolysis by these blocks. TATp-modified carriers were also tested for their ability to accumulate in EL-4, HeLa, and B16-F10 cells. Trypsin treatment of TATp-modified liposomes and micelles resulted in decreased uptake and cell interaction, as measured by fluorescence microscopy and fluorescence-activated cell sorting techniques. Furthermore, a decrease in the cytotoxicity of TATp-modified liposomes loaded with doxorubicin (Doxil) was observed following trypsin treatment. In conclusion, steric shielding of TATp is essential to ensure its in vivo therapeutic function. PMID:21438724

  11. Quantification of nortriptyline in plasma by HPLC and fluorescence detection.

    PubMed

    Almudever, Patricia; Peris, José-Esteban; Garrigues, Teresa; Diez, Octavio; Melero, Ana; Alós, Manuel

    2010-03-15

    A simple, sensitive and specific high-performance liquid chromatography method has been developed for the determination of nortriptyline (NT) in plasma samples. The assay involved derivatization with 9H-fluoren-9-ylmethyl chloroformate (Fmoc-Cl) and isocratic reversed-phase (C(18)) chromatography with fluorescence detection. The developed method required only 100 microl of plasma sample, deproteinized and derivatized in one step. Calibration curves were lineal over the concentration range of 5-5000 ng/ml. The derivatization reaction was performed at room temperature in 20 min and the obtained NT derivative was stable for at least 48 h at room temperature. The within-day and between-day relative standard deviation was below 8%. The limit of detection (LOD) was 2 ng/ml, and the lower limit of quantification (LLOQ) was established at 10 ng/ml. The method was applied on plasma collected from rats, at different time intervals, after intravenous administration of 0.5 mg of NT.

  12. Residue determination and levels of glyphosate in surface waters, sediments and soils associated with oil palm plantation in Tasik Chini, Pahang, Malaysia

    NASA Astrophysics Data System (ADS)

    Mardiana-Jansar, K.; Ismail, B. S.

    2014-09-01

    Levels of glyphosate and its main metabolite were determined in surface water, soil and sediment samples from an oil palm plantation area located at Tasik Chini, Pahang, Malaysia. The optimization analytical method has been developed for the determination of glyphosate herbicide and its metabolite amino-methyl-phosphonic acid (AMPA) in surface waters to a level of 0.1μg/L, while in sediments and soils to a level of 0.5μg/g with a good linearity in the calibration range of 1-100μg/L. The procedure involves a pre-columnderivatization step with 9-fluorenyl-methyl-chloroformate (FMOC-Cl) yielding highly fluorescent derivatives of the analytes which can be determined by HPLC with fluorescence detection. In the field, levels of glyphosate in surface waters ranges from not detected to 1.0mg/L, while in soils and sediments were from not detected to 6.0mg/kg. For AMPA, the residues in surface waters were between not detected to 2.0mg/L, while in soil and sediment samples were from not detected to 5mg/kg. This variation of glyphosate and AMPA levels depended directly on time of pesticide application and the season.

  13. 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.

  14. Spectrophotometric determination of arsenic by molybdenum blue method in zinc-lead concentrates and related smelter products after chloroform extraction of iodide complex.

    PubMed

    Rao, C S; Rajan, S C; Rao, N V

    1993-05-01

    The most popular and widely applied method for determination of arsenic in ore concentrates is by spectrophotometry of arsenomolybdic acid reduced to molybdenum blue. While applying this method, several authors have developed procedures which varied in the decomposition, separation of arsenic and in the final colour development. Data regarding interference from germanium is inadequate. The present paper describes a procedure, which combines the best features of the previous procedures and is simple, less time consuming and interference-free compared to earlier procedures. This method has been applied to zinc-lead concentrates and related smelter products.

  15. Birge-Sponer Estimation of the C-H Bond Dissociation Energy in Chloroform Using Infrared, Near-Infrared, and Visible Absorption Spectroscopy: An Experiment in Physical Chemistry

    ERIC Educational Resources Information Center

    Myrick, M. L.; Greer, A. E.; Nieuwland, A. A.; Priore, R. J.; Scaffidi, J.; Andreatta, Danielle; Colavita, Paula

    2008-01-01

    The fundamental and overtone vibrational absorption spectroscopy of the C-H unit in CHCl[subscript 3] is measured for transitions from the v = 0 energy level to v = 1 through v = 5 energy levels. The energies of the transitions exhibit a linearly-decreasing spacing between adjacent vibrational levels as the vibrational quantum number increases.…

  16. Global average concentration and trend for hydroxyl radicals deduced from ALE/GAGE trichloroethane (methyl chloroform) data for 1978-1990

    NASA Technical Reports Server (NTRS)

    Prinn, R.; Cunnold, D.; Simmonds, P.; Alyea, F.; Boldi, R.; Crawford, A.; Fraser, P.; Gutzler, D.; Hartley, D.; Rosen, R.

    1992-01-01

    An optimal estimation inversion scheme is utilized with atmospheric data and emission estimates to determined the globally averaged CH3CCl3 tropospheric lifetime and OH concentration. The data are taken from atmospheric measurements from surface stations of 1,1,1-trichloroethane and show an annual increase of 4.4 +/- 0.2 percent. Industrial emission estimates and a small oceanic loss rate are included, and the OH concentration for the same period (1978-1990) are incorporated at 1.0 +/- 0.8 percent/yr. The positive OH trend is consistent with theories regarding OH and ozone trends with respect to land use and global warming. Attention is given to the effects of the ENSO on the CH3CCl3 data and the assumption of continuing current industrial anthropogenic emissions. A novel tropical atmospheric tracer-transport mechanism is noted with respect to the CH3CCl3 data.

  17. Learning Chemistry from Good and (Why Not?) Problematic Results: Kinetics of the pH-Independent Hydrolysis of 4-Nitrophenyl Chloroformate

    ERIC Educational Resources Information Center

    El Seoud, Omar A.; Galgano, Paula D.; Are^as, Elizabeth P. G.; Moraes, Jamille M.

    2015-01-01

    The determination of kinetic data is central to reaction mechanism; science courses usually include experiments on chemical kinetics. Thanks to PC-controlled data acquisition and availability of software, the students calculate rate constants, whether the experiment has been done properly or not. This contrasts with their experience in, e.g.,…

  18. CHLOROFORM COMETABOLISM BY BUTANE-GROWN CF8, PSEUDOMONAS BUTANOVORA, MYCOBACTERIUM VACCAE JOB 5, AND METHANE-GROWN METHYLOSINUS TRICHOSPORIUM OB3B. (R825689C019)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  19. Regulation of Breast Carcinoma Growth and Neovacularization by Novel Peptide Sequences In Thromospondin

    DTIC Science & Technology

    1996-10-01

    were also synthesized using fmoc chemistry. Peptides were analyzed by reverse phase HPLC chromatography or gel permeation using a Superdex 75 HR 10...gel filtration (Fig. 15B). The major peak of the peptide comigrated with the purified monomer fraction from peptide 529 (Fig. 15C). Use of fmoc ...tBOC chemistry (Panel A), by tBOC chemistry using N-a-BOC-N-formyl-D-tryptophan (Panel B), or by FMOC chemistry (Panel D). Peptides were analyzed by

  20. Amino acid analysis by high-performance liquid chromatography after derivatization with 9-fluorenylmethyloxycarbonyl chloride Literature overview and further study.

    PubMed

    Jámbor, A; Molnár-Perl, I

    2009-04-10

    A literature overview is given of HPLC methods currently in use to determine amino acids as their 9-fluorenylmethyloxycarbonyl (FMOC) derivatives. On the basis of the detailed literature overview an exhaustive derivatization study was performed with 22 amino acids, applying photodiode array (DAD) and fluorescence (FL) detection simultaneously, in order to clear up the controversial points of FMOC derivatization. Model investigations have been carried out as a function of the reaction time and reaction conditions, such as the molar concentration of the reagent, the molar ratios of the reactants, the pH and the solvent composition of the reaction medium. Special emphasis was put (i) on the evaluation of the blank values of the reagents, as a function of the composition and that of the pH of the reaction medium, (ii) on the unambiguous quantitation of all amino acids, including the less reactive aspartic and glutamic acids, as well as on the formation and transformation of histidine and tyrosine, existing partly, as single (N-FMOC-histidine, N-FMOC-tyrosine), partly as double labeled species (N,NH-FMOC-histidine, N,O-FMOC-tyrosine). Reproducibilities of 22 amino acids, including both histidine and tyrosine derivatives, obtained under optimum derivatization conditions are presented (at 0.5mM FMOC concentration corresponding to the molar ratios of [FMOC]/[amino acids](T)=5.5/1 (note: the superscript 'T' means the total of amino acids), with acetonitrile containing reagents, at pH 9, derivatization time=20 min), and characterized with the relative standard deviation percentages of their responses (FMOC derivatives proved to be 1 pmol, except for serine, glycine, valine (2.5 pmol), for cystine, N,NH-FMOC-histidine and for the total of N-FMOC-tyrosine and N,O-FMOC-tyrosine (5 pmol) and for tryptophan (10 pmol).

  1. PSA Prodrug-Based Multimodality Agents for Imaging Metastatic Prostate Cancer

    DTIC Science & Technology

    2010-04-01

    Fmoc 2. piperidine 33 R = LQYQSKG-Mu 32 R = Leu 17 lepidine 29 27 30 31 Scheme 2. Notice that...Scheme 2. N N (CH2)nCH3 N N O NH 125I m R R = LQYQSKG-Mu or R = Leu 35 Scheme 3. 36 Fmoc Fmoc Fmoc Progress on this route is discussed below and...unable to prepare compound 27 but have prepared related compound 46 as shown in Scheme 5 but 46 was unreactive with lepidine. N OH Cl O O O O H2N

  2. Beta-Amino Acid Analogs of an Insect Neuropeptide Feature Potent Bioactivity and Resistance to Peptidase Hydrolysis

    DTIC Science & Technology

    2006-01-01

    AND METHODS Peptide Synthesis Insect kinin analogs were synthesized via Fmoc methodology on Rink Amide resin (Novabiochem, San Diego, CA) using Fmoc ...500–1,900) 100 a 95% confidence limit ( CL ) values in parentheses.10 b-Amino Acid Analogs of an Insect Neuropeptide 79 Biopolymers (Peptide Science) DOI

  3. Occurrence of the herbicide glyphosate and its metabolite AMPA in surface waters in Switzerland determined with on-line solid phase extraction LC-MS/MS.

    PubMed

    Poiger, Thomas; Buerge, Ignaz J; Bächli, Astrid; Müller, Markus D; Balmer, Marianne E

    2017-01-01

    Glyphosate is currently one of the most important herbicides worldwide. Its unique properties provide for a wide range of uses in agriculture but also in non-agricultural areas. At the same time, its zwitterionic nature prevents the inclusion in multi-residue analytical methods for environmental monitoring. Consequently, despite its extensive use, data on occurrence of glyphosate in the aquatic environment is still scarce. Based on existing methods, we developed a simplified procedure for the determination of glyphosate and its main metabolite aminomethylphosphonic acid (AMPA) in water samples using derivatization with fluorenylmethyl chloroformate FMOC-Cl, combined with on-line solid phase extraction and liquid chromatography-tandem mass spectrometry (LC-MS/MS) detection. This method was extensively tested on over 1000 samples of surface water, groundwater, and treated wastewater and proved to be simple, sensitive, and reliable. Limits of quantification of 0.005 μg/L were routinely achieved. Glyphosate and AMPA were detected in the vast majority of stream water samples in the area of Zurich, Switzerland, with median concentrations of 0.11 and 0.20 μg/L and 95th percentile concentrations of 2.1 and 2.6 μg/L, respectively. Stream water data and data from treated wastewater indicated that non-agricultural uses may significantly contribute to the overall loads of glyphosate and AMPA in surface waters. In the investigated groundwater samples, selected specifically because they had shown presence of other herbicides in previous monitoring programs, glyphosate and AMPA were generally not detected, except for two monitoring sites in Karst aquifers, indicating that these compounds show much less tendency for leaching.

  4. Adsorption on molecularly imprinted polymers of structural analogues of a template. Single-component adsorption isotherm data

    SciTech Connect

    Kim, Hyunjung; Guiochon, Georges A

    2005-10-01

    The equilibrium adsorption isotherms on two otherwise identical polymers, one imprinted with Fmoc-L-tryptophan (Fmoc-L-Trp) (MIP), the other nonimprinted (NIP), of compounds that are structural analogues of the template were acquired by frontal analysis (FA) in an acetonitrile/acetic acid (99/1 v/v) mobile phase, over a wide concentration range (from 0.005 to 50 mM). These analogues were Fmoc-L-tyrosine, Fmoc-L-serine, Fmoc-L-phenyalanine, Fmoc-glycine (Fmoc-Gly), Fmoc-L-tryptophan pentafluorophenyl ester (Fmoc-L-Trp(OPfp)), and their antipodes. These substrates have different numbers of functional groups able to interact with the 4-vinylpyridine groups of the polymer. For a given number of the functional groups, these substrates have different hydrophobicities of their side groups (as indicated by their partition coefficients (log P{sub ow}) in the octanol-water system (e.g., from 4.74 for Fmoc-Trp to 2.53 for Fmoc-Gly)). Statistical results from the fitting of the FA data to Langmuirian isotherm models, the calculation of the affinity energy distribution, and the comparison of calculated and experimental band profiles show that all these sets of FA data are best accounted for by a tri-Langmuir isotherm model, except for the data of Fmoc-L-Trp(OPfp) that are best modeled by a simple Langmuir isotherm. So, all compounds but Fmoc-L-Trp(OPfp) find three different types of adsorption sites on both the MIP and the NIP. The properties of these different types of sites were studied systematically. The results show that the affinity of the structural analogues for the NIP is controlled mostly by the number of the functional groups on the substrates and somewhat by the hydrophobicity of their side groups. These two factors control also the MIP affinity toward the enantiomers of the structural analogues that have a stereochemistry different from that of the template. In contrast, the affinity of the highest affinity sites of the MIP toward the enantiomers of these

  5. Calcium-Ion-Triggered Co-assembly of Peptide and Polysaccharide into a Hybrid Hydrogel for Drug Delivery

    NASA Astrophysics Data System (ADS)

    Xie, Yanyan; Zhao, Jun; Huang, Renliang; Qi, Wei; Wang, Yuefei; Su, Rongxin; He, Zhimin

    2016-04-01

    We report a new approach to constructing a peptide-polysaccharide hybrid hydrogel via the calcium-ion-triggered co-assembly of fluorenylmethyloxycarbonyl-diphenylalanine (Fmoc-FF) peptide and alginate. Calcium ions triggered the self-assembly of Fmoc-FF peptide into nanofibers with diameter of about 30 nm. Meanwhile, alginate was rapidly crosslinked by the calcium ions, leading to the formation of stable hybrid hydrogel beads. Compared to alginate or Fmoc-FF hydrogel alone, the hybrid Fmoc-FF/alginate hydrogel had much better stability in both water and a phosphate-buffered solution (PBS), probably because of the synergistic effect of noncovalent and ionic interactions. Furthermore, docetaxel was chosen as a drug model, and it was encapsulated by hydrogel beads to study the in vitro release behavior. The sustained and controlled docetaxel release was obtained by varying the concentration ratio between Fmoc-FF peptide and alginate.

  6. Evolving nanomaterials using enzyme-driven dynamic peptide libraries (eDPL).

    PubMed

    Das, Apurba K; Hirsth, Andrew R; Ulijn, Rein V

    2009-01-01

    This paper describes the application of dynamic combinatorial libraries (DCL) towards the discovery of self-assembling nanostructures based on aromatic peptide derivatives and the continuous enzymatic exchange of amino acid sequences. Ultimately, the most thermodynamically stable self-assembling structures will dominate the system. In this respect, a library of precursor components, based on N-fluorenyl-9-methoxycarbonyl (Fmoc)-amino acids (serine, S and threonine, T) and nucleophiles (leucine, L-; phenylalanine, F-; tyrosine, Y-; valine, V-; glycine, G-; alanine, A-OMe amino-acid esters) were investigated to produce Fmoc-dipeptide esters, denoted Fmoc-XY-OMe. Upon exposure to a protease (thermolysin), which catalyses peptide bond formation and hydrolysis under aqueous conditions at pH 8, dynamic libraries of self-assembling gelator species were generated. Depending on the molecular composition of the precursors present in the library different behaviours were observed. Single components, Fmoc-SF-OMe and Fmoc-TF-OMe, dominated over time in Fmoc-S/(L+F+Y+V+G+A)-OMe and Fmoc-T/ (L+F+Y+V+G+A)-OMe libraries. This represented > 80% of all peptide formed suggesting that a single component molecular structure dominates in these systems. In a competition experiment between Fmoc-(S+T)/F-OMe, conversions to each peptide corresponded directly with ratios of starting materials, implying that a bi-component nanostructure, where Fmoc-TF-OMe and Fmoc-SF-OMe are incorporated equally favourably, was formed. Several techniques including HPLC, LCMS and fluorescence spectroscopy were used to characterize library composition and molecular interactions within the self-selecting libraries. Fluorescence spectroscopy analysis suggests that the most stable peptide nanostructures show significant pi-pi intermolecular electronic communication. Overall, the paper demonstrates a novel evolution-based approach with self-selection and amplification of supramolecular peptide nanostructures from a

  7. Solvent extraction of silver picrate by 3m-crown-m ethers (m = 5, 6) and its mono-benzo-derivative from water into benzene or chloroform: elucidation of an extraction equilibrium using component equilibrium constants.

    PubMed

    Kudo, Yoshihiro; Usami, Jun; Katsuta, Shoichi; Takeda, Yasuyuki

    2004-03-10

    Ion-pair formation constant (K(AgPic) in mol(-1)dm(3)) of silver picrate (AgPic), those (K(AgLPic)) of its ion-pair complexes (AgLPic) with crown ethers (L) and complex formation constants (K(AgL)) of Ag(+) with L (15-crown-5 ether (15C5) and benzo-15C5) in water (w) were determined potentiometrically at 25 degrees C. Compounds used as L were 18-crown-6 ether (18C6), its benzo-derivative (B18C6) and the two 15C5 derivatives. Extraction constants (K(ex) in mol(-1)dm(3)) of AgPic with L (15C5, 18C6, B18C6) from acidic w-phases into either C(6)H(6) or CHCl(3) were recalculated from K(AgPic), K(AgL), K(AgLPic) and data opened in previous papers. Thus obtained K(ex) was divided into five component equilibrium constants containing K(AgL) and K(AgLPic) anew. Then, contributions of the component constants, K(AgL), K(AgLPic) and distribution constants of AgLPic between the w- and C(6)H(6)-phases, to K(ex) were discussed and compared with corresponding extraction systems of NaPic and KPic with18C6.

  8. EFFECT OF CHLOROFORM ON DICHLOROACETIC ACID AND TRICHLOROACETIC ACID-INDUCED HYPOMETHYLATION AND EXPRESSION OF THE C-MYC GENE AND ON THEIR PROMOTION OF LIVER AND KIDNEY TUMORS IN MICE. (R825384)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  9. EFFECT OF CHLOROFORM ON DICHLOROACETIC ACID AND TRICHLOROACETIC ACID-INDUCED HYPOMETHYLATION AND EXPRESSION OF THE C-MYC GENE AND ON THEIR PROMOTION OF LIVER AND KIDNEY TUMORS IN MICE. (R828083)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  10. Solvation properties of N-substituted cis and trans amides are not identical: significant enthalpy and entropy changes are revealed by the use of variable temperature 1H NMR in aqueous and chloroform solutions and ab initio calculations.

    PubMed

    Troganis, Anastassios N; Sicilia, Emilia; Barbarossou, Klimentini; Gerothanassis, Ioannis P; Russo, Nino

    2005-12-29

    The cis/trans conformational equilibrium of N-methyl formamide (NMF) and the sterically hindered tert-butylformamide (TBF) was investigated by the use of variable temperature gradient 1H NMR in aqueous solution and in the low dielectric constant and solvation ability solvent CDCl3 and various levels of first principles calculations. The trans isomer of NMF in aqueous solution is enthalpically favored relative to the cis (deltaH(o) = -5.79 +/- 0.18 kJ mol(-1)) with entropy differences at 298 K (298 x deltaS(o) = -0.23 +/- 0.17 kJ mol(-1)) playing a minor role. The experimental value of the enthalpy difference strongly decreases (deltaH(o) = -1.72 +/- 0.06 kJ mol(-1)), and the contribution of entropy at 298 K (298 x deltaS(o) = -1.87 +/- 0.06 kJ mol(-1)) increases in the case of the sterically hindered tert-butylformamide. The trans isomer of NMF in CDCl3 solution is enthalpically favored relative to the cis (deltaH(o) = -3.71 +/- 0.17 kJ mol(-1)) with entropy differences at 298 K (298 x deltaS(o) = 1.02 +/- 0.19 kJ mol(-1)) playing a minor role. In the sterically hindered tert-butylformamide, the trans isomer is enthalpically disfavored (deltaH(o) = 1.60 +/- 0.09 kJ mol(-1)) but is entropically favored (298 x deltaS(o) = 1.71 +/- 0.10 kJ mol(-1)). The results are compared with literature data of model peptides. It is concluded that, in amide bonds at 298 K and in the absence of strongly stabilizing sequence-specific inter-residue interactions involving side chains, the free energy difference of the cis/trans isomers and both the enthalpy and entropy contributions are strongly dependent on the N-alkyl substitution and the solvent. The significant decreasing enthalpic benefit of the trans isomer in CDCl3 compared to that in H2O, in the case of NMF and TBF, is partially offset by an adverse entropy contribution. This is in agreement with the general phenomenon of enthalpy versus entropy compensation. B3LY/6-311++G** and MP2/6-311++G** quantum chemical calculations confirm the stability orders of isomers and the deltaG decrease in going from water to CHCl3 as solvent. However, the absolute calculated values, especially for TBF, deviate significantly from the experimental values. Consideration of the solvent effects via the PCM approach on NMF x H2O and TBF x H2O supermolecules improves the agreement with the experimental results for TBF isomers, but not for NMF.

  11. Quantitation of amino acids in plasma by high performance liquid chromatography: simultaneous deproteinization and derivatization with 9-fluorenylmethyloxycarbonyl chloride.

    PubMed

    Jámbor, A; Molnár-Perl, I

    2009-08-21

    This paper, as a novelty to this field, presents the deproteinization and derivatization of plasma's free amino acids (PFAAs), simultaneously, in a single step, with the acetonitrile (ACN) containing 9-fluorenylmethyloxycarbonyl chloride (FMOC) reagent. Deproteinization and derivatization, were studied with 22 amino acids, applying photodiode array (DAD) and fluorescence (FL) detection, simultaneously. Model investigations have been carried out as a function of the FMOC concentration, reaction time and reaction conditions: with standard solutions, with human plasma samples in its initial condition and fortified with standard amino acids (excluding tryptophan because it co-elutes with the hydrolyzed FMOC). Reproducibilities of 22 amino acids, including both histidine and tyrosine derivatives, obtained under optimum derivatization conditions are presented (at 3.0 mM FMOC concentration, at pH 9; derivatization time - 20 min), and characterized with the relative standard deviation percentages of their responses (FMOC derivatives proved to be 2.5 pmol, except for cystine, ornithine (5 pmol) and for the total of tyrosines (N-FMOC-tyrosine and N,O-FMOC-tyrosine 10 pmol).

  12. Novel monodisperse PEGtide dendrons: design, fabrication and evaluation of mannose receptor-mediated macrophage targeting

    PubMed Central

    Gao, Jieming; Chen, Peiming; Singh, Yashveer; Zhang, Xiaoping; Szekely, Zoltan; Stein, Stanley; Sinko, Patrick J.

    2014-01-01

    Novel PEGtide dendrons of generation 1 through 5 (G1.0-5.0) containing alternating discrete poly(ethylene glycol) (dPEG) and amino acid/peptide moieties were designed and developed. To demonstrate their targeting utility as nanocarriers, PEGtide dendrons functionalized with mannose residues were developed and evaluated for macrophage targeting. PEGtide dendrons were synthesized using 9-fluorenylmethyloxycarbonyl (Fmoc) solid-phase peptide synthesis (SPPS) protocols. The N-α-Fmoc-N-ε-(5-carboxyfluorescein)-L-lysine (Fmoc-Lys(5-FAM)-OH) and monodisperse Fmoc-dPEG6-OH were sequentially coupled to Fmoc-β-Ala-resin to obtain the resin-bound intermediate Fmoc-dPEG6-Lys(5-FAM)-β-Ala (1). G1.0 dendrons were obtained by sequentially coupling Fmoc-Lys(Fmoc)-OH, Fmoc-β-Ala-OH, and Fmoc-dPEG6-OH to (1). Dendrons of higher generation, G2.0 to 5.0, were obtained by repeating the coupling cycles used for the synthesis of G1.0. Dendrons containing eight mannose residues (G3.0-Mannose8) were developed for mannose receptor (MR)-mediated macrophage targeting by conjugating α-D-mannopyranosylphenyl isothiocyanate to G3.0 dendrons. In the present study PEGtide dendrons up to G5.0 were synthesized. The molecular weights of the dendrons determined by MALDI-TOF were in agreement with calculated values. The hydrodynamic diameters measured using dynamic light scattering (DLS) ranged from 1 to 8 nm. Cell viability in the presence of G3.0 and G3.0-Mannose8 was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and was found to be statistically indistinguishable from that of untreated cells. G3.0-Mannose8 exhibited 12-fold higher uptake than unmodified G3.0 control dendrons in MR-expressing J774.E murine macrophage-like cells. Uptake was nearly completely inhibited in the presence of 10 mg/mL mannan, a mannose analog and known MR substrate. Confocal microscopy studies demonstrated the presence of significant intracellular punctate fluorescence

  13. An Improved D-α-Tocopherol-Based Nanocarrier for Targeted Delivery of Doxorubicin with Reversal of Multidrug Resistance

    PubMed Central

    Lu, Jianqin; Zhao, Wenchen; Liu, Hao; Marquez, Rebecca; Huang, Yixian; Zhang, Yifei; Li, Jiang; Xie, Wen; Venkataramanan, Raman; Xu, Liang; Li, Song

    2014-01-01

    Nanocarriers have recently emerged as an attractive platform for delivery of various types of therapeutics including anticancer agents. Previously, we developed an improved TPGS delivery system (PEG5K-VE2) which demonstrated improved colloidal stability and greater in vivo antitumor activity. Nevertheless, the application of this system is still limited by a relatively low drug loading capacity (DLC). In this study we report that incorporation of a fluorenylmethyloxycarbonyl (Fmoc) motif at the interfacial region of PEG5K-VE2 led to significant improvement of the system through the introduction of an additional mechanism of drug/carrier interaction. Doxorubicin (DOX) could be effectively loaded into PEG5K-Fmoc-VE2 micelles at a DLC of 39.9%, which compares favorably to most reported DOX nanoformulations. In addition, PEG5K-Fmoc-VE2/DOX mixed micelles showed more sustained release of DOX in comparison to the counterpart without Fmoc motif. MTT assay showed that PEG5K-Fmoc-VE2/DOX exerted significantly higher levels of cytotoxicity over DOX, Doxil as well as PEG5K-VE2/DOX in PC-3 and 4T1.2 cells. Cytotoxicity assay with NCI/ADR-RES, a drug resistant cell line, suggested that PEG5K-Fmoc-VE2 may have a potential to reverse the multidrug resistance, which was supported by its inhibition on P-gp ATPase. Pharmacokinetics (PK) and biodistribution studies showed an increased half-life in blood circulation and more effective tumor accumulation for DOX formulated in PEG5K-Fmoc-VE2 micelles. More importantly, DOX-loaded PEG5K-Fmoc-VE2 micelles showed an excellent safety profile with a MTD (~30 mg DOX/kg) that is about 3 times as much as that for free DOX. Finally, superior antitumor activity was demonstrated for PEG5K-Fmoc-VE2/DOX in both drug-sensitive (4T1.2 and PC-3) and drug-resistant (KB 8-5) tumor models compared to DOX, Doxil, and PEG5K-VE2/DOX. PMID:25456831

  14. New concept for long-acting insulin: spontaneous conversion of an inactive modified insulin to the active hormone in circulation: 9-fluorenylmethoxycarbonyl derivative of insulin.

    PubMed

    Gershonov, E; Shechter, Y; Fridkin, M

    1999-07-01

    Insulin is a short-lived species in the circulatory system. After binding to its receptor sites and transmission of its biological signals, bound insulin undergoes receptor-mediated endocytosis and consequent degradation. An inactive insulin derivative that is not recognized by the receptor has a longer circulation life, but obviously is biologically impotent. (Fmoc)2 insulin is an insulin derivative purified through high-performance liquid chromatography in which two 9-fluorenylmethoxycarbonyl (Fmoc) moieties are covalently linked to the (alpha-amino group of phenylalanine B1 and the epsilon-amino group of lysine B29. It has 1-2% of the biological potency and receptor binding capacity of the native hormone. After incubation, (Fmoc)2 insulin undergoes a time-dependent spontaneous conversion to fully active insulin in aqueous solution at 37 degrees C and a pH range of 7-8.5. At pH 7.4, the conversion proceeds slowly (t1/2 = 12 +/- 1 days) and biological activity is generated gradually. A single subcutaneous administration of (Fmoc)2 insulin to streptozocin-treated diabetic rats normalized their blood glucose levels and maintained the animals in an anabolic state over 2-3 days. A broad shallow peak of immunoreactive insulin was found to persist in circulation over this period. To confirm further that the long-acting effect of (Fmoc)2 insulin proceeds via slow release in the blood circulation itself, we administered native insulin, NPH insulin, or the (Fmoc)2 derivative intraperitoneally. The rats recovered from hypoglycemia at t1/2 = 8.0 +/- 0.3 and 10 +/- 0.4 h after administration of native and NPH insulin, respectively. In contrast, (Fmoc)2 insulin was active for a significantly longer time, with an extended onset of t1/2 = 26 +/- 1h, and a glucose-lowering effect even 40 h after administration. (Fmoc)2 insulin was also found to be more resistant to proteolysis. Finally, we found that (Fmoc)2 insulin does not induce antigenic effects. In summary, we present here a

  15. Anthelmintic activity of leaves of justicia beddomei.

    PubMed

    Srinivasa, U; Rao, J Venkateshwara; Krupanidhi, A M; Shanmukhappa, S

    2007-01-01

    Ethanolic and Chloroform extract of leaves of Justicia beddomei were evaluated separately for anthelmintic activity on adult Indian earthworms Pheretima posthuma, using Piperazine citrate as reference standard. The results indicated that ethanolic extract was more potent than the chloroform extract.

  16. Safety Case for Service Contracts

    SciTech Connect

    Martinez, Israel L.

    2014-07-01

    Safety cases developed for the Facilities Management and Operations Center (FMOC) are based on the requirements in MN471021, Work Planning and Control Criteria for Safe Design and Operations. The FMOC performs maintenance activities in various locations at Sandia National Laboratories, New Mexico (SNL/NM). SNL/NM consists of more than 6,000,000 square feet of buildings, structures, and site infrastructure on approximately 13,000 acres of land. The FMOC performs approximately 7500 service contract work orders a year to assist with operations and maintenance at the SNL/NM site and facilities. As part of the continual improvement process, this Safety Case will be reviewed and updated, as needed, or at a minimum every three years.

  17. Single-step analysis of low abundance phosphoamino acids via on-line sample preconcentration with chemical derivatization by capillary electrophoresis.

    PubMed

    Ptolemy, Adam S; Britz-McKibbin, Philip

    2005-09-01

    New strategies for rapid, sensitive and high-throughput analysis of low abundance metabolites in biological samples are required for future metabolomic research. In this report, a direct method for sub-micromolar analyses of phosphoamino acids was developed using on-line sample preconcentration with 9-fluorenylmethyloxycarbonyl chloride (FMOC) derivatization by capillary electrophoresis (CE) and UV detection. Analyte focusing by dynamic pH junction and FMOC labeling efficiency were influenced by several experimental factors including buffer pH, ionic strength, sample injection length and FMOC concentration. About a 200-fold enhancement in concentration sensitivity was achieved under optimal conditions relative to conventional off-line derivatization, as reflected by a detection limit (S/N approximately 3) of 0.1 microM. In-capillary sample preconcentration with chemical labeling by CE offers a unique single-step analytical platform for high-throughput screening of low abundance metabolites without intrinsic chromophores.

  18. Application of 2-chlorotrityl resin in solid phase synthesis of (Leu15)-gastrin I and unsulfated cholecystokinin octapeptide. Selective O-deprotection of tyrosine.

    PubMed

    Barlos, K; Gatos, D; Kapolos, S; Poulos, C; Schäfer, W; Yao, W Q

    1991-12-01

    The carboxyl terminal dipeptide amide, Fmoc-Asp-Phe-NH2, of gastrin and cholecystokinin (CCK) has been attached in high yield through its free side chain carboxyl group to the acid labile 2-chlorotrityl resin. The obtained peptide resin ester has been applied in the solid phase synthesis of partially protected (Leu15)-gastrin I utilising Fmoc-amino acids. Quantitative cleavage of this peptide from resin, with the t-butyl type side chain protection intact is achieved using mixtures of acetic acid/trifluoroethanol/dichloromethane. Under the same conditions complete detritylation of the tyrosine phenoxy function occurs simultaneously. Thus, the solid-phase synthesis of peptides selectively deprotected at the side chain of tyrosine is rendered possible by the use of 2-chlorotrityl resin and Fmoc-Tyr(Trt)-OH. The efficiency of this approach has been proved by the subsequent high-yield synthesis of three model peptides and the CCK-octapeptide.

  19. A Phosphorylated Prodrug for the Inhibition of Pin1

    PubMed Central

    Zhao, Song; Etzkorn, Felicia A.

    2008-01-01

    Fmoc-pSer-Ψ[(Z)CH=C]-Pro-(2)-N-(3)-ethylaminoindole 1, showed moderate inhibition towards the mitotic regulator, Pin1 (IC50 = 28.3μM). To improve the cell permeability, the charged phosphate was masked as the bis-pivaloyloxymethyl (POM) phosphate in Fmoc-(bisPOM)-pSer-Ψ[(Z)CH=C]-Pro-(2)-N-(3)-ethylaminoindole 2. Antiproliferative activity towards A2780 ovarian cancer cells of 1 (IC50 = 46.2 μM) was improved significantly in 2 (IC50 = 26.9 μM), comparable to the IC50 of 1 towards Pin1 enzymatic activity. PMID:17935986

  20. Synthesis of Peptides Containing C-Terminal Methyl Esters Using Trityl Side-Chain Anchoring: Application to the Synthesis of a-Factor and a-Factor Analogs

    PubMed Central

    Diaz-Rodriguez, Veronica; Mullen, Daniel G.; Ganusova, Elena; Becker, Jeffrey M.; Distefano, Mark D.

    2012-01-01

    A new cysteine anchoring method was developed for the synthesis of peptides containing C-terminal cysteine methyl esters. This method consists of attachment of Fmoc-Cys-OCH3 to either 2-ClTrt-Cl or Trt-Cl resins (via the side-chain thiol) followed by preparation of the desired peptide using Fmoc-based SPPS. We applied this method to the synthesis of the mating pheromone a-factor and a 5-FAM labeled a-factor analog. The peptides were obtained with high yield and purity and were shown to be bioactive in a growth arrest assay. PMID:23121562

  1. Development of orthogonally protected hypusine for solid-phase peptide synthesis.

    PubMed

    Song, Aimin; Tom, Jeffrey; Yu, Zhiyong; Pham, Victoria; Tan, Dajin; Zhang, Dengxiong; Fang, Guoyong; Yu, Tao; Deshayes, Kurt

    2015-04-03

    An orthogonally protected hypusine reagent was developed for solid-phase synthesis of hypusinated peptides using the Fmoc/t-Bu protection strategy. The reagent was synthesized in an overall yield of 27% after seven steps from Cbz-Lys-OBzl and (R)-3-hydroxypyrrolidin-2-one. The side-chain protecting groups (Boc and t-Bu) are fully compatible with standard Fmoc chemistry and can be readily removed during the peptide cleavage step. The utility of the reagent was demonstrated by solid-phase synthesis of hypusinated peptides.

  2. Synthesis of peptides containing C-terminal methyl esters using trityl side-chain anchoring: application to the synthesis of a-factor and a-factor analogs.

    PubMed

    Diaz-Rodriguez, Veronica; Mullen, Daniel G; Ganusova, Elena; Becker, Jeffrey M; Distefano, Mark D

    2012-11-16

    A new cysteine anchoring method was developed for the synthesis of peptides containing C-terminal cysteine methyl esters. This method consists of attachment of Fmoc-Cys-OCH(3) to either 2-ClTrt-Cl or Trt-Cl resins (via the side-chain thiol) followed by preparation of the desired peptide using Fmoc-based SPPS. We applied this method to the synthesis of the mating pheromone a-factor and a 5-FAM labeled a-factor analog. The peptides were obtained with high yield and purity and were shown to be bioactive in a growth arrest assay.

  3. Rapid solid-phase peptide synthesis using thermal and controlled microwave irradiation.

    PubMed

    Bacsa, Bernadett; Desai, Bimbisar; Dibó, Gábor; Kappe, C Oliver

    2006-10-01

    A rapid and efficient microwave-assisted solid-phase synthesis method is described for the preparation of the nonapeptide WDTVRISFK, using conventional Fmoc/Bu(t) orthogonal protection strategy. The synthesis protocol is based on the use of cycles of pulsed microwave irradiation with intermittent cooling of the reaction during the removal of the Fmoc protecting group and during the coupling. The desired nonapeptide was obtained in highest yield and purity by employing MicroKan technology. The chemical reactions were carried out in a single-mode microwave reactor, equipped with a fiber-optic probe to monitor the reaction temperature continuously.

  4. Synthesis of a Homologous Series of Side Chain Extended Orthogonally-Protected Aminooxy-Containing Amino Acids

    PubMed Central

    Liu, Fa; Thomas, Joshua; Burke, Terrence R.

    2008-01-01

    Practical methodology is reported for the synthesis of a homologous series of side chain extended amino acids containing aminooxy functionality bearing orthogonal protection suitable for Fmoc peptide synthesis. These reagents may be useful for the preparation of libraries containing fragments joined by peptide linkers. PMID:19122755

  5. Guanidino Groups Greatly Enhance the Action of Antimicrobial Peptidomimetics Against Bacterial Cytoplasmic Membranes

    DTIC Science & Technology

    2014-05-28

    Chen, R.J. Doerksen, J. Kaplan, P.J. Carroll , M.L. Klein, W.F. DeGrado, De novo design of biomimetic antimicrobial polymers, Proc. Natl. Acad. Sci. U. S...J.J. Diaz-Mochon, L. Bialy, M. Bradley , Full orthogonality between Dde and Fmoc: the direct synthesis of PNA–peptide conjugates, Org. Lett. 6 (2004

  6. Thermodynamic analysis of the heterogeneous binding sites of molecularly imprinted polymers

    SciTech Connect

    Kim, Hyunjung; Kaczmarski, Krzysztof; Guiochon, Georges A

    2005-11-01

    The thermodynamic interactions of two polymers, one Fmoc-L-Trp-imprinted (MIP), the other one an unimprinted reference (NIP), with the two Fmoc-tryptophan enantiomers were studied by frontal analysis, which allows accurate measurements of the adsorption isotherms. These isotherms were acquired at temperatures of 40, 50, 60, and 70 C, for sample concentrations ranging between 0.005 and 40 mM. The mobile phase used was acetonitrile with one percent acetic acid as an organic modifier. Within the measured concentration ranges, the tri-Langmuir isotherm model accounts best for the isotherm data of both enantiomers on the MIP, the bi-Langmuir model for the isotherm data of Fmoc-L-Trp on the NIP. These isotherm models were selected using three independent processes: statistical tests on the results from regression of the isotherm data to different isotherm models; calculation of the affinity energy distribution from the raw isotherm data; comparison of the experimental and the calculated band profiles. The isotherm parameters obtained from these best selected isotherm models showed that the enantiomeric selectivity does not change significantly with temperature, while the affinity of the substrates for both the MIP and the NIP decrease considerably with increasing temperatures. These temperature effects on the binding performance of the MIP were clarified by considering the thermodynamic functions (i.e., the standard molar Gibbs free energy, the standard molar entropy of adsorption, and the standard molar enthalpy of adsorption) for each identified type of adsorption sites, derived from the Van't Hoff equation. This showed that the entropy of transfer of Fmoc-L-Trp from the mobile to the MIP stationary phase is the dominant driving force for the selective adsorption of Fmoc-L-Trp onto the enantioselective binding sites. This entropy does not change significantly with increasing temperatures from 40 to 70 C.

  7. Lipid Removal from Human Serum Samples

    PubMed Central

    Castro, Arnold R.; Morrill, William E.; Pope, Victoria

    2000-01-01

    The efficacy of lipid removal from human serum samples obtained by using Cleanascite HC, a commercially available product, was compared to that obtained by the standard chloroform method. Separate samples of 21 frozen, banked human serum samples used in the preparation of samples for proficiency testing were treated with either Cleanascite HC or chloroform. The lipid content was measured before and after treatment. The total percentages of lipid removed ranged from 61 to 70% with Cleanascite HC and from 60 to 62% with chloroform. The advantage of Cleanascite HC over chloroform is based on the simplicity of the procedure with Cleanascite HC without the environmental concerns inherent in the use of chloroform. In 15 serum samples known to contain antibodies to treponemal and nontreponemal syphilis antigens, Cleanascite HC bound some immunoglobulin, but with only minimal loss of reactivity in the serologic tests for syphilis. Cleanascite HC is therefore an acceptable alternative to chloroform for lipid reduction in human serum samples. PMID:10702492

  8. [The first labor analgesia with drug was performed in late Meiji Period (1868-1912). Hypnosis also attracted attention as a method of labor analgesia in mid-Meiji Period].

    PubMed

    Okutomi, Toshiyuki

    2013-11-01

    Ether or chloroform, was in use for ambulatory surgery after 1861 in Japan. An inhalational anesthetic, especially chloroform, was administered for cesarean section in early Meiji Period (from 1868) up to 1897. According to an article in 1903, chloroform was recommended as a strategy for internal cephalic version. However, it is uncertain whether inhalational anesthetic had been utilized for vaginal deliveries before 1903. There is evidence that hypnosis had attracted attention as a method of labor analgesia around that time.

  9. New Non-Linear Optical Polymers

    DTIC Science & Technology

    1990-08-01

    filtration . Subsequent recrystallization from chloroform raised Tm to 99 -1000C. Dilute chloroform solution of II was passed through a silica gel permeation...were checked with Sadtler NMR Library. Furthermore, gel permeation chromotography was done with all dinitro compounds to ascertain their purity. Polymer...106’C. A dilute chloroform solution of I was passed through a silica gel permeation column. Only one peak, corresponding to I, was detected during a 15

  10. LIVER INJURY, LIVER PROTECTION, AND SULFUR METABOLISM

    PubMed Central

    Miller, L. L.; Whipple, G. H.

    1942-01-01

    Protein-depleted dogs are very susceptible to injurious agents—in particular, chloroform. Methionine given shortly before chloroform anesthesia will give complete protection against chloroform. Methionine (or cysteine plus choline) given 3 or 4 hours after chloroform anesthesia will give significant protection against the liver injury of chloroform anesthesia. Methionine given more than 4 hours after chloroform anesthesia gives no protection against liver injury. Choline alone given before chloroform gives no protection against liver injury. The protein-depleted dogs have livers which are deficient in both nitrogen and sulfur, but sulfur is depleted more than is the nitrogen. The N/S ratio therefore rises. Methionine or cystine feeding promptly makes up this liver sulfur deficit. Viable liver cells are necessary for this uptake of sulfur. Livers of fetuses in utero or of newborn pups tolerate a chloroform anesthesia which will cause fatal liver injury in adults. The nitrogen and sulfur values of these fetus or pup livers are within the high normal values for adults. Blood-forming cells are present in the fetus or pup livers during this period. When these blood islands are eliminated during the 3rd or 4th week of life, the liver then becomes normally susceptible to chloroform liver injury. Methionine or methionine-rich protein digests (e.g. casein) or various proteins by mouth or by vein should prove useful to protect the liver against certain types of injury and to aid in organ repair. PMID:19871248

  11. 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.

  12. Evaluating non-chlorinated solvents for welding applications

    SciTech Connect

    Mustaleski, T.M. Jr.

    1993-08-11

    There is interest in eliminating the use of chlorinated solvents such as methyl chloroform at the Oak Ridge Y-12 Plant for environmental reasons. Solvent 140 has been offered as an acceptable replacement. Methyl chloroform has frequently been used for the final cleaning of materials just prior to welding. Electron beam welds were made in an aluminum alloy to compare the potential contamination effect of Solvent 140 to that of methyl chloroform. Tests indicated that the Solvent 140 did not have an adverse effect on pumpdown time of electron beam welding equipment during normal handling. Solvent 140 resulted in significantly less weld porosity than; methyl chloroform in this test.

  13. Evaluating non-chlorinated solvents for welding applications

    NASA Astrophysics Data System (ADS)

    Mustaleski, T. M., Jr.

    1993-08-01

    There is interest in eliminating the use of chlorinated solvents such as methyl chloroform at the Oak Ridge Y-12 Plant for environmental reasons. Solvent 140 has been offered as an acceptable replacement. Methyl chloroform has frequently been used for the final cleaning of materials just prior to welding. Electron beam welds were made in an aluminum alloy to compare the potential contamination effect of Solvent 140 to that of methyl chloroform. Tests indicated that the Solvent 140 did not have an adverse effect on pump-down time of electron beam welding equipment during normal handling. Solvent 140 resulted in significantly less weld porosity than methyl chloroform in this test.

  14. MODELING VOLATILE ORGANIC COMPOUND PHARMACOKINETICS IN RAT PUPS

    EPA Science Inventory

    PBPK model predictions of internal dosimetry in young rats were compared to adult animals for benzene, chloroform (CHL), methylene chloride, methyl ethly ketone (MEK), perchloroethylene, and trichloroethylene.

  15. Antimicrobial activity of extracts of Terminalia catappa root.

    PubMed

    Pawar, S P; Pal, S C

    2002-06-01

    The effect against bacteria of petroleum ether (60-80 degrees C), chloroform and methanolic extract of dried root of Terminalia catappa Linn. (combrataceae) was employed by cup plate agar diffusion method. The chloroform extract showed prominent antimicrobial activity against S. aureus and E. coli as compared to other tested microorganisms, while petroleum ether extract was devoid of antimicrobial activity. The methanolic: extract exhibited MIC of 0.065 mg/ml against E. coli. and chloroform extract exhibited MIC of 0.4 mg/ml against S. aureus The chloroform has well as methanolic extracts showed good antimicrobial activity against Gram positive and Gram negative microorganisms.

  16. Assessing the utility of infrared spectroscopy as a structural diagnostic tool for β-sheets in self-assembling aromatic peptide amphiphiles.

    PubMed

    Fleming, Scott; Frederix, Pim W J M; Ramos Sasselli, Iván; Hunt, Neil T; Ulijn, Rein V; Tuttle, Tell

    2013-07-30

    β-Sheets are a commonly found structural motif in self-assembling aromatic peptide amphiphiles, and their characteristic "amide I" infrared (IR) absorption bands are routinely used to support the formation of supramolecular structure. In this paper, we assess the utility of IR spectroscopy as a structural diagnostic tool for this class of self-assembling systems. Using 9-fluorene-methyloxycarbonyl dialanine (Fmoc-AA) and the analogous 9-fluorene-methylcarbonyl dialanine (Fmc-AA) as examples, we show that the origin of the band around 1680-1695 cm(-1) in Fourier transform infrared (FTIR) spectra, which was previously assigned to an antiparallel β-sheet conformation, is in fact absorption of the stacked carbamate group in Fmoc-peptides. IR spectra from (13)C-labeled samples support our conclusions. In addition, DFT frequency calculations on small stacks of aromatic peptides help to rationalize these results in terms of the individual vibrational modes.

  17. Size reduction of CdSe/ZnS quantum dots by a peptidic amyloid supergelator.

    PubMed

    Zaman, Md Badruz; Bardelang, David; Prakesch, Michaël; Leek, Donald M; Naubron, Jean-Valère; Chan, Gordon; Wu, Xiaohua; Ripmeester, John A; Ratcliffe, Christopher I; Yu, Kui

    2012-03-01

    Anchoring of a self-assembling dipeptide on the surface of core/shell CdSe/ZnS quantum dots resulted in a competition between coordination of the surface atoms of the QDs and the strong tendency for the dipeptide to self-assemble in toluene. This resulted in a mild QD etching and in a corresponding increase in the band gap of the nanocrystals whose photoluminescent emission gradually turns blue with time. The FmocLeuLeuOH dipeptide supergelator self-assembles in fibrils in which the Fmoc groups are surrounded by the pendant isobutyl side chains of the leucine residues with vibrational circular dichroism (VCD) and liquid- and solid-state NMR attributes of twist anti-parallel β-sheets.

  18. Isotherm parameters and intraparticle mass transfer kinetics on molecularly imprinted polymers in acetonitrile/buffer mobile phases

    SciTech Connect

    Kim, Hyunjung; Kaczmarski, Krzysztof; Guiochon, Georges A

    2006-03-01

    The equilibrium isotherm and the intraparticle mass transfer kinetics of the enantiomers of the template were investigated on an Fmoc-L-tryptophan (Fmoc-L-Trp) imprinted polymer at different pHs and water concentrations in acetonitrile/aqueous buffer mobile phases. The equilibrium isotherm data were measured using frontal analysis at 25 {+-} 2 C. The adsorption energy distribution was found to be trimodal, with narrow modes. Consistent with this distribution, the adsorption data were modeled using a tri-Langmuir isotherm equation and the best estimates of the isotherm parameters were determined. The intraparticle mass transfer parameters were derived by comparing the profiles of experimental overloaded bands and the profiles calculated using the isotherm model and the lumped pore diffusion (POR) model of chromatography. These results showed that different adsorption and mass transfer mechanisms exist in mobile phases made of acetonitrile/aqueous buffer and of acetonitrile/acetic acid solutions.

  19. Synthesis of peptide .alpha.-thioesters

    DOEpatents

    Camarero, Julio A.; Mitchell, Alexander R.; De Yoreo, James J.

    2008-08-19

    Disclosed herein is a new method for the solid phase peptide synthesis (SPPS) of C-terminal peptide .alpha. thioesters using Fmoc/t-Bu chemistry. This method is based on the use of an aryl hydrazine linker, which is totally stable to conditions required for Fmoc-SPPS. When the peptide synthesis has been completed, activation of the linker is achieved by mild oxidation. The oxidation step converts the acyl-hydrazine group into a highly reactive acyl-diazene intermediate which reacts with an .alpha.-amino acid alkylthioester (H-AA-SR) to yield the corresponding peptide .alpha.-thioester in good yield. A variety of peptide thioesters, cyclic peptides and a fully functional Src homology 3 (SH3) protein domain have been successfully prepared.

  20. Synthesis and biological activity of homoarginine-containing opioid peptides.

    PubMed

    Izdebski, Jan; Kunce, Danuta; Schiller, Peter W; Chung, Nga N; Gers, Tomasz; Zelman, Monika; Grabek, Monika

    2007-01-01

    Two tris-alkoxycarbonyl homoarginine derivatives, Boc-Har{omega,omega'-[Z(2Br)]2}-OH and Boc-Har{omega,omega'-[Z(2Cl)]2}-OH, were prepared by guanidinylation of Boc-Lys-OH, and used for the synthesis of neo-endorphins and dynorphins. The results were compared with that obtained in the synthesis in which Boc-Lys(Fmoc)-OH was incorporated into the peptide chain, and after removing Fmoc protection, the resulting peptide-resin was guanidinylated with N,N'-[Z(2Br)]2- or N,N'-[Z(2Cl)]2-S-methylisourea. The peptides were tested in the guinea-pig ileum (GPI) and mouse vas deferens (MVD) assays. The results indicated that replacement of Arg by Har may be a good avenue for the design of biologically active peptides with increased resistance to degradation by trypsin-like enzymes.

  1. Influence of boric acid on electrospray ionization efficiency.

    PubMed

    Rebane, Riin; Herodes, Koit

    2012-01-01

    Liquid chromatography electrospray ionization mass-spectrometry (LC-ESI-MS) was used to analyze 9-fluorenylmethylmethoxycarbonyl chloride (Fmoc-Cl) and diethyl ethoxymethylenemalonate (Deemm) derivatives of three amino acids and five other compounds. Influence of boric acid on their ionization was investigated and dramatic impact on the signal was observed. The strongest signal suppression (6% of signal remains) was observed for the Deemm derivative of beta-Alanine (with ammonium acetate in eluent). With only formic acid as the eluent pH modifier, signal enhancement was observed, being largest for Fmoc-Cl derivative of Phenylalanine, 267%. Investigation of the influence of boric acid shows that it is a possible signal enhancer for LC-ESI-MS analysis.

  2. Comparison of the solid-phase fragment condensation and phase-change approaches in the synthesis of salmon I calcitonin.

    PubMed

    Gatos, D; Tzavara, C

    2001-02-01

    Salmon I calcitonin was synthesized using both phase-change and conventional solid-phase fragment condensation (SPFC) approaches, utilizing the Rink amide linker (Fmoc-amido-2,4-dimethoxybenzyl-4-phenoxyacetic acid) combined with 2-chlorotrityl resin and the Fmoc/tBu(Trt)-based protection scheme. Phase-change synthesis, performed by the selective detachment of the fully protected C-terminal 22-mer peptide-linker from the resin and subsequent condensation in solution with the N-terminal 1-10 fragment, gave a product of slightly less purity (85 vs. 92%) than the corresponding synthesis on the solid-phase. In both cases salmon I calcitonin was easily obtained in high purity.

  3. An efficient preparation of N-methyl-alpha-amino acids from N-nosyl-alpha-amino acid phenacyl esters.

    PubMed

    Leggio, Antonella; Belsito, Emilia Lucia; De Marco, Rosaria; Liguori, Angelo; Perri, Francesca; Viscomi, Maria Caterina

    2010-03-05

    In this paper we describe a simple and efficient solution-phase synthesis of N-methyl-N-nosyl-alpha-amino acids and N-Fmoc-N-methyl-alpha-amino acids. This represents a very important application in peptide synthesis to obtain N-methylated peptides in both solution and solid phase. The developed methodology involves the use of N-nosyl-alpha-amino acids with the carboxyl function protected as a phenacyl ester and the methylating reagent diazomethane. An important aspect of this synthetic strategy is the possibility to selectively deprotect the carboxyl function or alternatively both amino and carboxyl moieties by using the same reagent with a different molar excess and under mild conditions. Furthermore, the adopted procedure keeps unchanged the acid-sensitive side chain protecting groups used in Fmoc-based synthetic strategies.

  4. STUDIES ON THE BIOSYNTHESIS OF THE STEPHACIDINS AND NOTOAMIDES. TOTAL SYNTHESIS OF NOTOAMIDE S

    PubMed Central

    McAfoos, Timothy J.; Li, Shengying; Tsukamoto, Sachiko; Sherman, David H.

    2011-01-01

    Notoamide S has been suggested to be the final common precursor between two different Aspergillus sp. fungal strains before diverging to form enantiomerically opposite natural products (+)- and (−)-stephacidin A and (+)- and (−)-notoamide B. The synthesis of notoamide S comes from the coupling of N-Fmoc proline with a 6-hydroxy-7-prenyl-2-reverse prenyl tryptophan derivative that was synthesized via a late stage Claisen rearrangement from a 6-propargyl-2-reverse prenylated indole. PMID:21796227

  5. Single-Walled Carbon Nanotubes Targeted to the Tumor Vasculature for Breast Cancer Treatment

    DTIC Science & Technology

    2009-09-01

    Adsorption of a linker containing fluorene followed by reaction with the protein – This is a new method we developed that uses a linker that contains a... fluorene aromatic group for strong π-π binding to the graphite nanotube sidewall, polyethylene glycol (PEG) to increase the aqueous solubility of...Schematic of protein attachment to an SWNT using the Fmoc-amine-PEG- succinimidyl carboxy methyl ester linker. We have used fluorene in the linker

  6. Synthetic Lectins: New Tools for Detection and Management of Prostate Cancer

    DTIC Science & Technology

    2014-08-01

    the boronic acids do not interfere with the phenylisothiocyanate chemistry (Figure 5). Remarkably, the PBA does not appear to interfere with the...the mono and trivalent SL5 analogs are being synthesized on Rink-Amide resin using HBTU/Fmoc chemistry . The first amino acid attached to the resin...using thiol-selective maleimide chemistry (recall Cys is not used in the   Figure 7. Library design. A. Synthesis of carboxy benzoboroxole to

  7. Improved synthesis of C-terminal peptide thioesters on "safety-catch" resins using LiBr/THF.

    PubMed

    Quaderer, R; Hilvert, D

    2001-10-04

    [reaction: see text] The alkanesulfonamide "safety-catch" resin has proven useful for Fmoc-based synthesis of C-terminal peptide thioesters. We now report that the yield of isolated thioester can increase significantly when the cleavage reaction is carried out in 2 M LiBr/THF rather than DMF or THF. The largest effects are seen with problematic peptides that aggregate or form secondary structures on the resin.

  8. Solid-phase total synthesis of daptomycin and analogs.

    PubMed

    Lohani, Chuda Raj; Taylor, Robert; Palmer, Michael; Taylor, Scott D

    2015-02-06

    An entirely solid-phase synthesis of daptomycin, a cyclic lipodepsipeptide antibiotic currently in clinical use, was achieved using a combination of α-azido and Fmoc amino acids. This methodology was applied to the synthesis of several daptomycin analogs, one of which did not contain kynurenine or the synthetically challenging amino acid (2S,3R)-methylglutamate yet exhibited an MIC approaching that of daptomycin.

  9. Solution-phase automated synthesis of an α-amino aldehyde as a versatile intermediate

    PubMed Central

    Masui, Hisashi; Yosugi, Sae; Fuse, Shinichiro

    2017-01-01

    A solution-phase automated synthesis of the versatile synthetic intermediate, Garner’s aldehyde, was demonstrated. tert-Butoxycarbonyl (Boc) protection, acetal formation, and reduction of the ester to the corresponding aldehyde were performed utilizing our originally developed automated synthesizer, ChemKonzert. The developed procedure was also useful for the synthesis of Garner’s aldehyde analogues possessing fluorenylmethyloxycarbonyl (Fmoc) or benzyloxycarbonyl (Cbz) protection. PMID:28228851

  10. Preparation of Peptide p-Nitroanilides using an Aryl Hydrazine Solid Support

    SciTech Connect

    Kwon, Y; Welsh, K; Mitchell, A R; Camarero, J A

    2004-08-05

    Peptide p-nitroanilides are useful compounds for studying protease activity, however the poor nucleophilicity of p-nitroaniline makes their preparation difficult. We describe a new efficient approach for the Fmoc-based synthesis of peptide p-nitroanilides using an aryl hydrazine resin. Mild oxidation of the peptide hydrazide resin yields a highly reactive acyl diazene, which efficiently reacts with weak nucleophiles. We have prepared several peptide p-nitroanilides, including substrates for the Lethal Factor protease from B. anthracis.

  11. Ultratrace-level determination of glyphosate, aminomethylphosphonic acid and glufosinate in natural waters by solid-phase extraction followed by liquid chromatography-tandem mass spectrometry: performance tuning of derivatization, enrichment and detection.

    PubMed

    Hanke, Irene; Singer, Heinz; Hollender, Juliane

    2008-07-01

    A sensitive and robust analytical method for the quantification of glyphosate, aminomethylphosphonic acid (AMPA) and glufosinate in natural water has been developed on the basis of a derivatization with 9-fluorenylmethylchloroformate (FMOC-Cl), solid-phase extraction (SPE) and liquid chromatography followed by electrospray tandem mass spectrometry (LC-ESI-MS/MS). In order to maximize sensitivity, the derivatization was optimized regarding organic solvent content, amount of FMOC-Cl and reaction time. At an acetonitrile content of 10% a derivatization yield of 100% was reached within two hours in groundwater and surface water samples. After a twofold dilution the low acetonitrile content allowed solid-phase extraction of a sample of originally 80 mL over 200 mg Strata-X cartridges. In order to decrease the load of the LC column and mass spectrometer with derivatization by-products (e.g., 9-fluorenylmethanol FMOC-OH), a rinsing step was performed for the SPE cartridge with dichloromethane. Acidification of the sample and addition of EDTA was used to minimize complexation of the target compounds with metal ions in environmental samples. Due to the large sample volume and the complete FMOC-OH removal, limits of quantification of 0.7 ng/L, 0.8 ng/L and 2.3 ng/L were achieved in surface water for glyphosate, AMPA and glufosinate, respectively. The limits of detection were as low as 0.2 ng/L, 0.2 ng/L and 0.6 ng/L for glyphosate, AMPA and glufosinate, respectively. Surface water and ground water samples spiked at 2 ng/L showed recoveries of 91-107%.

  12. SPPS of protected peptidyl aminoalkyl amides.

    PubMed

    Karavoltsos, Manolis; Mourtas, Spyros; Gatos, Dimitrios; Barlos, Kleomenis

    2002-11-01

    Monophthaloyl diamines derived from naturally occurring amino acids were attached through their free amino functions to resins of the trityl type. The phthaloyl groups were removed by hydrazinolysis, and peptide chains were assembled using Fmoc/tBu-amino acids on the liberated amino functions. The peptidyl aminoalkyl amides obtained were cleaved from the resins by mild acidolysis, with the tBu-side chain protection remaining intact.

  13. Fluorophore Nanogold Particle-Based Optical Breast Cancer Locator

    DTIC Science & Technology

    2009-07-01

    cysteamine ) was chosen based on initial observations by Kang et al. (Kang and Hong, 2006) and subsequent observations by our collaborator Achilefu et al...Methoxy-activated, cysteamine -loaded polystyrene resin (Novabiochem) was reacted with the Fmoc-protected amino acids depicted in Scheme 1 following...Synthesis. Cysteamine hydrochloride was trityl protected and the resultant amine 4 was coupled to commercial PEG reagents 5a (n = 27

  14. Feasibility of Biomonitoring of Exposure to Permethrin Through Analysis of Long-Lived (Metabolite) Adducts to Proteins

    DTIC Science & Technology

    2007-09-01

    Cl2CA The synthesis was carried out on a PHB -S-TG resin (Rapp Polymere; 0.24 mmol/g resin) containing immobilized Boc-Lys(Fmoc) on a 36 µmol scale. The...was attempted to quantify adduct formation by using [14C] labelled 3-PBA glucuronides, obtained by combined chemical and enzymatic synthesis ...quantitated by using [14C] labelled 3-PBA glucuronide, which was obtained by enzymatic synthesis . The binding studies were thwarted by non-covalent

  15. 77 FR 37630 - National Oil and Hazardous Substances Pollution Contingency Plan; National Priorities List...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-22

    ... of concern (COCs) included benzene, chloroform, 1,2-dichloroethane, and the metals chromium and lead... Chemicals of concern Performance goals GROUNDWATER Benzene 1 g/L. Chloroform 0.19 g/L. 1.2 Dichloroethane 0... concentrations in excess of applicable 2L GWQS except 1.24 g/L Benzene in the MWD-002 groundwater sample....

  16. Ozonation of coal vitrinites of different metamorphism degrees in gas and liquid phases

    SciTech Connect

    S.A. Semenova; Y.F. Patrakov

    2007-02-15

    The comparative analysis of the dynamics of accumulation of oxygen-containing groups in coals of different metamorphism degrees under their ozonation in the gas phase and in chloroform has been performed. The coals of the middle stage of metamorphism demonstrate the highest reactivity. The ozonation in chloroform proceeds with a higher intensity.

  17. Cytotoxic effects of gutta-percha solvents.

    PubMed

    Barbosa, S V; Burkard, D H; Spångberg, L S

    1994-01-01

    Cytotoxicity of commonly used gutta-percha solvents was evaluated. Gutta-percha dissolved by chloroform, halothane, or turpentine was evaluated with the radiochromium release method using L929 mouse fibroblast cells. All solvents were toxic. Turpentine was most toxic followed by halothane and chloroform, which caused similar levels of cell injury.

  18. In vitro study on the softening of gutta-percha cones in endodontic retreatment.

    PubMed

    Pécora, J D; Spanó, J C; Barbin, E L

    1993-01-01

    Softening time of gutta-percha cones was studied in vitro using five chemical solvents: xylol, chloroform, turpentine, eucalyptol, and orange oil. An apparatus which reproduces the penetration force of an endodontic file was used on the sectioned roots of previously filled teeth. The most rapid chemical solvent of gutta-percha cones was chloroform and the slowest was eucalyptol.

  19. Evaluation of Gutta-percha solvents.

    PubMed

    Kaplowitz, G J

    1990-11-01

    Five solvents (rectified white turpentine, oil of melaleuca, eucalyptol, white pine oil, and pine needle oil) were compared with chloroform for their ability to dissolve gutta-percha. All solvents dissolved at least 50% of the gutta-percha in 15 min at 37 degrees C with chloroform and rectified white turpentine dissolving the gutta-percha completely.

  20. Antibacterial Activity of Leptadenia reticulata (Retz.) Wight. & Arn. (Asclepidaceae).

    PubMed

    Kalidass, C; Glory, M; Borgio, Francis; Manickam, V S

    2009-04-01

    Leptadenia reticulata of Asclepidaceae family is a shrub, originally property of Petroleum ether, Alcohol & Chloroform extract of L. reticulata. The antimicrobial testing was carried out by "Disc diffusion method". Amongst the tested three extracts, chloroform extract showed high antimicrobial activity against E. coli, alcoholic extract showed high antibacterial activity against Pseudomonas aeruginosa, while Petroleum ether extract showed antibacterial activity against Klebsilla pneumonae.

  1. Antimicrobial, antimycobacterial and antibiofilm properties of Couroupita guianensis Aubl. fruit extract

    PubMed Central

    2012-01-01

    Background Couroupita guianensis Aubl. (Lecythidaceae) is commonly called Ayahuma and the Cannonball tree. It is distributed in the tropical regions of northern South America and Southern Caribbean. It has several medicinal properties. It is used to treat hypertension, tumours, pain, inflammatory processes, cold, stomach ache, skin diseases, malaria, wounds and toothache. Methods The fruits of Couroupita guianensis were extracted with chloroform. Antimicrobial, antimycobacterial and antibiofilm forming activities of the chloroform extract were investigated. Quantitative estimation of Indirubin, one of the major constituent, was identified by HPLC. Results Chloroform extract showed good antimicrobial and antibiofilm forming activities; however it showed low antimycobacterial activity. The zones of inhibition by chloroform extract ranged from 0 to 26 mm. Chloroform extract showed effective antibiofilm activity against Pseudomonas aeruginosa starting from 2 mg/mL BIC, with 52% inhibition of biofilm formation. When the chloroform extract was subjected to HPLC-DAD analysis, along with Indirubin standard, in the same chromatographic conditions, it was found that Indirubin was one of the major compounds in this plant (0.0918% dry weight basis). Conclusions The chloroform extract showed good antimicrobial and antibiofilm properties. Chloroform extract can be evaluated further in drug development programmes. PMID:23206492

  2. Use of Designed Experiments in the "Green Ammunition" Program

    DTIC Science & Technology

    2001-08-13

    Excellence HEAVY METALS Primer Compositions Lead Styphnate Barium Nitrate Antimony Sulfide Green Ammunition Target Materials HEAVY METALS Projectile Slug Lead... Barium Peroxide Lead Dioxide Barium Nitrate ODCs Methyl Chloroform ODCs Casemouth Sealant Methyl Chloroform Committed To Excellence Non-Toxic Ammunition

  3. 40 CFR Table 2 to Subpart Ggg of... - Partially Soluble HAP

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... (mek) Ethylene oxide 1,4-Dichlorobenzene Hexachlorobenzene 2-Nitropropane Hexachlorobutadiene 4-Methyl-2-pentanone (MIBK) Hexachloroethane Acetaldehyde Methyl methacrylate Acrolein Methyl-t-butyl ether... GGG of Part 63—Partially Soluble HAP 1,1,1-Trichloroethane (methyl chloroform) Chloroform...

  4. 40 CFR Table 2 to Subpart Ggg of... - Partially Soluble HAP

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... (MIBK) Hexachloroethane Acetaldehyde Methyl methacrylate Acrolein Methyl-t-butyl ether Acrylonitrile...—Partially Soluble HAP 1,1,1-Trichloroethane (methyl chloroform) Chloroform 1,1,2,2-Tetrachloroethane...-Dibromoethane Dichloroethyl ether 1,2-Dichloroethane (ethylene dichloride) Dinitrophenol...

  5. 40 CFR Table 2 to Subpart Ggg of... - Partially Soluble HAP

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... (mek) Ethylene oxide 1,4-Dichlorobenzene Hexachlorobenzene 2-Nitropropane Hexachlorobutadiene 4-Methyl-2-pentanone (MIBK) Hexachloroethane Acetaldehyde Methyl methacrylate Acrolein Methyl-t-butyl ether... GGG of Part 63—Partially Soluble HAP 1,1,1-Trichloroethane (methyl chloroform) Chloroform...

  6. 40 CFR Table 2 to Subpart Ggg of... - Partially Soluble HAP

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... (MIBK) Hexachloroethane Acetaldehyde Methyl methacrylate Acrolein Methyl-t-butyl ether Acrylonitrile...—Partially Soluble HAP 1,1,1-Trichloroethane (methyl chloroform) Chloroform 1,1,2,2-Tetrachloroethane...-Dibromoethane Dichloroethyl ether 1,2-Dichloroethane (ethylene dichloride) Dinitrophenol...

  7. Ultrasound-induced gelation of fluorenyl-9-methoxycarbonyl-l-lysine(fluorenyl-9-methoxycarbonyl)-OH and its dipeptide derivatives showing very low minimum gelation concentrations.

    PubMed

    Geng, Huimin; Ye, Lin; Zhang, Ai-Ying; Shao, Ziqiang; Feng, Zeng-Guo

    2017-03-15

    Four l-Lysine(Lys)-l-glutamic acid(Glu) dipeptide derivatives (1-4) and their precursor-a single fluorenyl-9-methoxycarbonyl(Fmoc)-l-Lys(Fmoc)-OH amino acid (5) were demonstrated as gelators to gelate a variety of alcohols and aromatic solvents under the sonication conditions. Compared to the routine heating-cooling protocol, the ultrasound substantially brought down the minimum gelation concentrations (MGCs) of the resulting organogels. The Fourier transform infrared spectroscopy (FT-IR) and fluorescence studies revealed that the π-π stacking and hydrogen bonding act as major driving forces for the self-assembly of these lysine-based gelators into supramolecular fibrous three dimensional (3D) network, where the more the Fmoc protecting groups, the gelators are more responsive to ultrasound-stimulus and more conducive to an ordered molecular arrangement reinforcing the intermolecular forces. Moreover, the ultrasound-triggered organogels of 5 exhibited the thixotropic property. Upon imposing a mechanical shear, its gels with the fibrous 3D network structure were unraveled into sols. However, after standing quiescently over time, these sols returned to the gels showing a more ordered lamella-like packing structure as evidenced by scanning electron microscopy (SEM) and X-ray diffraction (XRD) analyses.

  8. Determination of chitosan with a modified acid hydrolysis and HPLC method.

    PubMed

    Li, Bo; Zhang, Jiali; Bu, Fen; Xia, Wenshui

    2013-01-25

    Acid hydrolysis and subsequent quantification of glucosamine (GlcN) are widely used for chitosan quantification. Degradation of GlcN during chitosan hydrolysis was the main reason for the decrease of recovery, which made the method improper for the quantification of chitosan. Ten milligram of chitosan hydrolyzed with 10 mL mixed acid solution of HCl-H₃PO₄ (75:25 in molar ratio) showed the highest recovery, significantly higher than HCl hydrolysis. Further study revealed that the optimum conditions involved the hydrolysis with HCl-H₃PO₄ (4.5:1.5M) for 24 h at 110 °C. The hydrolysate was neutralized and derived with 9-fluorenylmethoxycarbonyl chloride (FMOC-Cl) before HPLC quantification. The optimum ratio of FMOC-Cl:GlcN was 53:1, with excess FMOC-Cl induced by the high ionic strength of the solution. This quantification procedure was then validated and proved to be specific, with good linearity, accuracy, and precision, making it well-suited for the determination of chitosan.

  9. An immunostimulatory dual-functional nanocarrier that improves cancer immunochemotherapy

    PubMed Central

    Chen, Yichao; Xia, Rui; Huang, Yixian; Zhao, Wenchen; Li, Jiang; Zhang, Xiaolan; Wang, Pengcheng; Venkataramanan, Raman; Fan, Jie; Xie, Wen; Ma, Xiaochao; Lu, Binfeng; Li, Song

    2016-01-01

    Immunochemotherapy combines a chemotherapeutic agent with an immune-modulating agent and represents an attractive approach to improve cancer therapy. However, the success of immunochemotherapy is hampered by the lack of a strategy to effectively co-deliver the two therapeutics to the tumours. Here we report the development of a dual-functional, immunostimulatory nanomicellar carrier that is based on a prodrug conjugate of PEG with NLG919, an indoleamine 2,3-dioxygenase (IDO) inhibitor currently used for reversing tumour immune suppression. An Fmoc group, an effective drug-interactive motif, is also introduced into the carrier to improve the drug loading capacity and formulation stability. We show that PEG2k-Fmoc-NLG alone is effective in enhancing T-cell immune responses and exhibits significant antitumour activity in vivo. More importantly, systemic delivery of paclitaxel (PTX) using the PEG2k-Fmoc-NLG nanocarrier leads to a significantly improved antitumour response in both breast cancer and melanoma mouse models. PMID:27819653

  10. Design of strategies to study the metabolic profile of highly polar compounds in plasma by reversed-phase liquid chromatography-high resolution mass spectrometry.

    PubMed

    Sánchez-López, Elena; Crego, Antonio L; Marina, María Luisa

    2017-03-24

    Amino acids and related compounds are paramount analytes which are involved in numerous metabolic pathways. Most of these compounds are unable to be retained on Liquid Chromatography with Reversed-Phase stationary phases due to their high hydrophilic character. An interesting strategy is to reduce their polarity through their derivatization with a labelling reagent, such as the commercially available 9-fluorenylmethyloxycarbonyl (FMOC) which forms stable complexes with primary and secondary amine moieties rapidly. Although some derivatization reagents have been employed in the study of metabolic profiles, as far as we know, FMOC has never been employed for this purpose. In this work, it is demonstrated that the use of RP-LCMS(TOF) using a C18 column and FMOC as labelling agent enables the determination of a larger number of hydrophilic compounds (proteinogenic amino acids, non-proteinogenic amino acids, and biogenic amines) when compared to the use of a fully-wettable pentafluorophenyl column in fully-aqueous conditions (gradient starting in 0% of organic solvent) and HILIC column, both without using compound derivatization. Different strategies for plasma protein elimination were also carefully evaluated. Results revealed that ultrafiltration (UF) offered a lower variability from sample to sample when compared to the protein precipitation (PP) method (from 2 to 12 times lower variability found in UF). Additionally, UF preserved a larger number of possible compounds when compared to the PP approach: 4631 unique molecular features with UF, 666 unique molecular features with PP.

  11. Integrated work management system.

    SciTech Connect

    Williams, Edward J., Jr.; Henry, Karen Lynne

    2010-06-01

    Sandia National Laboratories develops technologies to: (1) sustain, modernize, and protect our nuclear arsenal (2) Prevent the spread of weapons of mass destruction; (3) Provide new capabilities to our armed forces; (4) Protect our national infrastructure; (5) Ensure the stability of our nation's energy and water supplies; and (6) Defend our nation against terrorist threats. We identified the need for a single overarching Integrated Workplace Management System (IWMS) that would enable us to focus on customer missions and improve FMOC processes. Our team selected highly configurable commercial-off-the-shelf (COTS) software with out-of-the-box workflow processes that integrate strategic planning, project management, facility assessments, and space management, and can interface with existing systems, such as Oracle, PeopleSoft, Maximo, Bentley, and FileNet. We selected the Integrated Workplace Management System (IWMS) from Tririga, Inc. Facility Management System (FMS) Benefits are: (1) Create a single reliable source for facility data; (2) Improve transparency with oversight organizations; (3) Streamline FMOC business processes with a single, integrated facility-management tool; (4) Give customers simple tools and real-time information; (5) Reduce indirect costs; (6) Replace approximately 30 FMOC systems and 60 homegrown tools (such as Microsoft Access databases); and (7) Integrate with FIMS.

  12. Sputtering Yields for Mixtures of Organic Materials Using Argon Gas Cluster Ions.

    PubMed

    Seah, M P; Havelund, R; Shard, A G; Gilmore, I S

    2015-10-22

    The sputtering yield volumes of binary mixtures of Irganox 1010 with either Irganox 1098 or Fmoc-pentafluoro-L-phenylalanine (FMOC) have been measured for 5 keV Ar2000(+) ions incident at 45° to the surface normal. The sputtering yields are determined from the doses to sputter through various compositions of 100 nm thick, intimately mixed, layers. Because of matrix effects, the profiles for secondary ions are distorted, and profile shifts in depth of 15 nm are observed leading to errors above 20% in the deduced sputtering yield. Secondary ions are selected to avoid this. The sputtering yield volumes for the mixtures are shown to be lower than those deduced from a linear interpolation from the pure materials. This is shown to be consistent with a simple model involving the changing energy absorbed for the sputtering of intimate mixtures. Evidence to support this comes from the secondary ion data for pairs of the different molecules. Both binary mixtures behave similarly, but matrix effects are stronger for the Irganox 1010/FMOC system.

  13. Structure elucidation of β-sitosterol with antibacterial activity from the root bark of Malva parviflora.

    PubMed

    Ododo, Mesfin Medihin; Choudhury, Manash Kumar; Dekebo, Ahmed Hussen

    2016-01-01

    The powder of root bark of Malva parviflora (Malvaceae) was successively extracted with petroleum ether (b.p. 60-80 °C), chloroform and ethanol. The chloroform extract showed antibacterial activity against Staphylococcus aureus and Escherichia coli, whereas the ethanolic extract showed antibacterial activity against only S. aureus. The chloroform extract, after column chromatographic separation on silica gel using petroleum ether:chloroform (3:1) as eluent, furnished 98 mg of white crystalline compound. The yield of the compound is 0.316 % (w/w). The compound has a melting point of 134-136 °C and the Rf value 0.56 in benzene:chloroform:acetone (1:15:1) on silica gel TLC. The compound was characterized as β-sitosterol by physical properties, chemical test, spectral analysis (FTIR, NMR and MS) and comparing the data obtained from the literature.

  14. Nanometer-Scale Water-Soluble Macrocycles from Nanometer-Sized Amino Acids

    PubMed Central

    Gothard, Chris M.

    2009-01-01

    This paper introduces the unnatural amino acids m-Abc2K and o-Abc2K as nanometersized building blocks for the creation of water-soluble macrocycles with well-defined shapes. m-Abc2K and o-Abc2K are homologues of the nanometer-sized amino acid Abc2K, which we recently introduced for the synthesis of water-soluble molecular rods of precise length. [J. Am. Chem. Soc. 2007, 129, 7272]. Abc2K is linear (180°), m-Abc2K creates a 120° angle, and o-Abc2K creates a 60° angle. m-Abc2K and o-Abc2K are derivatives of 3’-amino-[1,1’-biphenyl]-4-carboxylic acid and 2’-amino-[1,1’-biphenyl]-4-carboxylic acid, with two propyloxyammonium side chains for water solubility. m-Abc2K and o-Abc2K are prepared as Fmoc-protected derivatives Fmoc-m-Abc2K(Boc)-OH (1a) and Fmoc-o-Abc2K(Boc)-OH (1b). These derivatives can be used alone or in conjunction with Fmoc-Abc2K(Boc)-OH (1c) as ordinary amino acids in Fmoc-based solid-phase peptide synthesis. Building blocks 1a–c were used to synthesize macrocyclic “triangles” 9a–c, “parallelograms” 10a,b, and hexagonal “rings” 11a–d. The macrocycles range from a trimer to a dodecamer, with ring sizes from 24 to 114 atoms, and are 1–4 nm in size. Molecular modeling studies suggest that all the macrocycles except 10b should have well-defined triangle, parallelogram, and ring shapes if all of the amide linkages are trans and the ortho-alkoxy substituents are intramolecularly hydrogen bonded to the amide NH groups. The macrocycles have good water solubility and are readily characterized by standard analytical techniques, such as RP-HPLC, ESI-MS, and NMR spectroscopy. 1H and 13C NMR studies suggest that the macrocycles adopt conformations with all trans-amide linkages in CD3OD, that the “triangles” and “parallelograms” maintain these conformations in D2O, and that the “rings” collapse to form conformations with cis-amide linkages in D2O. PMID:20020731

  15. Phytochemical analysis, cytotoxic activity and constituents-activity relationships of the leaves of Cinnamomum iners (Reinw. ex Blume-Lauraceae).

    PubMed

    Ghalib, Raza Murad; Hashim, Rokiah; Sulaiman, Othman; Mehdi, Sayed Hasan; Anis, Zurida; Rahman, Syed Ziaur; Ahamed, B M Khadeer; Abdul Majid, Amin Malik Shah

    2012-01-01

    The leaves of Cinnamomum iners (Reinw. ex Blume-Lauraceae) have been refluxed successively with chloroform and alcohol to get chloroform extract and alcoholic extract. Both the extracts have been assayed for cytotoxicity against human colorectal tumour cells. The chloroform extract exhibited significant cytotoxicity with IC(50) 31 µg mL(-1) (p < 0.01). However, ethanol extract was found to be much less cytotoxic with IC(50) > 200 µg mL(-1). The chloroform extract has been further proceeded for chemical analysis by GC-TOFMS and 178 components were identified including acids, amines, amides, aldehydes, alcohols, esters, benzene derivatives, bicyclic compounds, terpenes, hydrocarbons, naphthalene derivatives, furan derivatives, azulenes, etc. Nine components representing 51.73% of the total chloroform extract were detected as major components. Caryophyllene (14.41%) and Eicosanoic acid ethyl ester (12.17%) are the most prominent components of the chloroform extract. β-Caryophyllene (14.41%) as most abundant compound supports potent cytotoxicity as shown by chloroform extract.

  16. 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

  17. Phytochemical analysis of Gymnema sylvestre and evaluation of its antimicrobial activity.

    PubMed

    Chodisetti, Bhuvaneswari; Rao, Kiranmayee; Giri, Archana

    2013-01-01

    Gymnema sylvestre (CS 149), known to be a rich source of saponins and other valuable phytochemicals, has been analysed for antimicrobial activity. The chloroform extracts of aerial and root parts of G. sylvestre exhibited higher antimicrobial activity as compared to diethyl ether and acetone. The root extracts of chloroform have shown competitive minimum inhibitory concentration and minimum bactericidal concentration values in the range of 0.04-1.28 mg mL(-1) and 0.08-2.56 mg/mL, respectively, towards the pathogens. The GC-MS analysis of chloroform extracts has shown the presence of compounds like eicosane, oleic acid, stigmasterol and vitamin E.

  18. [Biological activity of Penicillium sp. 10-51 exometabolites].

    PubMed

    Savchuk, Ia I; Zaĭchenko, A M; Tsyganenko, E S

    2012-01-01

    Silica gel column chromatography (silica gel "L" II kind of activity 100/160 mkm) of the chloroform extract from the cultural filtrate of Penicillium sp. 10-51 gave two fractions (chloroform and chloroform-acetone, 5:1) having biological activity. Recrystallization yielded two compounds. On the basis of physico-chemical and spectral data these compounds were identified as curvularin and hydroxycurvularin, which have a large spectrum of biological action as to bacteria, yeast, blue-green algae and phytopathogenic micromycetes.

  19. Antibacterial and cytotoxic activities of the sesquiterpene lactones cnicin and onopordopicrin.

    PubMed

    Bach, Sandra M; Fortuna, Mario A; Attarian, Rodgoun; de Trimarco, Juliana T; Catalán, César A N; Av-Gay, Yossef; Bach, Horacio

    2011-02-01

    The antimicrobial and cytotoxic activities of chloroform extracts from the weeds Centaurea tweediei and C. diffusa, and the main sesquiterpene lactones isolated from these species, onopordopicrin and cnicin, respectively, were assayed. Results show that the chloroform extracts from both Centaurea species possess antibacterial activities against a panel of Gram-positive and Gram-negative bacteria. Remarkable antibacterial activity against methicillin-resistant Staphylococcus aureus was also measured. Both the extracts and the purified sesquiterpene lactones show high cytotoxicity against human-derived macrophages. Despite this cytotoxicity, C. diffusa chloroform extract and cnicin are attractive candidates for evaluation as antibiotics in topical preparations against skin-associated pathogens.

  20. TREATMENT OF VOCS IN HIGH STRENGTH WASTES USING AN ANAEROBIC EXPANDED-BED GAS REACTOR

    EPA Science Inventory

    The potential of the expanded-bed granular activated carbon (GAC) anaerobic reactor in treating a high strength waste containing RCRA volatile organic compounds (VOCs) was studied. A total of six VOCs, methylene chloride, chlorobenzene, carbon tetrachloride, chloroform, toluene ...

  1. Antinociceptive activity of Helicteres isora.

    PubMed

    Venkatesh, Sama; Laxmi, K Sai; Reddy, B Madhava; Ramesh, M

    2007-02-01

    Helicteres isora root extracts were studied for antinociceptive activity on acetic acid-induced writhing test in mice, at a dose of 250 mg/kg. Petroleum ether, chloroform and aqueous ethanol extracts have shown significant activity.

  2. THE LIPID CONSTITUENTS OF WHOLE AND PAROTID SALIVA

    DTIC Science & Technology

    Chloroform: methanol extracts of whole and parotid saliva were subjected to paper chromatography to further characterize their lipid components. The...the sample materials. Whole and parotid saliva had similar nonphosphatides, but differed in their phospholipid composition. (Author)

  3. Photooxidation of groundwater contaminants: Trichloroethylene, trichloromethane, dibromochloropropane and benzene. A critical review. Technical report, January 1992-May 1994

    SciTech Connect

    Burrows, W.D.

    1994-05-01

    The literature on advanced oxidation, with special reference to photooxidation, has been reviewed for four Rocky Mountain Arsenal groundwater contaminants: trichloroethylene (TCE), trichloromethane (chloroform), benzene and 1,2-dibromo-d-chloropropane (DBCP or nemagon).

  4. GTE_PEMTB_P3B Parameters 17

    Atmospheric Science Data Center

    2013-02-19

    ... NMHC/Halocarbons/Alkyl Nitrates: Methyl Chloride F-12 F-114 F-11 HCFC-141B HCFC-134a HCFC-22 ... 2,3,4-trimethylpethane Methylene Chloride Chloroform Perchloroethylene HFC-134A HCFC22 HCFC142B ...

  5. GTE_PEMTB_DC8 Parameters 15

    Atmospheric Science Data Center

    2013-02-19

    ... NMHC/Halocarbons/Alkyl Nitrates: Methyl Chloride F-12 F-114 F-11 HCFC-141B HCFC-134a HCFC-22 ... 2,3,4-trimethylpethane Methylene Chloride Chloroform Perchloroethylene HFC-134A HCFC22 HCFC142B ...

  6. REMOVAL OF SYNTHETIC ORGANIC CHEMICAL CONTAMINANTS IN DRINKING WATER: RASCO, INC. ADVANCED SIMULTANEOUS OXIDATION PROCESS (ASOP)

    EPA Science Inventory

    The RASco, Inc. ASOP Drinking Water Treatment Module was tested at NSF’s Laboratory for the reduction of the following chemicals of concern: aldicarb, benzene, carbofuran, chloroform, dichlorvos, dicrotophos, methomyl, mevinphos, nicotine, oxamyl, paraquat, phorate, sodium fluor...

  7. PRN 93-5: Labeling Requirements of the Clean Air Act

    EPA Pesticide Factsheets

    A regulation under the Clean Air Act requires a warning statement on products (including pesticide products) manufactured with or containing Class I ozone-depleting substances, including chlorofluorocarbons, methyl chloroform and carbon tetrachloride.

  8. COMPETITIVE ADSORPTION OF VOCS AND BOM: THE ROLE OF MOLECULAR OXYGEN

    EPA Science Inventory

    In this study, the presence of background organic matter (BOM) was seen to reduce the adsorptive capacity of carbon for chloroform, chlorobenzene, and dibromochloropropane. Adsorption of these compounds was further reduced under oxic conditions. This additional reduction in cap...

  9. Phytochemical and termiticidal study of Lantana camara var. aculeata leaves.

    PubMed

    Verma, Rajesh K; Verma, Suman K

    2006-09-01

    Extracts of Lantana camara var. aculeata leaves were studied for their phytochemical constituents and termiticidal effects against adult termite workers. The 5% chloroform extract was found to be significantly effective against termite workers.

  10. EFFECTS OF DEFINED MIXTURES OF TRIHALOMETHANES AND HALOACETIC ACIDS ON PREGNANCY MAINTENANCE AND EYE DEVELOPMENT IN F 344 RATS

    EPA Science Inventory

    Although disinfection of drinking water is important for control of microbial contamination, it results in the formation of hundreds of disinfection by-products (DBPs). The most prevalent DBPs are trihalomethanes (THMs; chloroform, bromodichloromethane, chlorodibromomethane, bro...

  11. 40 CFR 52.2222 - Approval status.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    .... These compounds are as follows: methane, ethane, methyl chloroform (1,1,1-trichloroethane), CFC-113 (trichlorotrifluoroethane), methylene chloride, CFC 11 (trichlorofluoromethane), CFC-12 (dichlorodifluoromethane), CFC-22 (chlorodifluoromethane), FC-23 (trifluoromethane), CFC-114 (dichlorotetrafluoroethane), CFC-115...

  12. 40 CFR Appendix G to Part 403 - Pollutants Eligible for a Removal Credit

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... tetrachloride, Chlordane, Chloroform, Chloromethane, DDD, DDE, DDT, Dibromochloromethane, Dibutyl phthalate, 1,2... (total) 3 100 3 100 Copper 3 46 100 1400 DDD, DDE, DDT (Total) 1.2 2000 2000 2,4...

  13. The Impact of Mixture Composition, Mixing Ratio and Dose on the Interactions among the Four Trihalomethanes (THMs) Regulated in Drinking Water

    EPA Science Inventory

    Oxidizing disinfectants reduce microbial contamination but react with inorganic and organic materials in water forming disinfection byproducts (DBPs). The U.S. EPA regulates 4 THM DBPs (chloroform, CHCI3; bromodichloromethane, BDCM; chlorodibromomethane, CDBM; bromoform, CHBr3) a...

  14. 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

  15. Pregnancy loss and eye malformations in offspring of F344 rats following gestational exposure to mixtures of regulated trihalomethanes and haloacetic acids

    EPA Science Inventory

    Chlorination of drinking water results in the formation of hundreds of disinfection byproducts (DBPs), the most prevalent are trihalomethanes (THMs) and haloacetic acids (HAAs). Four THMs (chloroform, bromodichloromethane, chlorodibromomethane, bromoform) and five HAAs (chloroac...

  16. VISUALIZATION-BASED ANALYSIS FOR A MIXED-INHIBITION BINARY PBPK MODEL: DETERMINATION OF INHIBITION MECHANISM

    EPA Science Inventory

    A physiologically-based pharmacokinetic (PBPK) model incorporating mixed enzyme inhibition was used to determine mechanism of the metabolic interactions occurring during simultaneous inhalation exposures to the organic solvents chloroform and trichloroethylene (TCE).

    V...

  17. 40 CFR Table 7 to Subpart Vvvvvv... - Partially Soluble HAP

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 106990 23. Carbon disulfide 75150 24. Chlorobenzene 108907 25. Chloroethane (ethyl chloride) 75003 26. Chloroform 67663 27. Chloromethane 74873 28. Chloroprene 126998 29. Cumene 98828 30. Dichloroethyl ether 111444 31. Dinitrophenol 51285 32. Epichlorohydrin 106898 33. Ethyl acrylate 140885 34....

  18. 40 CFR Table 7 to Subpart Hhhhh of... - Partially Soluble Hazardous Air Pollutants

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 108907 26. Chloroethane (ethyl chloride) 75003 27. Chloroform 67663 28. Chloromethane 74873 29. Chloroprene 126998 30. Cumene 98828 31. Dichloroethyl ether 111444 32. Dinitrophenol 51285 33. Epichlorohydrin 106898 34. Ethyl acrylate 140885 35. Ethylbenzene 100414 36. Ethylene oxide 75218 37....

  19. ETV REPORT: REMOVAL OF CHEMICAL CONTAMINANTS IN DRINKING WATER — PALL/KINETICO PUREFECTA DRINKING WATER TREATMENT SYSTEM

    EPA Science Inventory

    The Pall/Kinetico Purefecta™ POU drinking water treatment system was tested for removal of aldicarb, benzene, cadmium, carbofuran, cesium, chloroform, dichlorvos, dicrotophos, fenamiphos, mercury, mevinphos, oxamyl, strontium, and strychnine. The Purefecta™ employs several compon...

  20. Biological Treatment of Water Disinfection Byproducts using Biotrickling Filter under Anoxic and Anaerobic Conditions

    EPA Science Inventory

    Major disinfection by-products (DBPs) from the chlorination process of drinking water include trihalomethanes (THMs) and haloacetic acides (HAA5). THMs mainly consist of chloroform, and other harsh chemicals. Prolonged consumptions of drinking water containing high levels of THMs...