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Sample records for column hplc separation

  1. Separation of kafirins on surface porous RP-HPLC columns

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Surface porous HPLC columns were investigated for the separation of kafarins, storage proteins of grain sorghum. Kafirins were successfully separated using C3, C8 and C18 surface porous stationary phases in less than 17 min. Separations using a monolithic C18 stationary phase were also developed ...

  2. SEPARATION OF OCTYLPHENOL POLYETHER ALCOHOLS SURFACTANTS BY CAPILLARY COLUMN SFC AND HPLC

    EPA Science Inventory

    Separation of nonionic octylphenol polyether alcohols (OPA) by supercritical fluid chromatography (SFC) and HPLC is described. Using a density programming and a 50-μm i.d. capillary column, a total of 18 group oligomers was separated. The effects of the operating parameters, such...

  3. Chiral HPLC Separation and Modeling of Four Stereomers of DL-Leucine-DL-Tryptophan Dipeptide on Amylose Chiral Column.

    PubMed

    Alajmi, Mohammed F; Hussain, Afzal; Suhail, Mohd; Mukhtar, Sofi Danish; Sahoo, Dibya Ranjan; Asnin, Leonid; Ali, Imran

    2016-09-01

    Chiral high-performance liquid chromatography (HPLC) separation and modeling of four stereomers of DL-leucine-tryptophan DL-dipeptide on AmyCoat-RP column are described. The mobile phase applied was ammonium acetate (10 mM)-methanol-acetonitrile (50:5:45, v/v). The flow rate of the mobile phases was 0.8 mL/min with UV detection at 230 nm. The values of retention factors for LL-, DD-, DL-, and LD- stereomers were 2.25, 3.60, 5.00, and 6.50, respectively. The values of separation and resolution factors were 1.60, 1.39, and 1.30 and 7.76, 8.05, and 7.19. The limits of detection and quantitation were ranging from 1.0-2.3 and 5.6-14.0 μg/mL. The simulation studies established the elution orders and the mechanism of chiral recognition. It was seen that π-π connections and hydrogen bondings were the main forces for enantiomeric resolution. The reported chiral HPLC method may be applied for the enantiomeric separation of DL-leucine-DL-tryptophan in unknown matrices. Chirality 28:642-648, 2016. © 2016 Wiley Periodicals, Inc. PMID:27474783

  4. Post column derivatisation analyses review. Is post-column derivatisation incompatible with modern HPLC columns?

    PubMed

    Jones, Andrew; Pravadali-Cekic, Sercan; Dennis, Gary R; Shalliker, R Andrew

    2015-08-19

    Post Column derivatisation (PCD) coupled with high performance liquid chromatography or ultra-high performance liquid chromatography is a powerful tool in the modern analytical laboratory, or at least it should be. One drawback with PCD techniques is the extra post-column dead volume due to reaction coils used to enable adequate reaction time and the mixing of reagents which causes peak broadening, hence a loss of separation power. This loss of efficiency is counter-productive to modern HPLC technologies, -such as UHPLC. We reviewed 87 PCD methods published from 2009 to 2014. We restricted our review to methods published between 2009 and 2014, because we were interested in the uptake of PCD methods in UHPLC environments. Our review focused on a range of system parameters including: column dimensions, stationary phase and particle size, as well as the geometry of the reaction loop. The most commonly used column in the methods investigated was not in fact a modern UHPLC version with sub-2-micron, (or even sub-3-micron) particles, but rather, work-house columns, such as, 250 × 4.6 mm i.d. columns packed with 5 μm C18 particles. Reaction loops were varied, even within the same type of analysis, but the majority of methods employed loop systems with volumes greater than 500 μL. A second part of this review illustrated briefly the effect of dead volume on column performance. The experiment evaluated the change in resolution and separation efficiency of some weak to moderately retained solutes on a 250 × 4.6 mm i.d. column packed with 5 μm particles. The data showed that reaction loops beyond 100 μL resulted in a very serious loss of performance. Our study concluded that practitioners of PCD methods largely avoid the use of UHPLC-type column formats, so yes, very much, PCD is incompatible with the modern HPLC column. PMID:26343427

  5. Synthesis and applications of monolithic HPLC columns

    NASA Astrophysics Data System (ADS)

    Liang, Chengdu

    Silica and carbon monolithic columns were synthesized and modified for liquid chromatography applications. Column configurations and cladding techniques were investigated in detail. Three novel approaches have been developed for the synthesis of bimodal porous rods. Out of these three methods, gel-casting was adopted for the synthesis of silica monoliths with ordered mesopores and uniform macropores; the use of colloidal templates and dual phase separation has been successfully implemented for the synthesis of carbon monoliths with well-controlled meso- and macro- porosities. The formation of mesopores in carbon materials has been further studied in the microphase separation of block copolymers. Electrochemical modification of carbon monoliths was discovered to be an efficient method for converting covalently bonded functionalities to carbon monoliths. N,N'-diethylaminobenzene has been attached to carbon surface for the separation of proteins and protein digests. The performances of carbon-based monolithic columns were studied intensely through frontal analysis and Van Deemter plot. Temperature and pressure effects were also investigated in carbon-based columns. The density of bonding on the modified carbon monoliths was characterized by thermogravimetric analysis.

  6. A new chiral derivatizing agent for the HPLC separation of α-amino acids on a standard reverse-phase column.

    PubMed

    Kotthaus, A F; Altenbach, H-J

    2011-02-01

    A new chiral derivatizing agent for α-amino acids is described which leads to diastereomers that can be separated by reverse-phase HPLC with direct detection by a diode array detector. The main advantage of the presented procedure is the fact that an excess of the derivatizing reagent can be employed as the product exhibits an absorption maximum at 360 nm, while the reagent has its absorption maximum at 260 nm. Therefore, it is possible to suppress the reagent signal by a detection wavelength of 400 nm leading to an easy and general method for the enantioseparation of a mixture of DL-amino acids and the determination of the enantiomeric purity of α-amino acid as exemplified by 16 different α-amino acids.

  7. Enantioselective separation and determination of adrafinil and modafinil on Chiralcel OJ-H column in rat serum and urine using solid-phase extraction followed by HPLC.

    PubMed

    Rao, R Nageswara; Shinde, Dhananjay D

    2009-08-01

    A simple and rapid normal-phase HPLC method for enantiospecific separation of a psychostimulant, adrafinil (ADL), and its metabolite modafinil (MDL) in rat serum and urine was developed. The separation was accomplished on a normal-phase polysaccharide stationary phase Chiralcel OJ-H using n-hexane-ethanol (62:38 v/v) as a mobile phase at a flow rate of 1.0 mL/min. Detection was carried out at 225 nm using a photo diode array (PDA) detector. The elution order of the enantiomers was determined by a polarimeter connected in series with the PDA. ADL and its metabolite were recovered from rat serum and urine by solid phase extraction using Oasis HLB cartridges and the mean recoveries were >or=80%. The enantiomers were eluted within 15 min without any interference from endogenous substances. The calibration curves were linear (r(2) > 0.998) in the concentration range of 1.20-500 microg/mL for ADL and MDL. The assay was specific, accurate, precise and reproducible (intra- and inter-day precisions RSDs <7.2%). ADL in rat serum was stable over three freeze-thaw cycles at ambient temperature for 4 h. The method was successfully applied to pharmacokinetic studies of adrafinil after an oral administration to rats. PMID:19353685

  8. Chiral separation of cathinone derivatives used as recreational drugs by HPLC-UV using a CHIRALPAK® AS-H column as stationary phase.

    PubMed

    Mohr, Stefan; Taschwer, Magdalena; Schmid, Martin G

    2012-06-01

    Cathinone derivatives gained high popularity on the recreational drugs market during the past 10 years. All these compounds are chiral, and the pharmacological potency of the enantiomers of these stimulants is supposed to differ. The goal of this research was to develop a reliable and easy-to-perform high-performance liquid chromatography ultraviolet method for the chiral separation of a set of 24 cathinone derivatives. A commercially available CHIRALPAK® AS-H column consisting of amylose tris [(S)-α-methylbenzylcarbamate] coated on 5-µm silica gel was found to be suitable to resolve a majority of the tested compounds. High-performance liquid chromatography measurements were performed in normal phase mode under isocratic conditions with a mobile phase consisting of hexane, isopropanol, and triethylamine at a flowrate of 1 ml/min. The ratio between hexane and isopropanol was optimized by means of three model substances. Under final conditions with a mobile phase of hexane, isopropanol, and triethylamine (97:3:0.1), 19 out of 24 compounds were successfully resolved into their enantiomers and detected at a wavelength of 254 nm. A correlation between the substituents of the nitrogen atom and the separation results are shown. Furthermore, enantiomer separation results of four cathinone derivatives were compared with the results of their amphetamine analogs.

  9. Separation of dansylated 17β-estradiol, 17α-estradiol, and estrone on a single HPLC column for simultaneous quantitation by LC-MS/MS.

    PubMed

    Szarka, Szabolcs; Nguyen, Vien; Prokai, Laszlo; Prokai-Tatrai, Katalin

    2013-04-01

    We show here that baseline separation of dansylated estrone, 17β-estradiol, and 17α-estradiol can be done, contrary to previous reports, within a short run time on a single RP-LC analytical column packed with particles bonded with phenyl-hexyl stationary phase. The chromatographic method coupled with isotope dilution tandem MS offers a simple assay enabling the simultaneous analysis of these analytes. The method employs (13)C-labeled estrogens as internal standards to eliminate potential matrix effects arising from the use of deuterated estrogens. The assay also offers adequate accuracy and sensitivity to be useful for biological samples. The practical applicability of the validated method is demonstrated by the quantitative analyses of in vivo samples obtained from rats treated with Premarin®. PMID:23371528

  10. Separation of dansylated 17β-estradiol, 17α-estradiol and estrone on a single HPLC column for simultaneous quantitation by LC-MS/MS

    PubMed Central

    Szarka, Szabolcs; Nguyen, Vien; Prokai, Laszlo; Prokai-Tatrai, Katalin

    2013-01-01

    We show here that baseline separation of dansylated estrone, 17β-estradiol and 17α-estradiol can be done, contrary to previous reports, within a short run time on a single RP-LC analytical column packed with particles bonded with phenyl-hexyl stationary phase. The chromatographic method coupled with isotope-dilution tandem MS offers a simple assay enabling the simultaneous analysis of these analytes. The method employs 13C-labeled estrogens as internal standards to eliminate potential matrix effects arising from the use of deuterated estrogens. The assay also offers adequate accuracy and sensitivity to be useful for biological samples. The practical applicability of the validated method is demonstrated by the quantitative analyses of in vivo samples obtained from rats treated with Premarin®. PMID:23371528

  11. HPLC SEPARATION OF CHIRAL ORGANOPHOSPHORUS PESTICIDES ON POLYSACCHARIDE CHIRAL STATIONARY PHASES

    EPA Science Inventory

    High-performance liquid chromatographic separation of the individual enantiomers of 12 organophosphorus pesticides (OPs) were obtained on polysaccharide chiral HPLC columns using an alkane-alcohol mobile phase. The OP pesticides were crotoxyphos, dialifor, dyfonate, fenamiphos, ...

  12. Development of an HPLC post-column antioxidant assay for Solidago canadensis radical scavengers.

    PubMed

    Marksa, Mindaugas; Radušienė, Jolita; Jakštas, Valdas; Ivanauskas, Liudas; Marksienė, Rūta

    2016-01-01

    The aim of this work was to modify and validate the post-column high-performance liquid chromatography (HPLC)-ABTS and DPPH methods for evaluating the antioxidant activity of the methanolic extracts of Solidago canadensis (Canadian goldenrod) leaves and flowers. Separation of the analytes was performed via the HPLC-PDA method on a YMC analytical column using a gradient elution program. Three compounds with antioxidant properties - chlorogenic acid, rutin and isoquercitrin - and two unidentified antioxidants were established. The research showed that the coil temperature regimes and loop length combinations influence the optimised post-column assay method for detecting the antioxidant activity of goldenrod radical scavengers. Investigations established that the temperature in the reaction coil was a substantial factor contributing to the signal strength of the analytes after reacting with the DPPH and ABTS radicals.

  13. Development of an HPLC post-column antioxidant assay for Solidago canadensis radical scavengers.

    PubMed

    Marksa, Mindaugas; Radušienė, Jolita; Jakštas, Valdas; Ivanauskas, Liudas; Marksienė, Rūta

    2016-01-01

    The aim of this work was to modify and validate the post-column high-performance liquid chromatography (HPLC)-ABTS and DPPH methods for evaluating the antioxidant activity of the methanolic extracts of Solidago canadensis (Canadian goldenrod) leaves and flowers. Separation of the analytes was performed via the HPLC-PDA method on a YMC analytical column using a gradient elution program. Three compounds with antioxidant properties - chlorogenic acid, rutin and isoquercitrin - and two unidentified antioxidants were established. The research showed that the coil temperature regimes and loop length combinations influence the optimised post-column assay method for detecting the antioxidant activity of goldenrod radical scavengers. Investigations established that the temperature in the reaction coil was a substantial factor contributing to the signal strength of the analytes after reacting with the DPPH and ABTS radicals. PMID:25835071

  14. Regenerated bacterial cellulose microfluidic column for glycoproteins separation.

    PubMed

    Chen, Chuntao; Zhu, Chunlin; Huang, Yang; Nie, Ying; Yang, Jiazhi; Shen, Ruiqi; Sun, Dongping

    2016-02-10

    To analysis and separate glycoproteins, a simple strategy to prepare regenerated bacterial cellulose (RBC) column with concanavalin A (Con A) lectin immobilized in microfluidic system was applied. RBC was filled into microchannel to fabricate RBC microcolumn after bacterial cellulose dissolved in NaOH-sulfourea water solution. Lectin Con A was covalently connected onto RBC matrix surface via Schiff-base formation. Lysozyme (non-glycoprotein) and transferrin (glycoprotein) were successfully separated based on their different affinities toward the immobilized Con A. Overall, the RBC microfluidic system presents great potential application in affinity chromatography of glycoproteins analysis, and this research represents a significant step to prepare bacterial cellulose (BC) as column packing material in microfluidic system. What is more, troublesome operations for lectin affinity chromatography were simplified by integrating the microfluidic chip onto a HPLC (High Performance Liquid Chromatography) system.

  15. Optimizing Chromatographic Separation: An Experiment Using an HPLC Simulator

    ERIC Educational Resources Information Center

    Shalliker, R. A.; Kayillo, S.; Dennis, G. R.

    2008-01-01

    Optimization of a chromatographic separation within the time constraints of a laboratory session is practically impossible. However, by employing a HPLC simulator, experiments can be designed that allow students to develop an appreciation of the complexities involved in optimization procedures. In the present exercise, a HPLC simulator from "JCE…

  16. Fast HPLC for quality control of Harpagophytum procumbens by using a monolithic silica column: method transfer from conventional particle-based silica column.

    PubMed

    Schmidt, Alexander H

    2005-05-01

    The applicability of a monolithic C18-bonded silica column for the rapid HPLC separation of ingredients in medicinal plants and their phytopharmaceutical preparations has been evaluated in the author's laboratory. In this presentation, an existing method for the determination of the iridoid glycoside harpagoside in Harpagophytum procumbens (Devil's Claw) was successfully transferred from a conventional particle-based C18 silica column to a monolithic silica column. The very high porosity of the stationary phase allows chromatography with a much lower backpressure than on conventional columns. Therefore, the flow rate could be easily increased from 0.8 mL/min (particle-based column) to 5 mL/min (monolithic column) and the run-time reduced from 30 to 5 min (that is a reduction about 85% !), without losing any chromatographic resolution of the compound of interest. The amount of harpagoside was measured with the original method on a conventional particle-based silica column and on the adapted method on a monolithic silica column. The statistical mean t-test showed no significant differences of the variances and the means indicating that the fast HPLC method is an acceptable alternative. The shorter analysis time makes the method very valuable for commercial quality control of Harpagophytum extracts and its pharmaceutical preparations. PMID:15909544

  17. Improvement in HPLC separation of acetic acid and levulinic acid in the profiling of biomass hydrolysate.

    PubMed

    Xie, Rui; Tu, Maobing; Wu, Yonnie; Adhikari, Sushil

    2011-04-01

    5-Hydroxymethylfurfural (HMF) and furfural could be separated by the Aminex HPX-87H column chromatography, however, the separation and quantification of acetic acid and levulinic acid in biomass hydrolysate have been difficult with this method. In present study, the HPLC separation of acetic acid and levulinic acid on Aminex HPX-87H column has been investigated by varying column temperature, flow rate, and sulfuric acid content in the mobile phase. The column temperature was found critical in resolving acetic acid and levulinic acid. The resolution for two acids increased dramatically from 0.42 to 1.86 when the column temperature was lowered from 60 to 30 °C. So did the capacity factors for levulinic acid that was increased from 1.20 to 1.44 as the column temperature dropped. The optimum column temperature for the separation was found at 45 °C. Variation in flow rate and sulfuric acid concentration improved not as much as the column temperature did.

  18. Miniaturized protein separation using a liquid chromatography column on a flexible substrate

    NASA Astrophysics Data System (ADS)

    Yang, Yongmo; Chae, Junseok

    2008-12-01

    We report a prototype protein separator that successfully miniaturizes existing technology for potential use in biocompatible health monitoring implants. The prototype is a liquid chromatography (LC) column (LC mini-column) fabricated on an inexpensive, flexible, biocompatible polydimethylsiloxane (PDMS) enclosure. The LC mini-column separates a mixture of proteins using size exclusion chromatography (SEC) with polydivinylbenzene beads (5-20 µm in diameter with 10 nm pore size). The LC mini-column is smaller than any commercially available LC column by a factor of ~11 000 and successfully separates denatured and native protein mixtures at ~71 psi of the applied fluidic pressure. Separated proteins are analyzed using NuPAGE-gel electrophoresis, high-performance liquid chromatography (HPLC) and an automated electrophoresis system. Quantitative HPLC results demonstrate successful separation based on intensity change: within 12 min, the intensity between large and small protein peaks changed by a factor of ~20. In further evaluation using the automated electrophoresis system, the plate height of the LC mini-column is between 36 µm and 100 µm. The prototype LC mini-column shows the potential for real-time health monitoring in applications that require inexpensive, flexible implant technology that can function effectively under non-laboratory conditions.

  19. Ultrafast UPLC-ESI-MS and HPLC with monolithic column for determination of principal flavor compounds in vanilla pods.

    PubMed

    Sharma, Upendra K; Sharma, Nandini; Sinha, Arun K; Kumar, Neeraj; Gupta, Ajai P

    2009-10-01

    In this study, two novel chromatographic methods based on monolithic column high-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography (UPLC) were developed for the ultrafast determination of principal flavor compounds namely vanillin, vanillic acid, p-hydroxybenzoic acid, and p-hydroxybenzaldehyde in ethanolic extracts of Vanilla planifolia pods. Good separation was achieved within 2.5 min using Chromolith RP18e column (100 mm x 4.6 mm) for HPLC and Acquity BEH C-18 (100 mm x 2.1 mm, 1.7 microm) column for UPLC. Both methods were compared in terms of total analysis time, mobile phase consumption, sensitivity, and validation parameters like precision, accuracy, LOD, and LOQ. Further, system suitability test data including resolution, capacity factor, theoretical plates, and tailing factor was determined for both the methods by ten replicate injections. Monolithic column based HPLC gave better results for most of the selected parameters while UPLC was found to be more eco-friendly with low mobile phase consumption and better sensitivity. Both methods may be used conveniently for the high throughput analysis of large number of samples in comparison to traditional particulate column.

  20. Determination of γ-Aminobutyric Acid (GABA) in Rambutan Fruit cv. Rongrian by HPLC-ELSD and Separation of GABA from Rambutan Fruit Using Dowex 50W-X8 Column.

    PubMed

    Meeploy, Maneerat; Deewatthanawong, Rujira

    2016-03-01

    A high-performance liquid chromatography method coupled with an evaporative light scattering detector (ELSD) was validated for the determination of γ-aminobutyric acid (GABA) in rambutan fruit without any sample pretreatment or derivatization. In the concentration range of 0.05-1.0 mg/mL GABA, the ELSD response was linear with a correlation coefficient (r) >0.999. Limit of detection and limit of quantitation were found to be 0.7 and 2.0 µg/mL, respectively. The method enabled the complete separation of GABA in the aqueous extract of rambutan flesh from the impurity peaks at 45.7 min. The recoveries of sample added GABA were obtained in the range of 92.0-99.3%. Intraday and interday relative standard deviations were <5.3%. Repeatability of the extraction process showed the acceptable precision. From the analysis of GABA content in rambutan flesh, 0.71 ± 0.23 mg of GABA was found in 1 g fresh weight. The recovery of GABA after passing through the Dowex 50W-X8 column was 96.65%. The analytical methodology could be potentially applied to the detection and quantification of GABA in other fruits and complex matrices when a sufficient quantity is available. PMID:26590236

  1. The flotation column as a froth separator

    SciTech Connect

    Schultz, C.W.; Mehta, R.K.; Bates, J.B. )

    1991-12-01

    The Mineral Resources Institute, The University of Alabama, has for the past three years been engaged in a program to develop a beneficiation system for eastern (Devonian) oil shales. One objective of that program was to evaluate advanced technologies for effecting a kerogen-mineral matter separation. Column flotation was among the advanced technologies selected for evaluation. One observation made in the course of optimization testing was that introducing the feed into the froth (above the pulp- froth interface) resulted in an improved combination of concentrate grade and kerogen recovery. This observation was reported in a previous paper. Because the practice of maintaining the pulp froth interface below the feed point is contrary to conventional practice, it was decided to subject the observation to a systematic series of tests. This paper describes a recent series of tests and the results that were obtained.

  2. Preliminary Study of High Resolution HPLC Analytical Method for Sedimentary Pigments Based on Coupled C8 Columns

    NASA Astrophysics Data System (ADS)

    Yao, P.; Yu, Z.; Deng, C.; Liu, S.; Zhao, J.

    2008-05-01

    The pigments in marine water columns can provide accurate estimates of community composition and abundance of phytoplankton. In addition, the sedimentary pigments, especially the derivatives of chlorophyll such as pyrophaeophytins, pyrophaeophorbides and steryl chlorin esters (SCEs) formed during early diagenesis can also provide information on the primary producer community and the changes in paleoproductivity. Accordingly, analysis of pigments and their derivatives is of great importance for oceanography, limnology and geochemistry. Many methods have been developed for the separation of chlorophylls, carotenoids and their derivatives derived from phytoplankton and water column samples using high-performance liquid chromatography (HPLC). Methods widely cited in the literatures include those developed by Wright et al. (1991) and Zapata et al. (2000). Both methods use reversed-phase columns, but C18 column was employed in Wright et al. (1991) and C8 column in Zapata et al. (2000). However, evident coelutions are observed in published works. This will particularly cause problematic identification and quantification in dealing with sedimentary pigments which are highly complex and often display a broad range in polarity. Clearly, it is necessary to improve the separation of the complex pigments if the information carried by the pigments is to be used fully. Coupled C18 columns were used in the HPLC method developed by Airs et al. (2001) for the analysis of complex pigment distributions. Improved chromatographic resolution, more pigment components and novel bacteriochlorophyll derivatives were obtained by this method. It indicates a new road for HPLC method development. C8 column has shorter carbon chains than that of C18 column and can provide less retention of apolar compounds which is of particular advantaged to hydrophobic chlorophyll a, b and their derivatives. That is one of the reasons why the C8 method developed by Zapata et al. (2000) is admittedly better than

  3. Fast and Universal HPLC Method for Determination of Permethrin in Formulations Using 1.8-µm Particle-Packed Column and Performance Comparison with Other Column Types

    PubMed Central

    Shishovska, Maja A.; Stefova, Marina T.

    2012-01-01

    An HPLC method has been developed for the fast separation and quantification of permethrin using C18 column packed with 1.8 µm particles. The method is specific with good resolution to degradation products and to other present components. It has acceptable validation results. The run time is 4.5 min (or may be within 1.6 min is rapid resolution mode) with an organic solvent consumption of 3.6 mL per run. The method has been applied to samples of formulations for various uses: mattress cleaner, shampoo, and veterinary powder. The performance of the applied column is compared with other common column types. The relationships between linear velocity of the mobile phase (u) and resolution factor (Rs), back-pressure (ΔP), and efficiency (H) are presented. The experimental data shows the advantages of 1.8-µm particle columns to be a significant reduction in solvent consumption (by factor of 4.4 and 1.5) and a reduction in run-time (by factor 4.7 and 1.5), and the weaknesses are a high back-pressure and lower efficiency. Finally, it has been shown that use of 1.8-µm particle packed columns with conventional HPLC systems is possible, but with limitations in mobile phase flow-rate. PMID:22291055

  4. The effect of re-dissolution solvents and HPLC columns on the analysis of mycosporine-like amino acids in the eulittoral macroalgae Prasiola crispa and Porphyra umbilicalis

    NASA Astrophysics Data System (ADS)

    Karsten, Ulf; Escoubeyrou, Karine; Charles, François

    2009-09-01

    Many macroalgal species that are regularly exposed to high solar radiation such as the eulittoral green alga Prasiola crispa and the red alga Porphyra umbilicalis synthesize and accumulate high concentrations of mycosporine-like amino acids (MAAs) as UV-sunscreen compounds. These substances are typically extracted with a widely used standard protocol following quantification by various high performance liquid chromatography (HPLC) techniques. However, further preparation steps prior to HPLC analysis as well as different HPLC column types have not been systematically checked regarding separation quality and reproducibility. Therefore pure methanol, distilled water and HPLC eluent were evaluated as re-dissolution solvent for dried Prasiola and Porphyra extracts, which were subsequently analyzed on three reversed-phase C8 and C18 HPLC columns. The data indicate that distilled water and the HPLC eluent gave almost identical peak patterns and MAA contents on the C8 and C18 columns. In contrast, the application of the widely used methanol led to double peaks or even the loss of specific peaks as well as to a strong decline in total MAA amounts ranging from about 35% of the maximum in P. crispa to 80% of the maximum in P. umbilicalis. Consequently, methanol should be avoided as re-dissolution solvent for the HPLC sample preparation. An improved protocol for the MAA analysis in macroalgae in combination with a reliable C18 column is suggested.

  5. Retention of [(18)F]fluoride on reversed phase HPLC columns.

    PubMed

    Ory, Dieter; Van den Brande, Jeroen; de Groot, Tjibbe; Serdons, Kim; Bex, Marva; Declercq, Lieven; Cleeren, Frederik; Ooms, Maarten; Van Laere, Koen; Verbruggen, Alfons; Bormans, Guy

    2015-01-01

    As [(18)F]fluoride is a starting reagent in the radiosynthesis of most fluorine-18 labeled positron emission tomography (PET) tracers, its chromatographic behavior on reversed phase (RP) HPLC columns is important for the purification performance and accuracy of RP HPLC quality control methods. We have investigated the chromatographic behavior and recovery of [(18)F]fluoride as a function of the type and brand of RP HPLC column, the pH and the composition of the mobile phase. Elution and elution profile of [(18)F]fluoride from six RP-HPLC columns (Waters XBridge C18 3 mm × 100 mm 3.5 μm; Grace Platinum EPS C18 4.6 mm × 100 mm, 3 μm; Waters XTerra C18 4.6 mm × 250 mm, 5 μm; Phenomenex C18 4.6 mm × 150 mm, 5 μm; Hamilton PRP-1 column 4.1 mm × 150 mm, 5 μm; Merck KGaA Chromolith Performance C18 3 mm × 100 mm) eluted with mobile phase composed of phosphate or acetate buffers (pH 2, 3, 4, 5, 7.3 and 9) and acetonitrile or ethanol as organic modifier were characterized. The elution profile was determined by on-line radioactivity measurement in the column eluate and recovery was calculated by comparison of radioactivity eluted with the HPLC column present or absent in the chromatographic flow path. Interestingly, [(18)F]fluoride recovery increased with increasing pH. At pH 3 all packed silica-based columns showed significant retention of fluorine-18, whereas almost no retention was observed on a polymeric PRP-1 column. However at pH 5, [(18)F]fluoride recovery was above 90% for each tested column. In addition, small differences were observed when changing the composition of the mobile phase. We therefore recommend to use a mobile phase with pH > 5 for silica based C18 columns for both quality control and semi-preparative HPLC of fluorine-18 labeled PET radiopharmaceuticals. If required a lower pH can be used in combination with a polymer based HPLC column.

  6. Direct coupling of microbore HPLC columns to MS systems

    NASA Technical Reports Server (NTRS)

    Mcnair, H. M.

    1985-01-01

    A detailed investigation using electron microscopy was conducted which examined the conditions of materials used in the construction of stable, high performance microbore liquid chromatography (LC) columns. Small details proved to be important. The effects of temperature on the elution of several homologous series used as probe compounds was examined in reverse phase systems. They showed that accessible temperature changes provide roughly half the increase in solvent strength that would be obtained going from a 100% aqueous to a 100% organic mobile phase, which is sufficient to warrant their use in many analyses requiring the use of gradients. Air circulation temperature control systems provide the easiest means of obtaining rapid, wide range changes in column temperature. However, slow heat transfer from the gas leads to thermal nonuniformity in the column and a decrease in resolution as the temperature program progresses.

  7. A rapid HPLC method for monitoring plasma levels of caffeine and theophylline using solid phase extraction columns.

    PubMed

    Pickard, C E; Stewart, A D; Hartley, R; Lucock, M D

    1986-07-01

    A simple HPLC method for the determination of caffeine and theophylline in plasma is described. Separation of theobromine, paraxanthine, theophylline, beta-hydroxyethyltheophylline and caffeine is obtained using a mobile phase of 1% acetic acid/methanol (83:17, v/v) and a Waters Associates NOVA-PAK C18 column protected by a Guard-PAK precolumn module containing a Guard-PAK CN cartridge. Rapid sample preparation is achieved by solid-phase extraction columns (Bond-Elut C18, 1 mL capacity) which provide excellent recovery values for both drugs. The cost per sample using this approach can be minimised by column regeneration and re-use. Results obtained for theophylline are in good agreement with values determined by other techniques.

  8. Isolation and identification of arctiin and arctigenin in leaves of burdock (Arctium lappa L.) by polyamide column chromatography in combination with HPLC-ESI/MS.

    PubMed

    Liu, Shiming; Chen, Kaoshan; Schliemann, Willibald; Strack, Dieter

    2005-01-01

    A simple method involving polyamide column chromatography in combination with HPLC-PAD and HPLC-ESI/MS for isolating and identifying two kinds of lignans, arctiin and arctigenin, in the leaves of burdock (Arctium lappa L.) has been established. After extraction of burdock leaves with 80% methanol, the aqueous phase of crude extracts was partitioned between water and chloroform and the aqueous phase was fractionated on a polyamide glass column. The fraction, eluting with 100% methanol, was concentrated and gave a white precipitate at 4 degrees C from which two main compounds were purified by semi-preparative HPLC. In comparison with the UV and ESI-MS spectra and the HPLC retention time of authentic standards, the compounds were determined to be arctiin and arctigenin. The extraction/separation technique was validated using an internal standard method.

  9. Separation of hydrophobic metabolites using monolithic silica column in high-performance liquid chromatography and supercritical fluid chromatography.

    PubMed

    Bamba, Takeshi; Fukusaki, Eiichiro

    2009-08-01

    Monolithic silica columns have very low back-pressures and offer several advantages over conventional columns packed with spherical particles, such as high separation efficiency and rapid analysis. In this review, we report the applicability of monolithic silica columns for the analysis of complex hydrophobic metabolites. We have used monolithic columns in HPLC and developed a separation technique for the high-speed and high-resolution analysis of the geometric analogs of natural polyprenols. We also used monolithic columns in supercritical fluid chromatography for the successful separation of the structural isomers of carotenoids after deciding the analytical conditions that were suitable for this separation and have developed a method for profiling biological samples containing complex matrices. We have proved that excellent resolution can be obtained by connecting a number of monolithic columns in series.

  10. Absorbance detector based on a deep UV light emitting diode for narrow-column HPLC.

    PubMed

    Bui, Duy Anh; Bomastyk, Benjamin; Hauser, Peter C

    2013-10-01

    A detector for miniaturized HPLC based on deep UV emitting diodes and UV photodiodes was constructed. The measurement is accomplished by the transverse passage of the radiation from the light-emitting diode (LED) through fused-silica tubing with an internal diameter of 250 μm. The optical cell allows flexible alignment of the LED, tubing, and photodiode for optimization of the light throughput and has an aperture to block stray light. A beam splitter was employed to direct part of the emitted light to a reference photodiode and the Lambert-Beer law was emulated with a log-ratio amplifier circuitry. The detector was tested with two LEDs with emission bands at 280 and 255 nm and showed noise levels as low as 0.25 and 0.22 mAU, respectively. The photometric device was employed successfully in separations using a column of 1 mm inner diameter in isocratic as well as gradient elution. Good linearities over three orders of magnitude in concentration were achieved, and the precision of the measurements was better than 1% in all cases. Detection down to the low micromolar range was possible. PMID:23893947

  11. Absorbance detector based on a deep UV light emitting diode for narrow-column HPLC.

    PubMed

    Bui, Duy Anh; Bomastyk, Benjamin; Hauser, Peter C

    2013-10-01

    A detector for miniaturized HPLC based on deep UV emitting diodes and UV photodiodes was constructed. The measurement is accomplished by the transverse passage of the radiation from the light-emitting diode (LED) through fused-silica tubing with an internal diameter of 250 μm. The optical cell allows flexible alignment of the LED, tubing, and photodiode for optimization of the light throughput and has an aperture to block stray light. A beam splitter was employed to direct part of the emitted light to a reference photodiode and the Lambert-Beer law was emulated with a log-ratio amplifier circuitry. The detector was tested with two LEDs with emission bands at 280 and 255 nm and showed noise levels as low as 0.25 and 0.22 mAU, respectively. The photometric device was employed successfully in separations using a column of 1 mm inner diameter in isocratic as well as gradient elution. Good linearities over three orders of magnitude in concentration were achieved, and the precision of the measurements was better than 1% in all cases. Detection down to the low micromolar range was possible.

  12. A sensitive post-column photochemical derivatization/fluorimetric detection system for HPLC determination of bisphosphonates.

    PubMed

    Pérez-Ruiz, Tomás; Martínez-Lozano, Carmen; García-Martínez, María Dolores

    2009-02-27

    A new reversed-phase ion-pair high-performance liquid chromatographic (HPLC) method has been developed for the determination of the following bisphosphonic acids: alendronic acid (ALEN), etidronic acid (ETID), ibandronic acid (IBAN) and risedronic acid (RISE). Separation was achieved on a C(18) column using a mixture of 50 mmol L(-1) borate buffer pH 9.0 containing 0.25 mmol L(-1) tetrabutylammonium chloride and 0.5 mmol L(-1) EDTA and acetonitrile (97:3) as the mobile phase. The sensitive detection of the above bisphosphonic acids was based on their oxidation to orthophosphate by the on-line peroxydisulfate-assisted photolysis followed by post-column reaction with molybdate to yield phosphomolybdate. This subsequently reacted with thiamine to generate thiochrome and, finally, the fluorescence of thiochrome was measured at 440 nm with excitation at 375 nm. The developed method is precise with a mean relative standard deviation of 1.3%, sensitive (with a detection limit at the nmol L(-1) level), accurate, specific, rapid (analysis time approximately 13 min) and inexpensive because to the low cost of the reagents. The assay was applied to the analysis of the four bisphosphonic acids in commercial dosage formulations, in which the excipients did not interfere with the determination. The method was also applied to the determination of etidronate, risedronate and ibandronate in human urine. Sample preparation involves precipitation of the analytes from urine along with endogenous phosphates such as calcium salts by addition of calcium chloride at alkaline pH and dissolution of the precipitate in 0.05 mol L(-1) ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid. PMID:19150069

  13. HILIC separation mechanisms of tetracyclines on amino bonded silica column

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Effects of mobile phase variations on the chromatographic separation on amino bonded silica column in hydrophilic interaction chromatography (HILIC) were investigated for four zwitterionic tetracyclines (TCs): oxytetracycline, doxycycline, chlortetracycline and tetracycline. A mixed-mode retention m...

  14. Preparation and Characterization of a Polymeric Monolithic Column for Use in High-Performance Liquid Chromatography (HPLC)

    ERIC Educational Resources Information Center

    Bindis, Michael P.; Bretz, Stacey Lowery; Danielson, Neil D.

    2011-01-01

    The high-performance liquid chromatography (HPLC) experiment, most often done in the undergraduate analytical instrumentation laboratory course, generally illustrates reversed-phase chromatography using a commercial C[subscript]18 silica column. To avoid the expense of periodic column replacement and introduce a choice of columns with different…

  15. Design and Prototype of an Automated Column-Switching HPLC System for Radiometabolite Analysis.

    PubMed

    Vasdev, Neil; Collier, Thomas Lee

    2016-01-01

    Column-switching high performance liquid chromatography (HPLC) is extensively used for the critical analysis of radiolabeled ligands and their metabolites in plasma. However, the lack of streamlined apparatus and consequently varying protocols remain as a challenge among positron emission tomography laboratories. We report here the prototype apparatus and implementation of a fully automated and simplified column-switching procedure to allow for the easy and automated determination of radioligands and their metabolites in up to 5 mL of plasma. The system has been used with conventional UV and coincidence radiation detectors, as well as with a single quadrupole mass spectrometer. PMID:27548189

  16. Design and Prototype of an Automated Column-Switching HPLC System for Radiometabolite Analysis

    PubMed Central

    Vasdev, Neil; Collier, Thomas Lee

    2016-01-01

    Column-switching high performance liquid chromatography (HPLC) is extensively used for the critical analysis of radiolabeled ligands and their metabolites in plasma. However, the lack of streamlined apparatus and consequently varying protocols remain as a challenge among positron emission tomography laboratories. We report here the prototype apparatus and implementation of a fully automated and simplified column-switching procedure to allow for the easy and automated determination of radioligands and their metabolites in up to 5 mL of plasma. The system has been used with conventional UV and coincidence radiation detectors, as well as with a single quadrupole mass spectrometer. PMID:27548189

  17. Enantiomeric determination of D-, L-lactate in diabetic rat urine using a column-switching HPLC

    NASA Astrophysics Data System (ADS)

    Chen, Chien-Ming; Tsai, Yih-Chiao; Fukushima, Takeshi; Imai, Kazuhiro; Lee, Jen-Ai

    2005-04-01

    A highly sensitive method for the determination of D-lactate in rat urine was developed by using a high-performance liquid chromatography (HPLC) with an octadecylsilica (ODS) connected to a chiral column. At first, (D+L)-lactate in the urine were derivatized with a fluorescent reagent, 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ), and separated on the ODS column and determined fluorimetrically at 547 nm with 491 nm of excitation wavelength. During the separation step on the ODS, the peak fraction of (D+L)-lactate derivative was introduced directly to an amylose tris (3,5-dimethylphenylcarbamate) (Chiralpak AD-RH) chiral column by changing the flow of the eluent via 6-port valve. Then, D-lactate derivative was separated enantiomerically from L-lactate derivative, and the enantiomeric ratio was determined from the chromatogram. The accuracy values for the determination of D-lactate in 20 μL of rat urine were 96.93% - 104.85%. The intra- and inter-day precision values were within 0.80% and 14.44%. The proposed method was applied to the urine of diabetic rats induced by intraperitoneal administration of streptozotocin, and the significant increases of D-lactate was observed in the diabetic rats as compared to the normal rats.

  18. METHOD TO TEST ISOTOPIC SEPARATION EFFICIENCY OF PALLADIUM PACKED COLUMNS

    SciTech Connect

    Heung, L; Gregory Staack, G; James Klein, J; William Jacobs, W

    2007-06-27

    The isotopic effect of palladium has been applied in different ways to separate hydrogen isotopes for many years. At Savannah River Site palladium deposited on kieselguhr (Pd/k) is used in a thermal cycling absorption process (TCAP) to purify tritium for over ten years. The need to design columns for different throughputs and the desire to advance the performance of TCAP created the need to evaluate different column designs and packing materials for their separation efficiency. In this work, columns with variations in length, diameter and metal foam use, were tested using an isotope displacement method. A simple computer model was also developed to calculate the number of theoretical separation stages using the test results. The effects of column diameter, metal foam and gas flow rate were identified.

  19. High efficiency, high temperature separations on silica based monolithic columns.

    PubMed

    Rogeberg, Magnus; Wilson, Steven Ray; Malerod, Helle; Lundanes, Elsa; Tanaka, Nobuo; Greibrokk, Tyge

    2011-10-14

    The effect of temperature on separation using reversed-phase monolithic columns has been investigated using a nano-LC pumping system for gradient separation of tryptic peptides with MS detection. A goal of this study was to find optimal conditions for high-speed separations. The chromatographic performance of the columns was evaluated by peak capacity and peak capacity per time unit. Column lengths ranging from 20 to 100 cm and intermediate gradient times from 10 to 30 min were investigated to assess the potential of these columns in a final step separation, e.g. after fractionation or specific sample preparation. Flow rates from 250 to 2000 nL/min and temperatures from 20 to 120°C were investigated. Temperature had a significant effect on fast separations, and a flow rate of 2000 nL/min and a temperature of 80°C gave the highest peak capacity per time unit. These settings produced 70% more protein identifications in a biological sample compared to a conventional packed column. Alternatively, an equal amount of protein identifications was obtained with a 40% reduction in run time compared to the conventional packed column.

  20. Separation of polyprenol and dolichol by monolithic silica capillary column chromatography.

    PubMed

    Bamba, Takeshi; Fukusaki, Eiiciro; Minakuchi, Hiroshi; Nakazawa, Yoshihisa; Kobayashi, Akio

    2005-10-01

    We attempted an analysis of naturally occurring polyprenol and dolichol using a monolithic silica capillary column in HPLC. First, the separation of the polyprenol mixture alone was performed using a 250 x 0.2 mm inner diameter (ID) octadecylsilyl (ODS)-monolithic silica capillary column. The resolution of the separation between octadecaprenol (prenol 18) and nonadecaprenol (prenol 19) exceeded by >or=2-fold the level recorded when using a conventional ODS-silica particle-packed column (250 x 4.6 mm ID) under the same elution conditions. Next, the mixture of the prenol type (polyprenol) and dolichol type (dihydropolyprenol) was subjected to this capillary HPLC system, and the separation of each homolog was successfully achieved. During the analysis of polyprenol fraction derived from Eucommia ulmoides leaves, dolichols were found as a single peak, including all-trans-polyprenol and cis-polyprenol previously identified. This sensitive high-resolution system is very useful for the analysis of compounds that are structurally close to polyprenols and dolichols and that have a low content.

  1. Column-switching HPLC determination of mexiletine using tris(bipyridine)ruthenium(III) electrogenerated chemiluminescence and precolumn derivatization with divinylsulfone.

    PubMed

    Kobayashi, Naoko; Miyamoto, Aoi; Uchikura, Kazuo

    2010-01-01

    A sensitive HPLC method combined with a column-switching system and tris(bipyridine)ruthenium(III) electrogenerated chemiluminescence (ECL) detection has been developed for the quantitative determination of mexiletine (MEX). MEX was derivatized by divinylsulfone (DVS) and then measured. The optimum conditions for the derivatization reaction were 10 µL of sample solutions, 40 mM DVS (pH 8.0), a reaction temperature of 50°C, and a reaction time of 15 min. The derivatized samples were cleaned up by an on-line pretreatment column. Also, after column-switching to the analytical column, the derivatized MEX was separated and detected. The calibration curves of MEX in human control serum showed good linear regression (r = 0.9996) from 0.008 to 6.56 µg ml(-1). The detection limit of MEX was 0.008 µg ml(-1) (S/N = 3). At a concentration of 2.0 µg ml(-1) MEX, the relative standard deviation (n = 5) was 0.98%. In this method the concentration of MEX in human control serum was readily measured, and this method was successfully applied to the time courses of the concentration of MEX in rabbit plasma after intravenous administration. The proposed method involved a simple and minimum sample-preparation procedure and a short run time (<20 min). Therefore it can be applied to routine therapeutic monitoring and pharmacokinetic studies of MEX. PMID:21157099

  2. Enantiomeric separation of asymmetric triacylglycerol by recycle high-performance liquid chromatography with chiral column.

    PubMed

    Nagai, Toshiharu; Mizobe, Hoyo; Otake, Ikuko; Ichioka, Kenji; Kojima, Koichi; Matsumoto, Yumiko; Gotoh, Naohiro; Kuroda, Ikuma; Wada, Shun

    2011-05-20

    In our previous studies, we employed recycle HPLC for the separation of triacylglycerol (TAG)-positional isomers (PIs). In this study, a recycle HPLC system equipped with a polysaccharide-based chiral column was applied to the enantiomeric separation of some asymmetric TAGs having straight-chain C16-C18 acyl residues. As a result, 1,2-dipalmitoyl-3-oleoyl-rac-glycerol (rac-PPO), 1,2-dioleoyl-3-palmitoyl-rac-glycerol (rac-OOP), and 1,2-dipalmitoyl-3-linoleoyl-rac-glycerol (rac-PPL) were resolved into their respective enantiomers. However, neither 1,2-dioleoyl-3-linoleoyl-rac-glycerol (rac-OOL), consisting of only unsaturated fatty acids, nor 1,2-dipalmitoyl-3-stearoyl-rac-glycerol (rac-PPS), consisting of only saturated fatty acids, was resolved. These results suggest that the asymmetric TAGs, used in this study, having both a palmitic acid moiety and an oleic acid (or a linoleic acid) moiety at the sn-1 or sn-3 positions are resolved by the chiral column. This new chiral separation method can be used in combination with atmospheric pressure chemical ionization mass spectrometry to determine the sn-OOP/sn-POO ratio in palm oil. This method is applicable for the chiral separation of asymmetric TAGs in palm oil.

  3. A rapid HPLC column switching method for sample preparation and determination of β-carotene in food supplements.

    PubMed

    Brabcová, Ivana; Hlaváčková, Markéta; Satínský, Dalibor; Solich, Petr

    2013-11-15

    A simple and automated HPLC column-switching method with rapid sample pretreatment has been developed for quantitative determination of β-carotene in food supplements. Commercially samples of food supplements were dissolved in chloroform with help of saponification with 1M solution of sodium hydroxide in ultrasound bath. A 20-min sample dissolution/extraction step was necessary before chromatography analysis to transfer β-carotene from solid state of food supplements preparations (capsules,tablets) to chloroform solution. Sample volume - 3μL of chloroform phase was directly injected into the HPLC system. Next on-line sample clean-up was achieved on the pretreatment precolumn Chromolith Guard Cartridge RP-18e (Merck), 10×4.6mm, with a washing mobile phase (methanol:water, 92:8, (v/v)) at a flow rate of 1.5mL/min. Valve switch to analytical column was set at 2.5min in a back-flush mode. After column switching to the analytical column Ascentis Express C-18, 30×4.6mm, particle size 2.7μm (Sigma Aldrich), the separation and determination of β-carotene in food supplements was performed using a mobile phase consisting of 100% methanol, column temperature at 60°C and flow rate 1.5mL/min. The detector was set at 450nm. Under the optimum chromatographic conditions standard calibration curve was measured with good linearity - correlation coefficient for β-carotene (r(2)=0.999014; n=6) between the peak areas and concentration of β-carotene 20-200μg/mL. Accuracy of the method defined as a mean recovery was in the range 96.66-102.40%. The intraday method precision was satisfactory at three concentration levels 20, 125 and 200μg/mL and relative standard deviations were in the range 0.90-1.02%. The chromatography method has shown high sample throughput during column-switching pretreatment process and analysis in one step in short time (6min) of the whole chromatographic analysis.

  4. Model of decision system for 13C Isotope Separation column

    NASA Astrophysics Data System (ADS)

    Boca, M. L.

    2015-11-01

    This paper presents the model of a decisional system for 13C Isotope Separation column, which is used to detect mission critical situation. The start model was a model of one distributed control system of critical situations that may arise in the operation of the distillation column. The research work it is proposed a model of decision system which implement a temperature sensor inside of liquid nitrogen level in the condenser. The condenser is a part of column where take place the cryogenic process using nitrogen liquid. The work temperature is very low about -192oC, and because the temperature can grow or go down more than 2 degrees is a very critical location inside the column. In this way the column has a deeply monitor and supervised and it take a decision in a proper time when the temperature is grow up or getting down and became a critical situation. For monitor and supervised it was used MatLAB SimuLink. The model, the decision system gives a signal to one sensor when something is wrong in the condenser which is the most critical place of the isotopic column. In this way it creates an alarm that something is getting wrong in the isotopic column.

  5. An automated HPLC method for the fractionation of polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins, and polychlorinated dibenzofurans in fish tissue on a porous graphitic carbon column

    USGS Publications Warehouse

    Echols, Kathy R.; Gale, Robert W.; Tillitt, Donald E.; Schwartz, Ted R.; O'Laughlin, Jerome

    1997-01-01

    The Ah (aryl-hydrocarbon) hydroxylase-receptor active polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) were fractionated by an automated high-performance liquid chromatography (HPLC) system using the Hypercarb™ porous graphitic carbon (PGC) column. This commercially available column was used to fractionate the di-, mono-, and non-ortho PCBs into three fractions for gas chromatography (GC)/electron capture detection analysis, and a fourth fraction containing the PCDDs/PCDFs for GC/mass spectrometry analysis. The recoveries of the PCBs ranged from 68 to 96%, and recoveries of the PCDDs/PCDFs ranged from 74 to 123%. The PGC column has the advantage of faster separations (110 min versus 446 min) and less solvent use (275 ml versus 1,100 ml) compared with automated fractionation of these compounds on activated carbon (PX-21), while still affording good separation of the classes. The PGC column may have an advantage over the pyrenyl-based HPLC method because it has a greater loading capacity (400 μg total PCBs versus 250 μg). Overall, the PGC is a standard column that provides reproducible fractionation of PCDD/PCDFs and PCBs for analytical measurement in environmental samples.

  6. Determination of antioxidants by a novel on-line HPLC-cupric reducing antioxidant capacity (CUPRAC) assay with post-column detection.

    PubMed

    Celik, Saliha Esin; Ozyürek, Mustafa; Güçlü, Kubilay; Apak, Reşat

    2010-07-26

    A novel on-line HPLC-cupric reducing antioxidant capacity (CUPRAC) method was developed for the selective determination of polyphenols (flavonoids, simple phenolic and hydroxycinnamic acids) in complex plant matrices. The method combines chromatographic separation, constituent analysis, and post-column identification of antioxidants in plant extracts. The separation of polyphenols was performed on a C18 column using gradient elution with two different mobile phase solutions, i.e., MeOH and 0.2% o-phosphoric acid. The HPLC-separated antioxidant polyphenols in the extracts react with copper(II)-neocuproine (Cu(II)-Nc) reagent in a post-column reaction coil to form a derivative. The reagent is reduced by antioxidants to the copper(I)-neocuproine (Cu(I)-Nc) chelate having maximum absorption at 450 nm. The negative peaks of antioxidant constituents were monitored by measuring the increase in absorbance due to Cu(I)-Nc. The detection limits of polyphenols at 450 nm (in the range of 0.17-3.46 microM) after post-column derivatization were comparable to those at 280 nm UV detection without derivatization. The developed method was successfully applied to the identification of antioxidant compounds in crude extracts of Camellia sinensis, Origanum marjorana and Mentha. The method is rapid, inexpensive, versatile, non-laborious, uses stable reagents, and enables the on-line qualitative and quantitative estimation of antioxidant constituents of complex plant samples.

  7. Bubble column apparatus for separating wax from catalyst slurry

    SciTech Connect

    Neathery, James K.; Davis, Burtron H.

    2004-07-13

    Novel methods and devices for production of liquid hydrocarbon products from gaseous reactants are disclosed. In one aspect, a method for separating a liquid hydrocarbon, typically a wax, from a catalyst containing slurry is provided, comprising passing the slurry through at least one downcomer extending from an overhead separation chamber and discharging into the bottom of a slurry bubble column reactor. The downcomer includes a cross-flow filtration element for separating a substantially particle-free liquid hydrocarbon for downstream processing. In another aspect, a method for promoting plug-flow movement in a recirculating slurry bubble column reactor is provided, comprising discharging the recirculating slurry into the reactor through at least one downcomer which terminates near the bottom of the reactor. Devices for accomplishing the above methods are also provided.

  8. The limits of the separation power of unidimensional column liquid chromatography.

    PubMed

    Guiochon, Georges

    2006-09-01

    The practical limit of the separation power of HPLC depends on time, money, and skill. That is it depends on the time available for the analysis, on the quality and performance of the pump and hardware and particularly on the maximum pressure at which the pump can deliver the mobile phase to the column, and on the temperature at which the column can be operated. It also depends on the properties of the packing material selected (e.g., its particle size, its pore geometry, and its connectivity) and on the packing method used since it affects the coefficients of the HETP equation. Finally, it depends on the thermal stability of the sample and the packing material. The complexity of the sample also plays an important role in that it determines whether the analysis should be made under isocratic, isothermal conditions, in gradient elution, in temperature programming, or with a combination of both types of programming. The various phenomena that affect column properties and separation performance are discussed. Past achievements suggest that columns providing efficiencies in excess of a million plates in less than 1 day are within the grasp of current technology. The possibility of further advances are considered.

  9. Optimization of separation and determination of moxifloxacin and its related substances by RP-HPLC.

    PubMed

    Djurdjevic, Predrag; Ciric, Andrija; Djurdjevic, Aleksandra; Stankov, Milena Jelikic

    2009-09-01

    A RP-HPLC method for the separation and determination of impurities of moxifloxacin, in its pharmaceutical forms as well as moxifloxacin degradation products, was developed with the aid of DryLab software and chemometric (response surface) approach. The separation of four synthesis-related impurities was achieved on a Waters C(18) XTerra column using a mobile phase of (water+triethylamine (2%, v/v)): acetonitrile=90:10 (v/v%); the pH of water phase being adjusted with phosphoric acid to 6.0. Flow rate of the mobile phase was 1.5 ml/min and UV detection at 290 nm was employed. The column was thermostated at 45 degrees C. The resolution between the two least resolved impurity peaks was in average, R(s,min) > 1.5. Method validation parameters indicate linear dynamic range 0.2-2.0 microg/ml with LOQ ca. 0.20 microg/ml and LOD ca. 0.05 microg/ml for all analytes. The method was applied for the impurities determination in drug tablets and infusion (Avelox), Bayer AG) and for degradation products determination in a stability study of moxifloxacin. The impurity content in the tablets and infusion was quantified as 0.1% of total drug. Two degradation products were noted under hydrolytic conditions. The method can also be used for rapid and accurate quantification of moxifloxacin hydrochloride in its tablets during stability testing.

  10. Systems for column-based separations, methods of forming packed columns, and methods of purifying sample components

    DOEpatents

    Egorov, Oleg B.; O'Hara, Matthew J.; Grate, Jay W.; Chandler, Darrell P.; Brockman, Fred J.; Bruckner-Lea, Cynthia J.

    2000-01-01

    The invention encompasses systems for column-based separations, methods of packing and unpacking columns and methods of separating components of samples. In one aspect, the invention includes a method of packing and unpacking a column chamber, comprising: a) packing a matrix material within a column chamber to form a packed column; and b) after the packing, unpacking the matrix material from the column chamber without moving the column chamber. In another aspect, the invention includes a system for column-based separations, comprising: a) a fluid passageway, the fluid passageway comprising a column chamber and a flow path in fluid communication with the column chamber, the flow path being obstructed by a retaining material permeable to a carrier fluid and impermeable to a column matrix material suspended in the carrier fluid, the flow path extending through the column chamber and through the retaining material, the flow path being configured to form a packed column within the column chamber when a suspension of the fluid and the column matrix material is flowed along the flow path; and b) the fluid passageway extending through a valve intermediate the column chamber and the retaining material.

  11. Systems For Column-Based Separations, Methods Of Forming Packed Columns, And Methods Of Purifying Sample Components.

    DOEpatents

    Egorov, Oleg B.; O'Hara, Matthew J.; Grate, Jay W.; Chandler, Darrell P.; Brockman, Fred J.; Bruckner-Lea, Cynthia J.

    2004-08-24

    The invention encompasses systems for column-based separations, methods of packing and unpacking columns and methods of separating components of samples. In one aspect, the invention includes a method of packing and unpacking a column chamber, comprising: a) packing a matrix material within a column chamber to form a packed column; and b) after the packing, unpacking the matrix material from the column chamber without moving the column chamber. In another aspect, the invention includes a system for column-based separations, comprising: a) a fluid passageway, the fluid passageway comprising a column chamber and a flow path in fluid communication with the column chamber, the flow path being obstructed by a retaining material permeable to a carrier fluid and impermeable to a column matrix material suspended in the carrier fluid, the flow path extending through the column chamber and through the retaining material, the flow path being configured to form a packed column within the column chamber when a suspension of the fluid and the column matrix material is flowed along the flow path; and b) the fluid passageway extending through a valve intermediate the column chamber and the retaining material.

  12. Systems For Column-Based Separations, Methods Of Forming Packed Columns, And Methods Of Purifying Sample Components

    DOEpatents

    Egorov, Oleg B.; O'Hara, Matthew J.; Grate, Jay W.; Chandler, Darrell P.; Brockman, Fred J.; Bruckner-Lea, Cynthia J.

    2006-02-21

    The invention encompasses systems for column-based separations, methods of packing and unpacking columns and methods of separating components of samples. In one aspect, the invention includes a method of packing and unpacking a column chamber, comprising: a) packing a matrix material within a column chamber to form a packed column; and b) after the packing, unpacking the matrix material from the column chamber without moving the column chamber. In another aspect, the invention includes a system for column-based separations, comprising: a) a fluid passageway, the fluid passageway comprising a column chamber and a flow path in fluid communication with the column chamber, the flow path being obstructed by a retaining material permeable to a carrier fluid and impermeable to a column matrix material suspended in the carrier fluid, the flow path extending through the column chamber and through the retaining material, the flow path being configured to form a packed column within the column chamber when a suspension of the fluid and the column matrix material is flowed along the flow path; and b) the fluid passageway extending through a valve intermediate the column chamber and the retaining material.

  13. Method for determination of aflatoxin M₁ in cheese and butter by HPLC using an immunoaffinity column.

    PubMed

    Sakuma, Hisako; Kamata, Yoichi; Sugita-Konishi, Yoshiko; Kawakami, Hiroshi

    2011-01-01

    A rapid, sensitive convenient method for determination of aflatoxin M₁ (AFM₁) in cheese and butter by HPLC was developed and validated. The method employs a safe extraction solution (mixture of acetonitrile, methanol and water) and an immunoaffinity column (IAC) for clean-up. Compared with the widely used method employing chloroform and a Florisil column, the IAC method has a short analytical time and there are no interference peaks. The limits of quantification (LOQ) of the IAC method were 0.12 and 0.14 µg/kg, while those of the Florisil column method were 0.47 and 0.23 µg/kg in cheese and buffer, respectively. The recovery and relative standard deviation (RSD) for cheese (spiked at 0.5 µg/kg) in the IAC method were 92% and 7%, respectively, while for the Florisil column method the corresponding values were 76% and 10%. The recovery and RSD for butter (spiked at 0.5 µg/kg) in the IAC method were 97% and 9%, and those in the Florisil method were 74% and 9%, respectively. In the IAC method, the values of in-house precision (n=2, day=5) of cheese and butter (spiked at 0.5 µg/kg) were 9% and 13%, respectively. The IAC method is superior to the Florisil column method in terms of safety, ease of handling, sensitivity and reliability. A survey of AFM₁ contamination in imported cheese and butter in Japan was conducted by the IAC method. AFM₁ was not detected in 60 samples of cheese and 30 samples of butter.

  14. Real-time separation of natural products by ultrafast 2D NMR coupled to on-line HPLC.

    PubMed

    Queiroz, Luiz H K; Queiroz, Darlene P K; Dhooghe, Liene; Ferreira, Antonio G; Giraudeau, Patrick

    2012-05-21

    Hyphenated HPLC-NMR is an extremely efficient analytical tool, which makes it possible to perform on-flow experiments where 1D NMR spectra are obtained in real time as the analytes are separated and eluted from the chromatographic column. However, it is incompatible with multidimensional NMR methods that form an indispensible tool for the study of complex mixtures. Recently, Frydman and co-workers have proposed an ultrafast 2D NMR approach, where a complete 2D NMR correlation can be recorded in a single scan, thus providing a solution to the irreversibility of hyphenated techniques. This paper presents the first implementation of on-line ultrafast HPLC-NMR. Ultrafast COSY spectra are acquired every 12 s in the course of a chromatographic run performed on a mixture of natural aromatic compounds. The results, obtained on a commercial HPLC-NMR setup, highlight the generality of the ultrafast HPLC-NMR methodology, thus opening the way to a number of applications in the numerous fields in which HPLC-NMR forms a routine analytical tool.

  15. [Application and improvement of aflatoxin analysis in foods using a multifunctional column and HPLC].

    PubMed

    Goda, Y; Akiyama, H; Otsuki, T; Fujii, A; Toyoda, M

    2001-02-01

    In an earlier report, we developed a rapid, sensitive and clean method consisting of non-chloroform extraction, clean-up on a commercial multifunctional cartridge column and HPLC with fluorescence detection for the analyses of aflatoxins. In this report, we applied this method to analyze aflatoxins in nuts, giant corn, cereals, spice and black teas. The method was effective for macadamia nuts, walnuts, hazelnuts, brazil nuts, giant corn, rice, wheat and buckwheat, and the recoveries of aflatoxins B1, B2, G1 and G2 spiked in them at the level of 10 ng/g were 85-106%. However, in the chromatograms of spices and black tea, many background peaks were observed. Therefore, we added a purification step with an affinity column to the clean-up of these samples with the multifunctional cartridge column. After the additional purification, most of the background peaks were gone. The recoveries of aflatoxins B1, B2 and G1 spiked at the level of 10 ng/g were 71-112% except for the case of B2 in white pepper (48%). The recoveries of G2 were 49-95%.

  16. [Application and improvement of aflatoxin analysis in foods using a multifunctional column and HPLC].

    PubMed

    Goda, Y; Akiyama, H; Otsuki, T; Fujii, A; Toyoda, M

    2001-02-01

    In an earlier report, we developed a rapid, sensitive and clean method consisting of non-chloroform extraction, clean-up on a commercial multifunctional cartridge column and HPLC with fluorescence detection for the analyses of aflatoxins. In this report, we applied this method to analyze aflatoxins in nuts, giant corn, cereals, spice and black teas. The method was effective for macadamia nuts, walnuts, hazelnuts, brazil nuts, giant corn, rice, wheat and buckwheat, and the recoveries of aflatoxins B1, B2, G1 and G2 spiked in them at the level of 10 ng/g were 85-106%. However, in the chromatograms of spices and black tea, many background peaks were observed. Therefore, we added a purification step with an affinity column to the clean-up of these samples with the multifunctional cartridge column. After the additional purification, most of the background peaks were gone. The recoveries of aflatoxins B1, B2 and G1 spiked at the level of 10 ng/g were 71-112% except for the case of B2 in white pepper (48%). The recoveries of G2 were 49-95%. PMID:11383158

  17. An optimized and validated RP-HPLC/UV detection method for simultaneous determination of all-trans-retinol (vitamin A) and alpha-tocopherol (vitamin E) in human serum: comparison of different particulate reversed-phase HPLC columns.

    PubMed

    Khan, Abad; Khan, Muhammad I; Iqbal, Zafar; Shah, Yasar; Ahmad, Lateef; Watson, David G

    2010-09-01

    A novel, simple and fast reversed-phase HPLC/UV method was developed, optimized for various chromatographic conditions, and validated according to international guidelines for simultaneous determination of all-trans-retinol and alpha-tocopherol in human serum using retinyl acetate as internal standard in the concentration of 0.5 microg/ml. A liquid-phase extraction was applied to the 250 microl of serum with n-hexane-dichloromethane mixture (70:30, v/v), in two steps, using ethanol-methanol mixture (95:5, v/v) for protein precipitation and BHT (butylated hydroxy toluene) as stabilizer for sample preparation. Both analytes were analyzed on Kromasil 100 C(18) column (150 mm x 4.6 mm, 5 microm), Brownlee analytical (Perkin Elmer) C(18) column (150 mm x 4.6 mm, 5 microm), and Supelco (Supelcosil) LC-18 column (150 mm x 3 mm, 3 microm), protected by a Perkin Elmer C(18) (30 mm x 4.6 mm, 10 microm; Norwalk, USA) pre-column guard cartridge, at 292 nm wavelength, using methanol-water (99:1, v/v), in isocratic mode as mobile phase applied at flow rate of 1.5 ml/min and 1 ml/min for both 5 microm and 3 microm columns, respectively. Complete separation of all the analytes was achieved in 3 and 6 min on 3 microm and 5 microm columns, respectively by injecting 20 microl of sample into the HPLC system by autosampler, keeping column oven temperature at 25 degrees C. Different particulate reversed-phase chromatographic columns were evaluated in order to select the best column in terms of sensitivity, selectivity, resolution and short run time of both the analytes and it was concluded that 3 microm columns are better to be used in clinical set up as well as in laboratories for the separation of these analytes in a shorter time as compared with 5 microm columns. The method was validated and applied for the analysis of all-trans-retinol and alpha-tocopherol in the serum of human volunteers.

  18. New trend in the LC separation analysis of pharmaceuticals--high performance separation by ultra high-performance liquid chromatography (UHPLC) with core-shell particle C18 columns--.

    PubMed

    Nishi, Hiroyuki; Nagamatsu, Kumi

    2014-01-01

    This article presents a mini-review of the recent results in the ultra high-performance liquid chromatography (UHPLC) separation of pharmaceuticals by our group. High performance UHPLC separation employing core-shell particle C18 columns was demonstrated. High performance (high theoretical plate number of approximately 20000/10 cm, low theoretical plate height of 5 μm) was obtained without any specific devices in the conventional HPLC apparatus, only through changing detector sampling times and the inner diameter of the connecting tube. High theoretical plate numbers with low column back pressure obtained by the core-shell particle columns enabled fast separation of the analytes. Methanol, which gives high column pressure drops in the reversed-phase mode HPLC compared with acetonitrile, can be used without any trouble. One analysis of the purity testing of diltiazem hydrochloride was performed within 100 s. One analysis in the photostability testing of mecobalamin (vitamin B12 analogue) was successful within 180 s.

  19. Tritium Isotope Separation Using Adsorption-Distillation Column

    SciTech Connect

    Fukada, Satoshi

    2005-07-15

    In order to miniaturize the height of a distillation tower for the detritiation of waste water from fusion reactors, two experiments were conducted: (1) liquid frontal chromatography of tritium water eluting through an adsorption column and (2) water distillation using a column packed with adsorbent particles. The height of the distillation tower depends on the height equivalent to a theoretical plate, HETP, and the equilibrium isotope separation factor, {alpha}{sub H-T}{sup equi}. The adsorption action improved not only HETP but also {alpha}{sub H-T}{sup equi}. Since the adsorption-distillation method proposed here can shorten the tower height with keeping advantages of the distillation, it may bring an excellent way for miniaturizing the distillation tower to detritiate a large amount of waste water from fusion reactors.

  20. Heterogeneous post-column immunoreaction detection using magnetized beads and a laboratory-constructed electromagnetic separator.

    PubMed

    Tang, Zhe; Karnes, H Thomas

    2003-01-01

    The nature of immune reactors allows development of quantitative analytical methods that are highly selective and can often be used directly with complex biological matrixes such as blood, plasma or urine. A major limitation of immunoassay is that antibodies are sometimes unable to discriminate structurally similar species such as drug metabolites and synthetic analogs. The problem associated with the lack of discrimination can be circumvented by coupling immunoassay with liquid chromatography post-column. The most commonly used separation method in post-column immunoreaction detection is the affinity column. Affinity columns may create undesired effects such as a compromise of the chromatographic separation efficiency, the requirement for an antibody with fast reaction kinetics and the need for flushing the column. This paper reports a post-column immunoreaction detection system coupled with a laboratory-constructed on-line magnetic separation flow chamber that is designed to overcome these problems. The system uses disposable magnetic beads as a solid-phase support for separation that can be easily removed from the system. The model analytes chosen for this study were digoxin and its metabolites due to the commercial availability of monoclonal antibodies for these compounds. Digoxin was separated using a chromatographic method prior to being interfaced through a liquid handler system to the immunoreactor. Compatibility of the HPLC mobile phase was determined to be acceptable with a mixing ratio of 1:3 between the LC fraction and immunoreagent solution. The dynamic range of the calibration curve in digoxin-spiked phosphate buffer was found to be 0.25-12 ng/ml and a quadratic fit was found to provide the best fit to the data with a correlation coefficient of 0.9974. The residual error for all standards was less than 15%. The percentage RSDs for the two controls, 2 and 10 ng/ml, were 6.88 and 4.82% (n = 6) and the percentage errors were 7.07 and -6.89% (n = 6

  1. A NEW HPLC METHOD FOR SEPARATION OF PHYTOPLANKTON PIGMENTS IN NATURAL SAMPLES

    EPA Science Inventory

    A new high-performance liquid chromatographic (HPLC) method was developed to analyze, in a single run, most polar and non-polar chlorophylls and carotenoids from marine phytoplankton. The method is based on a reverse-phase amide C16 (RP-amide C16) column and an elution gradient o...

  2. NEW COLUMN SEPARATION METHOD FOR EMERGENCY URINE SAMPLES

    SciTech Connect

    Maxwell, S; Brian Culligan, B

    2007-08-28

    The Savannah River Site Environmental Bioassay Lab participated in the 2007 NRIP Emergency Response program administered by the National Institute for Standards and Technology (NIST) in May, 2007. A new rapid column separation method was applied directly to the NRIP 2007 emergency urine samples, with only minimal sample preparation to reduce preparation time. Calcium phosphate precipitation, previously used to pre-concentrate actinides and Sr-90 in NRIP 2006 urine and water samples, was not used for the NRIP 2007 urine samples. Instead, the raw urine was acidified and passed directly through the stacked resin columns (TEVA+TRU+SR Resins) to separate the actinides and strontium from the NRIP urine samples more quickly. This improvement reduced sample preparation time for the NRIP 2007 emergency urine analyses significantly. This approach works well for small volume urine samples expected during an emergency response event. Based on initial feedback from NIST, the SRS Environmental Bioassay Lab had the most rapid analysis times for actinides and strontium-90 analyses for NRIP 2007 urine samples.

  3. Extraction, Separation, and Identification of Phenolic Compounds in Virgin Olive Oil by HPLC-DAD and HPLC-MS

    PubMed Central

    Tasioula-Margari, Maria; Tsabolatidou, Eleftheria

    2015-01-01

    The aim of this study was to evaluate the recovery of individual phenolic compounds extracted from virgin olive oil (VOO), from different Greek olive varieties. Sufficient recoveries (90%) of all individual phenolic compounds were obtained using methanol as an extraction solvent, acetonitrile for residue solubilization, and two washing steps with hexane. Moreover, in order to elucidate structural characteristics of phenolic compounds in VOO, high performance liquid chromatography with a diode array detector (HPLC-DAD) at 280 and 340 nm and HPLC coupled to electrospray ionization mass spectrometry (HPLC-ESI-MS) in the negative-ion mode were performed. The most abundant phenolic compounds were oleuropein derivatives with m/z 319 and 377 and ligstroside derivatives with m/z 303, 361. Lignans, such as 1-acetoxypinoresinol and pinoresinol were also present in substantial quantities in the phenolic fraction. However, pinoresinol was co-eluted with dialdehydic form of ligstroside aglycone (DAFLA) and it was not possible to be quantified separately. The phenolic extracts, obtained from different VOO samples, yielded similar HPLC profiles. Differences, however, were observed in the last part of the chromatogram, corresponding to isomers of the aldehydic form of ligstroside aglycone. Oxidized phenolic products, originating from secoiridoids, were also detected. PMID:26783843

  4. An Eco-Friendly Direct Injection HPLC Method for Methyldopa Determination in Serum by Mixed-Mode Chromatography Using a Single Protein-Coated Column.

    PubMed

    Emara, Samy; Masujima, Tsutomu; Zarad, Walaa; Kamal, Maha; Fouad, Marwa; El-Bagary, Ramzia

    2015-09-01

    A simple, rapid and environment-friendly direct injection HPLC method for the determination of methyldopa (MTD) in human serum has been developed and validated. The method was based on cleanup and separation of MTD from serum by mixed-mode liquid chromatography using a single protein-coated TSK gel ODS-80 TM analytical column (50 × 4.0 mm i.d., 5 µm). The protein-coated column exhibited excellent resolution, selectivity and functioned in two chromatographic modes: size-exclusion chromatography [i.e., solid-phase extraction (SPE) for serum proteins] and reversed-phase chromatography for the final separation of MTD. SPE and HPLC separation were carried out simultaneously with a green mobile phase consisting of acetate buffer (0.1 M, pH 2.4) at a flow rate of 1 mL/min and at room temperature (23 ± 1°C). The eluent was monitored at emission and excitation wavelengths of 320 and 270 nm, respectively. A calibration curve was linear over the range of 0.1-30 µg/mL with a detection limit of 0.027 µg/mL. This online SPE method was successfully applied to real samples obtained from patients receiving MTD therapy.

  5. Coal liquefaction process streams characterization and evaluation: High performance liquid chromatography (HPLC) of coal liquefaction process streams using normal-phase separation with uv diode array detection

    SciTech Connect

    Clifford, D.J.; McKinney, D.E.; Hou, Lei; Hatcher, P.G.

    1994-01-01

    This study demonstrated the considerable potential of using two-dimensional, high performance liquid chromatography (HPLC) with normal-phase separation and ultraviolet (UV) diode array detection for the examination of filtered process liquids and the 850{degrees}F{sup {minus}} distillate materials derived from direct coal liquefaction process streams. A commercially available HPLC column (Hypersil Green PAH-2) provided excellent separation of the complex mixture of polynuclear aromatic hydrocarbons (PAHs) found in coal-derived process streams process. Some characteristics of the samples delineated by separation could be attributed to processing parameters. Mass recovery of the process derived samples was low (5--50 wt %). Penn State believes, however, that, improved recovery can be achieved. High resolution mass spectrometry and gas chromatography/mass spectrometry (GC/MS) also were used in this study to characterize the samples and the HPLC fractions. The GC/MS technique was used to preliminarily examine the GC-elutable portion of the samples. The GC/MS data were compared with the data from the HPLC technique. The use of an ultraviolet detector in the HPLC work precludes detecting the aliphatic portion of the sample. The GC/MS allowed for identification and quantification of that portion of the samples. Further development of the 2-D HPLC analytical method as a process development tool appears justified based on the results of this project.

  6. A porous graphitized carbon column HPLC method for the quantification of paracetamol, pseudoephedrine, and chlorpheniramine in a pharmaceutical formulation.

    PubMed

    Kalogria, Eleni; Koupparis, Michael; Panderi, Irene

    2010-01-01

    A simple, rapid, and stability-indicating HPLC method has been developed, fully validated, and applied to the quantification of paracetamol, pseudoephedrine hydrochloride, and chlorpheniramine maleate in a pharmaceutical formulation, using hydrochlorothiazide as an internal standard. Chromatographic separation was achieved isocratically on an RP porous graphitized carbon analytical column (125 x 2.1 mm id, particle size 5 microm) using 5.0 mM ammonium acetate-acetonitrile (35 + 65, v/v) mobile phase at a flow rate of 0.50 mL/min. UV spectrophotometric detection at 220 nm was used. The method had linear calibration curves over the range of 30-70 microg/mL for paracetamol, 1.8-4.2 microg/mL for pseudoephedrine hydrochloride, and 120-280 ng/mL for chlorpheniramine maleate. The intraday and interday RSD values were less than 3.2% for all compounds, while the relative error was less than 2.9%. Accelerated stability studies performed under various stress conditions proved the selectivity of the method. The developed method was applied successfully to QC and content uniformity tests of commercial tablets.

  7. Greener liquid chromatography using a guard column with micellar mobile phase for separation of some pharmaceuticals and determination of parabens.

    PubMed

    Youngvises, Napaporn; Chaida, Thanatcha; Khonyoung, Supada; Kuppithayanant, Nattawan; Tiyapongpattana, Warawut; Itharat, Arunporn; Jakmunee, Jaroon

    2013-03-15

    In this research, a greener chromatography employing a short column, Zorbax SB C18 cartridge (12.5 × 4.6 mm, 5 μm) commonly used as a guard column in a reverse phase high performance liquid chromatography (RP-HPLC), was utilized as the analytical column in conjunction with a more eco-friendly micellar mobile phase of sodium dodecyl sulfate (SDS) for separation tertiary mixtures of local anesthetics and antihistamines; and binary mixture of colds drugs; and quaternary mixture of some parabens with different separation conditions. The chromatographic behavior of these analytes was studied to demonstrate separation efficiency of this guard column in a micellar mobile phase. Moreover, this column and SDS mobile phase was exploited for determination of parabens in 64 samples of cosmetic product, both those that were produced locally in the community and those that were commercially manufactured. Linear calibration graphs of the parabens as detected at 254 nm were obtained in the range of 1-100 μmol L(-1) with R(2)>0.9990. Percentage recoveries were 92.4-109.2 with %RSD<3, and the limit of detection and quantitation were 0.04-0.10 and 0.20-0.80 μmol L(-1), respectively. This analytical system is not only greener but also faster and employing simpler sample preparation than a conventional liquid chromatographic system. PMID:23598137

  8. Greener liquid chromatography using a guard column with micellar mobile phase for separation of some pharmaceuticals and determination of parabens.

    PubMed

    Youngvises, Napaporn; Chaida, Thanatcha; Khonyoung, Supada; Kuppithayanant, Nattawan; Tiyapongpattana, Warawut; Itharat, Arunporn; Jakmunee, Jaroon

    2013-03-15

    In this research, a greener chromatography employing a short column, Zorbax SB C18 cartridge (12.5 × 4.6 mm, 5 μm) commonly used as a guard column in a reverse phase high performance liquid chromatography (RP-HPLC), was utilized as the analytical column in conjunction with a more eco-friendly micellar mobile phase of sodium dodecyl sulfate (SDS) for separation tertiary mixtures of local anesthetics and antihistamines; and binary mixture of colds drugs; and quaternary mixture of some parabens with different separation conditions. The chromatographic behavior of these analytes was studied to demonstrate separation efficiency of this guard column in a micellar mobile phase. Moreover, this column and SDS mobile phase was exploited for determination of parabens in 64 samples of cosmetic product, both those that were produced locally in the community and those that were commercially manufactured. Linear calibration graphs of the parabens as detected at 254 nm were obtained in the range of 1-100 μmol L(-1) with R(2)>0.9990. Percentage recoveries were 92.4-109.2 with %RSD<3, and the limit of detection and quantitation were 0.04-0.10 and 0.20-0.80 μmol L(-1), respectively. This analytical system is not only greener but also faster and employing simpler sample preparation than a conventional liquid chromatographic system.

  9. Study on the Alkaloids in Tibetan Medicine Aconitum pendulum Busch by HPLC-MSn Combined with Column Chromatography.

    PubMed

    Wang, Beibei; Dong, Jie; Ji, Jiaojiao; Yuan, Jiang; Wang, Jiali; Wu, Jiarui; Tan, Peng; Liu, Yonggang

    2016-01-01

    A rapid, convenient and effective identification method of alkaloids was established and an attempt on isolating and analyzing the alkaloids in Aconitum pendulum Busch was conducted successfully. In this article, four high-content components including deoxyaconitine, benzoylaconine, aconine and neoline were isolated by using column chromatography. HPLC-MS(n)was employed to deduce the regulations of fragmentation of diterpenoid alkaloids which displayed a characteristic behavior of loss of CO(28u), CH3COOH(60u), CH3OH(32u), H2O(18u) and C6H5COOH(122u). Then, according to fragmentation regulation of mass spectrometry, 42 alkaloids were found inA. pendulum Among them, 38 compounds were identified and 29 alkaloids were reported for the first time for this herb. Therefore, this means that HPLC-MS(n)combined with column chromatography could work as an effective and reliable tool for rapid identification of the chemical components of herbal medicine.

  10. Study on the Alkaloids in Tibetan Medicine Aconitum pendulum Busch by HPLC-MSn Combined with Column Chromatography.

    PubMed

    Wang, Beibei; Dong, Jie; Ji, Jiaojiao; Yuan, Jiang; Wang, Jiali; Wu, Jiarui; Tan, Peng; Liu, Yonggang

    2016-01-01

    A rapid, convenient and effective identification method of alkaloids was established and an attempt on isolating and analyzing the alkaloids in Aconitum pendulum Busch was conducted successfully. In this article, four high-content components including deoxyaconitine, benzoylaconine, aconine and neoline were isolated by using column chromatography. HPLC-MS(n)was employed to deduce the regulations of fragmentation of diterpenoid alkaloids which displayed a characteristic behavior of loss of CO(28u), CH3COOH(60u), CH3OH(32u), H2O(18u) and C6H5COOH(122u). Then, according to fragmentation regulation of mass spectrometry, 42 alkaloids were found inA. pendulum Among them, 38 compounds were identified and 29 alkaloids were reported for the first time for this herb. Therefore, this means that HPLC-MS(n)combined with column chromatography could work as an effective and reliable tool for rapid identification of the chemical components of herbal medicine. PMID:26896350

  11. A new affinity-HPLC packing for protein separation: Cibacron blue attached uniform porous poly(HEMA-co-EDM) beads.

    PubMed

    Unsal, Ender; Durdu, Aysun; Elmas, Begum; Tuncel, Murvet; Tuncel, Ali

    2005-11-01

    In this study, a new affinity high-performance liquid chromatography (HPLC) stationary phase suitable for protein separation was synthesized. In the first stage of the synthesis, uniform porous poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate), poly(HEMA-co-EDM), beads 6.2 mum in size were obtained. Homogeneous distribution of hydroxyl groups in the bead interior was confirmed by confocal laser scanning microscopy. The plain poly(HEMA-co-EDM) particles gave very low non-specific protein adsorption with albumin. The selected dye ligand Cibacron blue F3G-A (CB F3G-A) was covalently linked onto the beads via hydroxyl groups. In the batch experiments, albumin adsorption up to 60 mg BSA/g particles was obtained with the CB F3G-A carrying poly(HEMA-co-EDM) beads. The affinity-HPLC of selected proteins (albumin and lysozyme) was investigated in a 25 mm x 4.0-mm inner diameter column packed with CB F3G-A carrying beads and both proteins were successfully resolved. By a single injection, 200 mug of protein was loaded and quantitatively eluted from the column. The protein recovery increased with increasing flow rate and salt concentration of the elution buffer and decreased with the increasing protein feed concentration. During the albumin elution, theoretical plate numbers up to 30,000 plates/m were achieved by increasing the salt concentration.

  12. Rigid porous polyacrylamide-based monolithic columns containing butyl methacrylate as a separation medium for the rapid hydrophobic interaction chromatography of proteins.

    PubMed

    Xie, S; Svec, F; Fréchet, J M

    1997-07-18

    Macroporous poly(acrylamide-co-butyl methacrylate-co-N,N'-methylenebisacrylamide) monoliths containing up to 15% butyl methacrylate units have been prepared by direct polymerization within the confines of HPLC columns. The hydrodynamic and chromatographic properties of these 50 mm x 8 mm I.D. columns-such as back pressure at different flow-rates, effect of percentage of hydrophobic component in the polymerization mixture, effect of salt concentration on the retention of proteins, dynamic loading capacity, and recovery-were determined under conditions typical of hydrophobic interaction chromatography. Using the monolithic column, five proteins were easily separated within only 3 min.

  13. [Automated pre-column derivatization with o-phthaldialdehyde (OPA)> A new RP-HPLC method for the determination of biogenic amines in food].

    PubMed

    Petridis, K D; Steinhart, H

    1995-09-01

    A simple, selective and highly sensitive HPLC method for the routine determination of the biogenic amines in food is presented. Sample preparation is based on a rapid amine extraction using 10% trichloroacetic acid and a cation exchange column for extract purification. For the RP-HPLC analysis OPA/2-mercaptoethanol is used for the pre-column derivatisation, followed by fluorescence detection (Ex 345 nm, Em 440 nm). The effects of several factors are discussed. A separation of 15 biogenic amines is achieved within 70 min. The recoveries for histamine, tyramine, putrescine, cadaverine, tryptamine and beta-phenylethylamine are higher than 95%. The detection limits lie between 0.1-0.5 pMol/injection (20 microliters), depending on the amine and a good linearity is achieved in the range from 0.5-500 pMol (r > 0.99). The method has been applied for the determination of biogenic amines in swiss cheese, salami, milk, beer and wine, the repeatability is very good.

  14. Parallel array of independent thermostats for column separations

    DOEpatents

    Foret, Frantisek; Karger, Barry L.

    2005-08-16

    A thermostat array including an array of two or more capillary columns (10) or two or more channels in a microfabricated device is disclosed. A heat conductive material (12) surrounded each individual column or channel in array, each individual column or channel being thermally insulated from every other individual column or channel. One or more independently controlled heating or cooling elements (14) is positioned adjacent to individual columns or channels within the heat conductive material, each heating or cooling element being connected to a source of heating or cooling, and one or more independently controlled temperature sensing elements (16) is positioned adjacent to the individual columns or channels within the heat conductive material. Each temperature sensing element is connected to a temperature controller.

  15. Ion chromatographic separation of inorganic ions using a combination of hydrophilic interaction chromatographic column and cation-exchange resin column.

    PubMed

    Arai, Kaori; Mori, Masanobu; Hironaga, Takahiro; Itabashi, Hideyuki; Tanaka, Kazuhiko

    2012-04-01

    A combination of hydrophilic interaction chromatographic (HILIC) column and a weakly acidic cation-exchange resin (WCX) column was used for simultaneous separation of inorganic anions and cations by ion chromatography (IC). Firstly, the capability of HILIC column for the separation of analyte ions was evaluated under acidic eluent conditions. The columns used were SeQuant ZIC-HILIC (ZIC-HILIC) with a sulfobetaine-zwitterion stationary phase (ZIC-HILIC) and Acclaim HILIC-10 with a diol stationary phase (HILIC-10). When using tartaric acid as the eluent, the HILIC columns indicated strong retentions for anions, based on ion-pair interaction. Especially, HILIC-10 could strongly retain anions compared with ZIC-HILIC. The selectivity for analyte anions of HILIC-10 with 5 mmol/L tartaric acid eluent was in the order of I(-) > NO3(-) > Br(-) > Cl(-) > H2PO4(-). However, since HILIC-10 could not separate analyte cations, a WCX column (TSKgel Super IC-A/C) was connected after the HILIC column in series. The combination column system of HILIC and WCX columns could successfully separate ten ions (Na+, NH4+, K+, Mg2+, Ca2+, H2PO4(-), Cl(-), Br(-), NO3(-) and I(-)) with elution of 4 mmol/L tartaric acid plus 8 mmol/L 18-crown-6. The relative standard deviations (RSDs) of analyte ions by the system were in the ranges of 0.02% - 0.05% in retention times and 0.18% - 5.3% in peak areas through three-time successive injections. The limits of detection at signal-to-noise ratio of 3 were 0.24 - 0.30 micromol/L for the cations and 0.31 - 1.2 micromol/L for the anions. This system was applied for the simultaneous determination of the cations and the anions in a vegetable juice sample with satisfactory results.

  16. The product composition regions of azeotropic distillation columns. 2. Separability in two-feed columns and entrainer selection

    SciTech Connect

    Wahnschafft, O.M.; Westerberg, A.W. . Engineering Design Research Center and Department of Chemical Engineering)

    1993-06-01

    A method to assess the product composition regions for distillation of ternary mixtures in single-feed distillation columns, introduced in the first paper of this series, is generalized to account for the effect of introducing multiple feeds of different trays. The method relies on so-called fixed point curves which are trajectories in the compositions space. These trajectories describe the possible compositions of pinch points in each column section as functions of the energy supplied to a column, i.e., for all conceivable values of the reflux ratio. Pinch point trajectories may be determined analytically or, for ternary mixtures, can be located graphically using residue curve maps. The authors carry out a mostly graphical analysis, using pinch point trajectories to establish separation feasibility ahead of design calculations. This analysis also provides information on the minimum entrainer supply for a specified separation and visualizes the phenomenon of the occurrence of a maximum reflux ratio for separation in a column with a separate, extractive agent feed. The analysis is analogous to that for single-feed columns, only the critical pinch trajectories may be those for the extractive column section between the feeds. This analogy suggests the notion of a generalized extractive distillation process, for which new entrainer selection criteria are proposed.

  17. Isolation of mono-caffeoylquinic acids from tobacco waste using continuous resin-based pre-separation and preparative HPLC.

    PubMed

    Wang, Jun; Lu, Dingqiang; Liang, Yao; Zhao, Hui; Luo, Min; Ling, Xiuquan; Ouyang, Pingkai

    2012-06-01

    Three isomers of mono-caffeoylquinic acid, specifically, 3-O-caffeoylquinic acid, 4-O-caffeoylquinic acid and 5-O-caffeoylquinic acid, were successfully isolated from a crude extract of tobacco (Nicotiana tobaccum L.) wastes using continuous resin-based pre-separation and preparative high-performance liquid chromatography (HPLC). The extract of tobacco wastes was continuously pre-separated by resin-based columns packed with D101 and XAD-4, yielding total mono-caffeoylquinic acids with a purity of 67.71% and a recovery rate of 90.06%. Variables affecting resolution and productivity of three mono-caffeoylquinic acid isomers in preparative HPLC (i.e. mobile-phase composition, pH, flow rate and loading amount) were studied. The optimum chromatographic conditions were determined to be a mobile phase consisting of 15% (v/v) methanol and aqueous acetic acid with a pH of 4.5, a flow rate of 4.0 mL/min, a loading amount of 4 mL and a detection wavelength of 360 nm. From 300 mg of loading sample, 56.3 mg of 3-O-caffeoylquinic acid, 92.8 mg of 5-O-caffeoylquinic acid and 73.1 mg of 4-O-caffeoylquinic acid were obtained in a single run, each with a purity of over 98% by HPLC. The structures of the isolated compounds were elucidated by ESI-MS, (1) H-NMR and (13) C-NMR spectral data.

  18. Improved high performance liquid chromatographic separation of anthocyanin compounds from grapes using a novel mixed-mode ion-exchange reversed-phase column.

    PubMed

    McCallum, Jason L; Yang, Raymond; Young, J Christopher; Strommer, Judith N; Tsao, Rong

    2007-04-27

    A novel mixed mode HPLC method using a column combining both ion-exchange and reversed-phase separation mechanisms has been developed to facilitate analysis of anthocyanins in grapes. Chromatographic performance and subsequent analysis of anthocyanidin diglucosides and acylated compounds are significantly improved using the new column, compared to those associated with conventional C18 reversed-phase methods. The mixed mode column produces a distinctive eluting pattern for the different anthocyanin subgroups, avoiding overlaps found with C18 columns. The enhanced chromatographic resolution provides nearly complete separation of 37 anthocyanin types, and permits detection of delphinidin 3-O-(6''-O-caffeoyl) beta-D-glucoside for the first time in extracts of skins from Concord grapes. PMID:17382950

  19. Synthesis of a mixed-model stationary phase derived from glutamine for HPLC separation of structurally different biologically active compounds: HILIC and reversed-phase applications.

    PubMed

    Aral, Tarık; Aral, Hayriye; Ziyadanoğulları, Berrin; Ziyadanoğulları, Recep

    2015-01-01

    A novel mixed-mode stationary phase was synthesised starting from N-Boc-glutamine, aniline and spherical silica gel (4 µm, 60 Å). The prepared stationary phase was characterized by IR and elemental analysis. The new stationary phase bears an embedded amide group into phenyl ring, highly polar a terminal amide group and non-polar groups (phenyl and alkyl groups). At first, this new mixed-mode stationary phase was used for HILIC separation of four nucleotides and five nucleosides. The effects of different separation conditions, such as pH value, mobile phase and temperature, on the separation process were investigated. The optimum separation for nucleotides was achieved using HILIC isocratic elution with aqueous mobile phase and acetonitrile with 20°C column temperature. Under these conditions, the four nucleotides could be separated and detected at 265 nm within 14 min. Five nucleosides were separated under HILIC isocratic elution with aqueous mobile phase containing pH=3.25 phosphate buffer (10mM) and acetonitrile with 20°C column temperature and detected at 265 nm within 14 min. Chromatographic parameters as retention factor, selectivity, theoretical plate number and peak asymmetry factor were calculated for the effect of temperature and water content in mobile phase on the separation process. The new column was also tested for nucleotides and nucleosides mixture and six analytes were separated in 10min. The chromatographic behaviours of these polar analytes on the new mixed-model stationary phase were compared with those of HILIC columns under similar conditions. Further, phytohormones and phenolic compounds were separated in order to see influence of the new stationary phase in reverse phase conditions. Eleven plant phytohormones were separated within 13 min using RP-HPLC gradient elution with aqueous mobile phase containing pH=2.5 phosphate buffer (10mM) and acetonitrile with 20°C column temperature and detected at 230 or 278 nm. The best separation

  20. TCAP HYDROGEN ISOTOPE SEPARATION USING PALLADIUM AND INVERSE COLUMNS

    SciTech Connect

    Heung, L.; Sessions, H.; Xiao, S.

    2010-08-31

    The Thermal Cycling Absorption Process (TCAP) was further studied with a new configuration. Previous configuration used a palladium packed column and a plug flow reverser (PFR). This new configuration uses an inverse column to replace the PFR. The goal was to further improve performance. Both configurations were experimentally tested. The results showed that the new configuration increased the throughput by a factor of more than 2.

  1. Analysis of amino acid composition in proteins of animal tissues and foods as pre-column o-phthaldialdehyde derivatives by HPLC with fluorescence detection.

    PubMed

    Dai, Zhaolai; Wu, Zhenlong; Jia, Sichao; Wu, Guoyao

    2014-08-01

    Studies of protein nutrition and biochemistry require reliable methods for analysis of amino acid (AA) composition in polypeptides of animal tissues and foods. Proteins are hydrolyzed by 6M HCl (110°C for 24h), 4.2M NaOH (105°C for 20 h), or proteases. Analytical techniques that require high-performance liquid chromatography (HPLC) include pre-column derivatization with 4-chloro-7-nitrobenzofurazan, 9-fluorenyl methylchloroformate, phenylisothiocyanate, naphthalene-2,3-dicarboxaldehyde, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate, and o-phthaldialdehyde (OPA). OPA reacts with primary AA (except cysteine or cystine) in the presence of 2-mercaptoethanol or 3-mercaptopropionic acid to form a highly fluorescent adduct. OPA also reacts with 4-amino-1-butanol and 4-aminobutane-1,3-diol produced from oxidation of proline and 4-hydroxyproline, respectively, in the presence of chloramine-T plus sodium borohydride at 60°C, or with S-carboxymethyl-cysteine formed from cysteine and iodoacetic acid at 25°C. Fluorescence of OPA derivatives is monitored at excitation and emission wavelengths of 340 and 455 nm, respectively. Detection limits are 50 fmol for AA. This technique offers the following advantages: simple procedures for preparation of samples, reagents, and mobile-phase solutions; rapid pre-column formation of OPA-AA derivatives and their efficient separation at room temperature (e.g., 20-25°C); high sensitivity of detection; easy automation on the HPLC apparatus; few interfering side reactions; a stable chromatography baseline for accurate integration of peak areas; and rapid regeneration of guard and analytical columns. Thus, the OPA method provides a useful tool to determine AA composition in proteins of animal tissues (e.g., skeletal muscle, liver, intestine, placenta, brain, and body homogenates) and foods (e.g., milk, corn grain, meat, and soybean meal).

  2. Performance evaluation of an aqueous-organic phase separator for post-column reactions in high-performance liquid chromatography, and its application to the enhanced detection of some basic drugs of abuse.

    PubMed

    Roy, I M; Jefferies, T M

    1990-01-01

    A phase separator is described that is suitable for post-column HPLC applications. It operates with commonly used HPLC eluents and immiscible organic solvents as long as the two phases remain immiscible. It is compatible with gradient elution systems. Separation efficiency is routinely better than 0.8, which ensures that analyte peak heights are about 95% of the maximum height under these conditions. An application for the detection of pethidine, cocaine, methadone, piritramide and dipipanone at 0.8-1.8 ng on-column loadings is described.

  3. Separation and identification of cis and trans isomers of 2-butene-1,4-diol and lafutidine by HPLC and LC-MS*

    PubMed Central

    Pan, Chun-xiu; Xu, Xiu-zhu; He, Hong-mei; Cai, Xiao-jun; Zhang, Xue-jun

    2005-01-01

    The cis and trans isomers separation of 2-butene-1,4-diol and lafutidine were studied by HPLC on two kinds of chiral columns: (S,S)-Whelk-O 1 and ChiraSpher. The isomers of 2-butene-1,4-diol can be separated on both chiral columns while the isomers of lafutidine can only be resolved on ChiraSpher column. The influence of different type and amount of mobile phase modifier on the isomers separation was extensively studied. The resolution of cis and trans isomers of 2-butene-1,4-diol was 2.61 on (S,S)-Whelk-O 1 column with hexane-ethanol (97:3, v/v) as the mobile phase. The resolution of lafutidine was 1.89 on ChiraSpher column with hexane-ethanol-THF-diethylamine (92:3:5:0.1, v/v/v/v) as the mobile phase. LC-MS methods were developed to identify the isomer peaks. PMID:15593397

  4. Separation and quantification of water buffalo milk protein fractions and genetic variants by RP-HPLC.

    PubMed

    Bonfatti, Valentina; Giantin, Mery; Rostellato, Roberta; Dacasto, Mauro; Carnier, Paolo

    2013-01-15

    A RP-HPLC method, developed for the separation and quantification of the most common genetic variants of bovine milk proteins, was successfully applied to the analysis of water buffalo milk. All the most common buffalo casein and whey proteins fractions, as well as their genetic variants, were detected and separated simultaneously in 40 min. Purified buffalo proteins were used as calibration standards and a total of 536 individual milk samples were analysed for protein composition. α(S1)-, α(S2)-, βγ-, and κ-casein were 32.2%, 15.8%, 36.5%, and 15.5%, respectively, of total casein content, whereas content of β-Lactoglobulin was approximately 1.3 times as high as that of α-Lactalbumin. The existence of a polymorphism of κ-casein was demonstrated in Mediterranean water buffalo and α(S1)- and κ-casein genetic variants were successfully detected by RP-HPLC.

  5. Improved micromachined column design and fluidic interconnects for programmed high-temperature gas chromatography separations.

    PubMed

    Gaddes, David; Westland, Jessica; Dorman, Frank L; Tadigadapa, Srinivas

    2014-07-01

    This work focuses on the development and experimental evaluation of micromachined chromatographic columns for use in a commercial gas chromatography (GC) system. A vespel/graphite ferrule based compression sealing technique is presented using which leak-proof fluidic interconnection between the inlet tubing and the microchannel was achieved. This sealing technique enabled separation at temperatures up to 350°C on a μGC column. This paper reports the first high-temperature separations in microfabricated chromatographic columns at these temperatures. A 2m microfabricated column using a double Archimedean spiral design with a square cross-section of 100μm×100μm has been developed using silicon microfabrication techniques. The microfabricated column was benchmarked against a 2m 100μm diameter commercial column and the performance between the two columns was evaluated in tests performed under identical conditions. High temperature separations of simulated distillation (ASTM2887) and polycyclic aromatic hydrocarbons (EPA8310) were performed using the μGC column in temperature programmed mode. The demonstrated μGC column along with the high temperature fixture offers one more solution toward potentially realizing a portable μGC device for the detection of semi-volatile environmental pollutants and explosives without the thermal limitations reported to date with μGC columns using epoxy based interconnect technology. PMID:24866564

  6. Improved micromachined column design and fluidic interconnects for programmed high-temperature gas chromatography separations.

    PubMed

    Gaddes, David; Westland, Jessica; Dorman, Frank L; Tadigadapa, Srinivas

    2014-07-01

    This work focuses on the development and experimental evaluation of micromachined chromatographic columns for use in a commercial gas chromatography (GC) system. A vespel/graphite ferrule based compression sealing technique is presented using which leak-proof fluidic interconnection between the inlet tubing and the microchannel was achieved. This sealing technique enabled separation at temperatures up to 350°C on a μGC column. This paper reports the first high-temperature separations in microfabricated chromatographic columns at these temperatures. A 2m microfabricated column using a double Archimedean spiral design with a square cross-section of 100μm×100μm has been developed using silicon microfabrication techniques. The microfabricated column was benchmarked against a 2m 100μm diameter commercial column and the performance between the two columns was evaluated in tests performed under identical conditions. High temperature separations of simulated distillation (ASTM2887) and polycyclic aromatic hydrocarbons (EPA8310) were performed using the μGC column in temperature programmed mode. The demonstrated μGC column along with the high temperature fixture offers one more solution toward potentially realizing a portable μGC device for the detection of semi-volatile environmental pollutants and explosives without the thermal limitations reported to date with μGC columns using epoxy based interconnect technology.

  7. Surface modification of polytetrafluoroethylene column for two-stationary phase separations by counter-current chromatography.

    PubMed

    Quan, Kai-jun; Huang, Xin-yi; Li, Xiao-ting; Wang, Gao-hong; Liu, Yan-juan; Duan, Wen-da; Di, Duo-long

    2015-11-27

    To improve the separation capability of CCC, a novel solid-liquid two-stationary phases CCC (ASP-CCC) column was prepared employing graphene oxide (GO) conjugated poly-dopamine (PD) coating (GO/PD) as auxiliary stationary phase (ASP). The results of Scanning electron microscopy (SEM), contact angle and X-ray photoelectron spectroscopy (XPS) indicated that nanostructured GO and PD were successfully grafted on the inner wall of the PTFE column. Three alkaloid compounds were selected as the target analytes to evaluate the performance of the novel column. Because of the intermolecular force (hydrogen bond, electrostatic interaction and π-π interaction) between the ASP and model compounds, three analytes were well separated with this novel ASP-CCC column. Additionally, the novel column exhibited higher stationary phase retention ratio, about 8%, than original column without changing the chromatographic condition. Furthermore, the eluotropic sequence of analytes on novel column was in accordance with that in the original column. This suggested that the novel column is a CCC column with auxiliary stationary phase (ASP) in its own right, and the present separation mode is the combination of partition chromatography and adsorption chromatography.

  8. Surface modification of polytetrafluoroethylene column for two-stationary phase separations by counter-current chromatography.

    PubMed

    Quan, Kai-jun; Huang, Xin-yi; Li, Xiao-ting; Wang, Gao-hong; Liu, Yan-juan; Duan, Wen-da; Di, Duo-long

    2015-11-27

    To improve the separation capability of CCC, a novel solid-liquid two-stationary phases CCC (ASP-CCC) column was prepared employing graphene oxide (GO) conjugated poly-dopamine (PD) coating (GO/PD) as auxiliary stationary phase (ASP). The results of Scanning electron microscopy (SEM), contact angle and X-ray photoelectron spectroscopy (XPS) indicated that nanostructured GO and PD were successfully grafted on the inner wall of the PTFE column. Three alkaloid compounds were selected as the target analytes to evaluate the performance of the novel column. Because of the intermolecular force (hydrogen bond, electrostatic interaction and π-π interaction) between the ASP and model compounds, three analytes were well separated with this novel ASP-CCC column. Additionally, the novel column exhibited higher stationary phase retention ratio, about 8%, than original column without changing the chromatographic condition. Furthermore, the eluotropic sequence of analytes on novel column was in accordance with that in the original column. This suggested that the novel column is a CCC column with auxiliary stationary phase (ASP) in its own right, and the present separation mode is the combination of partition chromatography and adsorption chromatography. PMID:26518492

  9. Automated HPLC separation of endohedral metallofullerene ScC{sub 2n} and YC{sub 2n} fractions

    SciTech Connect

    Stevenson, S.; Dorn, H.C.; Burbank, P.; Harich, K.; Haynes, J. Jr.; Kiang, C.H.; Salem, J.R.; DeVries, M.S.; Loosdrecht, P.H.M. van; Johnson, R.D.; Yannoni, C.S.; Bethune, D.S.

    1994-09-01

    We describe an automated HPLC separation of the endohedral metallofullerenes such as SCC{sub 2n} and YC{sub 2n} from empty-cage fullerenes utilizing two polystyrene chromatographic columns (500 and 1000 A) in series. Rapid separation of the metallofullerene fraction from the empty-cage fullerenes (e.g., C{sub 60}) under anaerobic conditions is achieved. For the isolated SCC{sub 2n} fraction, all even-carbon-membered species from Sc{sub 2}C{sub 74} to Sc{sub 2}C{sub 104} were identified by negative-ion chemical ionization mass spectrometry. In addition, Sc{sub 3}C{sub 82} was a prominent component of this fraction. For the separated YC{sub 2n} sample, the mass spectral data indicate the presence of YC{sub 82} and all even-carbon-numbered diyttrium species from Y{sub 2}C{sub 82} to Y{sub 2}C{sub 104}. 14 refs., 7 figs.

  10. A STUDY OF MULTISTAGE/MULTIFUNCTION COLUMN FOR FINE PARTICLE SEPARATION

    SciTech Connect

    Dr. Shiao-Hung Chiang

    1999-10-01

    A non-agitated multi-stage column was constructed and applied to wastewater treatment. Preliminary oil/water separation tests were performed. Excellent separation results verifies the multi-function feature of the multi-stage column. Hydrodynamic behavior is considered as the underlying cause for the separation performance. Therefore, a series of experiments were carried out to investigate the hydrodynamic parameters, including gas holdups and liquid circulating velocities. The experimental data will be used to create a mathematical model to simulate the multi-stage column process. The model will further shed light on the future scale-up of the MSTLFLO process.

  11. A Study of Multistage/Multifunction Column for Fine Particle Separation.

    SciTech Connect

    Chiang, S.

    1997-09-15

    A non-agitated multi-stage column was constructed and applied to wastewater treatment. Preliminary oil/water separation tests were performed. Excellent separation results verifies the multi-function feature of the multi-stage column. Hydrodynamic behavior is considered as the underlying cause for the separation performance. Therefore, a series of experiments were carried out to investigate the hydrodynamic parameters, including gas holdups and liquid circulating velocities. The experimental data will be used to create a mathematical model to simulate the multi-stage column process. The model will further shed light on the future scale-up of the MSTLFLO process.

  12. Process for the production of ultrahigh purity silane with recycle from separation columns

    NASA Technical Reports Server (NTRS)

    Coleman, Larry M. (Inventor)

    1982-01-01

    Tri- and dichlorosilanes formed by hydrogenation in the course of the reaction of metallurgical silicon, hydrogen and recycle silicon tetrachloride are employed as feed into a separation column arrangement of sequential separation columns and redistribution reactors which processes the feed into ultrahigh purity silane and recycle silicon tetrachloride. A slip stream is removed from the bottom of two sequential columns and added to the recycle silicon tetrachloride process stream causing impurities in the slip streams to be subjected to reactions in the hydrogenation step whereby waste materials can be formed and readily separated.

  13. Process for the production of ultrahigh purity silane with recycle from separation columns

    DOEpatents

    Coleman, Larry M.

    1982-07-20

    Tri- and dichlorosilanes formed by hydrogenation in the course of the reaction of metallurgical silicon, hydrogen and recycle silicon tetrachloride are employed as feed into a separation column arrangement of sequential separation columns and redistribution reactors which processes the feed into ultrahigh purity silane and recycle silicon tetrachloride. A slip stream is removed from the bottom of two sequential columns and added to the recycle silicon tetrachloride process stream causing impurities in the slip streams to be subjected to reactions in the hydrogenation step whereby waste materials can be formed and readily separated.

  14. Separation, identification, quantification, and method validation of anthocyanins in botanical supplement raw materials by HPLC and HPLC-MS.

    PubMed

    Chandra, A; Rana, J; Li, Y

    2001-08-01

    A method has been established and validated for identification and quantification of individual, as well as total, anthocyanins by HPLC and LC/ES-MS in botanical raw materials used in the herbal supplement industry. The anthocyanins were separated and identified on the basis of their respective M(+) (cation) using LC/ES-MS. Separated anthocyanins were individually calculated against one commercially available anthocyanin external standard (cyanidin-3-glucoside chloride) and expressed as its equivalents. Amounts of each anthocyanin calculated as external standard equivalent were then multiplied by a molecular-weight correction factor to afford their specific quantities. Experimental procedures and use of a molecular-weight correction factors are substantiated and validated using Balaton tart cherry and elderberry as templates. Cyanidin-3-glucoside chloride has been widely used in the botanical industry to calculate total anthocyanins. In our studies on tart cherry and elderberry, its use as external standard followed by use of molecular-weight correction factors should provide relatively accurate results for total anthocyanins, because of the presence of cyanidin as their major anthocyanidin backbone. The method proposed here is simple and has a direct sample preparation procedure without any solid-phase extraction. It enables selection and use of commercially available anthocyanins as external standards for quantification of specific anthocyanins in the sample matrix irrespective of their commercial availability as analytical standards. It can be used as a template and applied for similar quantification in several anthocyanin-containing raw materials for routine quality control procedures, thus providing consistency in analytical testing of botanical raw materials used for manufacturing efficacious and true-to-the-label nutritional supplements.

  15. Separation characteristics of multistage water/hydrogen exchange column for water detritiation in fusion reactors

    SciTech Connect

    Yamanishi, T.; Okuno, K.

    1995-10-01

    A simulation code of multistage chemical exchange columns has been developed. The sieve trays for liquid-vapor scrubbing and the catalyst beds for vapor-hydrogen exchange reactions are alternately piled within the column. The code deals with all the twelve molecular species of hydrogen gas and water; and is based on the Newton-Raphson method. The characteristics of the column were discussed from the calculated results by this code such as effects of temperature and pressure. Similar to the distillation columns, the phase flow rates within the column (hydrogen gas and water vapor) and product flow rates have large effects on the separation performance of the column. A control method of the column was also proposed from these calculated results. 9 refs., 5 figs., 4 tabs.

  16. The selection of suitable columns for a reversed-phase liquid chromatographic separation of beta-lactam antibiotics and related substances via chromatographic column parameters.

    PubMed

    Zhang, Wei-qing; Hu, Qiu-xin; Zhang, Xia; Li, Ya-ping; Wang, Ming-juan; Hu, Chang-qin

    2014-01-01

    The selection of RP-LC columns suitable for a particular analysis in official compendia is difficult as only a general description of the stationary phase in the description of a LC method is given. General methods to characterize RP-LC columns often assume that each of the column parameters is equally important. This can cause the user to select columns inappropriate for particular analyses. This paper focuses on the relationship between the critical peak pairs and the column parameters (H, S, A, B, and C) in the Snyder/Dolan column characterization methodology to find the key parameters influencing real separations. Some varieties of β-lactam antibiotics and their related compounds were used as test compounds. We found column parameter A to be the most important factor affecting their separation. Parameters B and C also played an important role in some separation processes. This indicated that the hydrogen bonding of column and solute can directly affect the separation of β-lactam antibiotics. Choosing columns for which column parameter A is near 0.1 can facilitate the ideal separations of impurities from β-lactam antibiotics. The most suitable column for any common pharmaceutical analysis could be selected easily if the key column parameters would be given in the description of the chromatographic method. For these reasons, key column parameters should be listed in the monographs of official compendia.

  17. Colorful Column Chromatography: A Classroom Demonstration of a Three-Component Separation

    ERIC Educational Resources Information Center

    Heumann, Lars V.

    2008-01-01

    A classroom demonstration detailing the procedure for the separation of a ternary mixture consisting of intensely colored compounds using silica gel column chromatography is described. The audience can follow the compounds during their passage through the column as individual, colored bands while learning about different tools and techniques used…

  18. Polydopamine assisted fabrication of titanium oxide nanoparticles modified column for proteins separation by capillary electrochromatography.

    PubMed

    Zhang, Yamin; Wang, Wentao; Ma, Xiangdong; Jia, Li

    2016-11-01

    Development of a simple method for preparation of stable open tubular (OT) columns for proteins separation by capillary electrochromatography is still challenging. In this work, the titanium oxide (TiO2) nanoparticles coated OT column was successfully prepared for separation of proteins by capillary electrochromatography. The polydopamine (PDA) film was first formed in the inner surface of a fused-silica capillary by the self-polymerization of dopamine under alkaline conditions. Then the TiO2 coating was deposited onto the surface of pre-modified capillary with PDA by a liquid phase deposition process. The plentifully active hydroxyl groups in PDA coating can chelate with Ti(4+) to boost the nucleation and growth of TiO2 film. The as-prepared TiO2 coated OT column was characterized by scanning electron microscopy and measurement of electroosmotic flow. Furthermore, the influence of liquid phase deposition time on the TiO2 coating was investigated. The TiO2 coated OT column was used for successful separation of two variants of β-lactoglobulin and eight glycoisoforms of ovalbumin. The column demonstrated good repeatability and stability. The relative standard deviations of migration times of proteins representing run-to-run, day-to-day, and column-to-column were less than 3.7%. Moreover, the application of the column was verified by successful separation of acidic proteins in egg white. PMID:27555440

  19. Rapid separation and highly sensitive detection methodology for sulfonamides in shrimp using a monolithic column coupled with BDD amperometric detection.

    PubMed

    Sangjarusvichai, Haruthai; Dungchai, Wijitar; Siangproh, Weena; Chailapakul, Orawon

    2009-09-15

    In this report, we aimed to extend our previous efforts toward the evaluation of sulfonamides (SAs) with a boron-doped diamond (BDD) electrode. We improved this method by reducing the analysis time using a monolithic column coupled with amperometric detection to determine seven sulfonamides (sulfaguanidine, sulfadiazine, sulfamethazine, sulfamonomethoxine, sulfamethoxazole, sulfadimethoxine and sulfaquinoxaline). Because of its rapid separation, low back-pressure and high separation efficiency compared to a particle-packed column, a monolithic column (100 mm x 4.6mm) was used for sulfonamide separation. Chromatographic separation was performed in less than 8 min. The analysis was carried out using phosphate buffer (0.1M, pH 3): acetonitrile: methanol in a ratio of 80:15:5 (v/v/v) as the mobile phase with a flow rate of 1.5 mL min(-1). The optimal detection potential using hydrodynamic voltammetry was found to be 1.2V versus Ag/AgCl. The method was applied to determine seven sulfonamides in shrimp after sample preparation by solid-phase extraction. The recoveries of the sulfonamides in spiked shrimp samples at 1.5, 5 and 10 microg g(-1) were in the range of 81.7 to 97.5% with a relative standard deviation (R.S.D.) between 1.0 and 4.6%. Our methodology produced results that were highly correlated with HPLC-MS data. Therefore, we propose a method that can be used for the rapid, selective and sensitive evaluation of sulfonamides in contaminated food. PMID:19615505

  20. New trend in the LC separation analysis of pharmaceuticals--high performance separation by ultra high-performance liquid chromatography (UHPLC) with core-shell particle C18 columns--.

    PubMed

    Nishi, Hiroyuki; Nagamatsu, Kumi

    2014-01-01

    This article presents a mini-review of the recent results in the ultra high-performance liquid chromatography (UHPLC) separation of pharmaceuticals by our group. High performance UHPLC separation employing core-shell particle C18 columns was demonstrated. High performance (high theoretical plate number of approximately 20000/10 cm, low theoretical plate height of 5 μm) was obtained without any specific devices in the conventional HPLC apparatus, only through changing detector sampling times and the inner diameter of the connecting tube. High theoretical plate numbers with low column back pressure obtained by the core-shell particle columns enabled fast separation of the analytes. Methanol, which gives high column pressure drops in the reversed-phase mode HPLC compared with acetonitrile, can be used without any trouble. One analysis of the purity testing of diltiazem hydrochloride was performed within 100 s. One analysis in the photostability testing of mecobalamin (vitamin B12 analogue) was successful within 180 s. PMID:24521905

  1. An HPLC-MS/MS method for the separation of α-retinyl esters from retinyl esters.

    PubMed

    Goetz, Hilary J; Kopec, Rachel E; Riedl, Ken M; Cooperstone, Jessica L; Narayanasamy, Sureshbabu; Curley, Robert W; Schwartz, Steven J

    2016-09-01

    Enzymatic cleavage of the nonsymmetric provitamin A carotenoid α-carotene results in one molecule of retinal (vitamin A), and one molecule of α-retinal, a biologically inactive analog of true vitamin A. Due to structural similarities, α-retinyl esters and vitamin A esters typically coelute, resulting in the overestimation of vitamin A originating from α-carotene. Herein, we present a set of tools to identify and separate α-retinol products from vitamin A. α-Retinyl palmitate (αRP) standard was synthesized from α-ionone following a Wittig-Horner approach. A high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method employing a C30 column was then developed to separate the species. Authentic standards of retinyl esters and the synthesized α-RP confirmed respective identities, while other α-retinyl esters (i.e. myristate, linoleate, oleate, and stearate) were evidenced by their pseudomolecular ions observed in electrospray ionization (ESI) mode, fragmentation, and elution order. For quantitation, an atmospheric pressure chemical ionization (APCI) source operated in positive ion mode was used, and retinol, the predominant in-source parent ion was selected and fragmented. The application of this method to a chylomicron-rich fraction of human plasma is demonstrated. This method can be used to better determine the quantity of vitamin A derived from foods containing α-carotene. PMID:27423669

  2. Rapid separation and analysis of six organic acids in Bayer liquors by RP-HPLC after solid-phase extraction.

    PubMed

    Xiao, Jian Bo; Chen, Xiao Qing; Jiang, Xin Yu; Wu, Sheng De

    2006-01-01

    A rapid revised phase high-performance liquid chromatographic (RP-HPLC) method for the determination of six organic acids in Bayer liquors is reported. Oxalic, tartaric, acetic, succinic, glutaric and butene dicarboxylic acid were separated and quantified in 10 min. First time repeatability, reproducibility and recoveries were determined out for these acids in Bayer liquors. The organic acids were removed from Bayer liquor by using a solid-phase extraction procedure with anion-exchange cartridges. The chromatographic separation was achieved with only one Kromasil RP-C18 column thermo stated at 25 degrees C. Organic acids were detected with a UV-vis detector (215 nm). The precision results showed that the relative standard deviations of the repeatability and reproducibility were < 2.80% and < 3.74%, respectively. The accuracy of the method was confirmed with an average recovery ranging between 85.2 and 107.3%. Under optimum conditions the detection limits ranged from 50 to 1000 mg/L.

  3. Determination of carotenoids in Taraxacum formosanum by HPLC-DAD-APCI-MS and preparation by column chromatography.

    PubMed

    Kao, T H; Loh, C H; Inbaraj, B Stephen; Chen, B H

    2012-07-01

    The objectives of this study were to determine the variety and content of carotenoids in Taraxacum formosanum, a traditional Chinese herb possessing vital biological activities, by developing an HPLC-DAD-APCI-MS method and a preparative column chromatographic method for carotenoid isolation. A total of 25 carotenoids were resolved within 66 min by employing a YMC C30 column and a gradient mobile phase of methanol-acetonitrile-water (79:14:7, v/v/v) and methylene chloride (100%) with flow rate at 1.0 mL/min and detection at 450 nm. All-trans-canthaxanthin was shown to be an appropriate internal standard for quantitation, with all-trans-β-carotene and its cis isomers present in largest amount (413.6 μg/g), followed by all-trans-violoxanthin and its cis isomers (209.5 μg/g), all-trans-lutein and its cis isomers (212.4 μg/g), all-trans-neoxanthin and its cis isomers (134.6 μg/g), antheraxanthin (16.5 μg/g), all-trans-β-cryptoxanthin and its cis isomers (5.8 μg/g), all-trans-zeaxanthin (3.6 μg/g) and neochrome (0.1 μg/g). For preparative chromatography, with a glass column containing 52 g of magnesium oxide-diatomaceous earth (1:3, w/w) as adsorbent, the carotenoid fraction was eluted with 300 mL of ethyl acetate with flow rate at 10 mL/min. Some more epoxides and cis isomers of carotenoids were generated during preparative column chromatography. Nevertheless, the carotenoids isolated from T. formosanum may be used as raw material for possible production of health food in the future.

  4. Carboxyl modified magnetic nanoparticles coated open tubular column for capillary electrochromatographic separation of biomolecules.

    PubMed

    Wang, Wentao; Xiao, Xing; Chen, Jia; Jia, Li

    2015-09-11

    Carboxyl modified magnetic nanoparticles (Fe3O4-COOH MNPs) coated open tubular (OT) columns were prepared for capillary electrochromatography. The Fe3O4-COOH MNPs coatings were constructed on the surface of positively charged poly(diallydimethylammonium chloride) (PDDA) modified capillaries through electrostatic self-assembly approach. The as-prepared PDDA@Fe3O4-COOH MNPs coated OT columns were characterized by scanning electron microscopy and electroosmotic flow measurement. The electrochromatographic characterization of the OT columns was evaluated by separation of amino acids, dipeptides and proteins. The influences of background solution pH, concentration, and organic modifier content on separation were investigated. The separation of these analytes was primarily based on the electrophoretic mechanism in combination with chromatographic mechanism. The Fe3O4-COOH MNPs coatings improved the separation resolution of these analytes due to their large surface area. Three variants of bovine serum albumin, two variants of β-lactoglobulin and nine glycoisoforms of ovalbumin were successfully separated. The relative standard deviations of migration times of analytes representing run-to-run, day-to-day and column-to-column were less than 4.3%. Furthermore, the feasibility of the PDDA@Fe3O4-COOH MNPs coated OT column was verified by successful separation of acidic proteins in egg white. PMID:26265004

  5. Simultaneous separation and determination of process-related substances and degradation products of venlafaxine by reversed-phase HPLC.

    PubMed

    Nageswara Rao, R; Narasa Raju, A

    2006-12-01

    A simple and rapid gradient RP HPLC method for simultaneous separation and determination of venlafaxine and its related substances in bulk drugs and pharmaceutical formulations has been developed. As many as four process impurities and one degradation product of venlafaxine have been separated on a Kromasil KR100-5C18 (4.6 mm x 250 mm; particle size 5 microm) column with gradient elution using 0.3% diethylamine buffer (pH 3.0) and ACN/methanol (90:10 v/v) as a mobile phase. The column was maintained at 40 degrees C and the eluents were monitored with photo diode array detection at 225 nm. The chromatographic behaviour of all the compounds was examined under variable compositions of different solvents, temperatures, buffer concentrations and pH. The method was validated in terms of accuracy, precision and linearity as per ICH guidelines. The inter- and intraday assay precision was < 4.02% (%RSD) and the recoveries were in the range of 96.19-101.14% with %RSD < 1.15%. The correlation coefficients (r2) for calibration curves of venlafaxine as well as impurities were in the range of 0.9942-0.9999. The proposed RP-LC method was successfully applied to the analysis of commercial formulations and the recoveries of venlafaxine were in the range of 99.32-100.67 with %RSD <0.58%. The method could be of use not only for rapid and routine evaluation of the quality of venlafaxine in bulk drug manufacturing units but also for the detection of its impurities in pharmaceutical formulations. Forced degradation of venlafaxine was carried out under thermal, photo, acidic, basic and peroxide conditions and the acid degradation products were characterized by ESI-MS/MS, 1H NMR and FT-IR spectral data.

  6. Reverse-phase h.p.l.c. separation, quantification and preparation of bilirubin and its conjugates from native bile. Quantitative analysis of the intact tetrapyrroles based on h.p.l.c. of their ethyl anthranilate azo derivatives.

    PubMed Central

    Spivak, W; Carey, M C

    1985-01-01

    We describe a facile and sensitive reverse-phase h.p.l.c. method for analytical separation of biliary bile pigments and direct quantification of unconjugated bilirubin (UCB) and its monoglucuronide (BMG) and diglucuronide (BDG) conjugates in bile. The method can be 'scaled up' for preparative isolation of pure BDG and BMG from pigment-enriched biles. We employed an Altex ultrasphere ODS column in the preparative steps and a Waters mu-Bondapak C18 column in the separatory and analytical procedures. Bile pigments were eluted with ammonium acetate buffer, pH 4.5, and a 20 min linear gradient of 60-100% (v/v) methanol at a flow rate of 2.0 ml/min for the preparative separations and 1.0 ml/min for the analytical separations. Bile pigments were eluted in order of decreasing polarity (glucuronide greater than glucose greater than xylose conjugates greater than UCB) and were chemically identified by t.l.c. of their respective ethyl anthranilate azo derivatives. Quantification of UCB was carried out by using a standard curve relating a range of h.p.l.c. integrated peak areas to concentrations of pure crystalline UCB. A pure crystalline ethyl anthranilate azo derivative of UCB (AZO . UCB) was employed as a single h.p.l.c. reference standard for quantification of BMG and BDG. We demonstrate that: separation and quantification of biliary bile pigments are rapid (approximately 25 min); bile pigment concentrations ranging from 1-500 microM can be determined 'on line' by using 5 microliters of bile without sample pretreatment; bilirubin conjugates can be obtained preparatively in milligram quantities without degradation or contamination by other components of bile. H.p.l.c. analyses of a series of mammalian biles show that biliary UCB concentrations generally range from 1 to 17 microM. These values are considerably lower than those estimated previously by t.l.c. BMG is the predominant, if not exclusive, bilirubin conjugate in the biles of a number of rodents (guinea pig, hamster

  7. Separation of algal cells from water by column flotation

    SciTech Connect

    Liu, J.C.; Chen, Y.M.; Ju, Y.H.

    1999-08-01

    The dispersed air flotation (DiAF) process was utilized to separate algal cells (Chlorella sp.) from water. Two types of collector, cationic N-cetyl-N,N,N-trimethylammonium bromide (CTAB) and anionic sodium dodecylsulfate (SDS), were used. It was observed that 20% of cell removal was achieved in the presence of 40 mg/L of SDS, and ca. 86% of the cells were removed at 40 mg/L of CTAB. Upon the addition of 10 mg/L of chitosan, over 90% of the cells were removed when SDS (20 mg/L) was used as the collector. Air flow rate affected cell flotation slightly. Optimum pH values for cell flotation were from 4.0 to 5.0. Flotation efficiency decreased at high ionic strength. The electrostatic interaction between collector and cell surface plays a critical role in the separation processes.

  8. Development and design of a multi-column experimental setup for Kr/Xe separation

    SciTech Connect

    Garn, Troy G.; Greenhalgh, Mitchell; Watson, Tony

    2014-12-01

    As a precursor to FY-15 Kr/Xe separation testing, design modifications to an existing experimental setup are warranted. The modifications would allow for multi-column testing to facilitate a Xe separation followed by a Kr separation using engineered form sorbents prepared using an INL patented process. A new cooling apparatus capable of achieving test temperatures to -40° C and able to house a newly designed Xe column was acquired. Modifications to the existing setup are being installed to allow for multi-column testing and gas constituent analyses using evacuated sample bombs. The new modifications will allow for independent temperature control for each column enabling a plethora of test conditions to be implemented. Sample analyses will be used to evaluate the Xe/Kr selectivity of the AgZ-PAN sorbent and determine the Kr purity of the effluent stream following Kr capture using the HZ-PAN sorbent.

  9. Azeotropic distillation in a middle vessel batch column. 2: Nonlinear separation boundaries

    SciTech Connect

    Cheong, W.; Barton, P.I.

    1999-04-01

    On the basis of the analytical tools developed for the middle vessel column (MVC) operated under limiting conditions, analysis of the qualitative dynamics of the MVC in separating an azeotropic mixture is extended to the more realistic case in which the separation boundaries are nonlinear. The differences between batch stripper pot composition boundaries and batch rectifier pot composition being able to cross these pot composition boundaries. On the basis of these insights, operating procedures are developed in which ternary azeotropic mixtures of acetone, benzene, and chloroform can be separated into their constituent pure components, a separation not achievable with either the batch stripper or the batch rectifier. The operating procedures suggested for separating the ternary azeotropic mixture of acetone, benzene, and chloroform in the MVC are then shown to be the time analogues of sequences of continuous distillation columns that achieve the same separation. On the basis of this space-time analogy, further analogies are developed between the MVC and a continuous column, and it is postulated that many complex separations currently achieved with sequences of continuous columns can also be achieved with a single MVC. Thus, the MVC represents the ultimate multipurpose solvent recovery technology, as it can handle, in a batch multipurpose mode. separations that will otherwise require a dedicated continuous distillation sequence. Finally, the characteristics of perfect MVC batch entrainers, which allow the complete separation of any azeotrope into its constituent pure components in a single MVC, are discussed.

  10. Silica-based monolithic column with evaporative light scattering detector for HPLC analysis of bacosides and apigenin in Bacopa monnieri.

    PubMed

    Bhandari, Pamita; Kumar, Neeraj; Singh, Bikram; Singh, Virendra; Kaur, Inderjeet

    2009-08-01

    A high performance liquid chromatographic method using a silica-based monolithic column coupled with evaporative light scattering detector (HPLC-ELSD) was developed and validated for simultaneous quantification of bacosides (bacoside A, bacopaside I, bacoside A(3), bacopaside II, bacopaside X, bacopasaponin C) and apigenin in Bacopa monnieri. The chromatographic resolution was achieved on a Chromolith RP-18 (100x4.6 mm) column with acetonitrile/water (30:70) as mobile phase in isocratic elution at a flow rate of 0.7 mL/min. The drift tube temperature of the ELSD was set to 95 degrees C, and the nitrogen flow rate was 2.0 SLM (standard liter per minute). The calibration curves revealed a good linear relationship (r(2) > 0.9988) within the test ranges. The detection limits (S/N = 3) and the quantification limits (S/N = 10) for the compounds were in the range of 0.54-6.06 and 1.61-18.78 microg/mL, respectively. Satisfactory average recovery was observed in the range of 95.8-99.0%. The method showed good reproducibility for the quantification of these compounds in B. monnieri with intra- and inter-day precision of less than 0.69 and 0.67%, respectively. The validated method was successfully applied to quantify analytes in nine accessions of B. monnieri and thus provides a new basis for overall quality assessment of B. monnieri.

  11. [Comparison of photometric, electrochemical and post-column fluorescence detection for the determination of flavonoids by HPLC].

    PubMed

    Saito, A; Sugisawa, A; Umegaki, K

    2001-06-01

    For the analysis of flavonoids by HPLC, we compared three different detection methods, namely UV, electrochemical detection and post-column chelation with aluminum followed by fluorescence detection. Ten flavonoids were used: apigenin, myricetin, luteorin, taxifolin, quercetin-3-O-sulfate, kaempferol, isorhamnetin, isoquercitrin, quercetin and rutin. Nine flavonoids except apigenin were efficiently detected by the electrochemical method with a detection limit of 0.025-0.05 pmol. Flavonols having free 3-hydroxyl and 4-keto oxygen formed a fluorescent complex by post-column chelation and were detected by the fluorescence method. The detection limit of the fluorescence method was 0.05-0.5 pmol. Nine flavonoids except taxifolin were detected by the UV method (absorbance at 370 nm), but the detection level was poor (5-10 pmol). Flavonols added to human plasma were recovered by solid phase extraction, and were analyzed using the three detection methods. Most of the flavonols were efficiently detected by the electrochemical and fluorescence methods, and the detection limits were similar to those of standard samples. PMID:11577389

  12. Separation and identification of phenolic compounds in canned artichoke by LC/DAD/ESI-MS using core-shell C18 column: a comparative study.

    PubMed

    Wu, Jianbing; Qian, Yongsheng; Mao, Peipei; Chen, Linyao; Lu, Yanbin; Wang, Huizhong

    2013-05-15

    Core-shell silica stationary phase was considered as a breakthrough in column technology in HPLC world. In this work, the chromatographic performance of core-shell column, made by fusing a 0.5μm porous silica layer onto 1.7μm nonporous silica cores, was compared with sub-2μm fully porous particle materials for separation and identification of phenolic compounds in canned artichoke heads. The anti-oxidant caffeoylquinic acids of artichoke extract was taken as representative for calculating the plate heights in a wide flow rate range and analyzed on the basis of the van Deemter and Knox equations. Theoretical Poppe plots were constructed for each column to compare their kinetic performance. Both phases gave similar minimum plate heights when using non-reduced coordinates. Meanwhile, the flat C-term of core-shell column provided the possibilities for applying high flow rates without significant loss in efficiency. In addition, the peak capacities of both columns were measured, at constant chromatographic linear velocity and intrinsic gradient steepness, in order to generate comparable retention window for the least and the most retained compounds. Finally, the core-shell column was successfully applied for separation and identification of 10 phenolic compounds in canned artichoke extracts by liquid chromatography-diode array detection-tandem mass spectrometry, exhibiting great potential in the field of food analysis. PMID:23266111

  13. Ethyl-bridged hybrid column as an efficient alternative for HPLC analysis of plasma amino acids by pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate.

    PubMed

    Castellanos, Mar; Van Eendenburg, Cecile Van; Gubern, Carme; Sanchez, Juan M

    2016-09-01

    Conventional C18 silica columns have proven to be useful for the analysis of amino acids (AA) from protein hydrolysates but undesirable peak overlapping is usually found when analyzing body fluids given that a large number of AAs are present in the samples. As an alternative to silica packings, an ethyl-bridged packing for reversed-phase liquid chromatography of derivatized AAs with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) has been evaluated. The new packing material improves the separation efficiency allowing better separations when analyzing biological fluids. Moreover, this packing has advantages for routine AA analysis, such as a decrease in the total running time and an increase in the life-time of the columns. The pH of the mobile phase has a significant effect on the elution behavior of the AQC hydrolysis product (AMQ) and on the AA derivatives. It is not possible to elute AMQ before detecting the first AA derivative, which requires an accurate adjustment of the pH in the range of 5.30-5.35 to obtain good separation and resolution for the most polar compounds. Under the conditions proposed, it is possible to separate all AAs except the Gly-Gln pair, which is not a problem when hydrolyzed samples are analyzed. The AMQ-Ser pair requires either the use of a different mobile phase pH for its baseline separation or the use of fluorescence detection. Two different procedures for protein removal from plasma samples have been evaluated, solvent precipitation and ultrafiltration (UF) and it has been found that UF gives better results as no significant losses of AAs were observed. The validation of the proposed method with UV detection gives method detection limits in the range of 8-12μM, with repeatability values<8% (n=6) and inter-day precision in plasma samples ranging from 4 to 13% (n=4).

  14. Ethyl-bridged hybrid column as an efficient alternative for HPLC analysis of plasma amino acids by pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate.

    PubMed

    Castellanos, Mar; Van Eendenburg, Cecile Van; Gubern, Carme; Sanchez, Juan M

    2016-09-01

    Conventional C18 silica columns have proven to be useful for the analysis of amino acids (AA) from protein hydrolysates but undesirable peak overlapping is usually found when analyzing body fluids given that a large number of AAs are present in the samples. As an alternative to silica packings, an ethyl-bridged packing for reversed-phase liquid chromatography of derivatized AAs with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) has been evaluated. The new packing material improves the separation efficiency allowing better separations when analyzing biological fluids. Moreover, this packing has advantages for routine AA analysis, such as a decrease in the total running time and an increase in the life-time of the columns. The pH of the mobile phase has a significant effect on the elution behavior of the AQC hydrolysis product (AMQ) and on the AA derivatives. It is not possible to elute AMQ before detecting the first AA derivative, which requires an accurate adjustment of the pH in the range of 5.30-5.35 to obtain good separation and resolution for the most polar compounds. Under the conditions proposed, it is possible to separate all AAs except the Gly-Gln pair, which is not a problem when hydrolyzed samples are analyzed. The AMQ-Ser pair requires either the use of a different mobile phase pH for its baseline separation or the use of fluorescence detection. Two different procedures for protein removal from plasma samples have been evaluated, solvent precipitation and ultrafiltration (UF) and it has been found that UF gives better results as no significant losses of AAs were observed. The validation of the proposed method with UV detection gives method detection limits in the range of 8-12μM, with repeatability values<8% (n=6) and inter-day precision in plasma samples ranging from 4 to 13% (n=4). PMID:27428457

  15. Transport of Cryptosporidium parvum Oocysts in Soil Columns following Applications of Raw and Separated Liquid Slurries

    PubMed Central

    Petersen, Heidi H.; Enemark, Heidi L.; Olsen, Annette; Amin, M. G. Mostofa

    2012-01-01

    The potential for the transport of viable Cryptosporidium parvum oocysts through soil to land drains and groundwater was studied using simulated rainfall and intact soil columns which were applied raw slurry or separated liquid slurry. Following irrigation and weekly samplings over a 4-week period, C. parvum oocysts were detected from all soil columns regardless of slurry type and application method, although recovery rates were low (<1%). Soil columns with injected liquid slurry leached 73 and 90% more oocysts compared to columns with injected and surface-applied raw slurries, respectively. Among leachate samples containing oocysts, 44/72 samples yielded viable oocysts as determined by a dye permeability assay (DAPI [4′,6′-diamidino-2-phenylindole]/propidium iodide) with the majority (41%) of viable oocysts found in leachate from soil columns with added liquid slurry. The number of viable oocysts was positively correlated (r = 0.63) with the total number of oocysts found. Destructively sampling of the soil columns showed that type of slurry and irrigation played a role in the vertical distribution of oocysts, with more oocysts recovered from soil columns added liquid slurry irrespective of the irrigation status. Further studies are needed to determine the effectiveness of different slurry separation technologies to remove oocysts and other pathogens, as well as whether the application of separated liquid slurry to agricultural land may represent higher risks for groundwater contamination compared to application of raw slurry. PMID:22706058

  16. Transport of Cryptosporidium parvum oocysts in soil columns following applications of raw and separated liquid slurries.

    PubMed

    Petersen, Heidi H; Enemark, Heidi L; Olsen, Annette; Amin, M G Mostofa; Dalsgaard, Anders

    2012-09-01

    The potential for the transport of viable Cryptosporidium parvum oocysts through soil to land drains and groundwater was studied using simulated rainfall and intact soil columns which were applied raw slurry or separated liquid slurry. Following irrigation and weekly samplings over a 4-week period, C. parvum oocysts were detected from all soil columns regardless of slurry type and application method, although recovery rates were low (<1%). Soil columns with injected liquid slurry leached 73 and 90% more oocysts compared to columns with injected and surface-applied raw slurries, respectively. Among leachate samples containing oocysts, 44/72 samples yielded viable oocysts as determined by a dye permeability assay (DAPI [4',6'-diamidino-2-phenylindole]/propidium iodide) with the majority (41%) of viable oocysts found in leachate from soil columns with added liquid slurry. The number of viable oocysts was positively correlated (r = 0.63) with the total number of oocysts found. Destructively sampling of the soil columns showed that type of slurry and irrigation played a role in the vertical distribution of oocysts, with more oocysts recovered from soil columns added liquid slurry irrespective of the irrigation status. Further studies are needed to determine the effectiveness of different slurry separation technologies to remove oocysts and other pathogens, as well as whether the application of separated liquid slurry to agricultural land may represent higher risks for groundwater contamination compared to application of raw slurry.

  17. Transport of Cryptosporidium parvum oocysts in soil columns following applications of raw and separated liquid slurries.

    PubMed

    Petersen, Heidi H; Enemark, Heidi L; Olsen, Annette; Amin, M G Mostofa; Dalsgaard, Anders

    2012-09-01

    The potential for the transport of viable Cryptosporidium parvum oocysts through soil to land drains and groundwater was studied using simulated rainfall and intact soil columns which were applied raw slurry or separated liquid slurry. Following irrigation and weekly samplings over a 4-week period, C. parvum oocysts were detected from all soil columns regardless of slurry type and application method, although recovery rates were low (<1%). Soil columns with injected liquid slurry leached 73 and 90% more oocysts compared to columns with injected and surface-applied raw slurries, respectively. Among leachate samples containing oocysts, 44/72 samples yielded viable oocysts as determined by a dye permeability assay (DAPI [4',6'-diamidino-2-phenylindole]/propidium iodide) with the majority (41%) of viable oocysts found in leachate from soil columns with added liquid slurry. The number of viable oocysts was positively correlated (r = 0.63) with the total number of oocysts found. Destructively sampling of the soil columns showed that type of slurry and irrigation played a role in the vertical distribution of oocysts, with more oocysts recovered from soil columns added liquid slurry irrespective of the irrigation status. Further studies are needed to determine the effectiveness of different slurry separation technologies to remove oocysts and other pathogens, as well as whether the application of separated liquid slurry to agricultural land may represent higher risks for groundwater contamination compared to application of raw slurry. PMID:22706058

  18. A convenient HPLC method for detection of okadaic acid analogs as 9-anthrylmethyl esters with automated sample cleanup by column switching.

    PubMed

    Uchida, Hajime; Watanabe, Ryuichi; Matsushima, Ryoji; Uchida, Naoyuki; Nagai, Hiroshi; Kamio, Michiya; Murata, Masakazu; Yasumoto, Takeshi; Suzuki, Toshiyuki

    2014-01-01

    A convenient HPLC-fluorometric detection (FLD) method for okadaic acid (OA) analogs as 9-anthrylmethyl esters was developed with the addition of column switching to simplify and automate cleanup. Methanol extracts of shellfish were first treated to hydrolyze OA esters and then reacted with 9-anthryldiazomethane (ADAM). ADAM derivatives of OA and dinophysistoxin-1 (DTX1) were subsequently determined by HPLC-FLD following automated column-switching cleanup. The LOD (S/N = 3) and LOQ (S/N = 10) of OA and DTX1 obtained from bivalves fortified with toxin in our method were approximately 2.6 and 8.6 ng/g whole meat, respectively. The recoveries of OA and DTX1 at all fortification levels of bivalve extracts ranged from 90 to 113%, with RSD values of 0.9-9.9%. The new method is applicable to the routine monitoring of OA analogs as an inexpensive and convenient alternative to HPLC/MS.

  19. Liquid-phase thermal diffusion isotope separation apparatus and method having tapered column

    DOEpatents

    Rutherford, William M.

    1988-05-24

    A thermal diffusion counterflow method and apparatus for separating isotopes in solution in which the solution is confined in a long, narrow, vertical slit which tapers from bottom to top. The variation in the width of the slit permits maintenance of a stable concentration distribution with relatively long columns, thus permitting isotopic separation superior to that obtainable in the prior art.

  20. Liquid-phase thermal diffusion isotope separation apparatus and method having tapered column

    DOEpatents

    Rutherford, W.M.

    1985-12-04

    A thermal diffusion counterflow method and apparatus for separating isotopes in solution in which the solution is confined in a long, narrow, vertical slit which tapers from bottom to top. The variation in the width of the slit permits maintenance of a stable concentration distribution with relatively long columns, thus permitting isotopic separation superior to that obtained in the prior art.

  1. Determination of antihyperglycemic biguanides in serum and urine using an ion-pair solid-phase extraction technique followed by HPLC-UV on a pentafluorophenylpropyl column and on an octadecyl column.

    PubMed

    Tahara, Kayoko; Yonemoto, Ayumi; Yoshiyama, Yuji; Nakamura, Toshiya; Aizawa, Masaaki; Fujita, Yoshikuni; Nishikawa, Takashi

    2006-11-01

    An HPLC-UV method was established for the determination of metformin and buformin in biological fluids. Metformin was not retained on particles packed in conventional solid-phase extraction cartridges; in contrast, buformin was retained too firmly and not eluted with a solvent for recovery. However, both drugs were retained on particles that had been treated with an ion-pair reagent of heptanesulfonate or dodecylsulfate and recovered almost completely. The recovered fraction was subjected to HPLC on a pentafluorophenylpropyl column which was suitable for the determination of both biguanides in serum and in urine. Limits of quantitation were low enough for clinical use, and reproducibility was high with an RSD of 0.9-2.3%. HPLC on a conventional octadecyl column was suitable only for the determination of buformin in serum since interfering peaks appeared on the chromatograms of urine samples. The method was applied to analysis of some clinical specimens.

  2. Separation and quantification of water buffalo milk protein fractions and genetic variants by RP-HPLC.

    PubMed

    Bonfatti, Valentina; Giantin, Mery; Rostellato, Roberta; Dacasto, Mauro; Carnier, Paolo

    2013-01-15

    A RP-HPLC method, developed for the separation and quantification of the most common genetic variants of bovine milk proteins, was successfully applied to the analysis of water buffalo milk. All the most common buffalo casein and whey proteins fractions, as well as their genetic variants, were detected and separated simultaneously in 40 min. Purified buffalo proteins were used as calibration standards and a total of 536 individual milk samples were analysed for protein composition. α(S1)-, α(S2)-, βγ-, and κ-casein were 32.2%, 15.8%, 36.5%, and 15.5%, respectively, of total casein content, whereas content of β-Lactoglobulin was approximately 1.3 times as high as that of α-Lactalbumin. The existence of a polymorphism of κ-casein was demonstrated in Mediterranean water buffalo and α(S1)- and κ-casein genetic variants were successfully detected by RP-HPLC. PMID:23122071

  3. Evaluation of a silicon oxynitride hydrophilic interaction liquid chromatography column in saccharide and glycoside separations.

    PubMed

    Wan, Huihui; Sheng, Qianying; Zhong, Hongmin; Guo, Xiujie; Fu, Qing; Liu, Yanfang; Xue, Xingya; Liang, Xinmiao

    2015-05-01

    The retention characteristics of a silicon oxynitride stationary phase for carbohydrate separation were studied in hydrophilic interaction chromatography mode. Four saccharides including mono-, di-, and trisaccharides were employed to investigate the effects of water content and buffer concentration in the mobile phase on hydrophilic interaction liquid chromatography retention. For the tested saccharides, the silicon oxynitride column demonstrated excellent performance in terms of separation efficiency, hydrophilicity, and interesting separation selectivity for carbohydrates compared to the bare silica stationary phase. Finally, the silicon oxynitride hydrophilic interaction liquid chromatography column was employed in the separation of complex samples of fructooligosaccharides, saponins, and steviol glycoside from natural products. The resulting chromatograms demonstrated good separation efficiency and longer retention compared with silica, which further confirmed the advantages and potential application of silicon oxynitride stationary phase for hydrophilic interaction liquid chromatography separation.

  4. Evaluation of fructooligosaccharides separation using a fixed-bed column packed with activated charcoal.

    PubMed

    Kuhn, Raquel Cristine; Mazutti, Marcio A; Albertini, Lilian Buoro; Filho, Francisco Maugeri

    2014-05-25

    Recent studies have shown that the chromatographic separation of mixtures of saccharides may be improved by making use of activated charcoal, a promising low cost material for the separation of sugars, including fructooligosaccharides. In this work, the development of a methodology to separate fructooligosaccharides from glucose, fructose and sucrose, using a fixed bed column packed with activated charcoal is proposed. The influence of temperature, eluant concentration and step gradients were evaluated to increase the separation efficiency and fructooligosaccharide purity. The final degree of fructooligosaccharide purification and separation efficiency were about 94% and 3.03 respectively, using ethanol gradient concentration ranging from 3.5% to 15% (v/v) at 40°C. The fixed bed column packed with the activated charcoal was shown to be a promising alternative for sugar separation, mainly those rich in fructooligosaccharides, leading to solutions of acceptable degrees of purification.

  5. Quantitative analysis of olanzapine in rat brain microdialysates by HPLC-MS/MS coupled with column-switching technique.

    PubMed

    Zheng, Qiaoling; Wang, Feng; Li, Huande; Xu, Ping; Tang, Huaibo; Li, Lanfang; Cheng, Rihua

    2012-09-15

    A rapid and sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method coupled with column-switching technique was developed for the determination of olanzapine in rat brain microdialysates. A C8 guard column was used to desalt the samples before analytical separation on a C18 column and detection with tandem mass spectrometry. The mobile phase consisted of methanol/acetonitrile/water (v/v/v, 22.5/22.2/55) was used for desalting and the mobile phase consisted of methanol/acetonitrile/water (v/v/v, 43/43/14) was for analytical separation, water in both mobile phases contained 0.1% ammonium acetate. The lower limit of quantification (LLOQ) for olanzapine was 0.085 ng/ml. The method was linear from LLOQ to 34 ng/ml with a coefficient of determination >0.998. Intra- and inter-day accuracy and precision were determined with variability less than 13.24% (R.S.D). This sensitive method was successfully applied to quantify the concentration of olanzapine in rat brain microdialysates. With this study, the effect of the alcohol extract of Schisandra sphenanthera Rehd. et Wils on the concentration of olanzapine in brain was investigated. PMID:22917592

  6. Development of a novel amide-silica stationary phase for the reversed-phase HPLC separation of different classes of phytohormones.

    PubMed

    Aral, Hayriye; Aral, Tarık; Ziyadanoğulları, Berrin; Ziyadanoğulları, Recep

    2013-11-15

    A novel amide-bonded silica stationary phase was prepared starting from N-Boc-phenylalanine, cyclohexylamine and spherical silica gel (4 µm, 60 Å). The amide ligand was synthesised with high yield. The resulting amide bonded stationary phase was characterised by SEM, IR and elemental analysis. The resulting selector bearing a polar amide group is used for the reversed-phase chromatography separation of different classes of thirteen phytohormones (plant hormones). The chromatographic behaviours of these analytes on the amide-silica stationary phase were compared with those of RP-C18 column under same conditions. The effects of different separation conditions, such as mobile phase, pH value, flow rate and temperature, on the separation and retention behaviours of the 13 phytohormones in this system were studied. The optimum separation was achieved using reversed-phase HPLC gradient elution with an aqueous mobile phase containing pH=6.85 potassium phosphate buffer (20 mM) and acetonitrile with a 22 °C column temperature. Under these experimental conditions, the 12 phytohormones could be separated and detected at 230 or 270 nm within 26 min.

  7. Polydopamine-coated open tubular column for the separation of proteins by capillary electrochromatography.

    PubMed

    Xiao, Xing; Wang, Wentao; Chen, Jia; Jia, Li

    2015-08-01

    The separation and determination of proteins in food is an important aspect in food industry. Inspired by the self-polymerization of dopamine under alkaline conditions and the natural adhesive properties of polydopamine, in this paper, a simple and economical method was developed for the preparation of polydopamine-coated open tubular column, in which ammonium persulfate was used as the source of oxygen to induce and facilitate the polymerization of dopamine to form polydopamine. In comparison with a naked fused-silica capillary, the direction and magnitude of the electro-osmotic flow of the as-prepared polydopamine-coated open tubular column could be manipulated by varying the pH values of background solutions due to the existence of amine and phenolic hydroxyl groups on polydopamine coating. The surface morphology of the polydopamine-coated open tubular column was studied by scanning electron microscopy, and the thickness of polydopamine coating was 106 nm. The performance of the polydopamine-coated open tubular column was validated by analysis of proteins. The relative standard deviations of migration times of proteins representing run-to-run, day-to-day, and column-to-column were less than 3.5%. In addition, the feasibility of the polydopamine-coated open tubular column for real samples was verified by the separation of proteins in chicken egg white and pure milk. PMID:26017540

  8. Multi-Column Experimental Test Bed for Xe/Kr Separation

    SciTech Connect

    Greenhalgh, Mitchell Randy; Garn, Troy Gerry; Welty, Amy Keil; Lyon, Kevin Lawrence; Watson, Tony Leroy

    2015-08-31

    Previous research studies have shown that INL-developed engineered form sorbents are capable of capturing both Kr and Xe from various composite gas streams. The previous experimental test bed provided single column testing for capacity evaluations over a broad temperature range. To advance research capabilities, the employment of an additional column to study selective capture of target species to provide a defined final gas composition for waste storage was warranted. The second column addition also allows for compositional analyses of the final gas product to provide for final storage determinations. The INL krypton capture system was modified by adding an additional adsorption column in order to create a multi-column test bed. The purpose of this modification was to investigate the separation of xenon from krypton supplied as a mixed gas feed. The extra column was placed in a Stirling Ultra-low Temperature Cooler, capable of controlling temperatures between 190 and 253K. Additional piping and valves were incorporated into the system to allow for a variety of flow path configurations. The new column was filled with the AgZ-PAN sorbent which was utilized as the capture medium for xenon while allowing the krypton to pass through. The xenon-free gas stream was then routed to the cryostat filled with the HZ-PAN sorbent to capture the krypton at 191K. Selectivities of xenon over krypton were determined using the new column to verify the system performance and to establish the operating conditions required for multi-column testing. Results of these evaluations verified that the system was operating as designed and also demonstrated that AgZ-PAN exhibits excellent selectivity for xenon over krypton in air at or near room temperature. Two separation tests were performed utilizing a feed gas consisting of 1000 ppmv xenon and 150 ppmv krypton with the balance being made up of air. The AgZ-PAN temperature was held at 295 or 253K while the HZ-PAN was held at 191K for both

  9. Evaluation of a column classification method using the separation of alfuzosin from its related substances.

    PubMed

    Szulfer, Jarosław; Plenis, Alina; Bączek, Tomasz

    2012-03-16

    The popularity and commercial availability of reversed-phase liquid chromatographic (RP-LC) stationary phases cause analysts to be often confronted with the problem of column selection. For this reason, general test methods to characterize RP-LC columns have been extensively studied since the 1970s. This paper focuses on correlating the column classification based on a method developed at the Katholieke Universiteit Leuven (KUL method) with the selectivity obtained for a real separation. The analysis of alfuzosin hydrochloride and related compounds was carried out according to the method prescribed in the European Pharmacopoeia (Ph. Eur.) monograph. This separation was performed on 36 new RP-LC stationary phases which had been previously characterized chromatographically. For deeper comparative analysis of KUL classification of the stationary RP-LC brands and their column performance in pharmaceutical practice two chemometric tools, such as principal component analysis (PCA) and cluster analysis (CA), have been used. It was shown that stationary phase classes closely related by KUL method gave comparable separation for alfuzosin and related compounds. Therefore, the column ranking system based on the evaluation of F-values can be considered as a helpful tool in the selection of a suitable column for pharmaceutical analyses.

  10. HPLC determination of capsaicinoids with cross-linked C18 column and buffer-free eluent.

    PubMed

    Daood, Hussein G; Halasz, Gábor; Palotás, Gábor; Palotás, Gabriella; Bodai, Zsolt; Helyes, Lajos

    2015-01-01

    A simple and efficient high-performance liquid chromatographic method was developed and validated for the separation and determination of capsaicin and its major dihydro- and homoderivatives in spice paprika products in 20 min with fluorescent and 35 min with mass-spectrometric detection. The separation was performed on reversed-phase chromatographic adsorbent of cross-linked endcapping with eluent consisting of 1:1 acetonitrile-water or acetonitrile-0.1% formic acid under isocratic conditions. Excellent separation of all the major and minor capsaicinoids with resolution index between 1.08 and 7.34 was achieved. The detection and quantification limits of capsaicinoids in standard material solutions were between 2 and 10 ng/mL. The lowest detectable amount of capsaicin, with fluorescent detection, was found to be <1 µg/g non-pungent spice paprika powder. The naturally occurring capsaicinoids could be distinguished from the non-capsaicinoids compounds appeared on liquid chromatography-fluorescence profile of extract from drastically processed paprika by applying mass spectroscopic detection. Hungarian spice paprika were evaluated as mild to very hot (capsaicinoid content: 334-1,660 µg/g) and chili products as very or extremely hot products (1,543-2,818 µg/g). PMID:24837232

  11. HPLC determination of capsaicinoids with cross-linked C18 column and buffer-free eluent.

    PubMed

    Daood, Hussein G; Halasz, Gábor; Palotás, Gábor; Palotás, Gabriella; Bodai, Zsolt; Helyes, Lajos

    2015-01-01

    A simple and efficient high-performance liquid chromatographic method was developed and validated for the separation and determination of capsaicin and its major dihydro- and homoderivatives in spice paprika products in 20 min with fluorescent and 35 min with mass-spectrometric detection. The separation was performed on reversed-phase chromatographic adsorbent of cross-linked endcapping with eluent consisting of 1:1 acetonitrile-water or acetonitrile-0.1% formic acid under isocratic conditions. Excellent separation of all the major and minor capsaicinoids with resolution index between 1.08 and 7.34 was achieved. The detection and quantification limits of capsaicinoids in standard material solutions were between 2 and 10 ng/mL. The lowest detectable amount of capsaicin, with fluorescent detection, was found to be <1 µg/g non-pungent spice paprika powder. The naturally occurring capsaicinoids could be distinguished from the non-capsaicinoids compounds appeared on liquid chromatography-fluorescence profile of extract from drastically processed paprika by applying mass spectroscopic detection. Hungarian spice paprika were evaluated as mild to very hot (capsaicinoid content: 334-1,660 µg/g) and chili products as very or extremely hot products (1,543-2,818 µg/g).

  12. Simultaneous separation and determination of fructose, sorbitol, glucose and sucrose in fruits by HPLC-ELSD.

    PubMed

    Ma, Chunmei; Sun, Zhen; Chen, Changbao; Zhang, Lili; Zhu, Shuhua

    2014-02-15

    A high-performance liquid chromatography (HPLC) method with evaporative light scattering detection (ELSD) was optimised for simultaneous determination of fructose, sorbitol, glucose and sucrose in fruits. The analysis was carried out on a Phenomenex Luna 5u NH₂ 100A column (250 mm × 4.60mm, 5 micron) with isocratic elution of acetonitrile:water (82.5:17.5, v/v). Drift tube temperature of the ELSD system was set to 82 °C and nitrogen flow rate was 2.0 L min⁻¹. The regression equation revealed good linear relationship (R = 0.9967-0.9989) within test ranges. The limits of detection (LOD) and quantification (LOQ) for four analytes (peach, apple, watermelon, and cherry fruits) were in the range of 0.07-0.27 and 0.22-0.91 mg L⁻¹, respectively. The proposed HPLC-ELSD method was validated for quantification of sugars in peach, apple, watermelon, and cherry fruits, and the results were satisfactory. The results showed that the contents of the four sugars varied among fruits. While fructose (5.79-104.01 mg g⁻¹) and glucose (9.25-99.62 mg g⁻¹) emerged as common sugars in the four fruits, sorbitol (8.70-19.13 mg g⁻¹) were only found in peach, apple and cherry fruits, and sucrose (15.82-106.39 mg g⁻¹) were in peach, apple and watermelon. There was not detectable sorbitol in watermelon and sucrose in cherry fruits, respectively.

  13. Semi-industrial experimental study on bauxite separation using a cell-column integration process

    NASA Astrophysics Data System (ADS)

    Zhang, Ning-ning; Zhou, Chang-chun; Cong, Long-fei; Cao, Wen-long; Zhou, You

    2016-01-01

    The cyclonic-static micro-bubble flotation column (FCSMC) is a highly efficient mineral processing equipment. In this study, a cell-column (FCSMC) integration process was investigated for the separation of bauxite and its feasibility was analyzed on a theoretical basis. The properties of low-grade bauxite ore from Henan Province, China were analyzed. Parameters such as reagent dosage, scraping bubble time, and pressure of the circulating pump during the sorting process were investigated and optimized to improve the flotation efficiency. On the basis of these parameters, continuous separation experiments were conducted. Bauxite concentrate with an aluminum-to-silicon (A/S) mass ratio of 6.37 and a 77.63wt% recovery rate were achieved via a flow sheet consisting of "fast flotation using a flotation cell, one roughing flotation and one cleaning flotation using flotation columns". Compared with the full-flotation-cells process, the cell-column integration process resulted in an increase of the A/S ratio by 0.41 and the recovery rate by 17.58wt%. Cell-column integration separation technology represents a new approach for the separation of middle-to-low-grade bauxite ore.

  14. Application of statistical design for the optimization of amino acid separation by reverse-phase HPLC.

    PubMed

    Gheshlaghi, R; Scharer, J M; Moo-Young, M; Douglas, P L

    2008-12-01

    Modified resolution and overall separation factors used to quantify the separation of complex chromatography systems are described. These factors were proven to be applicable to the optimization of amino acid resolution in reverse-phase (RP) HPLC chromatograms. To optimize precolumn derivatization with phenylisothiocyanate, a 2(5-1) fractional factorial design in triplicate was employed. The five independent variables for optimizing the overall separation factor were triethylamine content of the aqueous buffer, pH of the aqueous buffer, separation temperature, methanol/acetonitrile concentration ratio in the organic eluant, and mobile phase flow rate. Of these, triethylamine concentration and methanol/acetonitrile concentration ratio were the most important. The methodology captured the interaction between variables. Temperature appeared in the interaction terms; consequently, it was included in the hierarchic model. The preliminary model based on the factorial experiments was not able to explain the response curvature in the design space; therefore, a central composite design was used to provide a quadratic model. Constrained nonlinear programming was used for optimization purposes. The quadratic model predicted the optimal levels of the variables. In this study, the best levels of the five independent variables that provide the maximum modified resolution for each pair of consecutive amino acids appearing in the chromatograph were determined. These results are of utmost importance for accurate analysis of a subset of amino acids.

  15. Separative analyses of a chromatographic column packed with a core-shell adsorbent for lithium isotope separation

    SciTech Connect

    Sugiyama, T.; Sugura, K.; Enokida, Y.; Yamamoto, I.

    2015-03-15

    Lithium-6 is used as a blanket material for sufficient tritium production in DT fueled fusion reactors. A core-shell type adsorbent was proposed for lithium isotope separation by chromatography. The mass transfer model in a chromatographic column consisted of 4 steps, such as convection and dispersion in the column, transfer through liquid films, intra-particle diffusion and and adsorption or desorption at the local adsorption sites. A model was developed and concentration profiles and time variation in the column were numerically simulated. It became clear that core-shell type adsorbents with thin porous shell were saturated rapidly relatively to fully porous one and established a sharp edge of adsorption band. This is very important feature because lithium isotope separation requires long-distance development of adsorption band. The values of HETP (Height Equivalent of a Theoretical Plate) for core-shell adsorbent packed column were estimated by statistical moments of the step response curve. The value of HETP decreased with the thickness of the porous shell. A core-shell type adsorbent is, then, useful for lithium isotope separation. (authors)

  16. Evolutionary multi-objective optimization based comparison of multi-column chromatographic separation processes for a ternary separation.

    PubMed

    Heinonen, Jari; Kukkonen, Saku; Sainio, Tuomo

    2014-09-01

    Performance characteristics of two advanced multi-column chromatographic separation processes with discontinuous feed, Multi-Column Recycling Chromatogrphy (MCRC) and Japan Organo (JO), were investigated for a ternary separation using multi-objective optimization with an evolutionary algorithm. Conventional batch process was used as a reference. Fractionation of a concentrated acid hydrolysate of wood biomass into sulfuric acid, monosaccharide, and acetic acid fractions was used as a model system. Comparison of the separation processes was based on selected performance parameters in their optimized states. Flow rates and step durations were taken as decision variables whereas the column configuration and dimensions were fixed. The MCRC process was found to be considerably more efficient than the other processes with respect to eluent consumption. The batch process gave the highest productivity and the JO process the lowest. Both of the multi-column processes gave significantly higher monosaccharide yield than the batch process. When eluent consumption and monosaccharide yield are taken into account together with productivity, the MCRC process was found to be the most efficient in the studied case. PMID:25060000

  17. Simulation of a Novel Single-column Cryogenic Air Separation Process Using LNG Cold Energy

    NASA Astrophysics Data System (ADS)

    Jieyu, Zheng; Yanzhong, Li; Guangpeng, Li; Biao, Si

    In this paper, a novel single-column air separation process is proposed with the implementation of heat pump technique and introduction of LNG coldenergy. The proposed process is verifiedand optimized through simulation on the Aspen Hysys® platform. Simulation results reveal that thepower consumption per unit mass of liquid productis around 0.218 kWh/kg, and the total exergy efficiency of the systemis 0.575. According to the latest literatures, an energy saving of 39.1% is achieved compared with those using conventional double-column air separation units.The introduction of LNG cold energy is an effective way to increase the system efficiency.

  18. Development of an HPLC Method with an ODS Column to Determine Low Levels of Aspartame Diastereomers in Aspartame.

    PubMed

    Ohtsuki, Takashi; Nakamura, Ryoichiro; Kubo, Satoru; Otabe, Akira; Oobayashi, Yoko; Suzuki, Shoko; Yoshida, Mika; Yoshida, Mitsuya; Tatebe, Chiye; Sato, Kyoko; Akiyama, Hiroshi

    2016-01-01

    α-L-Aspartyl-D-phenylalanine methyl ester (L, D-APM) and α-D-aspartyl-L-phenylalanine methyl ester (D, L-APM) are diastereomers of aspartame (N-L-α-Aspartyl-L-phenylalanine-1-methyl ester, L, L-APM). The Joint FAO/WHO Expert Committee on Food Additives has set 0.04 wt% as the maximum permitted level of the sum of L, D-APM and D, L-APM in commercially available L, L-APM. In this study, we developed and validated a simple high-performance liquid chromatography (HPLC) method using an ODS column to determine L, D-APM and D, L-APM in L, L-APM. The limits of detection and quantification, respectively, of L, D-APM and D, L-APM were found to be 0.0012 wt% and 0.004 wt%. This method gave excellent accuracy, repeatability, and reproducibility in a recovery test performed on five different days. Moreover, the method was successfully applied to the determination of these diastereomers in commercial L, L-APM samples. Thus, the developed method is a simple, useful, and practical tool for determining L, D-APM and D, L-APM levels in L, L-APM.

  19. Development of an HPLC Method with an ODS Column to Determine Low Levels of Aspartame Diastereomers in Aspartame.

    PubMed

    Ohtsuki, Takashi; Nakamura, Ryoichiro; Kubo, Satoru; Otabe, Akira; Oobayashi, Yoko; Suzuki, Shoko; Yoshida, Mika; Yoshida, Mitsuya; Tatebe, Chiye; Sato, Kyoko; Akiyama, Hiroshi

    2016-01-01

    α-L-Aspartyl-D-phenylalanine methyl ester (L, D-APM) and α-D-aspartyl-L-phenylalanine methyl ester (D, L-APM) are diastereomers of aspartame (N-L-α-Aspartyl-L-phenylalanine-1-methyl ester, L, L-APM). The Joint FAO/WHO Expert Committee on Food Additives has set 0.04 wt% as the maximum permitted level of the sum of L, D-APM and D, L-APM in commercially available L, L-APM. In this study, we developed and validated a simple high-performance liquid chromatography (HPLC) method using an ODS column to determine L, D-APM and D, L-APM in L, L-APM. The limits of detection and quantification, respectively, of L, D-APM and D, L-APM were found to be 0.0012 wt% and 0.004 wt%. This method gave excellent accuracy, repeatability, and reproducibility in a recovery test performed on five different days. Moreover, the method was successfully applied to the determination of these diastereomers in commercial L, L-APM samples. Thus, the developed method is a simple, useful, and practical tool for determining L, D-APM and D, L-APM levels in L, L-APM. PMID:27015640

  20. Development of an HPLC Method with an ODS Column to Determine Low Levels of Aspartame Diastereomers in Aspartame

    PubMed Central

    Ohtsuki, Takashi; Nakamura, Ryoichiro; Kubo, Satoru; Otabe, Akira; Oobayashi, Yoko; Suzuki, Shoko; Yoshida, Mika; Yoshida, Mitsuya; Tatebe, Chiye; Sato, Kyoko; Akiyama, Hiroshi

    2016-01-01

    α-L-Aspartyl-D-phenylalanine methyl ester (L, D-APM) and α-D-aspartyl-L-phenylalanine methyl ester (D, L-APM) are diastereomers of aspartame (N-L-α-Aspartyl-L-phenylalanine-1-methyl ester, L, L-APM). The Joint FAO/WHO Expert Committee on Food Additives has set 0.04 wt% as the maximum permitted level of the sum of L, D-APM and D, L-APM in commercially available L, L-APM. In this study, we developed and validated a simple high-performance liquid chromatography (HPLC) method using an ODS column to determine L, D-APM and D, L-APM in L, L-APM. The limits of detection and quantification, respectively, of L, D-APM and D, L-APM were found to be 0.0012 wt% and 0.004 wt%. This method gave excellent accuracy, repeatability, and reproducibility in a recovery test performed on five different days. Moreover, the method was successfully applied to the determination of these diastereomers in commercial L, L-APM samples. Thus, the developed method is a simple, useful, and practical tool for determining L, D-APM and D, L-APM levels in L, L-APM. PMID:27015640

  1. Online pre-column derivatization with chromatographic separation to determine folic acid.

    PubMed

    Emara, Samy; Masujima, Tsutomu; Zarad, Walaa; Kamal, Maha; Ei-Bagary, Ramzia

    2013-07-01

    A simple, sensitive, and selective online pre-column derivatization high-performance liquid chromatographic method was developed and validated for the first time to determine trace levels of folic acid (FA). An oxidant cerium (IV) trihydroxyhydroperoxide packed reactor was used for pre-column oxidation and was combined by column switching with a C18 analytical column for sample enrichment and separation. The method was based on oxidative cleavage of FA into highly fluorescence products, 2-amino-4-hydroxypteridine-6-carboxaldehyde and the corresponding 2-amino-4-hydroxypteridine-6-carboxylic acid, during the flow of 0.04 M phosphate buffer (pH 3.5) containing the analyte through packed reactor at a flow rate of 0.2 mL/min and 40°C. The fluorescent products were enriched on the head of the analytical column for the final separation. The separation was performed at room temperature using a mobile phase consisting of phosphate buffer (0.04 M, pH 3.5) and acetonitrile (90:10, v/v). The eluents were monitored at emission and excitation wavelengths of 463 and 367 nm, respectively. The method showed excellent recovery, precision and accuracy with detection limits of 0.067 ng/mL from 500 µL of sample FA. The developed method was successfully applied to the determination of FA in pharmaceutical formulations and showed a recovery of 99.31% and a relative standard deviation of 1.72%.

  2. Recent advances in the preparation and application of monolithic capillary columns in separation science.

    PubMed

    Hong, Tingting; Yang, Xi; Xu, Yujing; Ji, Yibing

    2016-08-10

    Novel column technologies involving various materials and efficient reactions have been investigated for the fabrication of monolithic capillary columns in the field of analytical chemistry. In addition to the development of these miniaturized systems, a variety of microscale separation applications have achieved noteworthy results, providing a stepping stone for new types of chromatographic columns with improved efficiency and selectivity. Three novel strategies for the preparation of capillary monoliths, including ionic liquid-based approaches, nanoparticle-based approaches and "click chemistry", are highlighted in this review. Furthermore, we present the employment of state-of-the-art capillary monolithic stationary phases for enantioseparation, solid-phase microextraction, mixed-mode separation and immobilized enzyme reactors. The review concludes with recommendations for future studies and improvements in this field of research. PMID:27282747

  3. Recent advances in the preparation and application of monolithic capillary columns in separation science.

    PubMed

    Hong, Tingting; Yang, Xi; Xu, Yujing; Ji, Yibing

    2016-08-10

    Novel column technologies involving various materials and efficient reactions have been investigated for the fabrication of monolithic capillary columns in the field of analytical chemistry. In addition to the development of these miniaturized systems, a variety of microscale separation applications have achieved noteworthy results, providing a stepping stone for new types of chromatographic columns with improved efficiency and selectivity. Three novel strategies for the preparation of capillary monoliths, including ionic liquid-based approaches, nanoparticle-based approaches and "click chemistry", are highlighted in this review. Furthermore, we present the employment of state-of-the-art capillary monolithic stationary phases for enantioseparation, solid-phase microextraction, mixed-mode separation and immobilized enzyme reactors. The review concludes with recommendations for future studies and improvements in this field of research.

  4. Separation of the Carotenoid Bixin from Annatto Seeds Using Thin-Layer and Column Chromatography

    ERIC Educational Resources Information Center

    McCullagh, James V.; Ramos, Nicholas

    2008-01-01

    In this experiment the carotenoid bixin is isolated from annatto ("Bixa orellana") seeds using column chromatography. The experiment has several key advantages over previous pigment separation experiments. First, unlike other experiments significant quantities of the carotenoid (typically 20 to 25 mg) can be isolated from small quantities of plant…

  5. Analysis of the operation of a packed rectifying column in the process of binary mixture separation

    NASA Astrophysics Data System (ADS)

    Moshinskii, A. I.

    2013-09-01

    On the basis of a continuum mathematical model, the operation of a packed rectifying column in the stationary (continuous) and nonstationary (periodic) regimes in the process of binary mixture separation under certain conditions has been analyzed. We have considered examples of the realization of the above process, for which analytical expressions convenient for calculations have been obtained.

  6. Separation of {sup 32}P-postlabeled DNA adducts of polycyclic aromatic hydrocarbons and nitrated polycyclic aromatic hydrocarbons by HPLC

    SciTech Connect

    King, L.C.; Gallagher, J.E.; Lewtas, J.; George, M.

    1994-07-01

    The {sup 32}P-postlabeling assay, thin-layer chromatography, and reverse-phase high-pressure liquid chromatography (HPLC) were used to separate DNA adducts formed from 10 polycyclic aromatic hydrocarbons (PAHs) and 6 nitrated polycyclic aromatic hydrocarbons (NO{sub 2}-PAHs). The PAHs included benzo[j]fluoranthene, benzo[k]fluoranthene, indeno[1,2,3-cd]pyrene, benzo[a]pyrene, chrysene, 6-methylchrysene, 5-methylchrysene, and benz[a]anthracene. The NO{sub 2}-PAHs included 1-nitropyrene, 2-nitrofluoranthene, 3-nitrofluoranthene, 1,6-dinitropyrene, 1,3-dinitropyrene, and 1,8-dinitropyrene. Separation of seven of the major PAH-DNA adducts was achieved by an initial PAH HPLC gradient system. The major NO{sub 2}-PAH-DNA adducts were not all separated from each other using the initial PAH HPLC gradient but were clearly separated from the PAH-DNA adducts. A second NO{sub 2}-PAH HPLC gradient system was developed to separate NO{sub 2}-PAH-DNA adducts following one-dimensional TLC and HPLC analysis. HPLC profiles of NO{sub 2}-PAH-DNA adducts were compared using both adduct enhancement versions of the {sup 32}P-postlabeling assay to evaluate the use of this technique on HPLC to screen for the presence of NO{sub 2}-PAH-DNA adducts. To demonstrate the application of these separation methods to a complex mixture of DNA adducts, the chromatographic mobilities of the {sup 32}P-postlabeled DNA adduct standards (PAHs and NO{sub 2}-PAHs) were compared with those produced by a complex mixture of polycyclic organic matter (POM) extracted from diesel emission particles. The diesel-derived adducts did not elute with the identical retention time of any of the PAH or NO{sub 2}-PAH standards used in this study. HPLC analyses of the NO{sub 2}-PAH-derived adducts (butanol extracted) revealed the presence of multiple DNA adducts.

  7. Preparation and evaluation of packed capillary columns for the separation of nucleic acids by ion-pair reversed-phase high-performance liquid chromatography.

    PubMed

    Oberacher, H; Krajete, A; Parson, W; Huber, C G

    2000-09-29

    Oligonucleotides and double stranded DNA fragments were separated in 200 microm I.D. capillary columns packed with micropellicular, octadecylated, 2.1 microm poly(styrene-divinylbenzene) particles by ion-pair reversed-phase high-performance liquid chromatography (IP-RP-HPLC). Both the length and the diameter of the connecting capillaries (150 x 0.020 mm I.D.) as well as the detection volume (3 nl) had to be kept to a minimum in order to maintain the high efficiency of this chromatographic separation system with peak widths at half height in the range of a few seconds. Three different types of frits, namely sintered silica particles, sintered octadecylsilica particles, and monolithic poly(styrene-divinylbenzene) (PS-DVB) frits were evaluated with respect to their influence on chromatographic performance. Best performance for the separation of oligonucleotides and long DNA fragments was observed with the PS-DVB frits, whereas the short DNA fragments were optimally resolved in columns terminated by octadecylsilica frits. The maximum loading capacity of 60 x 0.20 mm I.D. columns ranged from 20 fmol (7.7 ng) for a 587 base pair DNA fragment to 500 fmol (2.4 ng) for a 16-mer oligonucleotide. Lower mass- and concentration detection limits in the low femtomol and low nanomol per liter range, respectively, make capillary IP-RP-HPLC with UV absorbance detection highly attractive for the separation and characterization of minute amounts of synthetic oligonucleotides, DNA restriction fragments, and short tandem repeat sequences amplified by polymerase chain reaction.

  8. Separation of bacteriochlorophyll homologues from green photosynthetic sulfur bacteria by reversed-phase HPLC.

    PubMed

    Borrego, C M; Garcia-Gil, L J

    1994-07-01

    A reversed-phase High Performance Liquid Cromatography (HPLC) method has been developed to accurately separate bacteriochlorophyllsc, d ande homologues in a reasonably short run time of 60 minutes. By using this method, two well-defined groups of bacteriochlorophyll homologue peaks can be discriminated. The first one consists of 4 peaks (min 24 to 30), which corresponds to the four main farnesyl homologues. The second peak subset is formed by a cluster of up to 10 minor peaks (min 33 to 40). These peaks can be related with series of several alcohol esters of the different chlorosome chlorophylls. The number of homologues was, however, quite variable depending on both, the bacteriochlorophyll and the bacterial species. The method hereby described, also provides a good separation of other photosynthetic pigments, either bacterial (Bacteriochlorophylla, chlorobactene, isorenieratene and okenone) or algal ones (Chlorophylla, Pheophytina and β-carotene). A preliminary screening of the homologue composition of several green photosynthetic bacterial species and isolates, has revealed different relative quantitative patterns. These differences seem to be related to physiological aspects rather than to taxonomic ones. The application of the method to the study of natural populations avoids the typical drawbacks on the pigment identification of overlapping eukaryotic and prokaryotic phototrophic microorganisms, giving further information about their physiological status.

  9. Pre-Column Derivatization HPLC Procedure for the Quantitation of Aluminium Chlorohydrate in Antiperspirant Creams Using Quercetin as Chromogenic Reagent.

    PubMed

    Kalogria, Eleni; Varvaresou, Athanasia; Papageorgiou, Spyridon; Protopapa, Evaggelia; Tsaknis, Ioannis; Matikas, Alexios; Panderi, Irene

    2014-01-01

    This article describes the development and validation of a selective high-performance liquid chromatography method that allows, after liquid-liquid extraction and pre-column derivatization reaction with quercetin, the quantification of aluminium chlorohydrate in antiperspirant creams. Chromatographic separation was achieved on an XTerra MS C18 analytical column (150 × 3.0 mm i.d., particle size 5 μm) using a mobile phase of acetonitrile:water (15:85, v/v) containing 0.08 % trifluoroacetic acid at a flow rate of 0.30 mL min(-1). Ultraviolet spectrophotometric detection at 415 nm was used. The assay was linear over a concentration range of 3.7-30.6 μg mL(-1) for aluminium with a limit of quantitation of 3.74 μg mL(-1). Quality control samples (4.4, 17.1 and 30.6 μg mL(-1)) in five replicates from five different runs of analysis demonstrated intra-assay precision (% coefficient of variation <3.8 %), inter-assay precision (% coefficient of variation <5.4 %) and an overall accuracy (% recovery) between 96 and 101 %. The method was used to quantify aluminium in antiperspirant creams containing 11.0, 13.0 and 16.0 % (w/w) aluminium chlorohydrate, respectively. PMID:25278619

  10. The Human HPLC Column

    ERIC Educational Resources Information Center

    Frantz, Kyle

    2007-01-01

    Initiatives in education reform emphasize inquiry-based active learning and real-world relevance to increase science literacy nationwide. Active teaching and learning approaches yield rapid intellectual development and may increase interest and motivation to learn science. Incorporating the topic of drug use with neuroscience, biology, psychology,…

  11. ENANTIOMER SEPARATION OF POLYCHLORINATED BIPHENYL ATROPISOMERS AND POLYCHLORINATED BIPHENYL RETENTION BEHAVIOR ON MODIFIED CYCLODEXTRIN CAPILLARY GAS CHROMATOGRAPHY COLUMNS

    EPA Science Inventory

    Seven commercially-available chiral capillary gas chromatography columns containing modified cyclodextrins were evaluated for their ability to separate enantiomers of the 19 stable chiral polychlorinated biphenyl (PCB) atropisomers, and for their ability to separate these enantio...

  12. Determination of the cis-trans isomerization barriers of L-alanyl-L-proline in aqueous solutions and at water/hydrophobic interfaces by on-line temperature-jump relaxation HPLC and dynamic on-column reaction HPLC.

    PubMed

    Shibukawa, Masami; Miyake, Ayaka; Eda, Sayaka; Saito, Shingo

    2015-09-15

    Proline cis-trans isomerization is known to play a key role in the rate-determining steps of protein folding. It is thus very important to understand the influence of environments, not only bulk solutions but also microenvironments such as interfaces, on the isomerization reaction of proline peptides. Here we present two HPLC methods for measurements of kinetic and equilibrium parameters for the isomerization reactions in bulk solutions and at liquid/solid interfaces. On-line temperature-jump relaxation HPLC (T-jump HPLC) allows the determination of forward and reverse rate constants of the isomerization in a bulk solution by monitoring the whole time course of conversion of pure isomers from both sides of the reaction, in contrast to other HPLC and capillary zone electrophoresis as well as spectrometric and calorimetric methods, which use a mixture of the isomers. We can then determine cis-trans isomerization barriers of the peptide at liquid/solid interfaces from the kinetic data obtained by dynamic on-column reaction HPLC and T-jump HPLC. We observed that the interconversion around the peptide bond for l-alanyl-l-proline (Ala-Pro) in water is accelerated at the surfaces of an alkyl-bonded silica and a poly(styrene-divinylbenzene) copolymer resin, and this is caused by a remarkable decrease in the enthalpy of activation. The molecular structures of the cis and trans forms of Ala-Pro estimated by quantum mechanics calculation reveal that an equilibrium shift toward the cis form as well as the rapid isomerization of Ala-Pro at the water/hydrophobic interfaces can be attributed to the lower polarity of the interfacial water at the surfaces of the hydrophobic materials compared to that of bulk water. PMID:26320351

  13. Determination of the cis-trans isomerization barriers of L-alanyl-L-proline in aqueous solutions and at water/hydrophobic interfaces by on-line temperature-jump relaxation HPLC and dynamic on-column reaction HPLC.

    PubMed

    Shibukawa, Masami; Miyake, Ayaka; Eda, Sayaka; Saito, Shingo

    2015-09-15

    Proline cis-trans isomerization is known to play a key role in the rate-determining steps of protein folding. It is thus very important to understand the influence of environments, not only bulk solutions but also microenvironments such as interfaces, on the isomerization reaction of proline peptides. Here we present two HPLC methods for measurements of kinetic and equilibrium parameters for the isomerization reactions in bulk solutions and at liquid/solid interfaces. On-line temperature-jump relaxation HPLC (T-jump HPLC) allows the determination of forward and reverse rate constants of the isomerization in a bulk solution by monitoring the whole time course of conversion of pure isomers from both sides of the reaction, in contrast to other HPLC and capillary zone electrophoresis as well as spectrometric and calorimetric methods, which use a mixture of the isomers. We can then determine cis-trans isomerization barriers of the peptide at liquid/solid interfaces from the kinetic data obtained by dynamic on-column reaction HPLC and T-jump HPLC. We observed that the interconversion around the peptide bond for l-alanyl-l-proline (Ala-Pro) in water is accelerated at the surfaces of an alkyl-bonded silica and a poly(styrene-divinylbenzene) copolymer resin, and this is caused by a remarkable decrease in the enthalpy of activation. The molecular structures of the cis and trans forms of Ala-Pro estimated by quantum mechanics calculation reveal that an equilibrium shift toward the cis form as well as the rapid isomerization of Ala-Pro at the water/hydrophobic interfaces can be attributed to the lower polarity of the interfacial water at the surfaces of the hydrophobic materials compared to that of bulk water.

  14. Molecularly imprinted SPE coupled with HPLC for the selective separation and enrichment of alkyl imidazolium ionic liquids in environmental water samples.

    PubMed

    Xia, Gao; Jing, Fan; Guifen, Zhu; Xiaolong, Wang; Jianji, Wang

    2013-10-01

    A novel 1-butyl-3-methylimidazolium chloride ionic liquid surface imprinted solid-phase sorbent was synthesized. The as-prepared material was characterized by SEM, Brunauer-Emmett-Teller surface area analysis and Fourier Transform IR measurements. Then its adsorption properties for alkyl imidazolium ionic liquids, including adsorption capacities, adsorption kinetics, and properties of selective separation and enrichment were studied in detail. It was shown that the ionic liquid surface imprinted polymer exhibited high selective recognition characteristics for the imidazolium chloride ionic liquids with short alkyl chains (C(n)mimCl, n = 2, 4, 6, 8) and the adsorption equilibrium was achieved within 25 min. Various parameters were optimized for the 1-butyl-3-methylimidazolium chloride ionic liquid surface imprinted polymer SPE column, such as flow rate, eluent solvent, selectivity, and reusability of the column. Then, the SPE column coupled with HPLC was used for the determination of alkyl imidazolium ionic liquids. Experimental results showed that the existence of their structural analogs and common concomitants in environmental matrices did not affect the enrichment of 1-butyl-3-methyl imidazolium chloride ionic liquid. The average recoveries of 1-butyl-3-methylimidazolium chloride ionic liquid in spiked water samples were in the range of 92.0-102.0% with the RSD lower than 5.8%.

  15. Separation of amino acids, peptides and corresponding Amadori compounds on a silica column at elevated temperature.

    PubMed

    Hao, Zhigang; Lu, Chih-Ying Joey; Xiao, Baiming; Weng, Naidong; Parker, Barry; Knapp, Michael; Ho, Chi-Tang

    2007-04-20

    Maillard reaction of glucose with amino acids and peptides has become a very important experimental model in the food flavor and pharmaceutical industries for better understanding the mechanism of food flavor generation and drug stability. Because of the amino acid and sugar functional groups present in their structures, most of the reaction components formed during the initial stages of Maillard reaction as well as the substrates are relatively polar. These compounds are poorly retained on a conventional reversed phase column. While polar stationary phases like HILIC column do provide better retention for these polar components, method selectivity could still be a challenge due to the structural similarity between these analytes. In this report, parameters such as pH, mobile phase composition and temperature were investigated using different brands of bare silica columns in order to separate glycine (G), diglycine (DG), triglycine (TG), and the corresponding Amadori compounds of glucose-glycine (GG), glucose-diglycine (GDG) and glucose-triglycine (GTG). An excellent separation for glycine, glycine peptides and their Amadori compounds was obtained on a bare silica column at an elevated temperature. PMID:17350634

  16. Novel separation method for highly sensitive speciation of cancerostatic platinum compounds by HPLC-ICP-MS.

    PubMed

    Hann, S; Stefánka, Zs; Lenz, K; Stingeder, G

    2005-01-01

    A high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS) method is presented for analysis of cisplatin, monoaquacisplatin, diaquacisplatin, carboplatin, and oxaliplatin in biological and environmental samples. Chromatographic separation was achieved on pentafluorophenylpropyl-functionalized silica gel. For cisplatin, carboplatin, and oxaliplatin limits of detection of 0.09, 0.10, and 0.15 microg L(-1), respectively, were calculated at m/z 194, using aqueous standard solutions. (3 microL injection volume). The method was utilized for model experiments studying the stability of carboplatin and oxaliplatin at different chloride concentrations simulating wastewater and surface water conditions. It was found that a high fraction of carboplatin is stable in ultrapure water and in solutions containing 1.5 mol L(-1) Cl-, whereas oxaliplatin degradation was increased by increasing the chloride concentration. In order to support the assessment of oxaliplatin eco-toxicology, the method was tested for speciation of patient urine. The urine sample contained more than 17 different reaction products, which demonstrates the extensive biotransformation of the compound. In a second step of the study the method was successfully evaluated for monitoring cancerostatic platinum compounds in hospital waste water.

  17. Comparison of high performance TLC and HPLC for separation and quantification of chlorogenic acid in green coffee bean extracts.

    PubMed

    Urakova, Irina N; Pozharitskaya, Olga N; Shikov, Alexander N; Kosman, Vera M; Makarov, Valery G

    2008-02-01

    Two chromatographic methods, high-performance TLC (HPTLC) and HPLC, were developed and used for separation and quantitative determination of chlorogenic acid in green coffee bean extracts. For HPTLC silica gel Kieselgel 60 F 254 plates with ethyl acetate/dichlormethane/formic acid/acetic acid/water (100:25:10:10:11, v/v/v/v/v) as mobile phase were used. Densitometric determination of chlorogenic acid by HPTLC was performed at 330 nm. A gradient RP HPLC method was carried out at 330 nm. All necessary validation tests for both methods were developed for their comparison. There were no statistically significant differences between HPLC and HPTLC for quantitative determination of chlorogenic acid according to the test of equality of the means.

  18. Separation analysis of macrolide antibiotics with good performance on a positively charged C18HCE column.

    PubMed

    Wei, Jie; Shen, Aijin; Yan, Jingyu; Jin, Gaowa; Yang, Bingcheng; Guo, Zhimou; Zhang, Feifang; Liang, Xinmiao

    2016-03-01

    The separation of basic macrolide antibiotics suffers from peak tailing and poor efficiency on traditional silica-based reversed-phase liquid chromatography columns. In this work, a C18HCE column with positively charged surface was applied to the separation of macrolides. Compared with an Acquity BEH C18 column, the C18HCE column exhibited superior performance in the aspect of peak shape and separation efficiency. The screening of mobile phase additives including formic acid, acetic acid and ammonium formate indicated that formic acid was preferable for providing symmetrical peak shapes. Moreover, the influence of formic acid content was investigated. Analysis speed and mass spectrometry compatibility were also taken into account when optimizing the separation conditions for liquid chromatography coupled with tandem mass spectrometry. The developed method was successfully utilized for the determination of macrolide residues in a honey sample. Azithromycin was chosen as the internal standard for the quantitation of spiramycin and tilmicosin, while roxithromycin was used for erythromycin, tylosin, clarithromycin, josamycin and acetylisovaleryltylosin. Good correlation coefficients (r(2) > 0.9938) for all macrolides were obtained. The intra-day and inter-day recoveries were 73.7-134.7% and 80.7-119.7% with relative standard deviations of 2.5-8.0% and 3.9-16.1%, respectively. Outstanding sensitivity with limits of quantitation (S/N ≥ 10) of 0.02-1 μg/kg and limits of detection (S/N ≥ 3) of 0.01-0.5 μg/kg were achieved. PMID:26782089

  19. [Establishment of the model for online monitoring of the column separation and purification process by near-infrared spectroscopy and determination of total ginsenosides in Folium Ginseng].

    PubMed

    Liu, Hua; Zhao, Xin; Qi, Tian; Qi, Yun-Peng; Fan, Guo-Rong

    2013-12-01

    A method was developed for online monitoring of the constituents of ginsenoside of Folium Ginseng in the column separation and purification process using near-infrared (NIR) spectroscopy technology. Determination method of ginsenoside Rg1, Re and Rb1 was developed by high performance liquid chromatography (HPLC). After collecting 40%-ethanol eluant, their NIR spectra were detected and the contents of Rg1, Re and Rb1 were determined by the above HPLC method. The quantitative analysis models of the above three compounds and the total ginsenosides were established using partial least squares (PLS). During modeling, coefficient of determination (R2) and root mean square errors of cross-validation (RMSECV) were regarded as the indexes to select optimal wave numbers and preprocessing methods. The optimal wave numbers of ginsenoside Rg1, Re, Rb1 and total ginsenosides were all in the range of 12 000. 8 approximately 7499.8 cm-1; R2 were 0.9887, 0. 9603, 0.9905 and 0.9701, respectively; RMSECV were 0.0597, 0.0722, 0.00488 and 0.0755, respectively. A lot of samples, collected during the column separation and purification process of Folium Ginseng extract, were used to validate the predicttion effect of quantitative analysis model of total ginsenosides. As a result, the correlation coefficient of NIR predicted value and HPLC value of total ginsenosides was 0.9928 and the mean prediction recovery was 100.52%, which indicated that the prediction effect of the developed model was satisfactory. This method was proved to be fast, convenient and precise. It can be used for assaying and quality control of total ginsenosides in manufacture.

  20. [Establishment of the model for online monitoring of the column separation and purification process by near-infrared spectroscopy and determination of total ginsenosides in Folium Ginseng].

    PubMed

    Liu, Hua; Zhao, Xin; Qi, Tian; Qi, Yun-Peng; Fan, Guo-Rong

    2013-12-01

    A method was developed for online monitoring of the constituents of ginsenoside of Folium Ginseng in the column separation and purification process using near-infrared (NIR) spectroscopy technology. Determination method of ginsenoside Rg1, Re and Rb1 was developed by high performance liquid chromatography (HPLC). After collecting 40%-ethanol eluant, their NIR spectra were detected and the contents of Rg1, Re and Rb1 were determined by the above HPLC method. The quantitative analysis models of the above three compounds and the total ginsenosides were established using partial least squares (PLS). During modeling, coefficient of determination (R2) and root mean square errors of cross-validation (RMSECV) were regarded as the indexes to select optimal wave numbers and preprocessing methods. The optimal wave numbers of ginsenoside Rg1, Re, Rb1 and total ginsenosides were all in the range of 12 000. 8 approximately 7499.8 cm-1; R2 were 0.9887, 0. 9603, 0.9905 and 0.9701, respectively; RMSECV were 0.0597, 0.0722, 0.00488 and 0.0755, respectively. A lot of samples, collected during the column separation and purification process of Folium Ginseng extract, were used to validate the predicttion effect of quantitative analysis model of total ginsenosides. As a result, the correlation coefficient of NIR predicted value and HPLC value of total ginsenosides was 0.9928 and the mean prediction recovery was 100.52%, which indicated that the prediction effect of the developed model was satisfactory. This method was proved to be fast, convenient and precise. It can be used for assaying and quality control of total ginsenosides in manufacture. PMID:24611375

  1. Evaluation of two commercial capillary columns for the enantioselective gas chromatographic separation of organophosphorus pesticides.

    PubMed

    Fidalgo-Used, Natalia; Blanco-González, Elisa; Sanz-Medel, Alfredo

    2006-12-15

    The separation of the enantiomers of 13 organophosphorus pesticides (OPPs) has been investigated by gas chromatography (GC) with flame ionisation detection (FID) using two different commercially available chiral columns, Chirasil-Val (l-valine-tert-butylamide) and CP-Chirasil-Dex CB (heptakis (2,3,6-tri-O-metil)-beta-cyclodextrin). Using the Chirasil-Val column no chiral resolution was obtained for the OPPs investigated under any tested experimental condition. The use of the CP-Chirasil-Dex CB stationary phase enabled good individual enantiomeric separation of two OPPs, ruelene and trichlorfon and partial separation of naled, chloretoxyphos, isophenphos and metamidophos. Also, the obtained chromatographic results showed that Chirasil-Dex could resolve enantiomers through the combination of different mechanism (e.g. formation of inclusion complexes and/or interactions outside the cyclodextrin cavity). Under optimised conditions, precision, linearity range and detection limits were evaluated for the enantiomers of ruelene and trichlorfon using CP-Chirasil-Dex CB column and electron capture detection (ECD). By using the GC-ECD method the enantiomers of these OPPs could be satisfactorily detected at very low concentration levels. The detection limits observed were 1.5ngmL(-1) and 11.5ngmL(-1) for the enantiomers of trichlorfon and ruelene, respectively. PMID:18970881

  2. Separation of proteins by cation-exchange sequential injection chromatography using a polymeric monolithic column.

    PubMed

    Masini, Jorge Cesar

    2016-02-01

    Since sequential injection chromatography (SIC) emerged in 2003, it has been used for separation of small molecules in diverse samples, but separations of high molar mass compounds such as proteins have not yet been described. In the present work a poly(glycidyl methacrylate-co-ethylene dimethacrylate) (GMA-co-EDMA) monolithic column was prepared by free radical polymerization inside a 2.1-mm-i.d. activated fused silica-lined stainless steel tubing and modified with iminodiacetic acid (IDA). The column was prepared from a mixture of 24% GMA, 16% EDMA, 20% cyclohexanol, and 40% 1-dodecanol (all% as w/w) containing 1% of azobisisobutyronitrile (AIBN) (in relation to monomers). Polymerization was done at 60 °C for 24 h. The polymer was modified with 1.0 M IDA (in 2 M Na2CO3, pH 10.5) at 80 °C for 16 h. Separation of myoglobin, ribonuclease A, cytochrome C, and lysozyme was achieved at pH 7.0 (20 mM KH2PO4/K2HPO4) using a salt gradient (NaCl). Myoglobin was not retained, and the other proteins were separated by a gradient of NaCl created inside the holding coil (4 m of 0.8-mm-i.d. PTFE tubing) by sequential aspiration of 750 and 700 μL of 0.2 and 0.1 M NaCl, respectively. As the flow was reversed toward the column (5 μL s(-1)) the interdispersion of these solutions created a reproducible gradient which separated the proteins in 10 min, with the following order of retention: ribonuclease A < cytochrome C < lysozyme. The elution order was consistent with a cation-exchange mechanism as the retention increased with the isoelectric points.

  3. [Simultaneous separation and detection of principal component isomer and related substances of raw material drug of ammonium glycyrrhizinate by RP-HPLC and structure confirmation].

    PubMed

    Zhao, Yan-Yan; Liu, Li-Yan; Han, Yuan-Yuan; Li, Yue-Qiu; Wang, Yan; Shi, Min-Jian

    2013-08-01

    A simple, fast and sensitive analytical method for the simultaneous separation and detection of 18alpha-glycyrrhizinic acid, 18beta-glycyrrhizinic acid, related substance A and related substance B by RP-HPLC and drug quality standard was established. The structures of principal component isomer and related substances of raw material drug of ammonium glycyrrhizinate have been confirmed. Reference European Pharmacopoeia EP7.0 version, British Pharmacopoeia 2012 version, National Drug Standards of China (WS 1-XG-2002), domestic and international interrelated literature were referred to select the composition of mobile phase. The experimental parameters including salt concentration, pH, addition quantities of organic solvent, column temperature and flow rate were optimized. Finally, the assay was conducted on a Durashell-C18 column (250 mm x 4.6 mm, 5 microm) with 0.01 mol x mL(-1) ammonium perchlorate (add ammonia to adjust the pH value to 8.2) -methanol (48 : 52) as mobile phase at the flow rate of 0.8 mL x min(-1), and the detection wavelength was set at 254 nm. The column temperature was 50 degrees C and the injection volume was 10 microL. The MS, NMR, UV and RP-HPLC were used to confirm the structures of principal component isomer and related substances of raw material drug of ammonium glycyrrhizinate. Under the optimized separation conditions, the calibration curves of 18 alpha-glycyrrhizinic acid, 18beta-glycyrrhizinic acid, related substance A and related substance B showed good linearity within the concentration of 0.50-100 microg x mL(-1) (r = 0.999 9). The detection limits for 18alpha-glycyrrhizinic acid, 18beta-glycyrrhizinic acid, related substance A and related substance B were 0.15, 0.10, 0.10, 0.15 microg x mL(-1) respectively. The method is sensitive, reproducible and the results are accurate and reliable. It can be used for chiral resolution of 18alpha-glycyrrhizinic acid, 18Pbeta-glycyrrhizinic acid, and detection content of principal component and

  4. Preparative isolation of alkaloids from Dactylicapnos scandens using pH-zone-refining counter-current chromatography by changing the length of the separation column.

    PubMed

    Wang, Xiao; Dong, Hongjing; Yang, Bin; Liu, Dahui; Duan, Wenjuan; Huang, Luqi

    2011-12-01

    pH-Zone-refining counter-current chromatography was successfully applied for the preparative separation of alkaloids from Dactylicapnos scandens. The two-phase solvent system was composed of petroleum ether-ethyl acetate-methanol-water (3:7:1:9, v/v), where 20 mM of triethylamine (TEA) was added to the upper phase as a retainer and 5 mM of hydrochloric acid (HCl) to the aqueous phase as an eluter. In this experiment, the apparatus with an adjustable length of the separation column was used for the separation of alkaloids from D. scandens and the resolution of the compounds can be remarkably improved by increasing the length of the separation column. As a result, 70 mg protopin, 30 mg (+) corydine, 120 mg (+) isocorydine and 40 mg (+) glaucine were obtained from 1.0 g of the crude extracts and each with 99.2%, 96.5%, 99.3%, 99.5% purity as determined by HPLC. The chemical structures of these compounds were confirmed by positive ESI-MS and (1)H NMR. PMID:22056347

  5. Preparative isolation of alkaloids from Dactylicapnos scandens using pH-zone-refining counter-current chromatography by changing the length of the separation column.

    PubMed

    Wang, Xiao; Dong, Hongjing; Yang, Bin; Liu, Dahui; Duan, Wenjuan; Huang, Luqi

    2011-12-01

    pH-Zone-refining counter-current chromatography was successfully applied for the preparative separation of alkaloids from Dactylicapnos scandens. The two-phase solvent system was composed of petroleum ether-ethyl acetate-methanol-water (3:7:1:9, v/v), where 20 mM of triethylamine (TEA) was added to the upper phase as a retainer and 5 mM of hydrochloric acid (HCl) to the aqueous phase as an eluter. In this experiment, the apparatus with an adjustable length of the separation column was used for the separation of alkaloids from D. scandens and the resolution of the compounds can be remarkably improved by increasing the length of the separation column. As a result, 70 mg protopin, 30 mg (+) corydine, 120 mg (+) isocorydine and 40 mg (+) glaucine were obtained from 1.0 g of the crude extracts and each with 99.2%, 96.5%, 99.3%, 99.5% purity as determined by HPLC. The chemical structures of these compounds were confirmed by positive ESI-MS and (1)H NMR.

  6. The fabrication of monolithic capillary column based on poly (bisphenol A epoxy vinyl ester resin-co-ethylene glycol dimethacrylate) and its applications for the separation of small molecules in high performance liquid chromatography.

    PubMed

    Niu, Wenjing; Wang, Lijuan; Bai, Ligai; Yang, Gengliang

    2013-07-01

    A new polymeric monolith was synthesized in fused-silica capillary by in situ polymerization technique. In the polymerization, bisphenol A epoxy vinyl ester resin (VER) was used as the functional monomer, ethylene glycol dimethacrylate (EDMA) as the crosslinking monomer, 1,4-butanediol, 1-propanol and water as the co-porogens, and azobisisobutyronitrile (AIBN) as the initiator. The conditions of polymerization have been optimized. Morphology of the prepared poly (VER-co-EDMA) monolith was investigated by the scanning electron microscopy (SEM); pore properties were assayed by mercury porosimetry and nitrogen adsorption. The optimized poly (VER-co-EDMA) monolith showed a uniform structure, good permeability and mechanical stability. Then, the column was used as the stationary phase of high performance liquid chromatography (HPLC) to separate the mixture of benzene derivatives. The best column efficiency achieved for phenol was 235790 theoretical plates per meter. Baseline separations of benzene derivatives and halogenated benzene compounds under optimized isocratic mode conditions were achieved with high column efficiency. The column showed good reproducibility: the relative standard deviation (RSD) values based on the retention times (n=3) for run-to-run, column-to-column and batch-to-batch were less than 0.98, 1.68, 5.48%, respectively. Compared with poly (BMA-co-EDMA) monolithic column, the proposed monolith exhibited more efficiency in the separation of small molecules. PMID:23726080

  7. Detection of roasted and ground coffee adulteration by HPLC and by amperometric and by post-column derivatization UV-Vis detection.

    PubMed

    Domingues, Diego S; Pauli, Elis D; de Abreu, Julia E M; Massura, Francys W; Cristiano, Valderi; Santos, Maria J; Nixdorf, Suzana L

    2014-03-01

    Coffee is one of the most consumed beverages in the world. Due to its commercial importance, the detection of impurities and foreign matters has been a constant concern in fraud verification, especially because it is difficult to percept adulterations with the naked eye in samples of roasted and ground coffee. In Brazil, the most common additions are roasted materials, such as husks, sticks, corn, wheat middling, soybean, and more recently - acai palm seeds. The performance and correlation of two chromatographic methods, HPLC-HPAEC-PAD and post-column derivatization HPLC-UV-Vis, were compared for carbohydrate analysis in coffee samples. To verify the correlation between the two methods, the principal component analysis for the same mix of triticale and acai seeds in different proportions with coffee was employed. The performance for detecting adulterations in roasted and ground coffee of the two methods was compared.

  8. Separation of uremic toxins from urine with resorcinarene-based ion chromatography columns.

    PubMed

    Panahi, Tayyebeh; Weaver, Douglas J; Lamb, John D; Harrison, Roger G

    2015-01-01

    People with chronic kidney disease suffer from uremic toxins which accumulate in their bodies. Detection and quantification of uremic toxins help diagnose kidney problems and start patient care. The aim of this research was to seek a new method to assist this diagnosis by trace level detection and separation of guanidine containing uremic toxins in water and urine. To detect and quantify the uremic toxins, new stationary phases for ion chromatography (IC) columns based on glutamic acid functionalized resorcinarenes bound to divinylbenzene macroporous resin were prepared. The new column packing material afforded separation of the five compounds: guanidinoacetic acid, guanidine, methylguanidine, creatinine, and guanidinobenzoic acid in 30min. Peak resolutions ranged from 7.6 to 1.3. Gradient elutions at ambient temperature with methanesulfonic acid (MSA) solution as eluent resulted in detection levels in water from 10 to 47ppb and in synthetic urine from 28 to 180ppb. Limits of quantification for the analytes using pulsed amperometric detection were 30-160ppb in water and 93-590ppb in urine. Trace levels of creatinine (1ppm) were detected in the urine of a healthy individual using the columns.

  9. Azeotropic distillation in a middle vessel batch column. 1: Model formulation and linear separation boundaries

    SciTech Connect

    Cheong, W.; Barton, P.I.

    1999-04-01

    A mathematical model for the middle vessel batch distillation column (MVC) is developed using the concept of warped time analysis and used to study the qualitative dynamics of the MVC when it is used to separate multicomponent azeotropic mixtures. A limiting analysis is then developed for a MVC with an infinite number of trays, operated under infinite reflux/reboil ratios, under the assumption of linear separation boundaries. It is determined that, under limiting conditions, the distillate product drawn from the MVC is given by the {alpha} limit set of the MVC still pot composition, while the bottoms product drawn from the MVC is given by the {omega} limit set of the MVC still pot composition. The net product composition is determined by taking a convex combination of the two products. The notions of steering the still pot composition, the vector cone of possible motion for the still pot composition, and the equivalency of the MVC to the combined operation of a batch rectifier and a stripper are also explored. The definition of batch distillation regions for the MVC operated at a given value of the middle vessel parameter {lambda}, and the bifurcation of these regions with the variation of {lambda}, are investigated. Lastly, a mathematical model incorporating the concept of warped time is developed for a multivessel column. The MVC can be viewed as a specific case of the multivessel column.

  10. Infrared small target and background separation via column-wise weighted robust principal component analysis

    NASA Astrophysics Data System (ADS)

    Dai, Yimian; Wu, Yiquan; Song, Yu

    2016-07-01

    When facing extremely complex infrared background, due to the defect of l1 norm based sparsity measure, the state-of-the-art infrared patch-image (IPI) model would be in a dilemma where either the dim targets are over-shrinked in the separation or the strong cloud edges remains in the target image. In order to suppress the strong edges while preserving the dim targets, a weighted infrared patch-image (WIPI) model is proposed, incorporating structural prior information into the process of infrared small target and background separation. Instead of adopting a global weight, we allocate adaptive weight to each column of the target patch-image according to its patch structure. Then the proposed WIPI model is converted to a column-wise weighted robust principal component analysis (CWRPCA) problem. In addition, a target unlikelihood coefficient is designed based on the steering kernel, serving as the adaptive weight for each column. Finally, in order to solve the CWPRCA problem, a solution algorithm is developed based on Alternating Direction Method (ADM). Detailed experiment results demonstrate that the proposed method has a significant improvement over the other nine classical or state-of-the-art methods in terms of subjective visual quality, quantitative evaluation indexes and convergence rate.

  11. 'Click Chemistry' in the preparation of porous polymer-basedparticulate stationary phases for mu-HPLC separation of peptides andproteins

    SciTech Connect

    Slater, Michael; Snauko, Marian; Svec, Frantisek; Frechet, JeanM.J.

    2006-01-02

    With the use of the copper(I)-catalyzed (3 + 2) azide-alkynecycloaddition, an element of "click chemistry," stationary phasescarrying long alkyl chains or soybean trypsin inhibitor have beenprepared for use in HPLC separations in the reversed-phase and affinitymodes, respectively. The ligands were attached via a triazole ring tosize monodisperse porous beads containing either alkyne or azide pendantfunctionalities. Alkyne-containing beads prepared by directcopolymerization of propargyl acrylate with ethylene dimethacrylate wereallowed to react with azidooctadecane to give a reversed-phase sorbent.Azide-functionalized beads were prepared by chemical modification ofglycidyl methacrylate particles. Subsequent reaction with a terminalaliphatic alkyne produced a reversed-phase sorbent similar to thatobtained from the alkyne beads. Soybean trypsin inhibitor wasfunctionalized with N-(4-pentynoyloxy) succinimide to carry alkyne groupsand then allowed to react with the azide-containing beads to produce anaffinity sorbent for trypsin. The performance of these stationary phaseswas demonstrated with the HPLC separations of a variety of peptides andproteins.

  12. Rapid tea catechins and caffeine determination by HPLC using microwave-assisted extraction and silica monolithic column.

    PubMed

    Rahim, A A; Nofrizal, S; Saad, Bahruddin

    2014-03-15

    A rapid reversed-phase high performance liquid chromatographic method using a monolithic column for the determination of eight catechin monomers and caffeine was developed. Using a mobile phase of water:acetonitrile:methanol (83:6:11) at a flow rate of 1.4 mL min(-1), the catechins and caffeine were isocratically separated in about 7 min. The limits of detection and quantification were in the range of 0.11-0.29 and 0.33-0.87 mg L(-1), respectively. Satisfactory recoveries were obtained (94.2-105.2 ± 1.8%) for all samples when spiked at three concentrations (5, 40 and 70 mg L(-1)). In combination with microwave-assisted extraction (MAE), the method was applied to the determination of the catechins and caffeine in eleven tea samples (6 green, 3 black and 2 oolong teas). Relatively high levels of caffeine were found in black tea, but higher levels of the catechins, especially epigallocatechin gallate (EGCG) were found in green teas.

  13. Simple automated liquid chromatographic system for splitless nano column gradient separations.

    PubMed

    Sesták, Jozef; Duša, Filip; Moravcová, Dana; Kahle, Vladislav

    2013-02-01

    A simple splitless gradient liquid chromatographic system for micro and nano column separations has been assembled and tested. It consists of an OEM programmable syringe pump equipped with a glass microsyringe and ten-port selector valve. Gradient of mobile phase was created in the syringe barrel due to turbulent mixing. Capability of suggested system to create various gradient profiles was verified using 50-μl, 100-μl, and 250-μl glass syringes. Acetone, thiourea, and uracil were tested as gradient markers and finally, uracil was proved to be an excellent way of water-acetonitrile gradient tracing. It was found that up to 80% of the total syringe volume is available as a linear gradient section. In context to micro and nano column chromatography, the best results were obtained using the 100-μl syringe. Separations were performed on the capillary monolithic column Chromolith CapRod RP-18e (150mm×0.1mm) and system performance was evaluated using a test mixture of six alkylphenones as well as tryptic digest of bovine serum albumin. Results proved that suggested system is able to create uniform gradients with high repeatability of retention times of test solutes (RSD<0.3%). Repeatability of injection of sample volumes in the range of 0.1-3μl was evaluated using on-column preconcentration technique which means that sample was diluted in mobile phase of low eluting strength. Repeatability of the peak areas was measured and statistically evaluated (RSD<5%).

  14. Suspension column for recovery and separation of substances using ultrasound-assisted retention of bead sorbents.

    PubMed

    Spivakov, Boris Ya; Shkinev, Valeriy M; Danilova, Tatiana V; Knyazkov, Nikolai N; Kurochkin, Vladimir E; Karandashev, Vasiliy K

    2012-12-15

    A novel approach to sorption recovery and separation of different substances is proposed which is based on the use of suspended bead sorbents instead of conventional packed beds of such sorbents. This makes it possible to employ small-sized beads which are trapped in a low-pressure column due to ultrasound-assisted retention, without any frits to hold the sorption material. A flow system including a separation mini-column, named herein a suspension column, has been developed and tested by the studies of solid phase extraction (SPE) of trace metals from bi-distilled water and sea water using a 150-μL column with a silica-based sorbent containing iminodiacetic groups (DIAPAK IDA) and having a grain size of 6 μm. The adsorption properties of DIAPAK IDA suspension (9.5mg) were evaluated through adsorption/desorption experiments, where the effect of solution pH and eluent on the SPE of trace metals were examined by ICP-MS or ICP-AES measurements. When sample solution was adjusted to pH 8.0 and 1 mol L(-1) nitric acid was used as eluent, very good recoveries of more than 90% were obtained for a number of elements in a single-step extraction. To demonstrate the versatility of the approach proposed and to show another advantage of ultrasonic field (acceleration of sorbate/sorbent interaction), a similar system was used for heterogeneous immunoassays of some antigens in ultrasonic field using agarose sorbents modified by corresponding antibodies. It has been shown that immunoglobulins, chlamidia, and brucellos bacteria can be quantitatively adsorbed on 15-μm sorbent (15 particles in 50 μL) and directly determined in a 50-μL mini-chamber using fluorescence detection. PMID:23182579

  15. Open tubular capillary column for the separation of cytochrome C tryptic digest in capillary electrochromatography.

    PubMed

    Ali, Faiz; Cheong, Won Jo

    2015-10-01

    A silica capillary of 50 μm internal diameter and 500 mm length (416 mm effective length) was chemically modified with 4-(trifluoromethoxy) phenyl isocyanate in the presence of dibutyl tin dichloride as catalyst. Sodium diethyl dithiocarbamate was reacted with the terminal halogen of the bound ligand to incorporate the initiator moiety, and in situ polymerization was performed using a monomer mixture of styrene, N-phenylacrylamide, and methacrylic acid. The resultant open tubular capillary column immobilized with the copolymer layer was used for the separation of tryptic digest of cytochrome C in capillary electrochromatography. The sample was well eluted and separated into many components. The elution patterns of tryptic digest of cytochrome C were studied with respect to pH and water content in the mobile phase. This preliminary study demonstrates that open tubular capillary electrochromatography columns with a modified copolymer layer composed of proper nonpolar and polar units fabricated by reversible addition-fragmentation transfer polymerization can be useful as separation media for proteomic analysis. PMID:26289407

  16. Robust Feedback Linearization Applied to a Separation Column for {sup 13}C

    SciTech Connect

    Dulf, Eva-Henrietta; Pop, Cristina-Ioana; Festila, Clement; Dulf, Francisc

    2009-03-05

    In the present developing plan to apply the cryogenic technology for the production of the {sup 13}C, an efficient and safe operation is a strong reason to conceive and to apply a modern computer based control strategy. The authors are concerned with the problem of developing effective and readily implemental techniques for modelling and control of the isotope separation plant. These columns are characterized by complex nonlinearities, with large time-delays. Furthermore, are subject to external disturbances, which are difficult to model. The present paper presents two models of the plant: a nonlinear model and a linear system obtained by robust feedback linearization.

  17. Comparison of hypercrosslinked polystyrene columns for the separation of nitrogen group-types in petroleum using High Performance Liquid Chromatography.

    PubMed

    Oro, Nicole E; Lucy, Charles A

    2010-10-01

    High performance liquid chromatography in a quasi-normal phase mode (QNP) is used to separate the nitrogen group-types (pyrrole and pyridine) that are found in petroleum. A new type of stationary phase, hypercrosslinked polystyrene, is used to achieve this separation. Three different hypercrosslinked polystyrene stationary phases are compared under quasi-normal phase mode; a commercial 5-HGN packing, and two hypercrosslinked phases on silica particles. The utility of the columns for petroleum-based separations was explored with the use of 21 analytical standards. Partial elucidation of adsorption retention mechanisms for the columns are shown, as well as a comparison of retention characteristics for the three columns. The silica particle column derived with toluene (HC-Tol) was found to have the best selectivity for nitrogen group-types and polycyclic aromatic hydrocarbons (PAHs), attaining a separation under gradient conditions in less than 30 min.

  18. High resolution capillary column development for selective separations in gas chromatography

    SciTech Connect

    Przybyciel, M.

    1985-01-01

    A review of techniques for the preparation of high resolution capillary columns for gas chromatography is presented. Surface roughing, surface deactivation, stationary phase coating, and stationary phase crosslinking are discussed. Criteria for the selection of GC stationary phases and procedures for column evaluation are presented. A method is proposed for the isolation and determination of crude oil contamination in tropical plants and sediments. The method uses Florisil (TM) chromatography for the simultaneous clean-up and fractionation of aliphatic and aromatic hydrocarbons. Crosslinked SE-54 fused silica capillary columns prepared in our laboratory were employed for all GC separations. Mass spectrometry was used to help locate and identify specific oil components despite the intense background of the chromatogram. Crude oil components were identified in extracts of mangrove plant samples collected from the Peck Slip oil spill site at Media Munda, Puerto Rico. Crude oil components were also identified in sediment samples from controlled oil spill of Prudhoe Bay oil at Laguna de Chiriqui, Panama.

  19. Analysis of sterigmatocystin in cereals, animal feed, seeds, beer and cheese by immunoaffinity column clean-up and HPLC and LC-MS/MS quantification.

    PubMed

    Marley, Elaine; Brown, Phyllis; Mackie, Jennifer; Donnelly, Carol; Wilcox, Joyce; Pietri, Amedeo; Macdonald, Susan

    2015-01-01

    A method is reported for the analysis of sterigmatocystin in various food and feed matrices using a commercial sterigmatocystin immunoaffinity column (IAC) for sample clean-up prior to HPLC analysis by UV with mass spectrometric detection (LC-MS/MS). Cereals (wheat, oats, rye, maize and rice), sunflower seeds and animal feed were spiked with sterigmatocystin at levels from 0.75 to 50 µg kg(-1) to establish method performance. Using acetonitrile/water extraction followed by IAC clean-up, and analysis by HPLC with detection at 325 nm, recoveries ranged from 68% to 106%, with repeatability from 4.2% to 17.5%. The limit of quantification with UV detection in these matrices was 1.5 µg kg(-1). For the analysis of beer and cheese the sample preparation prior to IAC clean-up was changed to accommodate the different properties of the matrix, prior to analysis by LC-MS/MS. For beer and cheese spiked at 5.0 µg kg(-1) the recoveries were 94% and 104%, and precision (RSDs) were 1.9% and 2.9% respectively. The limits of quantification by LC-MS/MS in beer and cheese were 0.02 and 0.6 µg kg(-1) respectively. The sterigmatocystin IAC was demonstrated to provide an efficient clean-up of various matrices to enable this mycotoxin to be determined by either HPLC with UV detection or LC-MS/MS.

  20. [Determination of dl-alpha-tocopherol acetate and dl-alpha-tocopherol in foods by HPLC using post-column photochemical reaction].

    PubMed

    Terada, Hisaya; Tamura, Yukio

    2004-12-01

    A reliable analytical method for the simultaneous determination of dl-alpha-tocopherol acetate and dl-alpha-tocopherol in foods was established by HPLC using post-column photochemical reaction with UV and fluorescence detection. For low-fat food such as fruit juice and vegetable sauce, the tocopherols were extracted with methanol containing 0.1% ascorbic acid and the extract solution was injected into the HPLC. For fatty foods such as butter and margarine, the tocopherols were extracted with a mixed solvent of acetonitrile-2-propanol (9:1) containing ascorbic acid. The extract was cleaned up using a Sep Pak plus C18 cartridge and the eluent from the cartridge was injected into the HPLC. The peaks corresponding to tocopherols on the chromatogram were confirmed by comparing their UV spectra with those of the standard mixture at lamp-on and lamp-off of the photochemical reactor. The recoveries of tocopherols from low-fat foods (orange juice and barbecue sauce) fortified at levels of 10 and 100 microg/kg each were 88.3 to 105.8% (RSD 0.5 to 6.0%) and those from the fatty foods (peanut butter and margarine) fortified at 100 microg/kg each were 57.1 to 88.3% (RSD 3.0 to 6.4%). The determination limits corresponded to 10 microg/kg of the tocopherols in the low-fat foods and 20 microg/kg in the fatty foods.

  1. Analysis of sterigmatocystin in cereals, animal feed, seeds, beer and cheese by immunoaffinity column clean-up and HPLC and LC-MS/MS quantification.

    PubMed

    Marley, Elaine; Brown, Phyllis; Mackie, Jennifer; Donnelly, Carol; Wilcox, Joyce; Pietri, Amedeo; Macdonald, Susan

    2015-01-01

    A method is reported for the analysis of sterigmatocystin in various food and feed matrices using a commercial sterigmatocystin immunoaffinity column (IAC) for sample clean-up prior to HPLC analysis by UV with mass spectrometric detection (LC-MS/MS). Cereals (wheat, oats, rye, maize and rice), sunflower seeds and animal feed were spiked with sterigmatocystin at levels from 0.75 to 50 µg kg(-1) to establish method performance. Using acetonitrile/water extraction followed by IAC clean-up, and analysis by HPLC with detection at 325 nm, recoveries ranged from 68% to 106%, with repeatability from 4.2% to 17.5%. The limit of quantification with UV detection in these matrices was 1.5 µg kg(-1). For the analysis of beer and cheese the sample preparation prior to IAC clean-up was changed to accommodate the different properties of the matrix, prior to analysis by LC-MS/MS. For beer and cheese spiked at 5.0 µg kg(-1) the recoveries were 94% and 104%, and precision (RSDs) were 1.9% and 2.9% respectively. The limits of quantification by LC-MS/MS in beer and cheese were 0.02 and 0.6 µg kg(-1) respectively. The sterigmatocystin IAC was demonstrated to provide an efficient clean-up of various matrices to enable this mycotoxin to be determined by either HPLC with UV detection or LC-MS/MS. PMID:26515281

  2. Preparative separation and purification of rosmarinic acid from perilla seed meal via combined column chromatography.

    PubMed

    Tang, Weizhuo; Sun, Baoshan; Zhao, Yuqing

    2014-02-01

    In this study, the preparative separation and purification of rosmarinic acid (RA) from perilla seed meal (PSM), which is a by-product of edible oil production, was achieved using combined column chromatography over macroporous and polyamide resins. To optimize the RA enrichment process, the performance and separation characteristics of nine selected macroporous resins with different chemical and physical properties were investigated. SP825 resin was the most effective: the content of RA increased from 0.27% in the original extract to 16.58% in the 50% ethanol fraction (a 61.4-fold increase). During further purification treatment on polyamide resin, 90.23% pure RA could be obtained in the 70% ethanol fraction. RA with a higher purity (>95%) could also be easily obtained using one crystallization operation. The proposed method is simple, easily operated, cost-effective, and environmentally friendly and is suitable for both large-scale RA production and waste management.

  3. Optimization of an improved single-column chromatographic process for the separation of enantiomers.

    PubMed

    Kazi, Monzure-Khoda; Medi, Bijan; Amanullah, Mohammad

    2012-03-30

    This work addresses optimization of an improved single-column chromatographic (ISCC) process for the separation of guaifenesin enantiomers. Conventional feed injection and fraction collection systems have been replaced with customized components facilitating simultaneous separation and online monitoring with the ultimate objective of application of an optimizing controller. Injection volume, cycle time, desorbent flow rate, feed concentration, and three cut intervals are considered as decision variables. A multi-objective optimization technique based on genetic algorithm (GA) is adopted to achieve maximum productivity and minimum desorbent requirement in the region constrained by product specifications and hardware limitations. The optimization results along with the contribution of decision variables are discussed using Pareto fronts that identify non-dominated solutions. Optimization results of a similar simulated moving bed process have also been included to facilitate comparison with a continuous chromatographic process. PMID:22364669

  4. Optimization of an improved single-column chromatographic process for the separation of enantiomers.

    PubMed

    Kazi, Monzure-Khoda; Medi, Bijan; Amanullah, Mohammad

    2012-03-30

    This work addresses optimization of an improved single-column chromatographic (ISCC) process for the separation of guaifenesin enantiomers. Conventional feed injection and fraction collection systems have been replaced with customized components facilitating simultaneous separation and online monitoring with the ultimate objective of application of an optimizing controller. Injection volume, cycle time, desorbent flow rate, feed concentration, and three cut intervals are considered as decision variables. A multi-objective optimization technique based on genetic algorithm (GA) is adopted to achieve maximum productivity and minimum desorbent requirement in the region constrained by product specifications and hardware limitations. The optimization results along with the contribution of decision variables are discussed using Pareto fronts that identify non-dominated solutions. Optimization results of a similar simulated moving bed process have also been included to facilitate comparison with a continuous chromatographic process.

  5. Separation of Be and Al for AMS using single-step column chromatography

    NASA Astrophysics Data System (ADS)

    Binnie, Steven A.; Dunai, Tibor J.; Voronina, Elena; Goral, Tomasz; Heinze, Stefan; Dewald, Alfred

    2015-10-01

    With the aim of simplifying AMS target preparation procedures for TCN measurements we tested a new extraction chromatography approach which couples an anion exchange resin (WBEC) to a chelating resin (Beryllium resin) to separate Be and Al from dissolved quartz samples. Results show that WBEC-Beryllium resin stacks can be used to provide high purity Be and Al separations using a combination of hydrochloric/oxalic and nitric acid elutions. 10Be and 26Al concentrations from quartz samples prepared using more standard procedures are compared with results from replicate samples prepared using the coupled WBEC-Beryllium resin approach and show good agreement. The new column procedure is performed in a single step, reducing sample preparation times relative to more traditional methods of TCN target production.

  6. Preparative separation and purification of rosmarinic acid from perilla seed meal via combined column chromatography.

    PubMed

    Tang, Weizhuo; Sun, Baoshan; Zhao, Yuqing

    2014-02-01

    In this study, the preparative separation and purification of rosmarinic acid (RA) from perilla seed meal (PSM), which is a by-product of edible oil production, was achieved using combined column chromatography over macroporous and polyamide resins. To optimize the RA enrichment process, the performance and separation characteristics of nine selected macroporous resins with different chemical and physical properties were investigated. SP825 resin was the most effective: the content of RA increased from 0.27% in the original extract to 16.58% in the 50% ethanol fraction (a 61.4-fold increase). During further purification treatment on polyamide resin, 90.23% pure RA could be obtained in the 70% ethanol fraction. RA with a higher purity (>95%) could also be easily obtained using one crystallization operation. The proposed method is simple, easily operated, cost-effective, and environmentally friendly and is suitable for both large-scale RA production and waste management. PMID:24381020

  7. Rapid separation of polysaccharides using a novel spiral coil column by high-speed countercurrent chromatography.

    PubMed

    Li, Weili; Wu, Tao

    2016-04-01

    The separation of polysaccharides is time consuming. We developed and optimized a type-J counter-current chromatography system with a novel tri-rotor spiral coil column for the rapid separation of polysaccharides. The optimal composition of an aqueous PEG1000/K2 HPO4 /KH2 PO4 system was found to be 14:16:14 w/w/w where the lower phase was the mobile phase. Optimal performance was achieved at a column rotational speed, temperature, and flow rate of 1200 rpm, 45°C, and 3.0 mL/min, respectively. The mobile phase was pumped from the inner terminal in a ''head-to-tail'' elution mode. Polysaccharide LCP-1 (10.7 mg) was successfully obtained in high purity in one step from 50.0 mg of a crude polysaccharide extracted from the lychee fruit (Litchi chinensis) within 100 min. LCP-1 possess a number-average molecular weight and weight-average molecular weight of 1.05 × 10(5) and 1.59 × 10(5) kDa, respectively. The monosaccharide composition consists of the molar ratio of glucose, galactose, and arabinose of 1.3:3.5:1.

  8. Modelling aspects regarding the control in 13C isotope separation column

    NASA Astrophysics Data System (ADS)

    Boca, M. L.

    2016-08-01

    Carbon represents the fourth most abundant chemical element in the world, having two stable and one radioactive isotope. The 13Carbon isotopes, with a natural abundance of 1.1%, plays an important role in numerous applications, such as the study of human metabolism changes, molecular structure studies, non-invasive respiratory tests, Alzheimer tests, air pollution and global warming effects on plants [9] A manufacturing control system manages the internal logistics in a production system and determines the routings of product instances, the assignment of workers and components, the starting of the processes on not-yet-finished product instances. Manufacturing control does not control the manufacturing processes themselves, but has to cope with the consequences of the processing results (e.g. the routing of products to a repair station). In this research it was fulfilled some UML (Unified Modelling Language) diagrams for modelling the C13 Isotope Separation column, implement in STARUML program. Being a critical process and needing a good control and supervising, the critical parameters in the column, temperature and pressure was control using some PLC (Programmable logic controller) and it was made some graphic analyze for this to observe some critical situation than can affect the separation process. The main parameters that need to be control are: -The liquid nitrogen (N2) level in the condenser. -The electrical power supplied to the boiler. -The vacuum pressure.

  9. Optimization of preparative separation and purification of total polyphenols from Sargassum tenerrimum by column chromatography

    NASA Astrophysics Data System (ADS)

    Haider, Samee; Li, Zhenxing; Lin, Hong; Jamil, Khalid

    2009-12-01

    Polyphenols from the ethanol extracts of Sargassum tenerrimum (ST) with potent antiallergic effects were studied to optimize separation process through column chromatography. The adsorption and desorption characteristics of three widely used adsorbents: macroporous resin, silica gel, and polyvinylpolypyrrolidone (PVPP), were critically evaluated respectively and studied for the optimization of preparative separation of polyphenols. Static operations on these adsorbents showed that macroporous resin had the best adsorption and desorption capability among the three adsorbents. Dynamic adsorption and desorption with macroporous resin packed column were also conducted to optimize the parameters such as: with the optimal values shown in brackets, the concentration of extract solution (4 times diluted), pH value (6-7), adsorption speed (3 BV h-1, bed volumes/per hour), concentration of ethanol (80%), elution speed (3 BV h-1) and elution volume (7 BV). The chromatographic process so optimized gave a purity of 62.43% from the crude polyphenols, providing a promising basis for large scale preparation of bioactive polyphenols upon further scaling up tests.

  10. SEPARATION AND PURIFICATION OF TWO MINOR COMPOUNDS FROM RADIX ISATIDIS BY INTEGRATIVE MPLC AND HSCCC WITH PREPARATIVE HPLC

    PubMed Central

    Liang, Zhenjie; Li, Bin; Liang, Yong; Su, Yaping; Ito, Yoichiro

    2014-01-01

    Radix isatidis has been widely used as a Chinese traditional medicine for its anti-virus and anticancer activities where the minor components may contribute to these beneficial pharmaceutical effects. In order to enrich the target minor compounds effectively and rapidly, extraction, medium-pressure liquid chromatography (MPLC), high-speed countercurrent chromatography (HSCCC) and preparative high-performance liquid chromatography (pre-HPLC) were integratively used for separation and purification of two target minor compounds indole-3-acetonitrile-6-O-β-D-glucopyranoside (target 1) and clemastanin B (target 2) in the present study. Radix isatidis was dried, pulverized and extracted with 50% methanol at room temperature, then concentrated and subjected to pretreatment with D-101 macroporous resin chromatography and extraction by MPLC. The first target compound was separated by MPLC at the purity raised to 70–80%, but without the second minor compounds which were irreversibly adsorbed by C18 solid support. Therefore, the second target compound in the crude extract was directly separated by HSCCC at purity of 80–90%. Finally these refined samples were further separated by pre-HPLC to obtain a high purity at 98–99%. The chemical structure identification of each target compound was carried out by IR, ESI-MS and 1H NMR. PMID:25745338

  11. Preparation and characterization of a molecularly imprinted monolithic column for pressure-assisted CEC separation of nitroimidazole drugs.

    PubMed

    Liao, Sulan; Wang, Xiaochun; Lin, Xucong; Xie, Zenghong

    2010-08-01

    A polymethacrylate-based molecularly imprinted monolithic column bearing mixed functional monomers, using non-covalent imprinting approach, was designed for the rapid separation of nitroimidazole compounds. The new monolithic column has been prepared via simple in situ polymerization of 2-hydroxyethyl methacrylate, dimethylaminoethyl methacrylate and ethylene dimethacrylate, using (S)-ornidazole ((S)-ONZ) as template in a binary porogenic mixture consisting of toluene and dodecanol. The composition of the polymerization mixture was systematically altered and optimized by altering the amount of monomers as well as the composition of the porogenic solvent. The column performance was evaluated in pressure-assisted CEC mode. Separation conditions such as pH, voltage, amount of organic modifier and salt concentration were studied. The optimized monolithic column resulted in excellent separation of a group of structurally related nitroimidazole drugs within 10 min in isocratic elution condition. Column efficiencies of 99 000, 80 000, 103 000, 60 000 and 99 000 plates/m were obtained for metronidazole, secnidazole, ronidazole, tinidazole and dimetridazole, respectively. Parallel experiments were carried out using molecularly imprinted and non-imprinted capillary columns. The separation might be the result of combined effects including hydrophobic, hydrogen bonding and the imprinting cavities on the (S)-ONZ-imprinted monolithic column. PMID:20661943

  12. Removal Characteristics of Immunoadsorption With the Immusorba TR-350 Column Using Conventional and Selective Plasma Separators.

    PubMed

    Ohkubo, Atsushi; Okado, Tomokazu; Miyamoto, Satoko; Goto, Keigo; Yamamoto, Motoki; Maeda, Takuma; Itagaki, Ayako; Seshima, Hiroshi; Kurashima, Naoki; Sohara, Eisei; Uchida, Shinichi; Rai, Tatemitsu

    2016-08-01

    In Japan, immunoadsorption (IA) is performed using a conventional plasma separator and Immusorba TR-350 column (TR-350) for the treatment of neurological immune diseases. By this method, TR-350 has the limited maximal capacity of the immunoglobulin G (IgG) adsorption, and fibrinogen (Fbg) is reduced remarkably. Evacure EC-4A10 (EC-4A) is a selective plasma separator and the sieving coefficients of IgG and Fbg using EC-4A were 0.5 and 0, respectively. Here, we investigated the removal characteristics of IgG and Fbg in IA by TR-350 using two different plasma membrane separators: conventional plasma separator (PE-IA) and EC-4A (EC-IA). In vitro filtration using plasma effluent was performed with a closed circuit. When the processed volume was 3 L, estimated removal amounts by PE-IA were 3172 mg for IgG and 3329 mg for Fbg, respectively. When the processed volume was 3 L, estimated removal amounts by EC-IA were 4946 mg and 1916 mg, respectively. EC-IA can be considered useful for the removal of IgG, including auto-antibodies, while retaining Fbg, thereby allowing even daily use. PMID:27523076

  13. [Near-infrared spectroscopy technology for online monitoring of the column separation and purification process of active components of Centella asiatica L. Urban].

    PubMed

    Liu, Hua; Ye, Xiao-Lan; Yang, Guang; Qi, Yun-Peng; Fan, Guo-Rong

    2013-01-01

    The present paper is to study and develop a method for online monitoring of the column separation and purification process of active components that are madecassoside and asiaticoside of Centella asiatica L. Urban using near-infrared (NIR) spectroscopy technology. After collecting 50%-ethanol eluant, we detected their NIR spectra and developed the high performance liquid chromatography (HPLC) assay method of active components. Then, partial least square (PLS) was used to develop linear correlation between their NIR spectra and contents. During modeling, correlation coefficient (R2) and root mean square errors of cross-validation (RMSECV) were regarded as the indexes to select optimal wavenumbers and preprocessing methods. The optimal wavenumbers of madecassoside and asiaticoside were in the range of 12 000.8-7 499.8 cm(-1) and 12 000.8-9 750.3 cm(-1), respectively; R2 were 96.44 and 96.07, respectively, and RMSECV were 0.084 80 and 0.000 99, respectively. The above developed model was used for online monitoring of the contents of madecassoside and asiaticoside during the column separation and purification process of Centella asiatica L. Urban. The predicted results were satisfactory. This method was proved to be fast, convenient and precise. It can be used in online monitoring and quality control of the manufacturing of madecassoside and asiaticoside.

  14. [Near-infrared spectroscopy technology for online monitoring of the column separation and purification process of active components of Centella asiatica L. Urban].

    PubMed

    Liu, Hua; Ye, Xiao-Lan; Yang, Guang; Qi, Yun-Peng; Fan, Guo-Rong

    2013-01-01

    The present paper is to study and develop a method for online monitoring of the column separation and purification process of active components that are madecassoside and asiaticoside of Centella asiatica L. Urban using near-infrared (NIR) spectroscopy technology. After collecting 50%-ethanol eluant, we detected their NIR spectra and developed the high performance liquid chromatography (HPLC) assay method of active components. Then, partial least square (PLS) was used to develop linear correlation between their NIR spectra and contents. During modeling, correlation coefficient (R2) and root mean square errors of cross-validation (RMSECV) were regarded as the indexes to select optimal wavenumbers and preprocessing methods. The optimal wavenumbers of madecassoside and asiaticoside were in the range of 12 000.8-7 499.8 cm(-1) and 12 000.8-9 750.3 cm(-1), respectively; R2 were 96.44 and 96.07, respectively, and RMSECV were 0.084 80 and 0.000 99, respectively. The above developed model was used for online monitoring of the contents of madecassoside and asiaticoside during the column separation and purification process of Centella asiatica L. Urban. The predicted results were satisfactory. This method was proved to be fast, convenient and precise. It can be used in online monitoring and quality control of the manufacturing of madecassoside and asiaticoside. PMID:23586234

  15. Non-HPLC rapid separation of metallofullerenes and empty cages with TiCl4 Lewis acid.

    PubMed

    Akiyama, Kazuhiko; Hamano, Tatsuyuki; Nakanishi, Yusuke; Takeuchi, Erina; Noda, Shoko; Wang, Zhiyong; Kubuki, Shiro; Shinohara, Hisanori

    2012-06-13

    Rapid and efficient separation/purification of pure metallofullerenes M(x)@C(n) (M = metal; x = 1, 2; n > 70) and carbide metallofullerenes of the type M(y)C(2)@C(n-2) (y = 2, 3, 4; n - 2 > 68) has been reported. The present method utilizes rapid and almost perfect preferential formation of TiCl(4) (generally known as a Lewis acid)-metallofullerene complexes, which easily decompose to provide pure metallofullerene powders by a simple water treatment. The present method enables one to separate the metallofullerenes up to >99% purity within 10 min without using any type of high-performance liquid chromatography (HPLC). It is found that the oxidation potentials of the metallofullerenes are crucial factors for efficient purification. The current separation/purification technique may open a brand-new era for inducing further applications and commercialization of endohedral metallofullerenes.

  16. Fabrication of zeolitic imidazolate framework-8-methacrylate monolith composite capillary columns for fast gas chromatographic separation of small molecules.

    PubMed

    Yusuf, Kareem; Badjah-Hadj-Ahmed, Ahmed Yacine; Aqel, Ahmad; ALOthman, Zeid Abdullah

    2015-08-01

    A composite zeolitic imidazolate framework-8 (ZIF-8) with a butyl methacrylate-co-ethylene dimethacrylate (BuMA-co-EDMA) monolithic capillary column (33.5cm long×250μm i.d.) was fabricated to enhance the separation efficiency of methacrylate monoliths toward small molecules using conventional low-pressure gas chromatography in comparison with a neat butyl methacrylate-co-ethylene dimethacrylate (BuMA-co-EDMA) monolithic capillary column (33.5cm long×250μm i.d.). The addition of 10mgmL(-1) ZIF-8 micro-particles increased the BET surface area of BuMA-co-EDMA by 3.4-fold. A fast separation of five linear alkanes in 36s with high resolution (Rs≥1.3) was performed using temperature program. Isothermal separation of the same sample also showed a high efficiency (3315platesm(-1) for octane) at 0.89min. Moreover, the column was able to separate skeletal isomers, such as iso-octane/octane and 2-methyl octane/nonane. In addition, an iso-butane/iso-butylene gas mixture was separated at ambient temperature. Comparison with an open tubular TR-5MS column (30m long×250μm i.d.) revealed the superiority of the composite column in separating the five-membered linear alkane mixture with 4-5 times increase in efficiency and a total separation time of 0.89min instead of 4.67min. A paint thinner sample was fully separated using the composite column in 2.43min with a good resolution (Rs≥0.89). The perfect combination between the polymeric monolith, with its high permeability, and ZIF-8, with its high surface area and flexible 0.34nm pore openings, led to the fast separation of small molecules with high efficiency and opened a new horizon in GC applications.

  17. Fabrication of zeolitic imidazolate framework-8-methacrylate monolith composite capillary columns for fast gas chromatographic separation of small molecules.

    PubMed

    Yusuf, Kareem; Badjah-Hadj-Ahmed, Ahmed Yacine; Aqel, Ahmad; ALOthman, Zeid Abdullah

    2015-08-01

    A composite zeolitic imidazolate framework-8 (ZIF-8) with a butyl methacrylate-co-ethylene dimethacrylate (BuMA-co-EDMA) monolithic capillary column (33.5cm long×250μm i.d.) was fabricated to enhance the separation efficiency of methacrylate monoliths toward small molecules using conventional low-pressure gas chromatography in comparison with a neat butyl methacrylate-co-ethylene dimethacrylate (BuMA-co-EDMA) monolithic capillary column (33.5cm long×250μm i.d.). The addition of 10mgmL(-1) ZIF-8 micro-particles increased the BET surface area of BuMA-co-EDMA by 3.4-fold. A fast separation of five linear alkanes in 36s with high resolution (Rs≥1.3) was performed using temperature program. Isothermal separation of the same sample also showed a high efficiency (3315platesm(-1) for octane) at 0.89min. Moreover, the column was able to separate skeletal isomers, such as iso-octane/octane and 2-methyl octane/nonane. In addition, an iso-butane/iso-butylene gas mixture was separated at ambient temperature. Comparison with an open tubular TR-5MS column (30m long×250μm i.d.) revealed the superiority of the composite column in separating the five-membered linear alkane mixture with 4-5 times increase in efficiency and a total separation time of 0.89min instead of 4.67min. A paint thinner sample was fully separated using the composite column in 2.43min with a good resolution (Rs≥0.89). The perfect combination between the polymeric monolith, with its high permeability, and ZIF-8, with its high surface area and flexible 0.34nm pore openings, led to the fast separation of small molecules with high efficiency and opened a new horizon in GC applications. PMID:26141277

  18. Optimal performance of single-column chromatography and simulated moving bed processes for the separation of optical isomers

    NASA Astrophysics Data System (ADS)

    Medi, Bijan; Kazi, Monzure-Khoda; Amanullah, Mohammad

    2013-06-01

    Chromatography has been established as the method of choice for the separation and purification of optically pure drugs which has a market size of about 250 billion USD. Single column chromatography (SCC) is commonly used in the development and testing phase of drug development while multi-column Simulated Moving Bed (SMB) chromatography is more suitable for large scale production due to its continuous nature. In this study, optimal performance of SCC and SMB processes for the separation of optical isomers under linear and overloaded separation conditions has been investigated. The performance indicators, namely productivity and desorbent requirement have been compared under geometric similarity for the separation of a mixture of guaifenesin, and Tröger's base enantiomers. SCC process has been analyzed under equilibrium assumption i.e., assuming infinite column efficiency, and zero dispersion, and its optimal performance parameters are compared with the optimal prediction of an SMB process by triangle theory. Simulation results obtained using actual experimental data indicate that SCC may compete with SMB in terms of productivity depending on the molecules to be separated. Besides, insights into the process performances in terms of degree of freedom and relationship between the optimal operating point and solubility limit of the optical isomers have been ascertained. This investigation enables appropriate selection of single or multi-column chromatographic processes based on column packing properties and isotherm parameters.

  19. Preparative isolation and purification of flavone compounds from sophora japonica L. by high-speed counter-current chromatography combined with macroporous resin column separation.

    PubMed

    Sun, Ailing; Sun, Qinghua; Liu, Renmin

    2007-05-01

    High-speed counter-current chromatography combined with macroporous resin column separation was applied to the isolation and purification of genistein-7,4'-di-O-beta-D-glucoside (I), genistein-7-O-beta-D-glucopyranoside-4'-O-[(alpha-L-rhamnopyransoyl)-(1-2)-beta-D-glucopyranoside] (II), kaempferol-3-O-beta-D-sophoroside(III), quercetin-3-O-beta-L-ramnopyranosyl-(1 - 6)-beta-D-glucopyranoside (IV), genistein-4'-beta-L-rhamnopyransoyl-(1 - 2)-alpha-D-glucopyranoside (V), and kaempferol-3-O-beta-L-ramnopyranosyl-(1 - 6)-beta-D-glucopyranoside (VI) from the Chinese medicinal herb Sophora japonica L. The crude extracts from the pericarps of Sophora japonica L. were pre-separated on a D-101 macroporous resin column and divided into two parts as sample 1 and sample 2. An 80-mg portion of sample 1 was separated by using n-butanol-acetic acid (1%) (5:5, v/v) as the two-phase solvent system and yielded 30.1 mg of compound I, 23.3 mg of compound II. A 120 mg portion of sample 2 was separated by using ethyl acetate-n-butanol-acetic acid (1%) (5:0.8:5, v/v) as the two-phase solvent system and yielded 5.5 mg of compound III, 31.7 mg of compound IV, 37.4 mg of compound V, and 6.2 mg of compound VI. The purities of compounds I, II, III, IV, V, and VI were 98.7, 98.2, 97.8, 98.5, 99.3, and 98.9%, respectively, as determined by HPLC. The chemical structures of these components were identified by 1H-NMR and 13C-NMR. PMID:17566335

  20. Aspects regarding at 13C isotope separation column control using Petri nets system

    NASA Astrophysics Data System (ADS)

    Boca, M. L.; Ciortea, M. E.

    2015-11-01

    This paper is intended to show that Petri nets can be also applicable in the chemical industry. It used linear programming, modeling underlying Petri nets, especially discrete event systems for isotopic separation, the purpose of considering and control events in real-time through graphical representations. In this paper it is simulate the control of 13C Isotope Separation column using Petri nets. The major problem with 13C comes from the difficulty of obtaining it and raising its natural fraction. Carbon isotopes can be obtained using many methods, one of them being the cryogenic distillation of carbon monoxide. Some few aspects regarding operating conditions and the construction of such cryogenic plants are known today, and even less information are available as far as the separation process modeling and control are concerned. In fact, the efficient control of the carbon monoxide distillation process represents a necessity for large-scale 13C production. Referring to a classic distillation process, some models for carbon isotope separation have been proposed, some based on mass, component and energy balance equations, some on the nonlinear wave theory or the Cohen equations. For modeling the system it was used Petri nets because in this case it is deal with discrete event systems. In use of the non-timed and with auxiliary times Petri model, the transport stream was divided into sections and these sections will be analyzed successively. Because of the complexity of the system and the large amount of calculations required it was not possible to analyze the system as a unitary whole. A first attempt to model the system as a unitary whole led to the blocking of the model during simulation, because of the large processing times.

  1. Recent Progress in Monolithic Silica Columns for High-Speed and High-Selectivity Separations

    NASA Astrophysics Data System (ADS)

    Ikegami, Tohru; Tanaka, Nobuo

    2016-06-01

    Monolithic silica columns have greater (through-pore size)/(skeleton size) ratios than particulate columns and fixed support structures in a column for chemical modification, resulting in high-efficiency columns and stationary phases. This review looks at how the size range of monolithic silica columns has been expanded, how high-efficiency monolithic silica columns have been realized, and how various methods of silica surface functionalization, leading to selective stationary phases, have been developed on monolithic silica supports, and provides information on the current status of these columns. Also discussed are the practical aspects of monolithic silica columns, including how their versatility can be improved by the preparation of small-sized structural features (sub-micron) and columns (1 mm ID or smaller) and by optimizing reaction conditions for in situ chemical modification with various restrictions, with an emphasis on recent research results for both topics.

  2. Recent Progress in Monolithic Silica Columns for High-Speed and High-Selectivity Separations.

    PubMed

    Ikegami, Tohru; Tanaka, Nobuo

    2016-06-12

    Monolithic silica columns have greater (through-pore size)/(skeleton size) ratios than particulate columns and fixed support structures in a column for chemical modification, resulting in high-efficiency columns and stationary phases. This review looks at how the size range of monolithic silica columns has been expanded, how high-efficiency monolithic silica columns have been realized, and how various methods of silica surface functionalization, leading to selective stationary phases, have been developed on monolithic silica supports, and provides information on the current status of these columns. Also discussed are the practical aspects of monolithic silica columns, including how their versatility can be improved by the preparation of small-sized structural features (sub-micron) and columns (1 mm ID or smaller) and by optimizing reaction conditions for in situ chemical modification with various restrictions, with an emphasis on recent research results for both topics. PMID:27306311

  3. Reverse-phase HPLC separation of hemp seed (Cannabis sativa L.) protein hydrolysate produced peptide fractions with enhanced antioxidant capacity.

    PubMed

    Girgih, Abraham T; Udenigwe, Chibuike C; Aluko, Rotimi E

    2013-03-01

    Hemp seed protein hydrolysate (HPH) was produced through simulated gastrointestinal tract (GIT) digestion of hemp seed protein isolate followed by partial purification and separation into eight peptide fractions by reverse-phase (RP)-HPLC. The peptide fractions exhibited higher oxygen radical absorbance capacity as well as scavenging of 2,2-diphenyl-1-picrylhydrazyl, superoxide and hydroxyl radicals when compared to HPH. Radical scavenging activities of the fractionated peptides increased as content of hydrophobic amino acids or elution time was increased, with the exception of hydroxyl radical scavenging that showed decreased trend. Glutathione (GSH), HPH and the RP-HPLC peptide fractions possessed low ferric ion reducing ability but all had strong (>60 %) metal chelating activities. Inhibition of linoleic acid oxidation by some of the HPH peptide fractions was higher at 1 mg/ml when compared to that observed at 0.1 mg/ml peptide concentration. Peptide separation resulted in higher concentration of some hydrophobic amino acids (especially proline, leucine and isoleucine) in the fractions (mainly F5 and F8) when compared to HPH. The elution time-dependent increased concentrations of the hydrophobic amino acids coupled with decreased levels of positively charged amino acids may have been responsible for the significantly higher (p < 0.05) antioxidant properties observed for some of the peptide fractions when compared to the unfractionated HPH. In conclusion, the antioxidant activity of HPH after simulated GIT digestion is mainly influenced by the amino acid composition of some of its peptides.

  4. Antimony speciation in soils: improving the detection limits using post-column pre-reduction hydride generation atomic fluorescence spectroscopy (HPLC/pre-reduction/HG-AFS).

    PubMed

    Quiroz, Waldo; Olivares, David; Bravo, Manuel; Feldmann, Jorg; Raab, Andrea

    2011-04-15

    HG-AFS is highly sensitive and low cost detection system and its use for antimony chemical speciation coupled to HPLC is gaining popularity. However speciation analysis in soils is strongly hampered because the most efficient extractant reported in the literature (oxalic acid) strongly inhibits the generation of SbH(3) by Sb(V), the major species in this kind of matrix, severely affecting its detection limits. The purpose of this research is to reduce the detection limit of Sb(V), by using a post column on-line reduction system with l-cysteine reagent (HPLC/pre-reduction/HG-AFS). The system was optimized by experimental design, optimum conditions found were 2% (w/v) and 10°C temperature coil. Detection limits of Sb(V) and Sb(III) in oxalic acid (0.25 mol L(-1)) were improved from 0.3 and 0.1 μg L(-1) to 0.07 and 0.07 μg L(-1), respectively. The methodology developed was applied to Chilean soils, where Sb(V) was the predominant species.

  5. Short communication: Amino trap column improves the separation of methylimidazoles, 5-hydroxymethyl-2-furaldehyde, and sugars in Maillard reaction.

    PubMed

    Xu, Xian-Bing; Liu, Ding-Bo; Yu, Shu-Juan; Zhao, Zhen-Gang; Yu, Pei

    2014-11-01

    A simultaneous analysis of methylimidazoles, reducing sugars, and 5-hydroxymethyl-2-furaldehyde in the Maillard reaction was improved by use of an amino trap column. Analysis was carried out by using high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) coupled with an amino trap column. The amino trap column was a useful tool to improve the separation of methylimidazoles, reducing sugars, and 5-hydroxymethyl-2-furaldehyde. This technique is useful for simultaneous analysis of methylimidazoles, reducing sugars, and 5-hydroxymethyl-2-furaldehyde in risk assessment for dairy products.

  6. Separation of cannabinoids on three different mixed-mode columns containing carbon/nanodiamond/amine-polymer superficially porous particles.

    PubMed

    Hung, Chuan-Hsi; Zukowski, Janusz; Jensen, David S; Miles, Andrew J; Sulak, Clayton; Dadson, Andrew E; Linford, Matthew R

    2015-09-01

    Three mixed-mode high-performance liquid chromatography columns packed with superficially porous carbon/nanodiamond/amine-polymer particles were used to separate mixtures of cannabinoids. Columns evaluated included: (i) reversed phase (C18 ), weak anion exchange, 4.6 × 33 mm, 3.6 μm, and 4.6 × 100 mm, 3.6 μm, (ii) reversed phase, strong anion exchange (quaternary amine), 4.6×33 mm, 3.6 μm, and (iii) hydrophilic interaction liquid chromatography, 4.6 × 150 mm, 3.6 μm. Different selectivities were achieved under various mobile phase and stationary phase conditions. Efficiencies and peak capacities were as high as 54 000 N/m and 56, respectively. The reversed phase mixed-mode column (C18 ) retained tetrahydrocannabinolic acid strongly under acidic conditions and weakly under basic conditions. Tetrahydrocannabinolic acid was retained strongly on the reversed phase, strong anion exchange mixed-mode column under basic polar organic mobile phase conditions. The hydrophilic interaction liquid chromatography column retained polar cannabinoids better than the (more) neutral ones under basic conditions. A longer reversed phase (C18 ) mixed-mode column (4.6 × 100 mm) showed better resolution for analytes (and a contaminant) than a shorter column. Fast separations were achieved in less than 5 min and sometimes 2 min. A real world sample (bubble hash extract) was also analyzed by gradient elution. PMID:26075936

  7. Screening Approach for Chiral Separation of β-Aminoketones by HPLC on Various Polysaccharide-Based Chiral Stationary Phases.

    PubMed

    Addadi, Khadidja; Sekkoum, Khaled; Belboukhari, Nasser; Cheriti, Abdelkrim; Aboul-Enein, Hassan Y

    2015-05-01

    Nine β-aminoketones were synthesized via Mannich reaction when benzaldehyde was condensed with some primary amines and acetophenone. The purified compounds were identified by using spectroscopic methods. The enantiomeric separation of these derivatives was carried out by high-performance liquid chromatography (HPLC) using several coated and immobilized polysaccharide stationary phases, namely, Chiralcel(®) OD-H, Chiralcel(®) OD, Chiralcel(®) OJ, Chiralpak(®) AD, Chiralpak(®) IA, and Chiralpak(®) IB using different mobile phases composed of n-hexane and alcohol mixed in various ratios or pure ethanol or isopropanol. The retention behavior and selectivity of these chiral stationary phases were examined in isocratic normal phase mode. The results indicate that cellulose derivatives have higher enantioselectivity than amylose derivatives for the separation of racemic β-amino ketones. PMID:25752940

  8. [Preparation of a reversed-phase capillary monolithic column and its application in the separation of polypeptide mixtures].

    PubMed

    Xie, Jingxin; Bi, Kaishun; Qian, Xiaohong; Zuang, Yangiun

    2009-03-01

    Capillary monolithic columns (75 microm i.d.) were prepared by the copolymerization of lauryl methacrylate as the basic monomer, ethylene dimethacrylate as the cross-linking agent and 1-dodecyl alcohol, 1,4-butanediol and dimethyl sulfoxide as the porogenic mixture. The synthetic stationary phases had better mechanical properties and chemical stabilities. A series of characterization and evaluations were performed on the capillary monolithic columns including the scanning electron microscope (SEM) images, the influences of pressure and the effects on the separation of peptide mixtures by changing the proportions of the porogen solution and cross-linking agent. The final prescription ciontained 15% (w/w) monomer, 15% (w/ w) cross-linking agent, and 70% (w/w) porogenic agent. Then the solution was heated at 70 1C for 24 h. The test of relationship between column length and back pressure showed that the capillary monolithic columns prepared have superior permeability, so a longer column can be used to improve the effects of separation. The prepared capillary monolithic columns are fitted on the nano-scale high performance liquid chromatography for the separation of tryptic digests of bovine serum albumin (BSA) and human plasma samples, and better results have been obtained.

  9. Peak capacity optimization of peptide separations in reversed-phase gradient elution chromatography: fixed column format.

    PubMed

    Wang, Xiaoli; Stoll, Dwight R; Schellinger, Adam P; Carr, Peter W

    2006-05-15

    The optimization of peak capacity in gradient elution RPLC is essential for the separation of multicomponent samples such as those encountered in proteomic research. In this work, we study the effect of gradient time (tG), flow rate (F), temperature (T), and final eluent strength (phi(final)) on the peak capacity of separations of peptides that are representative of the range in peptides found in a tryptic digest. We find that there are very strong interactions between the individual variables (e.g., flow rate and gradient time) which make the optimization quite complicated. On a given column, one should first set the gradient time to the longest tolerable and then set the temperature to the highest achievable with the instrument. Next, the flow rate should be optimized using a reasonable but arbitrary value of phi(final). Last, the final eluent strength should be adjusted so that the last solute elutes as close as possible to the gradient time. We also develop an easily implemented, highly efficient, and effective Monte Carlo search strategy to simultaneously optimize all the variables. We find that gradient steepness is an important parameter that influences peak capacity and an optimum range of gradient steepness exists in which the peak capacity is maximized.

  10. Separation of aromatic carboxylic acids using quaternary ammonium salts on reversed-phase HPLC. 1. Separation behavior of aromatic carboxylic acids

    SciTech Connect

    Kawamura, K.; Okuwaki, A.; Verheyen, T.; Perry, G.J.

    2006-02-15

    In order to develop separation processes and analytical methods for aromatic carboxylic acids for the coal oxidation products, the separation behavior of aromatic carboxylic acids on a reversed-phase HPLC using eluent containing quaternary ammonium salt has been investigated. The retention mechanism of aromatic carboxylic acids was discussed on the basis of both ion-pair partition model and ion-exchange model. The retention behavior of aromatic carboxylic acids possessing one (or two) carboxylic acid group(s) followed the ion-pair partition model, where linear free energy relationship was observed between the capacity factor and the extraction equilibrium constants of benzoic acid and naphthalene carboxylic acid. Besides, the retention behavior followed ion-exchange model with increasing the number of carboxylic acids, where the capacity factor of benzene polycarboxylic acids is proportional to the association constants between aromatic acids and quaternary ammonium ions calculated on the basis of an electrostatic interaction model.

  11. Simultaneous multiresponse optimization of an HPLC method to separate seven cephalosporins in plasma and amniotic fluid: application to validation and quantification of cefepime, cefixime and cefoperazone.

    PubMed

    Nemutlu, Emirhan; Kir, Sedef; Katlan, Doruk; Beksaç, M Sinan

    2009-11-15

    An HPLC method for the separation of seven cephalosporins [Cefepime (CEP), ceftazidime (CTA), ceftizaxime (CTI), ceftriaxone (CTR), cefotaxime (COT), cefixime (CIX) and cefoperazone (COP)] in human plasma and amniotic fluid has been developed. Optimization of the chromatographic method was performed in three steps: a series of initial experiments followed by two sets of experiments based on different experimental designs. The initial experiments were performed to decide the basic analytical requirements of the method. Then screening experiment fractional factorial design was used in order to decrease the number of parameters by eliminating parameters which having insignificant effect on responses. The parameters having significant effect were further optimized through a full factorial design. Having studied two responses (retention times and resolutions), a desirability function that assess the responses together, was used to find experimental conditions where the system generated desirable results. The desirable results were obtained with XTerra C18 (250 mm x 4.6mm, 5 microm i.d.) column, 40 mM phosphate buffer, pH 3.2, 18% MeOH, 0.85 mL min(-1) flow rate and 32 degrees C column temperature. Gradient elution with MeOH was applied. A simple and efficient solid-phase extraction was applied for the preparation of plasma and amniotic fluid samples. The validation parameters of the method were evaluated in accordance with ICH guideline. The method validated was applied to the analysis of CEP and COP in maternal venous, fetal venous and fetal arterial plasma, and to the analysis of CIX in maternal venous plasma and amniotic fluid.

  12. Coupled achiral/chiral column techniques in subcritical fluid chromatography for the separation of chiral and nonchiral compounds.

    PubMed

    Phinney, K W; Sander, L C; Wise, S A

    1998-06-01

    A multicolumn approach was developed to address the limited achiral selectivity of chiral stationary phases. Groups of structurally related compounds, including beta-blockers and 1,4-benzodiazepines, were separated using coupled achiral/chiral stationary phases under subcritical fluid conditions. The achiral selectivity of amino and cyano stationary phases was used to modify the resolution of compounds on a Chiralcel OD chiral stationary phase by combining the achiral and chiral columns in series. In the case of the benzodiazepines, separation of achiral compounds was performed concurrently with the enantioseparation of chiral molecules. The separation of components of a multidrug cough and cold medication was also demonstrated on a cyano column coupled with a Chiralpak AD chiral stationary phase. The use of modified carbon dioxide eluents eliminated the mobile phase incompatibility problems associated with column coupling in liquid chromatography and incorporated the high efficiency of sub- and supercritical fluid chromatography.

  13. Chromatographic separation of simulants of nerve and blister agents by combining one- and two-channel columns with different stationary phases.

    PubMed

    Yuan, Huan; Du, Xiaosong; Li, Yi; Zhao, Xulan; Xu, Ming

    2016-04-01

    A two-channel gas chromatography column and a single-channel column were made by deep reactive-ion etching technology. The two short columns were coated with different stationary phases, and then linked without a modulator. This is to aim at increasing the sample capacity and achieving a higher separation efficiency in complex environments. The results show that the capacity of the connected column is approximately 4 and 1.5 times larger than that of the single- and two-channel columns, respectively. The linked column was utilized to separate a six-component mixture, composed of three simulants of nerve and blister agents and three interfering vapors. The results demonstrate that the combined column has a remarkably higher separation efficiency than the individual columns, and an acceptable resolution is achieved although the total length of the linked column is only 1.5 m. PMID:26843525

  14. [The novel copolymer coated capillary columns of electrophoresis and their applications to separation of proteins].

    PubMed

    Lu, G; Gao, D; Gu, J; Fu, R; Li, F; Zhang, H

    1999-01-01

    The copolymer of acrylonitrile, methyl acrylate, hydroxy ethyl acrylate (ZB-004), the copolymer of acrylonitrile, methyl acrylate, hydroxy ethyl acrylate, acrylamide (ZB-014) and the copolymer of acrylonitrile, hydroxy ethyl acrylate (ZB-016) were coated on the inner surface of fused-silica capillaries by just filling the capillary with solutions containing these copolymers followed by flushing the capillary with nitrogen. The physically adsorbed layer can reduce both protein adsorption and electroosmotic flow in the pH range of 3-5. Electroosmotic flow decreased by raising the concentrations of the copolymers. Separation performance of ZB-004 layer is better than those of other two layers due to its low hydrophilicity, but with higher pH values, appreciable peak deformation and increase in electroosmosis were observed. The intra day and inter day migration reproducibility were investigated in terms of relative standard deviation (RSD) with four basic proteins at pH 4.0. The RSDs of the intra day migration times were less than 2%. The RSDs of the inter day migration times were less than 4%. At pH 5.0, the RSDs of the migration times in two ZB-004-coated capillaries made on two different days were less than 1%. Separation efficiencies of four basic proteins in a ZB-004-coated capillary which stored in a buffer (pH 4.0) for fifteen days after being used for 14 days decreased 15%. These coatings were stable and exhibited reproducible separations from intra day, inter day and inter column under acidic conditions.

  15. Selection of reversed-phase liquid chromatographic columns with diverse selectivity towards the potential separation of impurities in drugs.

    PubMed

    Van Gyseghem, E; Jimidar, M; Sneyers, R; Redlich, D; Verhoeven, E; Massart, D L; Vander Heyden, Y

    2004-07-01

    To select appropriate stationary phases from the continuously expanding supply of potentially suitable HPLC columns, the properties of 28 frequently applied stationary phases were determined by measuring several chromatographic parameters. From these results, based on chromatographic expertise, eight stationary phases with different properties and selectivities were selected. The aim of this study is to apply chemometric tools to evaluate the initially selected set of columns, i.e. a more systematic approach for making such a selection is examined. Starting from the information obtained on the 28 stationary phases, the re-evaluation was performed independently based on the chemometric techniques Pareto-optimality, principal component analysis (PCA), and Derringer's desirability functions. The aim was to select a set of efficient columns exhibiting large selectivity differences. The chemometrically selected stationary phases were divided in groups based on hydrophobicity, a critical retention-determining property in reversed-phase chromatography. This allowed to further reducing the selection to three columns. It is demonstrated that the selection by the chemometric approaches in general is fairly comparable with the initial selection.

  16. A fully automated and fast method using direct sample injection combined with fused-core column on-line SPE-HPLC for determination of ochratoxin A and citrinin in lager beers.

    PubMed

    Lhotská, Ivona; Šatínský, Dalibor; Havlíková, Lucie; Solich, Petr

    2016-05-01

    A new fast and sensitive method based on on-line solid-phase extraction on a fused-core precolumn coupled to liquid chromatography with fluorescence detection has been developed for ochratoxin A (OTA) and citrinin (CIT) determination in lager beer samples. Direct injection of 100 μL filtered beer samples into an on-line SPE-HPLC system enabled fast and effective sample extraction including separation in less than 6 min. Preconcentration of OTA and CIT from beer samples was performed on an Ascentis Express RP C18 guard column (5 × 4.6 mm), particle size 2.7 μm, with a mobile phase of methanol/0.5% aqueous acetic acid pH 2.8 (30:70, v/v) at a flow rate of 2.0 mL min(-1). The flow switch from extraction column to analytical column in back-flush mode was set at 2.0 min and the separation was performed on the fused-core column Ascentis Express Phenyl-Hexyl (100 × 4.6 mm), particle size 2.7 μm, with a mobile phase acetonitrile/0.5% aqueous acetic acid pH 2.8 in a gradient elution at a flow rate of 1.0 mL min(-1) and temperature of 50 °C. Fluorescence excitation/emission detection wavelengths were set at 335/497 nm. The accuracy of the method, defined as the mean recoveries of OTA and CIT from light and dark beer samples, was in the range 98.3-102.1%. The method showed high sensitivity owing to on-line preconcentration; LOQ values were found to be 10 and 20 ng L(-1) for OTA and CIT, respectively. The found values of OTA and CIT in all tested light, dark and wheat beer samples were significantly below the maximum tolerable limits (3.0 μg kg(-1) for OTA and 2000 μg kg(-1) for CIT) set by the European Union. PMID:26993307

  17. Tubular metal-organic framework-based capillary gas chromatography column for separation of alkanes and aromatic positional isomers.

    PubMed

    Fang, Zhi-Li; Zheng, Sheng-Run; Tan, Jing-Bo; Cai, Song-Liang; Fan, Jun; Yan, Xia; Zhang, Wei-Guang

    2013-04-12

    In this work, a tubular metal-organic framework, MOF-CJ3, with a large one-dimensional channel was chosen as stationary phase to prepare a capillary gas chromatographic column via a verified dynamic coating procedure. The column offered good separations of linear and branched alkanes, as well as aromatic positional isomers (ethylbenzene, xylene, cresol, hydroquinone, dichlorobenzene, bromobenzonitrile, chloronitrobenzene, and nitrotoluene) based on a combination of host-guest interactions and adsorption effects. Elution sequence of most of the analytes followed an increasing order of their boiling points, except for the separation of n-heptanes/isooctane, cresol, and hydroquinone isomers. Separation behavior of the column upon different organic substances may be related to the tubular pore structure of MOF-CJ3, in which the van der Waals forces between the alkanes and the hydrophobic inner surfaces might have great effect on separation of n-heptanes and isooctane, whereas the separation of cresol and hydroquinone isomers were affected by (OH⋯O) hydrogen bonds formed between the analytes and the 1,3,5-benzenetricarboxylate ligands on the pore wall. The effects of temperature on separation of aromatic positional isomers were investigated to elucidate entropy and enthalpy controlling of the separation process. PMID:23473507

  18. Tubular metal-organic framework-based capillary gas chromatography column for separation of alkanes and aromatic positional isomers.

    PubMed

    Fang, Zhi-Li; Zheng, Sheng-Run; Tan, Jing-Bo; Cai, Song-Liang; Fan, Jun; Yan, Xia; Zhang, Wei-Guang

    2013-04-12

    In this work, a tubular metal-organic framework, MOF-CJ3, with a large one-dimensional channel was chosen as stationary phase to prepare a capillary gas chromatographic column via a verified dynamic coating procedure. The column offered good separations of linear and branched alkanes, as well as aromatic positional isomers (ethylbenzene, xylene, cresol, hydroquinone, dichlorobenzene, bromobenzonitrile, chloronitrobenzene, and nitrotoluene) based on a combination of host-guest interactions and adsorption effects. Elution sequence of most of the analytes followed an increasing order of their boiling points, except for the separation of n-heptanes/isooctane, cresol, and hydroquinone isomers. Separation behavior of the column upon different organic substances may be related to the tubular pore structure of MOF-CJ3, in which the van der Waals forces between the alkanes and the hydrophobic inner surfaces might have great effect on separation of n-heptanes and isooctane, whereas the separation of cresol and hydroquinone isomers were affected by (OH⋯O) hydrogen bonds formed between the analytes and the 1,3,5-benzenetricarboxylate ligands on the pore wall. The effects of temperature on separation of aromatic positional isomers were investigated to elucidate entropy and enthalpy controlling of the separation process.

  19. Spino-olivary projections in the rat are anatomically separate from postsynaptic dorsal column projections.

    PubMed

    Flavell, Charlotte R; Cerminara, Nadia L; Apps, Richard; Lumb, Bridget M

    2014-06-15

    The gracile nucleus (GN) and lateral part of rostral dorsal accessory olive (rDAO) are important relays for indirect, postsynaptic dorsal column, and direct ascending pathways, respectively, that terminate as climbing fibers in the "hindlimb-receiving" parts of the C1 and C3 zones in the cerebellar cortex. While the spinal cells of origin of that project to GN and rDAO are from largely separate territories in the spinal cord, previous studies have indicated that there could be an area of overlap between these two populations in the medial dorsal horn. Given the access of these two ascending tracts to sensory (thalamic) versus sensorimotor (precerebellar) pathways, the present study therefore addresses the important question of whether or not individual neurons have the potential to contribute axons to both ascending pathways. A double-fluorescent tracer strategy was used in rats (red Retrobeads and Fluoro-Ruby or green Retrobeads and Fluoro-Emerald) to map the spatial distribution of cells of origin of the two projections in the lumbar spinal cord. The two pathways were found to receive input from almost entirely separate territories within the lumbar cord (levels L3-L5). GN predominantly receives input from lamina IV, while rDAO receives its input from three cell populations: medial laminae V-VI, lateral lamina V, and medial laminae VII-VIII. Cells that had axons that branched to supply both GN and rDAO represented only about 1% of either single-labeled cell population. Overall, the findings therefore suggest functional independence of the two ascending pathways. PMID:24357064

  20. Effects of the operation parameters on HILIC separation of flavonoids on zwitterionic column.

    PubMed

    Sentkowska, Aleksandra; Biesaga, Magdalena; Pyrzynska, Krystyna

    2013-10-15

    The hydrophilic interaction chromatography (HILIC) coupled to mass spectrometry was employed to study retention behavior of several flavonoids from their different groups using the polymeric zwitterionic stationary phase (ZIC-pHILIC). It contains sulfobetaine-bonded ligand with an inner positively charged quaternary ammonium and an outer negatively charged sulfonate functional groups. Two organic solvents - acetonitrile (ACN) and methanol (MeOH) - were compared as a component of mobile phase. Separation parameters such as a content of organic modifier, pH of an eluent and a column temperature were studied. Retention of flavonoids is controlled primarily by a partition between the mobile phase eluent and a water-enriched layer on the hydrophilic stationary phase with some contribution from hydrogen bonding formation. Using MeOH, in contrast to ACN, strongly retained compounds (myricetin, morin, rutin and quercetrin) could be eluted under isocratic conditions. A better sensitivity was achieved with MeOH as mobile phase component, particularly for quercetin, naringenin and kaempferol. The method was applied to the determination of flavonoids in fruit juices.

  1. Analogy between mission critical detection in distributed systems and 13C isotope separation column

    NASA Astrophysics Data System (ADS)

    Boca, Maria L.; Secara, Mihai

    2015-02-01

    Carbon represents the fourth most abundant chemical element in the world, having two stable and one radioactive isotope. The 13 Carbon isotopes, with a natural abundance of 1.1%, plays an important role in numerous applications, such as the study of human metabolism changes, molecular structure studies, non-invasive respiratory tests, Alzheimer tests, air pollution and global warming effects on plants [2]. Distributed systems are increasingly being applied in critical real-time applications and their complexity forces programmers to use design methods which guarantee correctness and increase the maintainability of the products. Objectoriented methodologies are widely used to cope with complexity in any kind of system, but most of them lack a formal foundation to allow the analysis and verification of designs, which is one of the main requirements for dealing with concurrent and reactive systems. This research is intended to make an analogy between two tips of industrial processes, one 13C Isotope Separation Column and other one distributed systems. We try to highlight detection of "mission critical "situations for this two processes and show with one is more critical and needs deeply supervisyon [1], [3].

  2. Determination for multiple mycotoxins in agricultural products using HPLC-MS/MS via a multiple antibody immunoaffinity column.

    PubMed

    Zhang, Zhaowei; Hu, Xiaofeng; Zhang, Qi; Li, Peiwu

    2016-05-15

    Mycotoxins usually found in agricultural products such as peanut, corn, and wheat, are a serious threat to human health and their detection requires multiplexed and sensitive analysis methods. Herein, a simultaneous determination for aflatoxin B1, B2, G1, G2, ochratoxin A, zearalanone and T-2 toxin was investigated using high performance liquid chromatography coupled with tandem mass spectrometry in a single run via a home-made multiple immunoaffinity column. Four monoclonal antibodies were produced in our lab against aflatoxins, ochratoxin A, zearalanone and T-2 toxin, respectively, then combined as a pool and bound to Sepharose-4B for affinity chromatography. Seven mycotoxins were effectively extracted from the agricultural product samples by using acetonitrile/water/acetic acid (80:19:1, v/v/v) Then, the extraction was cleanup by multiple immunoaffinity column. This method demonstrated a considerable linear range of 0.30-25, 0.12-20, 0.30-20, 0.12-20, 0.60-30, 0.30-25, and 1.2-40μgkg(-1)and lower limits of detection at 0.1, 0.04, 0.1, 0.04, 0.2, 0.1 and 0.4μgkg(-1) for AFB1, AFB2, AFG1, AFG2, OTA, ZEN and T-2, respectively, in comparison with previously reported methods, as well as excellent recoveries. The mIAC capacity for AFB1, AFB2, AFG1, AFG2, OTA, ZEN, and T-2 were 187, 181, 153, 151, 105, 130, 88ng, respectively. It was found that all of the 7 mycotoxins were present in 90 agricultural product samples. The proposed method meets the requirements for rapid sample preparation and highly sensitive identification of multiple mycotoxins in agricultural product and food safety. This method provides a promising alternative with high throughput and high sensitivity for rapid analysis of seven mycotoxins in the monitoring of food safety. PMID:26948441

  3. Enantiomeric separations of illicit drugs and controlled substances using cyclofructan-based (LARIHC) and cyclobond I 2000 RSP HPLC chiral stationary phases.

    PubMed

    Padivitage, Nilusha L T; Dodbiba, Edra; Breitbach, Zachary S; Armstrong, Daniel W

    2014-06-01

    Recently a novel class of chiral stationary phases (CSPs) based on cyclofructan (CF) has been developed. Cyclofructans are cyclic oligosaccharides that possess a crown ether core and pendent fructofuranose moieties. Herein, we evaluate the applicability of these novel CSPs for the enantiomeric separation of chiral illicit drugs and controlled substances directly without any derivatization. A set of 20 racemic compounds were used to evaluate these columns including 8 primary amines, 5 secondary amines, and 7 tertiary amines. Of the new cyclofructan-based LARIHC columns, 14 enantiomeric separations were obtained including 7 baseline and 7 partial separations. The LARIHC CF6-P column proved to be the most useful in separating illicit drugs and controlled substances accounting for 11 of the 14 optimized separations. The polar organic mode containing small amounts of methanol in acetonitrile was the most useful solvent system for the LARIHC CF6-P CSP. Furthermore, the LARIHC CF7-DMP CSP proved to be valuable for the separation of the tested chiral drugs resulting in four of the optimized enantiomeric separations, whereas the CF6-RN did not yield any optimum separations. The broad selectivity of the LARIHC CF7-DMP CSP is evident as it separated primary, secondary and tertiary amine containing chiral drugs. The compounds that were partially or un-separated using the cyclofructan based columns were screened with a Cyclobond I 2000 RSP column. This CSP provided three baseline and six partial separations.

  4. Preparation of a novel ionic hybrid stationary phase by non-covalent functionalization of single-walled carbon nanotubes with amino-derivatized silica gel for fast HPLC separation of aromatic compounds.

    PubMed

    Aral, Hayriye; Çelik, K Serdar; Aral, Tarık; Topal, Giray

    2016-03-01

    Single-walled carbon nanotubes (SWCNTs) were immobilized on spherical silica gel with a 4-μm average particle size and a 60-Å average pore size. The amino-derivatized silica gel was non-covalently coated with carboxylated SWCNTs to preserve the structure of the nanotubes and their physico-chemical properties. The novel ionic hybrid stationary phase was characterized by scanning electron microscopy (SEM), infra-red (IR) spectroscopy and elemental analysis, and then, it was used to fill an empty 150×4.6mm(2) high-performance liquid chromatography (HPLC) column. Chromatographic parameters, such as the theoretical plate number, retention factor and peak asymmetry factor, and analytical parameters, such as the limit of detection (LOD), limit of quantification (LOQ), linear range, calibration equation, and R(2) value, and quantitative analysis parameters were calculated for all of the analytes. Using different mobile phases, five different classes of aromatic hydrocarbons were separated in a very short analysis time of 4-8min. Furthermore, a high theoretical plate number (up to 25000) and an excellent peak asymmetry factor (1.0) were obtained. The results showed that the surface of the SWNTs had very strong interactions with aromatic groups, therefore providing high selectivity for the separation of different classes of aromatic compounds. This study indicates that SWCNTs enable the extension of the application range of the newly prepared stationary phases for the fast separation of aromatic compounds by HPLC.

  5. Mass spectrometry and NMR spectroscopy: modern high-end detectors for high resolution separation techniques--state of the art in natural product HPLC-MS, HPLC-NMR, and CE-MS hyphenations.

    PubMed

    Seger, Christoph; Sturm, Sonja; Stuppner, Hermann

    2013-07-01

    Current natural product research is unthinkable without the use of high resolution separation techniques as high performance liquid chromatography or capillary electrophoresis (HPLC or CE respectively) combined with mass spectrometers (MS) or nuclear magnetic resonance (NMR) spectrometers. These hyphenated instrumental analysis platforms (CE-MS, HPLC-MS or HPLC-NMR) are valuable tools for natural product de novo identification, as well as the authentication, distribution, and quantification of constituents in biogenic raw materials, natural medicines and biological materials obtained from model organisms, animals and humans. Moreover, metabolic profiling and metabolic fingerprinting applications can be addressed as well as pharmacodynamic and pharmacokinetic issues. This review provides an overview of latest technological developments, discusses the assets and drawbacks of the available hyphenation techniques, and describes typical analytical workflows.

  6. Validation of an HPLC analytical method coupled to a multifunctional clean-up column for the determination of deoxynivalenol.

    PubMed

    Sugita-Konsihi, Yoshiko; Tanaka, Toshitsugu; Tabata, Setsuko; Nakajima, Masahiro; Nouno, Masanori; Nakaie, Yoko; Chonan, Takao; Aoyagi, Mitsutoshi; Kibune, Nobuyuki; Mizuno, Kazutoshi; Ishikuro, Eiichi; Kanamaru, Naoki; Minamisawa, Masatoshi; Aita, Norio; Kushiro, Masayo; Tanaka, Kenji; Takatori, Kosuke

    2006-04-01

    To evaluate a method using a multifunctional clean-up column coupled with high performance liquid chromatography as an official analytical method for the determination of deoxynivalenol in wheat used as food or feed, an inter-laboratory study was performed in 12 laboratories using four naturally contaminated wheat samples and one spiked sample. The relative standard deviations for repeatability (RSDr) and reproducibility (RSDR) of naturally contaminated wheat were in the range 5.8-11.3% and 12.0-20.7%, respectively. The HORRAT was less than 1.0 in each sample. From the spiking test, the recovery rate, RSDr, RSDR and HORRAT value were 100.0%, 11.2%, 10.3% and 0.5, respectively. The limit of quantification is 0.10 mg/kg from the range obtained in a linear calibration. Thus, it should be useful as a sensitive and validated analytical method for the determination of deoxynivalenol in wheat intended for use in food and feed.

  7. Quantification of Five Clinically Important Amino Acids by HPLC-Triple TOF™ 5600 Based on Pre-column Double Derivatization Method.

    PubMed

    Deng, Shuang; Scott, David; Garg, Uttam

    2016-01-01

    Phenylalanine, tyrosine, glycine, cystine, and phosphoethanolamine are commonly measured amino acids in various physiological fluids to diagnose or follow-up various inborn errors of metabolism. The gold standard method for the amino acids quantitation has been ion exchange chromatography with ninhydrin post-column derivatization. However, this method is very laborious and time consuming. In recent years, liquid-chromatography mass spectrometry is being increasingly used for the assay of amino acids. Pre-column butyl derivatization with reverse phase chromatography has been widely used for mass spectrometry analysis of amino acids. Phosphoethanolamine is not butylated and cannot be measured by this method. Nevertheless, phosphoethanolamine can be dansyl-derivatized using dansyl chloride. We developed a double derivatization method by using butanol and dansyl chloride to derivatize carboxylic and amino groups separately, and then combining the derivatives to simultaneously measure these five amino acids using TOF-MS scan. Stable isotope-labeled internal standards were used. PMID:26602116

  8. Quantification of Five Clinically Important Amino Acids by HPLC-Triple TOF™ 5600 Based on Pre-column Double Derivatization Method.

    PubMed

    Deng, Shuang; Scott, David; Garg, Uttam

    2016-01-01

    Phenylalanine, tyrosine, glycine, cystine, and phosphoethanolamine are commonly measured amino acids in various physiological fluids to diagnose or follow-up various inborn errors of metabolism. The gold standard method for the amino acids quantitation has been ion exchange chromatography with ninhydrin post-column derivatization. However, this method is very laborious and time consuming. In recent years, liquid-chromatography mass spectrometry is being increasingly used for the assay of amino acids. Pre-column butyl derivatization with reverse phase chromatography has been widely used for mass spectrometry analysis of amino acids. Phosphoethanolamine is not butylated and cannot be measured by this method. Nevertheless, phosphoethanolamine can be dansyl-derivatized using dansyl chloride. We developed a double derivatization method by using butanol and dansyl chloride to derivatize carboxylic and amino groups separately, and then combining the derivatives to simultaneously measure these five amino acids using TOF-MS scan. Stable isotope-labeled internal standards were used.

  9. Miniaturized monolithic columns for the electrochromatographic separation and SERS detection of molecules of exobiological interest

    NASA Astrophysics Data System (ADS)

    Carbonnier, Benjamin; Guerrouache, Mohamed

    Development of miniaturized separation and detection media represents one of the major challenges in the field of modern analytical chemistry dedicated to space exploration. To date, gas chromatography-mass spectrometry has been selected as the method of choice for exobiology flight experiments for seeking for organic molecules and especially potential chemical indicators of life. [1] Liquid phase separation methods have also been developed with for instance, the so-called Mars Organic Analyzer (MOA) capillary electrophoresis (CE) microchip.[2] Although CE offers the advantages of easy miniaturization and high separation efficiency it suffers from a lack of selectivity towards a broad range of analytes with varied chemical nature. In this respect, we propose the use of capillary columns filled with monolithic stationary phases for the electrochromatographic separation of organic molecules of exobiology interest. Polymer monoliths have attracted a great deal of interest in analytical science over the last years as (electro)chromatographic stationary phases [3], immunosensors [4]. Beyond the intrinsic properties of monolithic polymers, i.e. fast mass transport between the monolithic support and the surrounding fluid and high permeability, other major advantages are their easy in situ preparation and tuning of surface functionality. Indeed, monoliths can be simply prepared through free radical copolymerization of a homogeneous mixture made of monomers, cross-linkers, porogenic solvents and initiator. UV-initiation process has been exploited to the synthesis of a discrete section of monolith as a flow-through active element within the confines of micro channels [5,6] while two-step strategies have been reported for the design of varied adsorbent starting with a generic monolith [7,8]. Although a nearly limitless range of monolithic supports can be prepared by this traditional method, the resulting monoliths exhibit unique function. In this contribution, we describe an

  10. High-performance liquid chromatography separation of unsaturated organic compounds by a monolithic silica column embedded with silver nanoparticles.

    PubMed

    Zhu, Yang; Morisato, Kei; Hasegawa, George; Moitra, Nirmalya; Kiyomura, Tsutomu; Kurata, Hiroki; Kanamori, Kazuyoshi; Nakanishi, Kazuki

    2015-08-01

    The optimization of a porous structure to ensure good separation performances is always a significant issue in high-performance liquid chromatography column design. Recently we reported the homogeneous embedment of Ag nanoparticles in periodic mesoporous silica monolith and the application of such Ag nanoparticles embedded silica monolith for the high-performance liquid chromatography separation of polyaromatic hydrocarbons. However, the separation performance remains to be improved and the retention mechanism as compared with the Ag ion high-performance liquid chromatography technique still needs to be clarified. In this research, Ag nanoparticles were introduced into a macro/mesoporous silica monolith with optimized pore parameters for high-performance liquid chromatography separations. Baseline separation of benzene, naphthalene, anthracene, and pyrene was achieved with the theoretical plate number for analyte naphthalene as 36,000 m(-1). Its separation function was further extended to cis/trans isomers of aromatic compounds where cis/trans stilbenes were chosen as a benchmark. Good separation of cis/trans-stilbene with separation factor as 7 and theoretical plate number as 76,000 m(-1) for cis-stilbene was obtained. The trans isomer, however, is retained more strongly, which contradicts the long- established retention rule of Ag ion chromatography. Such behavior of Ag nanoparticles embedded in a silica column can be attributed to the differences in the molecular geometric configuration of cis/trans stilbenes.

  11. HPLC method development for the online-coupling of chromatographic Perilla frutescens extract separation with xanthine oxidase enzymatic assay.

    PubMed

    Kaufmann, Christine M; Grassmann, Johanna; Letzel, Thomas

    2016-05-30

    Enzyme-regulatory effects of compounds contained in complex mixtures can be unveiled by coupling a continuous-flow enzyme assay to a chromatographic separation. A temperature-elevated separation was developed and the performance was tested using Perilla frutescens plant extracts of various polarity (water, methanol, ethanol/water). Owning to the need of maintaining sufficient enzymatic activity, only low organic solvent concentrations can be added to the mobile phase. Hence, to broaden the spectrum of eluting compounds, two different organic solvents and various contents were tested. The chromatographic performance and elution was further improved by the application of a moderate temperature gradient to the column. By taking the effect of eluent composition as well as calculated logD values and molecular structure of known extract compounds into account, unknown features were tentatively assigned. The method used allowed the successful observation of an enzymatic inhibition caused by P. frutescens extract. PMID:26986639

  12. Research on the separation properties of empty-column gas chromatography (EC-GC) and conditions for simulated distillation (SIMDIS).

    PubMed

    Boczkaj, Grzegorz; Kamiński, Marian

    2013-10-01

    Previous studies have revealed it is possible to separate a high-boiling mixture by gas chromatography in empty fused-silica capillary tubing rather than in columns coated with stationary phase. Chromatographic separation occurs solely on the basis of the different boiling points of the substances separated. The high similarity of such separations to those in classic distillation seems advantageous when gas chromatography is used for simulated distillation. This paper presents results from further research on the separation properties of empty fused silica tubing. The efficiency of this chromatographic system has been examined. The usefulness of such conditions has been studied for simulated distillation, i.e. to determine the boiling-point distribution of complex mixtures, mainly petroleum fractions and products, on the basis of their retention relative to reference substances. The results obtained by use of empty-column gas chromatography (EC-GC) and by use of classical simulated distillation columns have been compared for solutes of different polarity. Studies revealed boiling points determined by EC-GC were more accurate than those obtained by the standard method of simulated distillation.

  13. Research on the separation properties of empty-column gas chromatography (EC-GC) and conditions for simulated distillation (SIMDIS).

    PubMed

    Boczkaj, Grzegorz; Kamiński, Marian

    2013-10-01

    Previous studies have revealed it is possible to separate a high-boiling mixture by gas chromatography in empty fused-silica capillary tubing rather than in columns coated with stationary phase. Chromatographic separation occurs solely on the basis of the different boiling points of the substances separated. The high similarity of such separations to those in classic distillation seems advantageous when gas chromatography is used for simulated distillation. This paper presents results from further research on the separation properties of empty fused silica tubing. The efficiency of this chromatographic system has been examined. The usefulness of such conditions has been studied for simulated distillation, i.e. to determine the boiling-point distribution of complex mixtures, mainly petroleum fractions and products, on the basis of their retention relative to reference substances. The results obtained by use of empty-column gas chromatography (EC-GC) and by use of classical simulated distillation columns have been compared for solutes of different polarity. Studies revealed boiling points determined by EC-GC were more accurate than those obtained by the standard method of simulated distillation. PMID:23925798

  14. Control of selectivity via nanochemistry: monolithic capillary column containing hydroxyapatite nanoparticles for separation of proteins and enrichment of phosphopeptides.

    PubMed

    Krenkova, Jana; Lacher, Nathan A; Svec, Frantisek

    2010-10-01

    New monolithic capillary columns with embedded commercial hydroxyapatite nanoparticles have been developed and used for protein separation and selective enrichment of phosphopeptides. The rod-shaped hydroxyapatite nanoparticles were incorporated into the poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) monolith by simply admixing them in the polymerization mixture followed by in situ polymerization. The effect of percentages of monomers and hydroxyapatite nanoparticles in the polymerization mixture on the performance of the monolithic column was explored in detail. We found that the loading capacity of the monolith is on par with other hydroxyapatite separation media. However, the speed at which these columns can be used is higher due to the fast mass transport. The function of the monolithic columns was demonstrated with the separations of a model mixture of proteins including ovalbumin, myoglobin, lysozyme, and cytochrome c as well as a monoclonal antibody and its aggregates with protein A. Selective enrichment and MALDI/MS characterization of phosphopeptides fished-out from complex peptide mixtures of ovalbumin, α-casein, and β-casein digests were also achieved using the hydroxyapatite monolith.

  15. Simultaneous determination of four aflatoxins and ochratoxin A in ginger and related products by HPLC with fluorescence detection after immunoaffinity column clean-up and postcolumn photochemical derivatization.

    PubMed

    Wen, Jing; Kong, Weijun; Wang, Jian; Yang, Meihua

    2013-12-01

    Ginger, a widely used spice and traditional Chinese medicine, is prone to be contaminated by mycotoxins. A simple, sensitive, and reproducible method based on immunoaffinity column clean-up coupled with HPLC and on-line postcolumn photochemical derivatization with fluorescence detection was developed for the simultaneous determination of aflatoxins (AFs) B1 , B2 , G1 , G2 , and ochratoxin A (OTA) in 25 batches of gingers and related products marketed in China for the first time. The samples were first extracted by ultrasonication with methanol/water (80:20, v/v) and then cleaned up with immunoaffinity columns for analysis. Under the optimized conditions, the LODs and LOQs for the five mycotoxins were 0.03-0.3 and 0.1-0.9 μg/kg, respectively. The average recoveries ranged from 81.3-100.8% for AFs and from 88.6-99.5% for OTA at three spiking levels. Good linearity was observed for the analytes with correlation coefficients all >0.9995. All moldy gingers were contaminated with at least one kind of the five investigated mycotoxins, while none of them were found in normal gingers. Ginger powder samples were contaminated slightly with the contamination levels below the LOQs, while ginger tea bags were mainly contaminated by OTA at 1.05-1.19 μg/kg and ginger black tea bags were mainly contaminated by AFs at 3.37-5.76 μg/kg. All the contamination levels were below the legally allowable limits.

  16. Separation of 2,3-butylene glycol and acetoin in fermented cheese whey permeate by liquid column chromatography

    SciTech Connect

    Lippi, M.S.

    1987-01-01

    While use of 2,3-butylene glycol could relieve pressure on consumption of petroleum-derived feedstocks, the economics of producing 2,3-butylene glycol by fermentation are still cost prohibitive. One of the main reasons for this is the high cost of recovering the 2,3-butylene glycol from the aqueous fermentation broth. The research presented here involves utilizing a low cost liquid column chromatographic operation for separating 2,3-butylene glycol and acetoin (another major by-product of the fermentation), in fermented cheese whey permeate. The procedure involves prewashing the column with an inexpensive solvent (aqueous sodium borate solution), and eluting samples with distilled and deionized water. Plain tap water was also shown to work equally well as the eluent. Separating 2,3-butylene glycol into the water eluent should improve the economics of the recovery process. The lower boiling water can be evaporated and distilled leaving the high boiling 2,3-butylene glycol (boiling point of 183 C). Steam generation and equipment specifications would be reduced thereby decreasing both capital and maintenance expenditures. Studies were performed and parameters were optimized on a laboratory scale and then scaled-up. Best results on the lab-scale was that a 54 ml separation was obtained from a 100 ml sample of the two compounds on a column 15 cm by 2.6 cm. Best results on the larger column showed that a one liter sample of ultrafiltered fermented cheese whey permeate containing 900 micrograms/ml of 2,3-butylene glycol and 300 micrograms/ml of acetoin was completely separated on a 20 cm by 11.4 cm column bed of Dowex 1-X8 anion-exchange resin.

  17. Evaluating the interactions of organic compounds with multi-walled carbon nanotubes by self-packed HPLC column and linear solvation energy relationship.

    PubMed

    Chu, Yingqian; Li, Xuehua; Xie, Hongbin; Fu, Zhiqiang; Yang, Xianhai; Qiao, Xianliang; Cai, Xiyun; Chen, Jingwen

    2013-12-15

    Understanding the interactions between organic pollutants and carbon nanotubes (CNTs) is critical for fate assessment of both CNTs and organic pollutants. In this study, the chromatographic approach was introduced based on CNTs as stationary phase for the evaluation of such interactions. The pristine multi-walled carbon nanotubes (MWCNTs) were packed into columns of high-pressure liquid chromatography (HPLC) and the retention factors (k') were determined to characterize the adsorption affinity of organic compounds onto MWCNTs. Nine compounds were tested. The results showed that their lnk' values followed the order: benzene < toluene < phenol < chlorobenzene < bromobenzene < aniline < sulfamethoxazole < sulfadiazine ≈ sulfadimidine. The linear solvation energy relationship (LSER) theory was adopted to correlate lnk' with the molecular solvatochromic parameters. We found that lnk' of the studied compounds correlate positively with molecular polarizability (E) significantly, suggesting that the π-/n-electrons-dependent polarizable interactions play a major role for the adsorption. Moreover, the thermodynamic parameters calculated from van't Hoff equations revealed that the interactions between the compounds and MWCNTs were spontaneous and exothermic processes.

  18. Hollow fiber-based liquid-liquid-liquid microextraction followed by flow injection analysis using column-less HPLC for the determination of phenazopyridine in plasma and urine.

    PubMed

    Saraji, Mohammad; Bidgoli, Ali Akbar Hajialiakbari; Farajmand, Bahman

    2011-07-01

    Hollow fiber-based liquid-liquid-liquid microextraction (HF-LLLME) followed by flow injection analysis and diode array detection (FIA-DAD) was applied as a simple and sensitive quantitative method for the determination of phenazopyridine in urine and plasma samples. Flow injection system included a conventional HPLC system (without a chromatographic column) and a diode array detector. The extraction of phenazopyridine was carried out using diphenyl ether as the organic phase for filling the pores of the hollow fiber wall, and 0.1 M H(2)SO(4) solution as acceptor phase in the lumen of the fiber. The factors affecting the HF-LLLME and flow injection analysis including type of organic solvent, pH of donor phase, extraction temperature, extraction time, stirring rate, and pH of mobile phase were investigated and the optimal extraction conditions were established. With the consumption of 5 mL of sample solution, the enrichment factor was about 230. The limit of detection was 0.5 μg/L with inter- and intra-day precision being (RSD%) 6.9 and 4.9, respectively. Excellent linearity was found between 5 and 200 μg/L.

  19. Improved Separation of Complex Polycyclic Aromatic Hydrocarbon Mixtures Using Novel Column Combinations in GC×GC/ToF-MS

    PubMed Central

    Manzano, Carlos; Hoh, Eunha; Simonich, Staci L. Massey

    2012-01-01

    Complex mixtures of polycyclic aromatic hydrocarbons (PAHs) are difficult to resolve because of the high degree of overlap in compound vapor pressures, boiling points and mass spectral fragmentation patterns. The objective of this research was to improve the separation of complex PAH mixtures (including 97 different parent, alkyl-, nitro-, oxy-, thio-, chloro-, bromo-, and high molecular weight PAHs) using GC×GC/ToF-MS by maximizing the orthogonality of different GC column combinations and improving the separation of PAHs from the sample matrix interferences, including unresolved complex mixtures (UCM). Four different combinations of non-polar, polar, liquid crystal and nano-stationary phase columns were tested. Each column combination was optimized and evaluated for orthogonality using a method based on conditional entropy that considers the quantitative peak distribution in the entire two-dimensional space. Finally, an atmospheric particulate matter with diameter < 2.5 µm (PM2.5) sample from Beijing, China, a soil sample from St. Maries Creosote Superfund Site, and a sediment sample from the Portland Harbor Superfund Site were analyzed for complex mixtures of PAHs. The highest chromatographic resolution, lowest synentropy, highest orthogonality and lowest interference from UCM were achieved using a 10 m × 0.15 mm × 0.10 µm LC-50 liquid crystal column in the first dimension and a 1.2 m × 0.10 mm × 0.10 µm NSP-35 nano-stationary phase column in the second dimension. In addition, the use of this column combination in GC×GC/ToF-MS resulted in significantly shorter analysis times (176 min) for complex PAH mixtures compared to one-dimensional GC/MS (257 min), as well as potentially reduced sample preparation time. PMID:22769970

  20. [Rapid determination of 137Cs in environmental samples--purification of 137Cs by ammonium molybdophosphate column separation].

    PubMed

    Nonaka, N; Sato, K; Higuchi, H; Hamaguchi, H

    1976-10-01

    A rapid method for the determination of 137Cs in environmental samples was proposed. The principal technic employed in this study is based on column separation of 137Cs using ammonium molybdophosphate mixed with glass fiber to eliminate contribution of natural radionuclides such as 40K and 87Rb. The separation of cesium from potassium and rubidium was performed by the elution with 0.5m ammonium nitrate solution. The time required for separation of cesium was five hours as compared with the conventional cation exchange separation which required thirteen hours. The chemical yield of cesium carrier was normally more than 90 percent. The results obtained were compared with that by the conventional methods using Bio-Rex cation exchange separation and the good agreement between the two methods was obtained. PMID:1037401

  1. Separation of aromatic carboxylic acids using quaternary ammonium salts on reversed-phase HPLC. 2. Application for the analysis of Loy Yang coal oxidation products

    SciTech Connect

    Kawamura, K.; Okuwaki, A.; Verheyen, T.V.; Perry, G.J.

    2006-07-01

    In order to develop separation processes and analytical methods for aromatic carboxylic acids for the coal oxidation products, the separation behavior of aromatic carboxylic acids on a reversed-phase HPLC using eluent containing quaternary ammonium salt was optimized using the solvent gradient method. This method was applied for the analysis of Loy Yang coal oxidation products. It was confirmed that the analytical data using this method were consistent with those determined using gas chromatography.

  2. Countercurrent Chromatographic Separation of Proteins Using an Eccentric Coiled Column with Synchronous and Nonsynchronous Type-J Planetary Motions

    PubMed Central

    SHINOMIYA, Kazufusa; YOSHIDA, Kazunori; TOKURA, Koji; TSUKIDATE, Etsuhiro; YANAGIDAIRA, Kazuhiro; ITO, Yoichiro

    2015-01-01

    Protein separation was performed using the high-speed counter-current chromatograph (HSCCC) at both synchronous and nonsynchronous type-J planetary motions. The partition efficiency was evaluated with two different column configurations, eccentric coil and toroidal coil, on the separation of a set of stable protein samples including cytochrome C, myoglobin and lysozyme with a polymer phase system composed of 12.5% (w/w) polyethylene glycol 1000 and 12.5% (w/w) dibasic potassium phosphate. Better peak resolution was obtained by the eccentric coil than by the toroidal coil using either lower or upper phase as the mobile phase. The peak resolution was further improved using the eccentric coil by the nonsynchronous type-J planetary motion with the combination of 1066 rpm of column rotation and 1000 rpm of revolution. PMID:25765276

  3. HPLC separation of human serum albumin isoforms based on their isoelectric points.

    PubMed

    Turell, Lucía; Botti, Horacio; Bonilla, Lucía; Torres, María José; Schopfer, Francisco; Freeman, Bruce A; Armas, Larissa; Ricciardi, Alejandro; Alvarez, Beatriz; Radi, Rafael

    2014-01-01

    Human serum albumin (HSA) is the most abundant protein in plasma. Cys34, the only free Cys residue, is the predominant plasma thiol and a relevant sacrificial antioxidant. Both in vivo circulating HSA and pharmaceutical preparations are heterogeneous with respect to the oxidation state of Cys34. In this work, we developed an external pH gradient chromatofocusing procedure that allows the analysis of the oxidation status of HSA in human plasma and biopharmaceutical products based on the different apparent isoelectric points and chemical properties of the redox isoforms. Specifically, reduced-mercury blocked HSA (HSA-SHg(+)), HSA with Cys34 oxidized to sulfenic acid (HSA-SOH) and HSA oxidized to sulfinate anion (HSA-SO2(-)) can be separated with resolutions of 1.4 and 3.1 (first and last pair) and hence quantified and purified. In addition, an N-terminally degraded isoform (HSA3-585) in different redox states can be resolved as well. Confirmation of the identity of the chromatofocusing isolated isoforms was achieved by high resolution whole protein MS. It is proposed that the chromatofocusing procedure can be used to produce more exact and complete descriptions of the redox status of HSA in vivo and in vitro. Finally, the scalability capabilities of the chromatofocusing procedure allow for the preparation of highly pure standards of several redox isoforms of HSA.

  4. HPLC separation of human serum albumin isoforms based on their isoelectric points.

    PubMed

    Turell, Lucía; Botti, Horacio; Bonilla, Lucía; Torres, María José; Schopfer, Francisco; Freeman, Bruce A; Armas, Larissa; Ricciardi, Alejandro; Alvarez, Beatriz; Radi, Rafael

    2014-01-01

    Human serum albumin (HSA) is the most abundant protein in plasma. Cys34, the only free Cys residue, is the predominant plasma thiol and a relevant sacrificial antioxidant. Both in vivo circulating HSA and pharmaceutical preparations are heterogeneous with respect to the oxidation state of Cys34. In this work, we developed an external pH gradient chromatofocusing procedure that allows the analysis of the oxidation status of HSA in human plasma and biopharmaceutical products based on the different apparent isoelectric points and chemical properties of the redox isoforms. Specifically, reduced-mercury blocked HSA (HSA-SHg(+)), HSA with Cys34 oxidized to sulfenic acid (HSA-SOH) and HSA oxidized to sulfinate anion (HSA-SO2(-)) can be separated with resolutions of 1.4 and 3.1 (first and last pair) and hence quantified and purified. In addition, an N-terminally degraded isoform (HSA3-585) in different redox states can be resolved as well. Confirmation of the identity of the chromatofocusing isolated isoforms was achieved by high resolution whole protein MS. It is proposed that the chromatofocusing procedure can be used to produce more exact and complete descriptions of the redox status of HSA in vivo and in vitro. Finally, the scalability capabilities of the chromatofocusing procedure allow for the preparation of highly pure standards of several redox isoforms of HSA. PMID:24316526

  5. Separation and characterization of oxaliplatin dinucleotides from DNA using HPLC-ESI ion trap mass spectrometry.

    PubMed

    Mowaka, Shereen; Linscheid, Michael

    2008-11-01

    Oxaliplatin is a third-generation platinum complex, and has a broad spectrum of antitumor activity. Such platinum complexes with the DACH carrier ligand have recently received increasing attention since they show efficacy against cisplatin-resistant cell lines. As the foremost indication of antitumor activity of platinum drugs is the formation of adducts with genomic DNA, calf thymus DNA-oxaliplatin adducts were the major target in this study. Calf thymus DNA was incubated with oxaliplatin, resulting in the formation of a large number of platinum-DNA adducts. Treated DNA was digested into the dinucleotides with a combination of enzymes, namely, benzonase, alkaline phosphatase, and nuclease S1. Using a high-performance liquid chromatography, we carried out the separation of individual platinum-DNA adducts which were concurrently identified using electrospray ionization ion trap mass spectrometry (MS). Both 1,2-intrastrand and 1,2-interstrand cross-linked adducts were found; however, those of the intrastrand nature have a considerably higher abundance than those of the interstrand cross-links. Among them, d(GpG)-oxaliplatin was the most abundant bifuctional adduct. To a lesser extent, a few monofunctional adducts were detected as well. MS(n) experiments served to ascertain the detailed structures of oxaliplatin adducts of dinucleoside monophosphates and of dinucleotides.

  6. Detailed characterization of the kinetic performance of first and second generation silica monolithic columns for reversed-phase chromatography separations.

    PubMed

    Cabooter, Deirdre; Broeckhoven, Ken; Sterken, Roman; Vanmessen, Alison; Vandendael, Isabelle; Nakanishi, Kazuki; Deridder, Sander; Desmet, Gert

    2014-01-17

    The kinetic performance of commercially available first generation and prototype second generation silica monoliths has been investigated for 2.0mm and 3.0-3.2mm inner diameter columns. It is demonstrated that the altered sol-gel process employed for the production of second generation monoliths results in structures with a smaller characteristic size leading to an improved peak shape and higher efficiencies. The permeability of the columns however, decreases significantly due to the smaller throughpore and skeleton sizes. Scanning electron microscopy pictures suggest the first generation monoliths have cylindrical skeleton branches, whereas the second generation monoliths rather have skeleton branches that resemble a single chain of spherical globules. Using recently established correlations for the flow resistance of cylindrical and globule chain type monolithic structures, it is demonstrated that the higher flow resistance of the second generation monoliths can be entirely attributed to their smaller skeleton sizes, which is also evident from the external porosity that is largely the same for both monolith generations (ɛe∼0.65). The recorded van Deemter plots show a clear improvement in efficiency for the second generation monoliths (minimal plate heights of 13.6-14.1μm for the first and 6.5-8.2μm for the second generation, when assessing the plate count using the Foley-Dorsey method). The corresponding kinetic plots, however, indicate that the much reduced permeability of the second generation monoliths results in kinetic performances (time needed to achieve a given efficiency) which are only better than those of the first generation for plate counts up to N∼45,000. For more complex samples (N≥50,000), the first generation monoliths can intrinsically still provide faster analysis due to their high permeability. It is also demonstrated that - despite the improved efficiency of the second generation monoliths in the practical range of separations (N=10

  7. Comparative analysis of astaxanthin and its esters in the mutant E1 of Haematococcus pluvialis and other green algae by HPLC with a C30 column.

    PubMed

    Peng, Juan; Xiang, WenZhou; Tang, QuanMing; Sun, Ni; Chen, Feng; Yuan, JianPing

    2008-12-01

    A gradient reversed-phase high-performance liquid chromatography (HPLC) method using a C30 column was developed for the simultaneous determination of astaxanthin, astaxanthin monoesters and astaxanthin diesters in the green algae Chlorococcum sp., Chlorella zofingiensis, Haematococcus pluvialis and the mutant E1, which was obtained from the mutagenesis of H. pluvialis by exposure to UV-irradiation and ethyl methanesulphonate (EMS) with subsequent screening using nicotine. The results showed that the contents of total astaxanthins including free astaxanthin and astaxanthin esters ranged from 1.4 to 30.9 mg/g dry biomass in these green algae. The lower total astaxanthin levels (< 2 mg/g dry biomass) were detected in the green algae Chlorococcum sp. and C. zofingiensis. The higher total astaxanthin levels (>16 mg/g dry biomass) were found in the green alga H. pluvialis and its mutant E1. It is notable that the mutant E1 is found to have considerably higher amounts of total astaxanthin (30.9 mg/g) as compared to the wild strain of H. pluvialis (16.1 mg/g). This indicates that UV-irradiation and EMS compound mutagenesis with subsequent screening using nicotine is an effective method for breeding of a high-producing astaxanthin strain of H. pluvialis. In addition, the green alga C. zofingiensis had a remarkably higher percentage of astaxanthin diesters (76.3% of total astaxanthins) and a remarkably lower percentage of astaxanthin monoesters (18.0% of total astaxanthins) in comparison with H. pluvialis (35.5% for diesters and 60.9% for monoesters), the mutant E1 (49.1% and 48.1%) and Chlorococcum sp. (18.0% and 58.6%).

  8. A Study of Multistage/Multifunction Column for Fine Particle Separation.

    SciTech Connect

    Chiang, S.

    1997-09-15

    Hydrodynamic tests were continued in this quarter. Liquid circulation velocities are the characteristic parameters in the multistage column. Conductivity tracer response method has been set up for liquid circulation velocities measurement. The period of dampened sinusoidal conductivity signals can be clearly identified and then converted into linear and superficial liquid velocities.

  9. HPLC Determination of Taurine in Sports Drinks

    NASA Astrophysics Data System (ADS)

    Orth, Dale L.

    2001-06-01

    The amino acid taurine (2-aminoethanesulfonic acid) is present as a nutritional supplement in many sports drinks. An experiment, suitable for a junior-senior level instrumental analysis course, is described to measure the amount of taurine in these sports drinks. A pre-column derivatization with Sanger's reagent, 2,4-dinitrofluorobenzene, is followed by an HPLC separation utilizing a gradient elution, and detection at 360 nm.

  10. Separation and purification of epigallocatechin-3-gallate (EGCG) from green tea using combined macroporous resin and polyamide column chromatography.

    PubMed

    Jin, Xin; Liu, Mingyan; Chen, Zaixing; Mao, Ruikun; Xiao, Qinghuan; Gao, Hua; Wei, Minjie

    2015-10-01

    Epigallocatechin-3-gallate (EGCG) is a major bioactive ingredient of green tea that produces beneficial neuroprotective effects. In this paper, to optimize the EGCG enrichment, thirteen macroporous resins with different chemical and physical properties were systemically evaluated. Among the thirteen tested resins, the H-bond resin HPD826 exhibited best adsorption/desorption capabilities and desorption ratio, as well as weakest affinity for caffeine. The absorption of EGCG on the HPD826 resin followed the pseudo-second-order kinetics and Langmuir isotherm model. The separation parameters of EGCG were optimized by dynamic adsorption/desorption experiments with the HPD826 resin column. Under the optimal condition, the content of EGCG in the 30% ethanol eluent increased by 5.8-fold from 7.7% to 44.6%, with the recovery yield of 72.1%. After further purification on a polyamide column, EGCG with 74.8% purity was obtained in the 40-50% ethanol fraction with a recovery rate of 88.4%. In addition, EGCG with 95.1% purity could be easily obtained after one-step crystallization in distilled water. Our study suggests that the combined macroporous resin and polyamide column chromatography is a simple method for large-scale separation and purification of EGCG from natural plants for food and pharmaceutical applications.

  11. Separation and purification of epigallocatechin-3-gallate (EGCG) from green tea using combined macroporous resin and polyamide column chromatography.

    PubMed

    Jin, Xin; Liu, Mingyan; Chen, Zaixing; Mao, Ruikun; Xiao, Qinghuan; Gao, Hua; Wei, Minjie

    2015-10-01

    Epigallocatechin-3-gallate (EGCG) is a major bioactive ingredient of green tea that produces beneficial neuroprotective effects. In this paper, to optimize the EGCG enrichment, thirteen macroporous resins with different chemical and physical properties were systemically evaluated. Among the thirteen tested resins, the H-bond resin HPD826 exhibited best adsorption/desorption capabilities and desorption ratio, as well as weakest affinity for caffeine. The absorption of EGCG on the HPD826 resin followed the pseudo-second-order kinetics and Langmuir isotherm model. The separation parameters of EGCG were optimized by dynamic adsorption/desorption experiments with the HPD826 resin column. Under the optimal condition, the content of EGCG in the 30% ethanol eluent increased by 5.8-fold from 7.7% to 44.6%, with the recovery yield of 72.1%. After further purification on a polyamide column, EGCG with 74.8% purity was obtained in the 40-50% ethanol fraction with a recovery rate of 88.4%. In addition, EGCG with 95.1% purity could be easily obtained after one-step crystallization in distilled water. Our study suggests that the combined macroporous resin and polyamide column chromatography is a simple method for large-scale separation and purification of EGCG from natural plants for food and pharmaceutical applications. PMID:26319304

  12. Protein separation and characterization by np-RP-HPLC followed by intact MALDI-TOF mass spectrometry and peptide mass mapping analyses.

    PubMed

    Dauly, Claire; Perlman, David H; Costello, Catherine E; McComb, Mark E

    2006-07-01

    Because of their complexity, the separation of intact proteins from complex mixtures is an important step to comparative proteomics and the identification and characterization of the proteins by mass spectrometry (MS). In the study reported, we evaluated the use of nonporous-reversed-phase (np-RP)-HPLC for intact protein separation prior to MS analyses. The separation system was characterized and compared to 1D-SDS-PAGE electrophoresis in terms of resolution and sensitivity. We demonstrate that np-RP-HPLC protein separation is highly reproducible and provides intact protein fractions which can be directly analyzed by MALDI-TOF-MS for intact molecular weight determination. An in-well digestion protocol was developed, allowing for rapid protein identification by peptide mass fingerprinting (PMF) and resulted in comparable or improved peptide recovery compared with in-gel digestion. The np-RP sensitivity of detection by UV absorbance at 214 nm for intact proteins was at the low ng level and the sensitivity of peptide analysis by MALDI-TOF-MS was in the 10-50 fmol level. A membrane protein fraction was characterized to demonstrate application of this methodology. Among the identified proteins, multiple forms of vimentin were observed. Overall, we demonstrate that np-RP-HPLC followed by MALDI-TOF-MS allows for rapid, sensitive, and reproducible protein fractionation and very specific protein characterization by integration of PMF analysis with MS intact molecular weight information.

  13. LIQUID CHROMATOGRAPHIC SEPARATION OF THE ENANTIOMERS OF TRANS-CHLORDANE, CIS-CHLORDANE, HEPTACHLOR, HEPTACHLOR EPOXIDE AND ALPHA-HEXACHLOROCYCLOHEXANE WITH APPLICATION TO SMALL-SCALE PREPARATIVE SEPARATION

    EPA Science Inventory

    Analytical high-performance liquid chromatographic separations of the individual enantiomers of five polychlorinated compounds were obtained on polysaccharide stereoselective HPLC columns. The enantiomers of the pesticides trans-chlordane, cis-chlordane and heptachlor were separa...

  14. Gas chromatographic separation of fatty acid esters of cholesterol and phytosterols on an ionic liquid capillary column.

    PubMed

    Hammann, Simon; Vetter, Walter

    2015-12-15

    Steryl esters are high molecular weight compounds (600-700g/mol) regularly present as a minor lipid class in animal and plant lipids. Different sterol backbones (e.g., cholesterol, β-sitosterol and brassicasterol) which can be esterified with various fatty acids can result in highly complex steryl ester patterns in food samples. The gas chromatographic (GC) analysis of intact steryl esters is challenging, since high elution temperatures are required for their elution. On nonpolar GC phases, steryl esters with fatty acids with differing degree of unsaturation (e.g., oleate and linoleate) cannot be separated and there are only few polar columns available with sufficient temperature stability. In this study, we used gas chromatography with mass spectrometry (GC/MS) and analyzed intact steryl esters on a commercial room temperature ionic liquid (RTIL) column which was shortened to a length of 12m. The column separated the steryl esters both by total carbon number and by degree of unsaturation of the fatty acid. For instance, cholesteryl esters with stearic acid (18:0), oleic acid (18:1n-9), linoleic acid (18:2n-6) and α-linolenic acid (18:3n-3) could be resolved (R≥1.3) from each other. By analysis of synthesized standard substances, the elution orders for different steryl backbones and different fatty acids on a given sterol backbone could be determined. Analysis of spreads and plant oils allowed to determine retention times for 37 steryl esters, although a few co-elutions were observed. The ionic liquid column proved to be well-suited for the analysis of intact steryl esters.

  15. Comparative study between extraction techniques and column separation for the quantification of sinigrin and total isothiocyanates in mustard seed.

    PubMed

    Cools, Katherine; Terry, Leon A

    2012-07-15

    Glucosinolates are β-thioglycosides which are found naturally in Cruciferae including the genus Brassica. When enzymatically hydrolysed, glucosinolates yield isothiocyanates and give a pungent taste. Both glucosinolates and isothiocyanates have been linked with anticancer activity as well as antifungal and antibacterial properties and therefore the quantification of these compounds is scientifically important. A wide range of literature exists on glucosinolates, however the extraction and quantification procedures differ greatly resulting in discrepancies between studies. The aim of this study was therefore to compare the most popular extraction procedures to identify the most efficacious method and whether each extraction can also be used for the quantification of total isothiocyanates. Four extraction techniques were compared for the quantification of sinigrin from mustard cv. Centennial (Brassica juncea L.) seed; boiling water, boiling 50% (v/v) aqueous acetonitrile, boiling 100% methanol and 70% (v/v) aqueous methanol at 70 °C. Prior to injection into the HPLC, the extractions which involved solvents (acetonitrile or methanol) were freeze-dried and resuspended in water. To identify whether the same extract could be used to measure total isothiocyanates, a dichloromethane extraction was carried out on the sinigrin extracts. For the quantification of sinigrin alone, boiling 50% (v/v) acetonitrile was found to be the most efficacious extraction solvent of the four tested yielding 15% more sinigrin than the water extraction. However, the removal of the acetonitrile by freeze-drying had a negative impact on the isothiocyanate content. Quantification of both sinigrin and total isothiocyanates was possible when the sinigrin was extracted using boiling water. Two columns were compared for the quantification of sinigrin revealing the Zorbax Eclipse to be the best column using this particular method. PMID:22743340

  16. Separation of beta-human chorionic gonadotropin and immunoglobulin G by a miniaturized size exclusion chromatography column

    NASA Astrophysics Data System (ADS)

    Yang, Yongmo; Chae, Junseok

    2009-04-01

    This report describes a miniaturized size exclusion chromatography column that effectively preseparates raw samples for medical point-of-care testing (POCT) devices. The minicolumn is constructed of polydimethylsiloxane fabricated on a glass slide. The minicolumn separates 300 ng/ml of beta-human chorionic gonadotropin (β-hCG) from an immunoglobulin G (IgG)-rich solution (100 μg/ml) in 7.7 min, with 2.23 resolution and 0.018 mm plate height. The complete analyte discrimination shows potential for the sample preparation stage of POCT devices for cancer screening, prognosis, and monitoring.

  17. Monolithic poly(N-vinylcarbazole-co-1,4-divinylbenzene) capillary columns for the separation of biomolecules.

    PubMed

    Koeck, Rainer; Bakry, Rania; Tessadri, Richard; Bonn, Guenther K

    2013-09-01

    Monolithic capillary columns were prepared by thermally initiated free radical copolymerization of N-vinylcarbazole (NVC) and 1,4-divinylbenzene (DVB) within the confines of 200 and 100 μm i.d. fused silica capillaries. The reaction was carried out under the influence of inert micro-(toluene) and macroporogen (1-decanol) and α,α'-azoisobutyronitrile (AIBN) as a free radical initiator. The material proved high mechanical stability applying water and acetonitrile as mobile phases. The morphological and porous properties were studied by scanning electron microscopy (SEM), nitrogen sorption (BET) and mercury intrusion porosimetry (MIP). The homogeneity of the copolymerization process was confirmed by elemental analysis and monomer conversion measurements. The newly developed NVC/DVB monolithic supports showed high separation efficiency towards biomolecules, applying reversed-phase (RP) and ion-pair reversed-phase (IP-RP) separation modes, which is exemplified by the separations of peptides, proteins and oligonucleotides. Furthermore the maximum loading capacity was evaluated. The chromatographic performance under isocratic elution was determined in terms of theoretical plate number and plate height, where up to 41,000 plates per column and a minimum plate height value of 1.7 μm were achieved, applying oligonucleotide separations. In gradient elution mode, peak capacities of 96 and 127 were achieved within a gradient time window of 60 min for protein and oligonucleotide separations, respectively. The material proved to have high permeability, good repeatability of the fabrication process and high surface areas in the range of 120-160 m(2) g(-1). PMID:23799449

  18. "Supermarket Column Chromatography of Leaf Pigments" Revisited: Simple and Ecofriendly Separation of Plant Carotenoids, Chlorophylls, and Flavonoids from Green and Red Leaves

    ERIC Educational Resources Information Center

    Dias, Alice M.; Ferreira, Maria La Salete

    2015-01-01

    A simple and ecofriendly procedure was developed in order to prepare extracts from red and green leaves. This procedure enables the separation of yellow, green, and red band pigments and optimizes the previously reported baking soda "supermarket column". The same extract also led to a novel and colorful potato starch column, which can…

  19. Preparation of quaternary amine monolithic column for strong anion-exchange chromatography and its application to the separation of Enterovirus 71.

    PubMed

    Gu, Huimin; Yin, Dezhong; Ren, Jie; Zhang, Baoliang; Zhang, Qiuyu

    2016-10-15

    Large size virion is unable to diffuse into pores of conventional porous chromatography particles. Therefore, separation of virion by conventional column-packing materials is not quite efficient. To solve this problem, a monolithic column with large convective pores and quaternary amine groups was prepared and was applied to separate Enterovirus 71 (EV71, ≈5700-6000kDa). Cross-section, pore structure, hydrodynamic performance, adsorption property and dynamic binding capacity of prepared monolithic column were determined. Double-pore structures, macropore at 2472nm and mesopore at 5-60nm, were formed. The porosity was up to 63.3%, which enable higher permeability and lower back pressure of the monolithic column than commercial UNO™ Q1 column. Based on the breakthrough curves, the loading capacity of bovine serum albumin was calculated to be 42.0mg per column. In addition, prepared quaternary amine monolithic column was proved to be suitable for the separation of protein mixture by strong anion-exchange chromatography. As a practical application, prepared monolith column presents excellent performance to the separation of EV71 from virus-proteins mixture.

  20. Characterization and classification of stationary phases in HPLC and SFC – a review.

    PubMed

    Galea, Charlene; Mangelings, Debby; Vander Heyden, Yvan

    2015-07-30

    Packed column supercritical fluid chromatography (pSFC) is an attractive technique in drug discovery related analysis because it offers several advantages over the more commonly used high-performance liquid chromatography (HPLC) technique. The environmental-friendly CO2 mobile phase, the high-throughput capacity, the increased efficiency and the lower operational costs give SFC additional benefits over HPLC in analysis related to drug development. The latter technique is well established and has been used for decades in the pharmaceutical industry. On the other hand, SFC is still in its infancy, even though the technique has been known for decades and researchers are still discovering the possibilities and limitations of this technique. Column characterization aims to obtain a quantitative understanding of the properties of a column that influence the selectivity of a separation. Columns have been extensively characterized in HPLC using several approaches. However, limited column characterization has been done in SFC. Determining column properties allows the rapid selection of appropriate columns for method development for a particular application. Hence, they also show which columns will provide similar or dissimilar separations. The aim of this review is to compare and contrast approaches used to characterize stationary phases in HPLC and SFC, and to highlight points that still may need to be researched further. PMID:26320631

  1. Continuous countercurrent membrane column for the separation of solute/solvent and solvent/solvent systems

    DOEpatents

    Nerad, Bruce A.; Krantz, William B.

    1988-01-01

    A reverse osmosis membrane process or hybrid membrane - complementary separator process for producing enriched product or waste streams from concentrated and dilute feed streams for both solvent/solvent and solute/solvent systems is described.

  2. HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC SEPARATION OF THE ENANTIOMERS OF ORGANOPHOSPHORUS PESTICIDES ON POLYSACCHARIDE CHIRAL STATIONARY PHASES

    EPA Science Inventory

    High-performance liquid chromatographic separation of the individual enantiomers of 12 organophosphorus pesticides (OPs) was obtained on polysaccharide enantioselective HPLC columns using alkane-alcohol mobile phase. The OP pesticides were crotoxyphos, dialifor, fonofos, fenamiph...

  3. On-line nano-HPLC/ESI QTOF MS and tandem MS for separation, detection, and structural elucidation of human erythrocytes neutral glycosphingolipid mixture.

    PubMed

    Kirsch, Stephan; Zarei, Mostafa; Cindrić, Mario; Müthing, Johannes; Bindila, Laura; Peter-Katalinić, Jasna

    2008-06-15

    A superior approach involving nano-high-performance liquid chromatography (nano-HPLC) in on-line conjunction to electrospray ionization quadrupole time-of-flight mass spectrometry (ESI QTOF MS) and tandem MS for screening and structural characterization of complex mixtures of neutral glycosphingolipids (GSLs) is here described. Neutral GSLs purified from human erythrocytes were efficiently separated according to the differences in carbohydrate chain length by an optimized nano-HPLC protocol and flow-through detected by ESI QTOF MS at the low femtomole level. Additionally, GSL species were accurately distinguished from the accompanying lipids in the mixture, thus permitting the determination of detailed structural characteristics by data-dependent analysis for identification of GSL constitution within single experiments. An alternative nano-HPLC/ESI QTOF MS approach was designed for dissection of unsaturation/saturation degree of the ceramide moieties defining the hydrophobic portion of GSLs and subsequent localization by nano-HPLC/ESI QTOF MS/MS of the -CH=CH- within the ceramide regions. The method is fast, highly sensitive, and high-throughput amenable and is highlighted as a new and valuable analytical dimension in glycolipidomics.

  4. High-performance liquid chromatography separation of phthalate acid esters with a MIL-53(Al)-packed column.

    PubMed

    Shu, Lun; Chen, Sha; Zhao, Wei-Wei; Bai, Yan; Ma, Xing-Chen; Li, Xiao-Xin; Li, Jian-Rong; Somsundaran, P

    2016-08-01

    In this study, a MIL-53(Al)-packed column was successfully prepared and firstly applied to separate phthalate acid esters (butyl benzyl phthalate, di-n-butyl phthalate, diethyl phthalate, bis(2-ethylhexyl) phthalate, and dimethyl phthalate). Their baseline separation could be achieved within 12 min with a mobile phase of methanol/H2 O ratio at 92:8, and the temperature and flow rate was 40°C and 0.6 mL/min, respectively. The stacking effect and electrostatic force were the key factors in the separation. Moreover, there was a substantial linear relation between the peak height, peak area, and the analyte mass, and the relative standard deviations of retention time, peak height, peak area, and half peak width for five replicate separations of the analytes were within the ranges 0.31-0.88%, 0.72-1.52%, 1.33-1.53%, and 0.46-0.95%, respectively. The results of the calculation of the thermodynamics parameters showed that the separation of phthalate acid esters was controlled by both enthalpy change (ΔH) and entropy change (ΔS). PMID:27357380

  5. Comprehensive overview of recent preparation and application trends of various open tubular capillary columns in separation science.

    PubMed

    Cheong, Won Jo; Ali, Faiz; Kim, Yune Sung; Lee, Jin Wook

    2013-09-20

    Open tubular (OT) capillary columns have been increasingly used in a variety of fields of separation science such as CEC, LC, and SPE. Especially their application in CEC has attracted a lot of attention for their outstanding separation performance. Various forms of OT stationary phase materials have been employed such as in-situ prepared polymers, molecular imprinted polymers (MIPs), brush ligands, host ligands, block copolymers, aptamers, carbon nanotubes, polysaccharides, proteins, tentacles, nanoparticles, monoliths, and polyelectrolyte multi-layers. They have been prepared either in the chemically bound format or physically adsorbed format. Sol-gel technologies and nanoparticles have been sometimes involved in their preparation. There have been also some unique miscellaneous studies, for example, adopting preferentially adsorbed mobile phase components as stationary phases. In this review, recent progresses since mostly 2007 will be critically discussed in detail with some summarized descriptions for the work before the date. PMID:23948434

  6. Study of the enantiomeric separation of oxfendazole and cetirizine using subcritical fluid chromatography on an amylose-based column.

    PubMed

    Toribio, L; del Nozal, M J; Bernal, J L; Cristofol, C; Alonso, C

    2006-07-21

    The enantiomeric separation of cetirizine and oxfendazole on a Chiralpak AD column using subcritical fluid chromatography has been studied in this work. The enantioseparation of cetirizine was only possible when 2-propanol was used as a modifier, obtaining better results in presence of the additives triethylamine (TEA) and trifluoroacetic acid (TFAA). On the contrary, 2-propanol provided the lowest enantioresolutions for oxfendazole, in this case the best results in terms of high resolution and short analysis time were obtained with ethanol. The study of the temperature effect revealed that in the case of cetirizine using 2-propanol, and oxfendazole using methanol, the separation was enthalpy-driven and the isoelution temperature was above the working range. Using ethanol or 2-propanol, the results showed that the oxfendazole enantioseparation was entropically driven and the isoelution temperatures were below the range studied.

  7. Dielectrophoretic Assembly of Semiconducting Carbon Nanotubes Separated and Enriched by Spin Column Chromatography and Its Application to Gas Sensing

    NASA Astrophysics Data System (ADS)

    Nakano, Michihiko; Fujioka, Masahiro; Mai, Kaori; Watanabe, Hideaki; Martin, Yul; Suehiro, Junya

    2012-04-01

    The present authors have previously demonstrated the electrokinetic fabrication of a single-walled carbon nanotube (SWCNT) gas sensor by employing dielectrophoresis. Because this method employs mass-produced SWCNTs, it can realize cheaper and more flexible SWCNT gas sensor fabrication than that based on the on-site synthesis of SWCNTs. In this study, a new protocol was proposed and tested for the separation and enrichment of semiconducting SWCNTs, aiming to improve the SWCNT gas sensor sensitivity. The protocol employed a spin column filled with size-exclusion dextran-based gel beads as well as two surfactants (sodium dodecyl sulfate and sodium deoxycholate), which had different affinities to metallic and semiconducting SWCNTs. The separation and enrichment of the semiconducting SWCNTs were confirmed by measuring their optical and electrical properties. The CNT gas sensor fabricated using enriched semiconducting SWCNTs was highly sensitive to nitrogen dioxide (NO2) gas, - more sensitive by 10 times than that fabricated using the pristine SWCNT mixture.

  8. Property evaluations and application for separation of small molecules of a nanodiamond-polymer composite monolithic column.

    PubMed

    Wang, Fengqing; Wei, Aile; Wang, Xixi; Liu, Haiyan; Bai, Ligai; Yan, Hongyuan

    2016-07-01

    A nanodiamond-polymer composite monolithic column was first prepared successfully with modified nanodiamond (ND) as monomer, ethylene glycol dimethacrylate (EDMA) as cross-linker, 1-dodecanol as porogenic agent and benzoyl peroxide/dimethylacetamide (BPO/DMA) as initiator at 35°C for 2.5h. There was a sharp increase of specific surface area with ND added about 22 times from 0mg (3.90m(2)/g) to 7mg (81.2m(2)/g) determined with BET. Characterizations including scanning electron microscopy (SEM), fourier-transform infrared spectra (FIRT) and mercury intrusion porosimetry (MIP) were used to determine the microstructure, group composition, pore size distribution (≃1.56μm) and porosity (≃0.7484μm) of the monolith. An excellent column stability was confirmed by permeability (1.258x10(-10)cm(2)) and good linearity (R(2)=0.998) corresponding to backpressures measured at different flow rates. The highest swelling ability of the composite was about (5%) and classical RPLC of the column obtained occurred with the acetonitrile concentration increasing from 20% to 85% in the mobile phase, above which another retention model of normal-phase chromatography appeared. The items of the eddy dispersion and the absorption-release kinetics were the decisional factors of the composite column compared with the factors of longitudinal diffusion, and the skeleton-eluent mass transfer resistance due to the finite diffusivity. Good separation of neutral and basic small molecules was obtained (24,350 plates/m for neutral molecules and 22,300 plates/m for basic ones) with the hydrophobicity retention mechanism, but not for the acidic ones except to regulate the pH of the mobile phase. PMID:27154670

  9. Separation by column chromatography of cells active in delayed-onset hypersensitivities.

    PubMed Central

    Godfrey, H P; Gell, P G

    1976-01-01

    Lymph node cells from guinea-pigs contact sensitive to 1-thiocyamo-2,4-dinitrobenzene have been fractionated by affinity chromatography over modified polyacrylamide beads. Cells mediating lymphokine release in response to active sensitizer were depleted only by chromatography over dinitrophyenyl (DNP) containing substrates and could be specifically eluted with DNP-glycine. DNP rosette-forming cells (RFC) were equally well depleted by chromatography using either DNP or trinitrophenyl containing materials but could not be eluted from the columns by DNP-glycine. While the antigen receptors of cells mediating the release of macrophage agglutination factor in response to DNP-containing antigens and of DNP-RFC were found to be hapten-specific, their specificity was shown to differ using chromatography over trinitrophenyl containing polyacrylamide. PMID:776818

  10. Preparative separation of gallocatechin gallate from Camellia ptilophylla using macroporous resins followed by sephadex LH-20 column chromatography.

    PubMed

    Li, Kaikai; Zhou, Xuelin; Liu, Cheuk-Lun; Yang, Xiaorong; Han, Xiaoqiang; Shi, Xianggang; Song, Xiaohong; Ye, Chuangxing; Ko, Chun-hay

    2016-02-01

    Gallocatechin gallate (GCG) possesses multiple potential biological activities. However, the content of GCG in traditional green tea is too low which limits its in-depth pharmacological research and application. In the present study, a simple, efficient and environment-friendly chromatographic separation method was developed for preparative enrichment and separation of GCG from cocoa tea (Camellia ptilophylla) which contains high content of GCG. In the first step, the adsorption properties of selected resins were evaluated, and XAD-7HP resin was chosen by its adsorption and desorption properties for GCG. In order to maximize column efficiency for GCG collection, the operating parameters (e.g., flow rate, ethanol concentration, and bed height) were optimized. We found that the best combination was the feed concentration at 20mg/mL, flow rate at 0.75 BV/h and the ratio of diameter to bed heights as 1:12. Under these conditions, the purity of GCG was 45% with a recovery of 89%. In order to obtain pure target, a second step was established using column chromatography with sephadex LH-20 gel and 55% ethanol-water solution as eluent. After this step, the purity of the GCG was 91% with a recovery of 68% finally.

  11. Continuous separation of cells of high osteoblastic differentiation potential from mesenchymal stem cells on an antibody-immobilized column.

    PubMed

    Mahara, Atsushi; Yamaoka, Tetsuji

    2010-05-01

    Here, we report that two distinctive cell populations with osteoblastic differentiation ability were found in adherent cell populations from bone marrow. Mesenchymal stem cells (MSCs) were conventionally isolated by using adherent property of bone marrow cells onto a plastic culture dish. MSCs enriched on the basis of their adherent property were considered phenotypically and functionally heterogeneous. We developed a ligand-immobilized surface for separating subpopulation of adherent cells derived from bone marrow by the cell rolling process. We successfully isolate two cell populations with high differentiation ability for osteoblasts in adherent bone marrow cells by using the anti-CD34 antibody-immobilized column. The antibody was covalently conjugated with polyacrylic acid and introduced onto the inner surface of a silicone tube. When cell suspension of MSCs was injected into the antibody-immobilized column, different cell populations were isolated. After the cultivation of isolated cells in the osteoblastic differentiation medium for 1 week, few sub-populations were strongly induced to form osteoblastic cells. This study revealed that the ligand-immobilized surface can be used to continually separate cell populations under a labeling-free condition. PMID:20185169

  12. Acrylate ester-based monolithic columns for capillary electrochromatography separation of triacylglycerols in vegetable oils.

    PubMed

    Lerma-García, M J; Vergara-Barberán, M; Herrero-Martínez, J M; Simó-Alfonso, E F

    2011-10-21

    A simple and reliable method for the evaluation of triacylglycerols (TAGs) in vegetable oils by capillary electrochromatography (CEC) with UV-Vis detection, using octadecyl acrylate (ODA) ester-based monolithic columns, has been developed. The percentages of the porogenic solvents in the polymerization mixture, and the mobile phase composition, were optimized. The optimum monolith was obtained at the following ratios: 40:60% (wt/wt) monomers/porogens, 60:40% (wt/wt) ODA/1,3-butanediol diacrylate and 23:77% (wt/wt) 1,4-butanediol/1-propanol (14 wt% 1,4-butanediol in the polymerization mixture). A satisfactory resolution between TAGs was achieved in less than 12 min with a 65:35 (v/v) acetonitrile/2-propanol mixture containing 5 mM ammonium acetate. The method was applied to the analysis of TAGs of vegetable oil samples. Using linear discriminant analysis of the CEC TAG profiles, the vegetable oils belonging to six different botanical origins (corn, extra virgin olive, hazelnut, peanut, soybean and sunflower) were correctly classified with an excellent resolution among all the categories.

  13. Analysis of cocaine, benzoylecgonine, ecgonine methyl ester, ethylcocaine and norcocaine in human urine using HPLC with post-column ion-pair extraction and fluorescence detection.

    PubMed

    Roy, I M; Jefferies, T M; Threadgill, M D; Dewar, G H

    1992-01-01

    The measurement of cocaine and its major metabolites has been achieved by an HPLC method that compensates for their different solubilities and detection properties. Although ecgonine methyl ester is a major metabolite it is generally not measured by HPLC because it is poorly detectable by UV, and its water solubility makes recovery from urine difficult. Using modified solid-phase extraction procedures recoveries of 85% for ecgonine methyl ester, 97% for cocaine, 106% for benzoylecgonine and 80% for ethylcocaine have been obtained from urine. Increased chromatographic retention and detection sensitivity has been obtained by formation of the t-butyldimethylsilyl derivative of ecgonine methyl ester which was found to be stable in the HPLC mobile phase for at least 1 week. Alkylation of norcocaine and benzoylecgonine has improved their detection sensitivity and also chromatographic resolution. All calibrations were linear over the range 200-1000 ng ml-1 in urine with correlation coefficients > 0.99.

  14. Separation of the Components of a Commercial Analgesic Tablet: A Two-Week Sequence Comparing Purification by Two-Base Extraction and Column Chromatography

    ERIC Educational Resources Information Center

    Revell, Kevin D.

    2011-01-01

    A new laboratory experiment is described in which students compare two benchtop separation methods to isolate the three active components of the commercial analgesic Excedrin. In the two-week sequence, aspirin, acetaminophen, and caffeine are separated using either a two-base liquid-liquid extraction or silica column chromatography. Students then…

  15. Semi-micro reversed-phase liquid chromatography for the separation of alkyl benzenes and proteins exploiting methacrylate- and polystyrene-based monolithic columns.

    PubMed

    Masini, Jorge Cesar

    2016-05-01

    Monolithic columns were synthesized inside 1.02 mm internal diameter fused-silica lined stainless-steel tubing. Styrene and butyl, hexyl, lauryl, and glycidyl methacrylates were the functional monomers. Ethylene glycol dimethacrylate and divinylbenzene were the crosslinkers. The glycidyl methacrylate polymer was modified with gold nanoparticles and dodecanethiol (C12 ). The separation of alkylbenzenes was investigated by isocratic elution in 60:40 v/v acetonitrile/water. The columns based on polystyrene-co-divinylbenzene and poly(glycidyl methacrylate)-co-ethylene glycol dimethacrylate modified with dodecanethiol did not provide any separation of alkyl benzenes. The poly(hexyl methacrylate)-co-ethylene glycol dimethacrylate and poly(lauryl methacrylate)-co-ethylene glycol dimethacrylate columns separated the alkyl benzenes with plate heights between 30 and 60 μm (50 μL min(-1) and 60°C). Similar efficiency was achieved in the poly(butyl methacrylate)-co-ethylene glycol dimethacrylate column, but only at 10 μL min(-1) (0.22 mm s(-1) ). Backpressures varied from 0.38 MPa in the hexyl methacrylate to 13.4 MPa in lauryl methacrylate columns (50 μL min(-1) and 60°C). Separation of proteins was achieved in all columns with different efficiencies. At 100 μL min(-1) and 60°C, the lauryl methacrylate columns provided the best separation, but their low permeability prevented high flow rates. Flow rates up to 500 μL min(-1) were possible in the styrene, butyl and hexyl methacrylate columns.

  16. [Determination of six main components in compound theophylline tablet by convolution curve method after prior separation by column partition chromatography

    NASA Technical Reports Server (NTRS)

    Zhang, S. Y.; Wang, G. F.; Wu, Y. T.; Baldwin, K. M. (Principal Investigator)

    1993-01-01

    On a partition chromatographic column in which the support is Kieselguhr and the stationary phase is sulfuric acid solution (2 mol/L), three components of compound theophylline tablet were simultaneously eluted by chloroform and three other components were simultaneously eluted by ammonia-saturated chloroform. The two mixtures were determined by computer-aided convolution curve method separately. The corresponding average recovery and relative standard deviation of the six components were as follows: 101.6, 1.46% for caffeine; 99.7, 0.10% for phenacetin; 100.9, 1.31% for phenobarbitone; 100.2, 0.81% for theophylline; 99.9, 0.81% for theobromine and 100.8, 0.48% for aminopyrine.

  17. Hypercrosslinking: New approach to porous polymer monolithic capillary columns with large surface area for the highly efficient separation of small molecules

    PubMed Central

    Urban, Jiri; Svec, Frantisek; Fréchet, Jean M.J.

    2010-01-01

    Monolithic polymers with an unprecedented surface area of over 600 m2/g have been prepared from a poly(styrene-co-vinylbenzyl chloride-co-divinylbenzene) precursor monolith that was swollen in 1,2-dichloroethane and hypercrosslinked via Friedel-Crafts reaction catalyzed by ferric chloride. Both the composition of the reaction mixture used for the preparation of the precursor monolith and the conditions of the hypercrosslinking reaction have been varied using mathematical design of experiments and the optimized system validated. Hypercrosslinked monolithic capillary columns contain an array of small pores that make the column ideally suited for the high efficiency isocratic separations of small molecules such as uracil and alkylbenzenes with column efficiencies reproducibly exceeding 60,000 plates/m for retained compounds. The separation process could be accelerated while also improving peak shape through the use of higher temperatures and a ternary mobile phase consisting of acetonitrile, tetrahydrofuran, and water. As a result, seven compounds were well separated in less than 2 min. These columns also facilitate separations of peptide mixtures such as a tryptic digest of cytochrome c using a gradient elution mode which affords a sequence coverage of 93%. A 65 cm long hypercrosslinked capillary column used in size exclusion mode with tetrahydrofuran as the mobile phase afforded almost baseline separation of toluene and five polystyrene standards. PMID:21092973

  18. Comparison of separations of fatty acids from fish products using a 30-m Supelcowax-10 and a 100-m SP-2560 column.

    PubMed

    Santercole, Viviana; Delmonte, Pierluigi; Kramer, John K G

    2012-03-01

    Commercial fish oils and foods containing fish may contain trans and/or isomerized fatty acids (FA) produced during processing or as part of prepared foods. The current American Oil Chemists' Society (AOCS) official method for marine oils (method Ce 1i-07) is based on separation by use of poly(ethylene glycol) (PEG) columns, for example Supelcowax-10 or equivalent, which do not resolve most unsaturated FA geometric isomers. Highly polar 100-m cyanopropyl siloxane (CPS) columns, for example SP-2560 and CP Sil 88 are recommended for separation of geometric FA isomers. Complementary separations were achieved by use of two different elution temperature programs with the same CPS column. This study is the first direct comparison of the separations achieved by use of 30-m Supelcowax-10 and 100-m SP-2560 columns for fatty acid methyl esters (FAME) prepared from the same fish oil and fish muscle sample. To simplify the identification of the FA in these fish samples, FA were fractionated on the basis of the number and type of double bonds by silver-ion solid-phase extraction (Ag⁺-SPE) before GC analysis. The results showed that a combination of the three GC separations was necessary to resolve and identify most of the unsaturated FA, FA isomers, and other components of fish products, for example phytanic and phytenic acids. Equivalent chain length (ECL) values of most FAME in fish were calculated from the separations achieved by use of both GC columns; the values obtained were shown to be consistent with previously reported values for the Supelcowax-10 column. ECL values were also calculated for the FA separated on the SP-2560 column. The calculated ECL values were equally valid under isothermal and temperature-programmed elution GC conditions, and were valuable for confirmation of the identity of several unsaturated FAME in the fish samples. When analyzing commercially prepared fish foods, deodorized marine oils, or foods fortified with marine oils it is strongly

  19. Scale-up protein separation on stainless steel wide bore toroidal columns in the type-J counter-current chromatography.

    PubMed

    Guan, Yue Hugh; Hewitson, Peter; van den Heuvel, Remco N A M; Zhao, Yan; Siebers, Rick P G; Zhuang, Ying-Ping; Sutherland, Ian

    2015-12-11

    Manufacturing high-value added biotech biopharmaceutical products (e.g. therapeutic proteins) requires quick-to-develop, GMP-compliant, easy-to-scale and cost effective preparatory chromatography technologies. In this work, we describe the construction and testing of a set of 5-mm inner diameter stainless steel toroidal columns for use on commercially available preparatory scale synchronous J-type counter-current chromatography (CCC) machinery. We used a 20.2m long column with an aqueous two-phase system containing 14% (w/w) PEG1000 and 14% (w/w) potassium phosphate at pH 7, and tested a sample loading of 5% column volume and a mobile phase flow rate of 20ml/min. We then satisfactorily demonstrated the potential for a weekly protein separation and preparation throughput of ca. 11g based on a normal weekly routine for separating a pair of model proteins by making five stacked injections on a single portion of stationary phase with no stripping. Compared to our previous 1.6mm bore PTFE toroidal column, the present columns enlarged the nominal column processing throughput by nearly 10. For an ideal model protein injection modality, we observed a scaling up factor of at least 21. The 2 scales of protein separation and purification steps were realized on the same commercial CCC device.

  20. CZE separation of amitrol and triazine herbicides in environmental water samples with acid-assisted on-column preconcentration.

    PubMed

    Arribas, Alberto Sánchez; Moreno, Mónica; Bermejo, Esperanza; Zapardiel, Antonio; Chicharro, Manuel

    2011-01-01

    A simple analytical scheme for the detection and quantification of amitrol and triazine herbicides (atrazine, ametryn and atraton) and degradation product (2-hydroxyatrazine) in environmental water samples by CZE is reported. On-column preconcentration of analytes from untreated water samples (mineral, spring, tap and river water) is accomplished by introducing an acid plug (200 mM citrate of pH 2.0) after the sample and then proceeding with the CZE separation, using 100 mM formiate buffer of pH 3.5 as running buffer and 25.0 KV as separation voltage. UV detection at 200 nm provides LODs from 50 to 300 nM in untreated samples and they were lowered tenfold by sample preconcentration by evaporation. Calculated recoveries were typically higher than 90%. Minimal detectable concentration of the electroactive amitrol could be decreased about 20-fold when electrochemical detection was employed by monitoring the amperometric signal at +800 mV using a carbon paste electrode (LOD of 9.6 nM, 0.81 μg/L, versus 170 nM, 14.3 μg/L, using amperometric and UV detection, respectively) in untreated water samples.

  1. Quality by design approach for the separation of naproxcinod and its related substances by fused core particle technology column.

    PubMed

    Inugala, Ugandar Reddy; Pothuraju, Nageswara Rao; Vangala, Ranga Reddy

    2013-01-01

    This paper describes the development of a rapid, novel, stability-indicating gradient reversed-phase high-performance liquid chromatographic method and associated system suitability parameters for the analysis of naproxcinod in the presence of its related substances and degradents using a quality-by-design approach. All of the factors that affect the separation of naproxcinod and its impurities and their mutual interactions were investigated and robustness of the method was ensured. The method was developed using an Ascentis Express C8 150 × 4.6 mm, 2.7 µm column with a mobile phase containing a gradient mixture of two solvents. The eluted compounds were monitored at 230 nm, the run time was 20 min within which naproxcinod and its eight impurities were satisfactorily separated. Naproxcinod was subjected to the stress conditions of oxidative, acid, base, hydrolytic, thermal and photolytic degradation. Naproxcinod was found to degrade significantly in acidic and basic conditions and to be stable in thermal, photolytic, oxidative and aqueous degradation conditions. The degradation products were satisfactorily resolved from the primary peak and its impurities, proving the stability-indicating power of the method. The developed method was validated as per International Conference on Harmonization guidelines with respect to specificity, linearity, limit of detection, limit of quantification, accuracy, precision and robustness.

  2. CZE separation of amitrol and triazine herbicides in environmental water samples with acid-assisted on-column preconcentration.

    PubMed

    Arribas, Alberto Sánchez; Moreno, Mónica; Bermejo, Esperanza; Zapardiel, Antonio; Chicharro, Manuel

    2011-01-01

    A simple analytical scheme for the detection and quantification of amitrol and triazine herbicides (atrazine, ametryn and atraton) and degradation product (2-hydroxyatrazine) in environmental water samples by CZE is reported. On-column preconcentration of analytes from untreated water samples (mineral, spring, tap and river water) is accomplished by introducing an acid plug (200 mM citrate of pH 2.0) after the sample and then proceeding with the CZE separation, using 100 mM formiate buffer of pH 3.5 as running buffer and 25.0 KV as separation voltage. UV detection at 200 nm provides LODs from 50 to 300 nM in untreated samples and they were lowered tenfold by sample preconcentration by evaporation. Calculated recoveries were typically higher than 90%. Minimal detectable concentration of the electroactive amitrol could be decreased about 20-fold when electrochemical detection was employed by monitoring the amperometric signal at +800 mV using a carbon paste electrode (LOD of 9.6 nM, 0.81 μg/L, versus 170 nM, 14.3 μg/L, using amperometric and UV detection, respectively) in untreated water samples. PMID:21254126

  3. Column properties and flow profiles of a flat, wide column for high-pressure liquid chromatography

    SciTech Connect

    Mriziq, Khaled S; Guiochon, Georges A

    2008-01-01

    The design and the construction of a pressurized, flat, wide column for high-performance liquid chromatography (HPLC) are described. This apparatus, which is derived from instruments that implement over-pressured thin layer chromatography, can carry out only uni-dimensional chromatographic separations. However, it is intended to be the first step in the development of more powerful instruments that will be able to carry out two-dimensional chromatographic separations, in which case, the first separation would be a space-based separation, LC{sup x}, taking place along one side of the bed and the second separation would be a time-based separation, LC{sup t}, as in classical HPLC but proceeding along the flat column, not along a tube. The apparatus described consists of a pressurization chamber made of a Plexiglas block and a column chamber made of stainless steel. These two chambers are separated by a thin Mylar membrane. The column chamber is a cavity which is filled with a thick layer (ca. 1 mm) of the stationary phase. Suitable solvent inlet and outlet ports are located on two opposite sides of the sorbent layer. The design allows the preparation of a homogenous sorbent layer suitable to be used as a chromatographic column, the achievement of effective seals of the stationary phase layer against the chamber edges, and the homogenous flow of the mobile phase along the chamber. The entire width of the sorbent layer area can be used to develop separations or elute samples. The reproducible performance of the apparatus is demonstrated by the chromatographic separations of different dyes. This instrument is essentially designed for testing detector arrays to be used in a two-dimensional LC{sup x} x LC{sup t} instrument. The further development of two-dimension separation chromatographs based on the apparatus described is sketched.

  4. HPLC Separation of the (S,S)- and (R,S)- forms of S-Adenosyl-L-methionine

    PubMed Central

    Zhang, Jianyu; Klinman, Judith P.

    2015-01-01

    S-Adenosyl-L-methionine, an important biological cofactor, exists in two chiral forms, (S,S)- and (R,S)-, only the former of which is biologically active. Herein, we develop a chromatographic method to obtain pure (S,S)-AdoMet using a single C18 column. PMID:25681113

  5. A multidimensional micro gas chromatograph employing a parallel separation multi-column chip and stop-flow μGC × μGCs configuration.

    PubMed

    Chen, Bo-Xun; Hung, Te-Yu; Jian, Rih-Sheng; Lu, Chia-Jung

    2013-04-01

    A dual-chip, multidimensional micro gas chromatographic module was designed, built and evaluated. Column chips were fabricated on a silicon wafer with an etched rectangular channel 100 μm (width) × 250 μm (depth) using a deep reactive ion etching (DRIE) process. The column chip for the first GC dimension was 3 m long and was coated with polydimethylsiloxane (DB-1) as the stationary phase. The columns on the second dimensional chip were etched with the same width and depth as the first chip, but the flow channel was split into three parallel columns, 1 m long, on the same sized silicon chip (i.e., 3 cm × 3 cm). These three parallel columns on the second chip were coated with polyethylene oxide (DB-Wax), trifluoropropylpolymethylsilicone (OV-210) and cyanopropylmethylphenylmethylpolysilicone (OV-225), accordingly, in order to provide diversified chromatographic retention. These two chips were connected via a stop-flow configuration to simultaneously generate multiple two-dimensional gas chromatograms for every analysis. This stop-flow μGC × μGCs design allowed the first column to function as a pre-separator and as a sequencing injector for the second parallel-separation chip. Fifteen volatile organic compounds with boiling points that ranged from 80-131 °C with various functional groups were tested using this μGC × μGCs module. Three discrete 2-D chromatograms were generated simultaneously, which demonstrated the advantages of simultaneously combining GC × GC with parallel separation GCs in microchip chromatography. The total traveling length in the column was only 4 m for each eluted peak and fully resolved separation was achieved through the cross reference among triplet 2-D chromatograms. PMID:23381092

  6. Polyacrylamide-based monolithic capillary column with coating of cellulose tris(3,5-dimethylphenyl-carbamate) for enantiomer separation in capillary electrochromatography.

    PubMed

    Dong, Xiaoli; Wu, Ren'an; Dong, Jing; Wu, Minghuo; Zhu, Yan; Zou, Hanfa

    2008-02-01

    A hydrophilic chiral capillary monolithic column for enantiomer separation in CEC was prepared by coating cellulose tris(3,5-dimethylphenyl-carbamate) (CDMPC) on porous hydrophilic poly(acrylamide-co-N,N'-methylene-bisacrylamide) (poly(AA-co-MBA)) monolithic matrix with confine of a fused-silica capillary. The coating conditions were optimized to obtain a stable and reproducible chiral stationary phase for CEC. The effect of organic modifier of ACN in aqueous mobile phase for the enantiomer separation by CEC was investigated, and the significant influence of ACN on the enantioresolution and electrochromatographic retention was observed. Twelve pairs of enantiomers including acidic, neutral, and basic analytes were tested and nine pairs of them were baseline-enantioresolved with acidic and basic aqueous mobile phases. A good within-column repeatability in retention time (RSD = 2.4%) and resolution (RSD = 3.2%) was obtained by consecutive injections of a neutral compound, benzoin, on a prepared chiral monolithic column, while the between-column repeatability in retention time (RSD = 6.4%) and resolution (RSD = 9.6%) was observed by column-to-column examination. The prepared monolithic stationary phase showed good stability in either acidic or basic mobile phase.

  7. Polyacrylamide-based monolithic capillary column with coating of cellulose tris(3,5-dimethylphenyl-carbamate) for enantiomer separation in capillary electrochromatography.

    PubMed

    Dong, Xiaoli; Wu, Ren'an; Dong, Jing; Wu, Minghuo; Zhu, Yan; Zou, Hanfa

    2008-02-01

    A hydrophilic chiral capillary monolithic column for enantiomer separation in CEC was prepared by coating cellulose tris(3,5-dimethylphenyl-carbamate) (CDMPC) on porous hydrophilic poly(acrylamide-co-N,N'-methylene-bisacrylamide) (poly(AA-co-MBA)) monolithic matrix with confine of a fused-silica capillary. The coating conditions were optimized to obtain a stable and reproducible chiral stationary phase for CEC. The effect of organic modifier of ACN in aqueous mobile phase for the enantiomer separation by CEC was investigated, and the significant influence of ACN on the enantioresolution and electrochromatographic retention was observed. Twelve pairs of enantiomers including acidic, neutral, and basic analytes were tested and nine pairs of them were baseline-enantioresolved with acidic and basic aqueous mobile phases. A good within-column repeatability in retention time (RSD = 2.4%) and resolution (RSD = 3.2%) was obtained by consecutive injections of a neutral compound, benzoin, on a prepared chiral monolithic column, while the between-column repeatability in retention time (RSD = 6.4%) and resolution (RSD = 9.6%) was observed by column-to-column examination. The prepared monolithic stationary phase showed good stability in either acidic or basic mobile phase. PMID:18219649

  8. Stationary phases in the screening of drug/impurity profiles and in their separation method development: identification of columns with different and similar selectivities.

    PubMed

    Van Gyseghem, E; Jimidar, M; Sneyers, R; De Smet, M; Verhoeven, E; Vander Heyden, Y

    2006-06-01

    The classification or characterization of stationary phases based on chromatographic parameters, in general, requires different test solutes/mixtures and several mobile phases. To simplify the classification/characterization of reversed-phase liquid chromatographic columns, to be used in separating drug/impurity profiles, a new test procedure was proposed. It consists of injecting two mixtures of relatively similar active substances applying a standard gradient. The aim was to evaluate from this approach the selectivity differences and overall separation quality of newly tested columns compared to that in an earlier selected set of eight stationary phases. The selectivity differences of the columns were evaluated by correlation coefficient-based weighted-average-linkage dendrograms and color maps. Derringer's desirability functions were used to rank similar stationary phases according to their overall separation quality. Four columns of 27 examined were, for instance, considered different from the earlier selected eight and could be added to the selection. A number of tested stationary phases might be considered as alternatives for some from the initial set. For three columns the newly tested stationary phases did not contain alternatives.

  9. One-step column chromatographic extraction with gradient elution followed by automatic separation of volatiles, flavonoids and polysaccharides from Citrus grandis.

    PubMed

    Han, Han-Bing; Li, Hui; Hao, Rui-Lin; Chen, Ya-Fei; Ni, He; Li, Hai-Hang

    2014-02-15

    Citrus grandis Tomentosa is widely used in traditional Chinese medicine and health foods. Its functional components include volatiles, flavonoids and polysaccharides which cannot be effectively extracted through traditional methods. A column chromatographic extraction with gradient elution was developed for one-step extraction of all bioactive substances from C. grandis. Dried material was loaded into a column with petroleum ether: ethanol (8:2, PE) and sequentially eluted with 2-fold PE, 3-fold ethanol: water (6:4) and 8-fold water. The elutes was separated into an ether fraction containing volatiles and an ethanol-water fraction containing flavonoids and polysaccharides. The later was separated into flavonoids and polysaccharides by 80% ethanol precipitation of polysaccharides. Through this procedure, volatiles, flavonoids and polysaccharides in C. grandis were simultaneously extracted at 98% extraction rates and simply separated at higher than 95% recovery rates. The method provides a simple and high-efficient extraction and separation of wide range bioactive substances.

  10. Comparison of antioxidant and antiproliferative activity between Kunlun Chrysanthemum flowers polysaccharides (KCCP) and fraction PII separated by column chromatography.

    PubMed

    Jing, Siqun; Chai, Wenjie; Guo, Gai; Zhang, Xiaoming; Dai, Jun; Yan, Liang-Jun

    2016-04-15

    The aim of the present study was to compare the antioxidant and antiproliferative effects on cancer cells between Kunlun Chrysanthemum flowers polysaccharides (KCCP) and its fraction PII that were separated by Biologic low pressure (LP) chromatography system followed by DEAE cellulose column chromatography. Results of in vitro experiments showed that the reducing power and the scavenging capacity of KCCP towards hydroxyl radicals (OH) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals increased in a concentration dependent manner and were stronger than that of fraction PII. Results of the antiproliferative effect of KCCP and fraction PII on cervical cancer HeLa cells, esophagus cancer Eca109 cells, and mouse ascites hepatomas H22 cells indicated that both KCCP and its fraction PII possessed inhibitory activity on all the tested cancer cells at a dose- and time-dependent manner, with KCCP showing higher inhibitory activity than that of fraction PII. The present study demonstrates that KCCP and its fraction PII have antioxidant properties that may help fight cancers. PMID:26809376

  11. A simplified mathematical model of the cryogenic distillation with application to the (13C) isotope separation column

    NASA Astrophysics Data System (ADS)

    Neaga, A. O.; Festila, C.; Dulf, E. H.; Both, R.; Szelitzky, T.; Gligan, M.

    2012-02-01

    The isotope (13C) has a widespread application in many fields such as chemistry, physics, medicine, etc. To obtain a high concentration in isotope of interest, in our case (13C), it is used the method of cryogenic distillation of carbon monoxide (CO) which is based on the difference between the vapor pressure of (12C16O) and (13C16O) at the temperature of liquid nitrogen. Isotopic separation plant, used to obtain the isotope (13C), is a complex installation, with many inputs and outputs, rather difficult to control. Due to this reason, from the point of view of automation, it is needed a simplified mathematical model. This model can be determined only with some presumption and simplification assumptions. Using the physical laws, the hydrodynamic part of the process and the mass balance will be described by partial differential equations. In order to design a controller for the column, it is needed a transfer function or a statespace realization of the plant, which is the main contribution of the present work. Implementing this mathematical model will be the key element for describing and understanding the operation of the plant and for future development of process control strategies.

  12. Separation and preconcentration of trace manganese from various samples with Amberlyst 36 column and determination by flame atomic absorption spectrometry.

    PubMed

    Kendüzler, Erdal; Türker, A Rehber; Yalçınkaya, Ozcan

    2006-06-15

    This work assesses the potential of a new adsorptive material, Amberlyst 36, for the separation and preconcentration of trace manganese(II) from various media. It is based on the sorption of manganese(II) ions onto a column filled with Amberlyst 36 cation exchange resin, followed by the elution with 5mL of 3mol/L nitric acid and determination by flame atomic absorption spectrometry (FAAS) without interference of the matrix. Different factors including pH of sample solution, sample volume, amount of resin, flow rate of sample solution, volume and concentration of eluent, and matrix effects for preconcentration were investigated. Good relative standard deviation (3%) and high recovery (>95%) at 100mug/L and high enrichment factor (200) and low analytical detection limit (0.245mug/L) were obtained. The adsorption equilibrium was described well by the Langmuir isotherm model with maximum adsorption capacity of 88mg/g of manganese on the resin. The method was applied for the manganese determination by FAAS in tap water, commercial natural drinking water, commercial treated drinking water and commercial tea bag sample. The accuracy of the method is confirmed by analyzing the certified reference material (tea leaves GBW 07605). The results demonstrated good agreement with the certified values. PMID:18970645

  13. Quest for organic polymer-based monolithic columns affording enhanced efficiency in high performance liquid chromatography separations of small molecules in isocratic mode.

    PubMed

    Svec, Frantisek

    2012-03-01

    The separations of small molecules using columns containing porous polymer monoliths invented two decades ago went a long way from the very modest beginnings to the current capillary columns with efficiencies approaching those featured by their silica-based counterparts. This review article presents a variety of techniques that have been used to form capillary formats of monolithic columns with enhanced separation performance in isocratic elutions. The following text first describes the traditional approaches used for the preparation of efficient monoliths comprising variations in polymerization conditions including temperature as well as composition of monomers and porogenic solvents. Encouraging results of these experiments fueled research of completely new preparation methods such as polymerization to an incomplete conversion, use of single crosslinker, hypercrosslinking, and incorporation of carbon nanotubes that are described in the second part of the text. PMID:21816401

  14. Use of zirconium(IV) arsenophosphate columns for cation exchange separation of metal ions interfering in the spectrophotometric determination of uranium with sodium diethyl dithiocarbamate

    SciTech Connect

    Varshney, K.G.; Agrawal, S.; Anwar, S.; Varshney, K.

    1985-01-01

    A simple cation exchange method has been developed for the quantitative separation of uranium from some metal ions which generally interfere in its spectrophotometric determination using sodium diethyl dithiocarbamate as a reagent. The method requires only a single bed operation and enables a satisfactory (Error + or - separation of uranium (UO/sub 2/ (II)) up to 1080 ..mu..g from ten metal ions on a 2 g column of zirconium (IV) arsenophosphate cation exchanger in H(I) form.

  15. Comparative studies on performance of CCC and preparative RP-HPLC in separation and purification of steroid saponins from Dioscorea zingiberensis C.H.Wright

    PubMed Central

    Zhang, Xinxin; Liang, Jinru; Zhang, Yongmin; Liu, Jianli; Sun, Wenji; Ito, Yoichiro

    2015-01-01

    Steroid saponins from Dioscorea zingiberensis C.H.Wright were separated for the first time using two chromatographic methods for comparison: counter-current chromatography (CCC) coupled with evaporative light scattering detector (ELSD) and preparative reversed phase high-performance liquid chromatography (RP-HPLC) with an ultraviolet detector. Ethyl acetate-n-butanol-methanol-water (4:1:2:4, v/v) was chosen as the two-phase solvent system for CCC, while the acetonitrile-water (25:75 for the first step and15:85 for the second step, v/v) was used as the mobile phase in the preparative RP-HPLC. The following five steroid saponins were purified by theses two chromatographic methods, in one-step operation by CCC and by two-step operation in preparative RP-HPLC: 1) 26-O-β-D- glucopyranosyl-(25R)-furost-5-en-3β, 22ζ, 26-triol-3-O-[β-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl-(1→4)-α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside (compound A), 2) 26-O-β-D-glucopyranosyl-(25R)-furost-5-en-3β, 22ζ, 4) 26-triol-3-O-[β-D-glucopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)]-β-D-glucopyranoside (compound B), 3) 26-O-β-D-glucopyranosyl-(25R)-furost-5-en-3β, 22ζ, 26-triol-3-O-[α-L-rhamnopyranosyl-(1→4)]-β-D-glucopyranoside (compound C), 4) 26-O-β-D-glucopyranosyl-(25R)-furost-5, 20(22)-diene-3β, 26-diol-3-O-{α-L-rhamnopyranosyl-(1→4)-[β-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl-(1→2)]}-β-D-glucopyranoside (compound D) and 5) 26-O-β-D-glucopyranosyl-(25R)-furost-5, 20(22)-diene-3β, 26-diol-3-O-[β-D-glucopyranosyl-(1→4)-α-L-rhamnopyranosy-(1→2)]-β-D-glucopyranoside (compound E). The purities of these five steroid saponins separated by both methods were over 95%, and structural identification of these compounds was performed by ESI-MS, and 13C NMR. Comparison of these two established approaches revealed that CCC required a longer separation time but with less solvent consumption, whereas preparative RP-HPLC gave a shorter separation time but

  16. Influence of the crosslinker type on the chromatographic properties of hydrophilic sulfoalkylbetaine-type monolithic columns.

    PubMed

    Liu, Chusheng; Chen, Weijia; Yuan, Guangxin; Xiao, Yao; Crommen, Jacques; Xu, Shihai; Jiang, Zhengjin

    2014-12-19

    In order to investigate the effects of the crosslinker on the separation performance of polar zwitterionic sulfoalkylbetaine-type monolithic columns, three crosslinkers, i.e. 1,4-bis(acryloyl)piperazine (PDA), ethylene dimethacrylate (EDMA) and N,N'-methylenebisacrylamide (MBA), were copolymerized with the hydrophilic monomer N,N-dimethyl-N-acryloyloxyethyl-N-(3-sulfopropyl)ammonium betaine (SPDA). The chromatographic properties of the three hydrophilic sulfoalkylbetaine-type monolithic columns, including column efficiency, permeability, porosity and separation mechanism, were systematically compared using scanning electron microscopy or micro-HPLC. Good selectivity in micro-HPLC separations was achieved on all three monolithic columns. The results indicate that the polarity of sulfoalkylbetaine-type monolithic columns may be related to the polarity of the crosslinker, which further affects column selectivity and efficiency. A particularly high column efficiency (100,000 plates/m) was obtained on the novel poly(SPDA-co-PDA) monolithic column at a linear velocity of 1mm/s using thiourea as test analyte. A higher resolution was also observed for nucleobases, nucleosides and hydrophilic organic acids on this novel poly(SPDA-co-PDA) monolithic column compared to the other two columns. PMID:25464999

  17. Rapid screening and identification of compounds with DNA-binding activity from Folium Citri Reticulatae using on-line HPLC-DAD-MS(n) coupled with a post column fluorescence detection system.

    PubMed

    Fu, Qingrong; Zhang, Cangman; Lin, Zongtao; Sun, Hongyang; Liang, Yi; Jiang, Haixiu; Song, Zhiling; Wang, Hong; Chen, Shizhong

    2016-02-01

    To study the interactions between natural compounds and deoxyribonucleic acid (DNA), a method has been established combining a high-performance liquid chromatography-diode array detector-multi-stage mass spectrometer with a fluorescence detector (HPLC-DAD-MS(n)-FLD). The FLD was used to monitor fluorescence intensity of the ethidium bromide-DNA (EB-DNA) complex when a compound separated by HPLC was introduced. This novel method was used to simultaneously obtain the HPLC fingerprint, UV spectra, MS(n) fragments and DNA-binding activity profile of various components in Folium Citri Reticulatae. As a result, 35 compounds were identified, of which 25 were found in the extract of Folium Citri Reticulatae for the first time, and 33 compounds showed DNA-binding activities, with the most active being feruloylhexaric and p-coumaroylhexaric acids. In addition, the precision, stability and reproducibility of this method were validated by two positive controls, quercetin and hesperidin. This new on-line method is accurate, precise and reliable for further high-throughput screening of DNA-binding compounds from food samples and other complex matrices.

  18. The separation of flavonoids from Pongamia pinnata using combination columns in high-speed counter-current chromatography with a three-phase solvent system.

    PubMed

    Yin, Hao; Zhang, Si; Long, Lijuan; Yin, Hang; Tian, Xinpeng; Luo, Xiongming; Nan, Haihan; He, Sha

    2013-11-01

    The mangrove plant Pongamia pinnata (Leguminosae) is well known as a plant pesticide. Previous studies have indicated that the flavonoids are responsible of the biological activities of the plant. A new high-speed counter-current chromatography (HSCCC) method for the separation of three flavonoids, karanjin (1), pinnatin (2), and pongaflavone (3), from P. pinnata was developed in the present study. The lower and intermediate phase (LP and IP) of a new three-phase solvent system, n-hexane-acetonitrile-dichloromethane-water, at a volume ratio of 5:5:1:5, were used as the stationary phases, while the upper phase (UP) was used as the mobile phase, and the volume ratio between the stationary phases in the CCC column could be tuned by varying the initial pumped volume ratio of the stationary phases. The CCC columns containing all three phases of the solvent system were considered combination columns. According to the theories of combination column, it is possible to optimize the retention time of the target compounds by varying the volume ratio of the stationary phases in the HSCCC combination columns, as well as the suitable volume ratios of the stationary phases for the separation of the target compounds were predicted from the partition coefficients of the compounds in the three-phase solvent system. Then, three HSCCC separations using the combination columns with initial pumped LP:IP volume ratios of 1:0, 0.9:0.1, and 0.7:0.3 were performed separately based on the prediction. Three target compounds were prepared with high purity when the initial pumped volume ratio of the stationary phases was 0.9:0.1. The baseline separation of compounds 2 and 3 was achieved on the combination column with an initial pumped volume ratio of 0.7:0.3. Furthermore, the three experiments clearly demonstrated that the retentions and resolutions of the target compounds increased with an increasing volume ratio of IP, which is consistent with the prediction for the retention times for the

  19. The separation of flavonoids from Pongamia pinnata using combination columns in high-speed counter-current chromatography with a three-phase solvent system.

    PubMed

    Yin, Hao; Zhang, Si; Long, Lijuan; Yin, Hang; Tian, Xinpeng; Luo, Xiongming; Nan, Haihan; He, Sha

    2013-11-01

    The mangrove plant Pongamia pinnata (Leguminosae) is well known as a plant pesticide. Previous studies have indicated that the flavonoids are responsible of the biological activities of the plant. A new high-speed counter-current chromatography (HSCCC) method for the separation of three flavonoids, karanjin (1), pinnatin (2), and pongaflavone (3), from P. pinnata was developed in the present study. The lower and intermediate phase (LP and IP) of a new three-phase solvent system, n-hexane-acetonitrile-dichloromethane-water, at a volume ratio of 5:5:1:5, were used as the stationary phases, while the upper phase (UP) was used as the mobile phase, and the volume ratio between the stationary phases in the CCC column could be tuned by varying the initial pumped volume ratio of the stationary phases. The CCC columns containing all three phases of the solvent system were considered combination columns. According to the theories of combination column, it is possible to optimize the retention time of the target compounds by varying the volume ratio of the stationary phases in the HSCCC combination columns, as well as the suitable volume ratios of the stationary phases for the separation of the target compounds were predicted from the partition coefficients of the compounds in the three-phase solvent system. Then, three HSCCC separations using the combination columns with initial pumped LP:IP volume ratios of 1:0, 0.9:0.1, and 0.7:0.3 were performed separately based on the prediction. Three target compounds were prepared with high purity when the initial pumped volume ratio of the stationary phases was 0.9:0.1. The baseline separation of compounds 2 and 3 was achieved on the combination column with an initial pumped volume ratio of 0.7:0.3. Furthermore, the three experiments clearly demonstrated that the retentions and resolutions of the target compounds increased with an increasing volume ratio of IP, which is consistent with the prediction for the retention times for the

  20. Separation and purification of bioactive botrallin and TMC-264 by a combination of HSCCC and semi-preparative HPLC from endophytic fungus Hyalodendriella sp. Ponipodef12.

    PubMed

    Mao, Ziling; Luo, Ruiya; Luo, Haiyu; Tian, Jin; Liu, Hongwei; Yue, Yang; Wang, Mingan; Peng, Youliang; Zhou, Ligang

    2014-09-01

    Two dibenzo-α-pyrones, botrallin (1) and TMC-264 (2) were preparatively separated from crude ethyl acetate extract of the endophytic fungus Hyalodendriella sp. Ponipodef12, which was isolated from the hybrid 'Neva' of Populus deltoides Marsh × P. nigra L. using a combination of high-speed counter-current chromatography (HSCCC) and semi-preparative HPLC. Botrallin (1) with 74.73% of purity and TMC-264 (2) with 82.29% of purity were obtained through HSCCC by employing a solvent system containing n-hexane-ethyl acetate-methanol-water at a volume ratio of 1.2:1.0:0.9:1.0. It was the first time for TMC-264 (2) to be isolated from this fungus. TMC-264 (2) showed strong antimicrobial and antinematodal activity, and botrallin (1) exhibited moderate inhibitory activity on acetylcholinesterase.

  1. High-separation efficiency micro-fabricated multi-capillary gas chromatographic columns for simulants of the nerve agents and blister agents

    NASA Astrophysics Data System (ADS)

    Li, Yi; Du, Xiaosong; Wang, Yang; Tai, Huiling; Qiu, Dong; Lin, Qinghao; Jiang, Yadong

    2014-05-01

    To achieve both high speed and separation efficiency in the separation of a mixture of nerve and blister agent simulants, a high-aspect-ratio micro-fabricated multi-capillary column (MCC, a 50-cm-long, 450-μm-deep, and 60-μm-wide four-capillary column) was fabricated by the application of the microelectromechanical system (MEMS) techniques. Mixtures of chemical warfare agent (CWA) simulants - dimethyl methylphosphonate (DMMP), triethyl phosphate (TEP), and methyl salicylate - were used as samples. The fabricated MCC allowed for the separation of all the components of the gaseous mixture within 24 s, even when the difference in boiling point was 4°C, as in the case of TEP and methyl salicylate. Furthermore, interfering agents - dichloromethane, ethanol, and toluene - were also included in the subsequent gaseous mixture samples. The boiling point of these six components ranged from 78°C to 219°C. All six components were clearly separated within 70 s. This study is the first to report the clear separation of gas mixtures of components with close boiling points. The column efficiency was experimentally determined to be 12,810 plates/m.

  2. High-separation efficiency micro-fabricated multi-capillary gas chromatographic columns for simulants of the nerve agents and blister agents.

    PubMed

    Li, Yi; Du, Xiaosong; Wang, Yang; Tai, Huiling; Qiu, Dong; Lin, Qinghao; Jiang, Yadong

    2014-01-01

    To achieve both high speed and separation efficiency in the separation of a mixture of nerve and blister agent simulants, a high-aspect-ratio micro-fabricated multi-capillary column (MCC, a 50-cm-long, 450-μm-deep, and 60-μm-wide four-capillary column) was fabricated by the application of the microelectromechanical system (MEMS) techniques. Mixtures of chemical warfare agent (CWA) simulants - dimethyl methylphosphonate (DMMP), triethyl phosphate (TEP), and methyl salicylate - were used as samples. The fabricated MCC allowed for the separation of all the components of the gaseous mixture within 24 s, even when the difference in boiling point was 4°C, as in the case of TEP and methyl salicylate. Furthermore, interfering agents - dichloromethane, ethanol, and toluene - were also included in the subsequent gaseous mixture samples. The boiling point of these six components ranged from 78°C to 219°C. All six components were clearly separated within 70 s. This study is the first to report the clear separation of gas mixtures of components with close boiling points. The column efficiency was experimentally determined to be 12,810 plates/m. PMID:24899869

  3. A reproducible and high-throughput HPLC/MS method to separate sarcosine from α- and β-alanine and to quantify sarcosine in human serum and urine.

    PubMed

    Meyer, Tamra E; Fox, Stephen D; Issaq, Haleem J; Xu, Xia; Chu, Lisa W; Veenstra, Timothy D; Hsing, Ann W

    2011-07-15

    While sarcosine was recently identified as a potential urine biomarker for prostate cancer, further studies have cast doubt on its utility to diagnose this condition. The inconsistent results may be due to the fact that alanine and sarcosine coelute on an HPLC reversed-phase column and the mass spectrometer cannot differentiate between the two isomers, since the same parent/product ions are generally used to measure them. In this study, we developed a high-throughput liquid chromatography-mass spectrometry (LC-MS) method that resolves sarcosine from alanine isomers, allowing its accurate quantification in human serum and urine. Assay reproducibility was determined using the coefficient of variation (CV) and intraclass correlation coefficient (ICC) in serum aliquots from 10 subjects and urine aliquots from 20 subjects across multiple analytic runs. Paired serum/urine samples from 42 subjects were used to evaluate sarcosine serum/urine correlation. Both urine and serum assays gave high sensitivity (limit of quantitation of 5 ng/mL) and reproducibility (serum assay, intra- and interassay CVs < 3% and ICCs > 99%; urine assay, intra-assay CV = 7.7% and ICC = 98.2% and interassay CV = 12.3% and ICC = 94.2%). In conclusion, this high-throughput LC-MS method is able to resolve sarcosine from α- and β-alanine and is useful for quantifying sarcosine in serum and urine samples.

  4. Ion-Exclusion High-Performance Liquid Chromatography of Aliphatic Organic Acids Using a Surfactant-Modified C18 Column.

    PubMed

    Fasciano, Jennifer M; Mansour, Fotouh R; Danielson, Neil D

    2016-07-01

    Ion exclusion chromatography (IELC) of short chain aliphatic carboxylic acids is normally done using a cation exchange column under standard HPLC conditions but not in the ultra-HPLC (UHPLC) mode. A novel IELC method for the separation of this class of carboxylic acids by either HPLC or UHPLC utilizing a C18 column dynamically modified with sodium dodecyl sulfate has been developed. The sample capacity is estimated to be near 10 mM for a 20 µL injection or 0.2 µmol using a 150 × 4.6 mm column. The optimum mobile phase determined for three standard mixtures of organic acids is 1.84 mM sulfuric acid at pH 2.43 and a flow rate of 0.6 mL/min. Under optimized conditions, a HPLC separation of four aliphatic carboxylic acids such as tartaric, malonic, lactic and acetic can be achieved in under 4 min and in <2 min in the UHPLC mode at 2.1 mL/min. A variety of fruit juice and soft drink samples are analyzed. Stability of the column as measured by the retention order of maleic and fumaric acid is estimated to be ∼4,000 column volumes using HPLC and 600 by UHPLC. Reproducible chromatograms are achieved over at least a 2-month period. This study shows that the utility of a C18 column can be easily extended when needed to IELC under either standard or UHPLC conditions.

  5. Methods and applications of HPLC-AMS

    NASA Astrophysics Data System (ADS)

    Buchholz, Bruce A.; Dueker, Stephen R.; Lin, Yumei; Clifford, Andrew J.; Vogel, John S.

    2000-10-01

    Pharmacokinetics of physiologic doses of nutrients, pesticides, and herbicides can easily be traced in humans using a 14C-labeled compound. Basic kinetics can be monitored in blood or urine by measuring the elevation in the 14C content above the control predose tissue and converting to equivalents of the parent compound. High performance liquid chromatography (HPLC) is an excellent method for the chemical separation of complex mixtures whose profiles afford estimation of biochemical pathways of metabolism. Compounds elute from the HPLC systems with characteristic retention times and can be collected in fractions that can then be graphitized for AMS measurement. Unknowns are tentatively identified by co-elution with known standards and chemical tests that reveal functional groupings. Metabolites are quantified with the 14C signal. Thoroughly accounting for the carbon inventory in the LC solvents, ion-pairing agents, samples, and carriers adds some complexity to the analysis. In most cases the total carbon inventory is dominated by carrier. Baseline background and stability need to be carefully monitored. Limits of quantitation near 10 amol of 14C per HPLC fraction are typically achieved. Baselines are maintained by limiting injected 14C activity <0.17 Bq (4.5 pCi) on the HPLC column.

  6. Performance of single particle fritted capillary columns in electrochromatography.

    PubMed

    Zhang, Bo; Liu, Qing; Yang, Lijun; Wang, Qiuquan

    2013-01-11

    Development of capillary electrochromatography (CEC) largely depends on column technology. The past ten years or so have seen a great number of CEC works performed on monolithic columns, due to simplicity and robustness in column fabrication. Monolithic columns eliminate the issue of column fritting, which conventionally made particle-packed capillary columns fragile and introduced nonuniformity to the chromatographic bed. The particulate packing material, however, is still a popular type of stationary phase widely used in CEC, as the rich library of HPLC packing material provides a wide range of choices for chromatographic separations performed in electrodriven mode. In this study, we investigated a purely physical fritting method, single particle fritting technology, to immobilize particulate chromatographic material inside capillary tube in a sinter-free manner to produce robust capillary columns. Single particle fritted columns present significantly improved column-to-column reproducibility (n=10) in peak efficiency, retention factor, peak area and asymmetry (%RSD=5.4, 7.7, 6.2 and 6.1, respectively, at 26 kV), enabling their practical application in high throughput parallel analysis using multiple columns.

  7. Monolithic metal-organic framework MIL-53(Al)-polymethacrylate composite column for the reversed-phase capillary liquid chromatography separation of small aromatics.

    PubMed

    Yusuf, Kareem; Badjah-Hadj-Ahmed, Ahmed Yacine; Aqel, Ahmad; ALOthman, Zeid Abdullah

    2016-03-01

    A monolithic capillary column containing a composite of metal-organic framework MIL-53(Al) incorporated into hexyl methacrylate-co-ethylene dimethacrylate was prepared to enhance the separation of mixtures of small aromatic compounds by using capillary liquid chromatography. The addition of 10 mg/mL MIL-53(Al) microparticles increased the micropore content in the monolithic matrix and increased the Brunauer-Emmett-Teller surface area from 26.92 to 85.12 m(2) /g. The presence of 1,4-benzenedicarboxylate moieties within the structure of MIL-53(Al) as an organic linker greatly influenced the separation of aromatic mixtures through π-π interactions. High-resolution separation was obtained for a series of alkylbenzenes (with resolution factors in the range 0.96-1.75) in less than 8 min, with 14 710 plates/m efficiency for propylbenzene, using a binary polar mobile phase of water/acetonitrile in isocratic mode. A reversed-phase separation mechanism was indicated by the increased retention factor and resolution as the water percentage in the mobile phase increased. A stability study on the composite column showed excellent mechanical stability under various conditions. The higher resolution and faster separation observed at increased temperature indicated an exothermic separation, whereas the negative values for the free energy change of transfer indicated a spontaneous process.

  8. Oligomers matrix-assisted dispersion of high content of carbon nanotubes into monolithic column for online separation and enrichment of proteins from complex biological samples.

    PubMed

    Zhou, Chanyuan; Du, Zhuo; Li, Gongke; Zhang, Yukui; Cai, Zongwei

    2013-10-01

    In this work, a new oligomer matrix-assisted dispersion (OMAD) method for the preparation of homogeneous dispersion of multi-walled carbon nanotubes (MWNTs) incorporated monolithic column was developed. Oligomers matrix as a scaffold could allow MWNTs to entangle with it instead of self-aggregation, so the MWNTs remain in the polymer network followed by in situ self-solidification. The OMAD method not only greatly enlarged the BET surface area of MWNTs incorporated monolithic column from 13.8 m(2) g(-1) to 85.5 m(2) g(-1) without a significant effect on the surface chemistry of the MWNTs, but also improved the dispersion of MWNTs making its content up to 5 wt% (with respect to monomers). The synthesized materials combine the favorable attributes of both high permeability and large surface area, making them excellent candidates for on-line separation and enrichment of proteins. The oligomer matrix-assisted dispersion MWNTs incorporated monolithic columns (OMAD-MMC) exhibited higher enrichment factors and the adsorption capacity is about 5-fold for basic proteins compared with MWNTs incorporated monolithic columns (MMC) prepared by the conventional in situ polymerization. The practical application of OMAD-MMC was proven by selective extraction of hemoglobin in human whole blood samples with SDS-PAGE. On the basis of the results, OMAD as a simple and effective method for dispersion high content MWNTs into monolithic columns shows great promise.

  9. Oligomers matrix-assisted dispersion of high content of carbon nanotubes into monolithic column for online separation and enrichment of proteins from complex biological samples.

    PubMed

    Zhou, Chanyuan; Du, Zhuo; Li, Gongke; Zhang, Yukui; Cai, Zongwei

    2013-10-01

    In this work, a new oligomer matrix-assisted dispersion (OMAD) method for the preparation of homogeneous dispersion of multi-walled carbon nanotubes (MWNTs) incorporated monolithic column was developed. Oligomers matrix as a scaffold could allow MWNTs to entangle with it instead of self-aggregation, so the MWNTs remain in the polymer network followed by in situ self-solidification. The OMAD method not only greatly enlarged the BET surface area of MWNTs incorporated monolithic column from 13.8 m(2) g(-1) to 85.5 m(2) g(-1) without a significant effect on the surface chemistry of the MWNTs, but also improved the dispersion of MWNTs making its content up to 5 wt% (with respect to monomers). The synthesized materials combine the favorable attributes of both high permeability and large surface area, making them excellent candidates for on-line separation and enrichment of proteins. The oligomer matrix-assisted dispersion MWNTs incorporated monolithic columns (OMAD-MMC) exhibited higher enrichment factors and the adsorption capacity is about 5-fold for basic proteins compared with MWNTs incorporated monolithic columns (MMC) prepared by the conventional in situ polymerization. The practical application of OMAD-MMC was proven by selective extraction of hemoglobin in human whole blood samples with SDS-PAGE. On the basis of the results, OMAD as a simple and effective method for dispersion high content MWNTs into monolithic columns shows great promise. PMID:23917344

  10. Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns.

    PubMed

    Jones, Andrew; Pravadali-Cekic, Sercan; Hua, Stanley; Kocic, Danijela; Camenzuli, Michelle; Dennis, Gary; Shalliker, Andrew

    2016-04-26

    A protocol for the use of reaction flow high performance liquid chromatography columns for methods employing post column derivatization (PCD) is presented. A major difficulty in adapting PCD to modern HPLC systems and columns is the need for large volume reaction coils that enable reagent mixing and then the derivatization reaction to take place. This large post column dead volume leads to band broadening, which results in a loss of observed separation efficiency and indeed detection in sensitivity. In reaction flow post column derivatization (RF-PCD) the derivatization reagent(s) are pumped against the flow of mobile phase into either one or two of the outer ports of the reaction flow column where it is mixed with column effluent inside a frit housed within the column end fitting. This technique allows for more efficient mixing of the column effluent and derivatization reagent(s) meaning that the volume of the reaction loops can be minimized or even eliminated altogether. It has been found that RF-PCD methods perform better than conventional PCD methods in terms of observed separation efficiency and signal to noise ratio. A further advantage of RF-PCD techniques is the ability to monitor effluent coming from the central port in its underivatized state. RF-PCD has currently been trialed on a relatively small range of post column reactions, however, there is currently no reason to suggest that RF-PCD could not be adapted to any existing one or two component (as long as both reagents are added at the same time) post column derivatization reaction.

  11. Separation of Peptides on HALO 2-Micron Particles.

    PubMed

    Mant, Colin T; Hodges, Robert S

    2016-01-01

    Reversed-phase high-performance liquid chromatography (RP-HPLC) is of fundamental importance to the isolation and separation of peptides, proteins, and other biomolecules. Hence, there is a continuing high demand for the development of RP-HPLC stationary-phase materials with enhanced separation efficiency. HALO packing materials began the revolution in "core-shell" technology with the advantages of faster separations, higher resolution and peak capacity, high temperature stability, and rugged reliable performance compared to traditional HPLC and UHPLC. These materials are characterized by a solid core surrounded by a thin layer of porous material, and represent a technology for the future with continuing refinements. Such refinements are aided via the use of designed synthetic peptide standards during stationary-phase development. Concomitantly, such standards also enable the researcher to monitor RP-HPLC column performance and develop optimized separation protocols for peptides from a wide array of sources. © 2016 by John Wiley & Sons, Inc. PMID:27479502

  12. Effect of polyethylene glycol on pore structure and separation efficiency of silica-based monolithic capillary columns.

    PubMed

    Hara, Takeshi; Desmet, Gert; Baron, Gino V; Minakuchi, Hiroyoshi; Eeltink, Sebastiaan

    2016-04-15

    Monolithic silica materials (first unclad monolith rods, then monolithic capillary columns) were prepared using various amounts of polyethylene glycols (PEGs) with different molecular weight (MW). The monolith rods were used to examine the mesoporosity by argon physisorption technique, and the macroporosity by mercury intrusion porosimetry. Subsequently, silica-based monolithic capillary columns with an inner diameter of 100 μm were produced using the same preparation conditions as used for the rods. The results obtained with the monolith rods showed the following important findings: (1) it is feasible to fabricate monolithic silica rods possessing macropore size of 0.5-1.4 μm by tuning the amount of PEGs (independently of the MW), whereas the macropore volume and the mesoporosity remain similar. (2) the smallest macropore size (0.5 μm) rod prepared with PEG having a MW=20,000g/mol provided a narrower macropore size distribution than with PEG with MW=10,000g/mol. The monolithic capillary columns produced with the different PEG type showed similar retention factors for hexylbenzene (k=2.3-2.4) and similar t0-based column permeability (Kv0=2.3-2.4×10(-14)m(2)) in 20:80% (v/v) water:methanol, as expected from the results obtained with the monolith rods. The column prepared with PEG of MW=20,000g/mol gave a plate height of H=4.0 μm for hexylbenzene at an optimal linear velocity of u0=2.6mm/s in 20:80% (v/v) water containing 0.1% formic acid:acetonitrile. To the best of our knowledge, this is the lowest plate height ever recorded for a monolithic column. Comparing the kinetic performance at 30MPa shows that the best monolithic silica column obtained in the present study performs better than the second-generation monolithic silica columns developed up till now in the practically most relevant range of plate numbers (N≤40,000). In this range, the performance is now similar to that of 2.7 μm core-shell particle columns. PMID:26976349

  13. Recent advances in SPE-chiral-HPLC methods for enantiomeric separation of chiral drugs in biological samples.

    PubMed

    Ali, Imran; Alam, Syed Dilshad; Al-Othman, Zeid A; Farooqi, Javed A

    2013-08-01

    In medical practices, the determination of enantiomeric ratio of the chiral drugs is very important for their activities, bioavailabilities and biodegradation. Only homochiral medication is safe for humans. The chiral analysis in biological samples is the first and most important step. The present article describes the technical strategies of the enantiomeric resolution of racemic drugs in biological samples. Attempts have been made to describe sample preparation by solid-phase extraction and enantiomeric resolution by chiral high-performance liquid chromatography. Various chiral stationary phases used in chiral separations of racemic drugs have been described. Efforts are also made to discuss the chiral recognition mechanism and future perspectives of chiral analyses in biological samples.

  14. On-line SPE-UHPLC method using fused core columns for extraction and separation of nine illegal dyes in chilli-containing spices.

    PubMed

    Khalikova, Maria A; Satínský, Dalibor; Smidrkalová, Tereza; Solich, Petr

    2014-12-01

    The presented work describes the development of a simple, fast and effective on-line SPE-UHPLC-UV/vis method using fused core particle columns for extraction, separation and quantitative analysis of the nine illegal dyes, most frequently found in chilli-containing spices. The red dyes Sudan I-IV, Sudan Red 7B, Sudan Red G, Sudan Orange G, Para Red, and Methyl Red were separated and analyzed in less than 9 min without labor-consuming pretreatment procedure. The chromatographic separation was performed on Ascentis Express RP-Amide column with gradient elution using mixture of acetonitrile and water, as a mobile phase at a flow rate of 1.0 mL min(-1) and 55°C of temperature. As SPE sorbent for cleanup and pre-concentration of illegal dyes short guard fused core column Ascentis Express F5 was used. The applicability of proposed method was proven for three different chilli-containing commercial samples. Recoveries for all compounds were between 90% and 108% and relative standard deviation ranged from 1% to 4% for within- and from 2% to 6% for between-day. Limits of detection showed lower values than required by European Union regulations and were in the range of 3.3-10.3 µg L(-1) for standard solutions, 5.6-235.6 µg kg(-1) for chilli-containing spices.

  15. Synthesis and characterization of a multimode stationary phase: Congo red derivatized silica in nano-flow HPLC.

    PubMed

    Zhang, Yi; Zhang, Yan; Wang, Guan; Chen, Wujuan; He, Pingang; Wang, Qingjiang

    2016-02-01

    A novel Congo red (CR) derivatized silica stationary phase was prepared and packed into a fused silica capillary tube for nano-flow HPLC. A variety of analytes including poly-aromatic hydrocarbons, parabens, acids, sulfonamides, bases, and nucleosides were successfully separated using the CR. In comparison with commercial ODS columns, this new stationary phase has a different separation mechanism (hydrophobically-assisted ion-exchange), which was evident in the separation of benzoic acid derivatives and sulfonamides. The successful application of CR-bonded silica stationary phase in the HILIC and PALC modes demonstrates the effectiveness of this potential chromatographic material in nano flow HPLC.

  16. Synthesis and characterization of a multimode stationary phase: Congo red derivatized silica in nano-flow HPLC.

    PubMed

    Zhang, Yi; Zhang, Yan; Wang, Guan; Chen, Wujuan; He, Pingang; Wang, Qingjiang

    2016-02-01

    A novel Congo red (CR) derivatized silica stationary phase was prepared and packed into a fused silica capillary tube for nano-flow HPLC. A variety of analytes including poly-aromatic hydrocarbons, parabens, acids, sulfonamides, bases, and nucleosides were successfully separated using the CR. In comparison with commercial ODS columns, this new stationary phase has a different separation mechanism (hydrophobically-assisted ion-exchange), which was evident in the separation of benzoic acid derivatives and sulfonamides. The successful application of CR-bonded silica stationary phase in the HILIC and PALC modes demonstrates the effectiveness of this potential chromatographic material in nano flow HPLC. PMID:26646316

  17. Determination of thimerosal in pharmaceutical industry effluents and river waters by HPLC coupled to atomic fluorescence spectrometry through post-column UV-assisted vapor generation.

    PubMed

    Acosta, Gimena; Spisso, Adrián; Fernández, Liliana P; Martinez, Luis D; Pacheco, Pablo H; Gil, Raúl A

    2015-03-15

    A high performance liquid chromatography coupled with atomic fluorescence spectrometry method for the determination of thimerosal (sodium ethylmercury thiosalicylate, C9H9HgNaO2S), ethylmercury, and inorganic mercury is proposed. Mercury vapor is generated by the post-column reduction of mercury species in formic acid media using UV-radiation. Thimerosal is quantitatively converted to Hg(II) followed by the reduction of Hg(II) to Hg(0). This method is applied to the determination of thimerosal (THM), ethylmercury (EtHg) and inorganic Hg in samples of a pharmaceutical industry effluent, and in waters of the San Luis River situated in the west side of San Luis city (Middle West, Argentine) where the effluents are dumped. The limit of detections, calculated on the basis of the 3σ criterion, where 0.09, 0.09 and 0.07 μg L(-1) for THM, EtHg(II) and for Hg(II), respectively. Linearity was attained from levels close to the detection limit up to at least 100 μg L(-1).

  18. A compact gas chromatograph and pre-column concentration system for enhanced in-field separation of levoglucosan and other polar organic compounds.

    PubMed

    Cropper, Paul M; Goates, Steven R; Hansen, Jaron C

    2015-10-23

    Portable and compact instruments for separating and detecting organic compounds are needed in the field for environmental studies. This is especially the case for pollution studies as in-field detection of organic compounds helps identify sources of pollution. Here we describe the development of a compact GC and simple pre-concentrator coupled to a MS detector. This simple system can easily be incorporated into portable instrumentation. Combining the pre-concentrator and compact column has the advantage of decoupling separation from manual injection and enhances separation of environmentally relevant polar organic compounds, such as levoglucosan. A detection limit of 2.2 ng was obtained for levoglucosan. This simple design has the potential to expand the use of gas chromatography as a routine in-field separation technique. PMID:26410183

  19. A compact gas chromatograph and pre-column concentration system for enhanced in-field separation of levoglucosan and other polar organic compounds.

    PubMed

    Cropper, Paul M; Goates, Steven R; Hansen, Jaron C

    2015-10-23

    Portable and compact instruments for separating and detecting organic compounds are needed in the field for environmental studies. This is especially the case for pollution studies as in-field detection of organic compounds helps identify sources of pollution. Here we describe the development of a compact GC and simple pre-concentrator coupled to a MS detector. This simple system can easily be incorporated into portable instrumentation. Combining the pre-concentrator and compact column has the advantage of decoupling separation from manual injection and enhances separation of environmentally relevant polar organic compounds, such as levoglucosan. A detection limit of 2.2 ng was obtained for levoglucosan. This simple design has the potential to expand the use of gas chromatography as a routine in-field separation technique.

  20. Simultaneous separation of water- and fat-soluble vitamins in isocratic pressure-assisted capillary electrochromatography using a methacrylate-based monolithic column.

    PubMed

    Yamada, Hiroki; Kitagawa, Shinya; Ohtani, Hajime

    2013-06-01

    A method of simultaneous separation of water- and fat-soluble vitamins using pressure-assisted CEC with a methacrylate-based capillary monolithic column was developed. In the proposed method, water-soluble vitamins were mainly separated electrophoretically, while fat soluble-ones were separated chromatographically by the interaction with a methacrylate-based monolith. A mixture of six water-soluble and four fat-soluble vitamins was separated simultaneously within 20 min with an isocratic elution using 1 M formic acid (pH 1.9)/acetonitrile (30:70, v/v) containing 10 mM ammonium formate as a mobile phase. When the method was applied to a commercial multivitamin tablet and a spiked one, the vitamins were successfully analyzed, and no influence of the matrix contained in the tablet was observed.

  1. Computational fluid dynamics simulations yielding guidelines for the ideal internal structure of monolithic liquid chromatography columns.

    PubMed

    Gzil, P; Baron, G V; Desmet, G

    2003-04-01

    A theoretical calculation of the separation performance of a (hypothetical) micro-structured monolithic LC column is presented, confirming that the polydispersity effect in parallel bundle columns can theoretically be eliminated to a very large extent by radially redistributing the mobile phase fluid at regular intervals. It is demonstrated that the flow can be redistributed in such a way that the advantage coming from the suppression of the polydispersity effect largely exceeds the losses caused by the additional pressure-drop and band broadening. The presently considered micro-structured column would allow to perform N > 100,000 plate separations in a few hundred of seconds, i.e., about an order of magnitude faster than the best possible packed bed and monolithic HPLC columns, while offering the same mass loadability. This clearly demonstrates that the currently available LC columns are still far away from the absolute resolution limit of the ideal, fully optimised LC column.

  2. Kinetic performance evaluation and perspectives of contemporary packed column capillary electrochromatography.

    PubMed

    De Smet, Seppe; Lynen, Frederic

    2014-08-15

    Capillary electrochromatography (CEC) is in essence a highly efficient and fast separation technique but practical constraints limit the current performance, robustness and routine implementation of the technique. In this work the kinetic performance limit (KPL) curve was used to evaluate commercial packed column CEC; this firstly in order to assess the broader applicability of the kinetic plot approach in electrodriven chromatographic techniques, and secondly to allow a more general unbiased comparison with HPLC performance. Evaluations were performed with a mixture of well retained and electrophoretically neutral phenones, to allow the observation of only chromatographic processes. Initial CEC retention time irreproducibility issues were solved by applying high acetonitrile content (80%) in the mobile phase, and solute retention was increased by increasing the phenone chain length. Comparison was performed with HPLC, with a column packed with an identical stationary phase to allow measurement of the performance under optimal conditions, and not with μ-LC on the CEC column as extra column peak broadening phenomena would thereby negatively affect the μ-LC performance. This comparison demonstrated that current HPLC performance largely outcompetes what is achievable with contemporary packed column CEC. Interestingly, significantly improved CEC performance could be obtained at lower temperatures (10°C) indicating a persistent degree of joule heating phenomena taking place in the contemporary packed column (100μm) CEC approach. Effective suppression of the latter opens possibilities for increasing the applicable voltage and outperforming HPLC and UHPLC.

  3. [Online enrichment ability of restricted-access column coupled with high performance liquid chromatography by column switching technique for benazepril hydrochloride].

    PubMed

    Zhang, Xiaohui; Wang, Rong; Xie, Hua; Yin, Qiang; Li, Xiaoyun; Jia, Zhengping; Wu, Xiaoyu; Zhang, Juanhong; Li, Wenbin

    2013-05-01

    The online enrichment ability of the restricted-access media (RAM) column coupled with high performance liquid chromatography by column switching technique for benazepril hydrochloride in plasma was studied. The RAM-HPLC system consisted of an RAM column as enrichment column and a C18 column as analytical column coupled via the column switching technique. The effects of the injection volume on the peak area and the systematic pressure were studied. When the injection volume was less than 100 microL, the peak area increased with the increase of the injection volume. However, when the injection volume was more than 80 microL, the pressure of whole system increased obviously. In order to protect the whole system, 80 microL was chosen as the maximum injection volume. The peak areas of ordinary injection and the large volume injection showed a good linear relationship. The enrichment ability of RAM-HPLC system was satisfactory. The system was successfully used for the separation and detection of the trace benazepril hydrochloride in rat plasma after its administration. The sensitivity of HPLC can be improved by RAM pre-enrichment. It is a simple and economic measurement method.

  4. Development and validation of an HPLC/UV assay for separation and quantification of peptide antigens from a liposomal vaccine delivery platform.

    PubMed

    Penwell, Andrea; Sharp, Kendall; Mansour, Marc; Sammatur, Leeladhar

    2012-07-01

    The development and validation of an HPLC method for the quantification of eight peptide antigens from the therapeutic cancer vaccine DPX-0907 is described. The antigens were formulated in DepoVax™, a patented liposomal vaccine delivery platform used in a phase 1 study for breast, ovarian, and prostate cancers. A gradient reversed-phase method with UV detection was optimized for separating and quantifying the peptide mixture. Several extraction methods investigated to extract the peptides from the lipids led to poor recovery of one or more of the peptides. A simple, reproducible, and high-recovery extraction procedure for the simultaneous quantification of hydrophilic and hydrophobic peptides was discovered using a liquid-liquid extraction with water-saturated n-butanol and sodium bicarbonate (0.1 M). The method was found to be specific, linear, accurate, precise, and reliable within the range of 50-150% of the nominal concentration for DPX-0907. The validated method was successfully applied to the assay of peptide content in pre-clinical and clinical batches of DPX-0907. PMID:22516680

  5. Comparison of microbial communities in Lake Tahoe surface sample with Tonga Trench water column samples using High Pressure Liquid Chromatography - Electrospray Ionization - Mass Spectroscopy (HPLC - ESI - MS) and Global Natural Products Social Molecular Network (GNPS)

    NASA Astrophysics Data System (ADS)

    Belmonte, M. A.

    2015-12-01

    Intact polar lipids (IPLs) are lipids composed of a head group, a glycerol, and a fatty acid chain that make up the lipid bilayer of cell membranes in living cells; and the varying head groups can be indicative of the type of microbes present in the environment (Van Mooy 2010). So by distinguishing and identifying the IPL distribution in an environment one can make inferences about the microbial communities in the said environment. In this study, we used High Pressure Liquid Chromatography-Electrospray Ionization- Mass Spectroscopy (HPLC-ESI-MS) and Global Natural Products Social Molecular Networking (GNPS) to compare the IPL distributions of two oligotrophic environments: surface waters of Lake Tahoe in the Sierra Nevada Mountains, and the water column of the Tonga Trench in the South Pacific. We hypothesized that the similar nutrient dynamics of the two oligotrophic environments would result in similar eukaryotic and prokaryotic communities, which would be reflected in the IPL composition of suspended particulate organic matter (POM). For simplicity we focused on the classes of IPLs most commonly observed in the marine environment: phosphotidylglycerol (PG), phosphotidylethanolamine (PE), diacylglyceryl-trimethyl-homoserine (DGTS), diacylglyceryl-hydroxymethyl-trimethylalanine (DGTA), sulfoquinovosyldiacylglycerol (SQDG), monoglycosyldiacylglycerol (MGDG) and diglycosyldiacylglycerol (DGDG). Our results showed that all of the marine IPLs of interest were present in Lake Tahoe which confirms that there are many of the same microbial communities in the fresh waters of Lake Tahoe and the salt waters Tonga Trench.

  6. Monolithic poly (SPE-co-BVPE) capillary columns as a novel hydrophilic interaction liquid chromatography stationary phase for the separation of polar analytes.

    PubMed

    Foo, Hsiao Ching; Heaton, James; Smith, Norman W; Stanley, Shawn

    2012-10-15

    A novel hydrophilic interaction liquid chromatography (HILIC) stationary phase was prepared by the co-polymerisation of zwitterionic N,N'-dimethyl-N-methacryloxyethyl-N-(3-sulfopropyl) ammonium betaine (SPE) and the crosslinker 1,2-bis(p-vinylphenyl) ethane (BVPE) in the presence of the porogens, toluene and methanol. Monolithic columns were produced by carrying out the α,α'-azoisobutyronitrile (AIBN) initiated reaction for 1, 2, 4, 8 and 12 h inside a 200 μm i.d. fused silica capillary at 75°C (water bath). The optimum polymerisation time was shown to be 2 h, as this resulted in good porosity, due to enlarged flow-channels and the presence of a higher proportion of mesopores provided a relatively larger surface area than the other columns. The chromatographic properties of the optimised poly (SPE-co-BVPE) monolithic column were evaluated with test mixtures containing both basic and neutral compounds in the HILIC gradient separation mode. This produced relatively sharp peaks (average peak width at half height=0.1 min) with average asymmetry factors of 1.4 and baseline resolution was obtained for all the compounds. Using the isocratic separation of the test mixture, the number of theoretical plates (N) per metre calculated was between 26,888 and 35,930 by using average values obtained for triplicate injections of the compounds thiourea, toluene and acrylamide.

  7. Application of an efficient strategy based on liquid-liquid extraction, high-speed counter-current chromatography, and preparative HPLC for the rapid enrichment, separation, and purification of four anthraquinones from Rheum tanguticum.

    PubMed

    Chen, Tao; Liu, Yongling; Zou, Denglang; Chen, Chen; You, Jinmao; Zhou, Guoying; Sun, Jing; Li, Yulin

    2014-01-01

    This study presents an efficient strategy based on liquid-liquid extraction, high-speed counter-current chromatography, and preparative HPLC for the rapid enrichment, separation, and purification of four anthraquinones from Rheum tanguticum. A new solvent system composed of petroleum ether/ethyl acetate/water (4:2:1, v/v/v) was developed for the liquid-liquid extraction of the crude extract from R. tanguticum. As a result, emodin, aloe-emodin, physcion, and chrysophanol were greatly enriched in the organic layer. In addition, an efficient method was successfully established to separate and purify the above anthraquinones by high-speed counter-current chromatography and preparative HPLC. This study supplies a new alternative method for the rapid enrichment, separation, and purification of emodin, aloe-emodin, physcione, and chrysophanol.

  8. Chiral separation of cathinone and amphetamine derivatives by HPLC/UV using sulfated ß-cyclodextrin as chiral mobile phase additive.

    PubMed

    Taschwer, Magdalena; Seidl, Yvonne; Mohr, Stefan; Schmid, Martin G

    2014-08-01

    In the last years the identification of new legal and illegal highs has become a huge challenge for the police and prosecution authorities. In an analytical context, only a few analytical methods are available to identify these new substances. Moreover, many of these recreational drugs are chiral and it is supposed that the enantiomers differ in their pharmacological potency. Since nonenantioselective synthesis is easier and cheaper, they are mainly sold as racemic mixtures. The goal of this research work was to develop an inexpensive method for the chiral separation of cathinones and amphetamines. This should help to discover if the substances are sold as racemic mixtures and give further information about their quality as well as their origin. Chiral separation of a set of 6 amphetamine and 25 cathinone derivatives, mainly purchased from various Internet shops, is presented. A LiChrospher 100 RP-18e, 250 x 4 mm, 5 µm served as the stationary phase. The chiral mobile phase consisted of methanol, water, and sulfated ß-cyclodextrin. Measurements were performed under isocratic conditions in reversed phase mode using UV detection. Four model compounds of the two substance classes were used to optimize the mobile phase. Under final conditions (methanol:water 2.5:97.5 + 2% sulfated ß-cyclodextrin) enantiomers of amphetamine and five derivatives were baseline separated within 23 min. In all, 17 cathinones were completely or partially chirally separated. However, as only 3 of 25 cathinones were baseline resolved, the application of this method is limited for cathinone analogs. Additionally, the results were compared with an RP-8e column.

  9. Separation of alkanes and aromatic compounds by packed column gas chromatography using functionalized multi-walled carbon nanotubes as stationary phases.

    PubMed

    Speltini, Andrea; Merli, Daniele; Quartarone, Eliana; Profumo, Antonella

    2010-04-23

    In the present work, we show a novel application of pristine and functionalized Multi-Walled Carbon Nanotubes (MWCNTs) as stationary phase in low-cost packed columns for the gas chromatographic separation of alkanes and aromatic hydrocarbons. The MWCNTs were deeply investigated by means of physical and chemical methods, like thermal analysis, IR and atomic force microscopy, and Inverse Gas Chromatography (IGC) in order to correlate the adsorption process and surface properties with the material purity level and functionalization degree. The derivatization process of the pristine nanotubes was a key factor to achieve a successful separation of both the light n-alkanes (C3-C5) and the related isomers (C4-C5 branched alkanes). Satisfactory results were similarly obtained in the case of separation of aromatic hydrocarbons (BTX).

  10. Retinoid quantification by HPLC/MS(n)

    NASA Technical Reports Server (NTRS)

    McCaffery, Peter; Evans, James; Koul, Omanand; Volpert, Amy; Reid, Kevin; Ullman, M. David

    2002-01-01

    Retinoic acid (RA) mediates most of the biological effects of vitamin A that are essential for vertebrate survival. It acts through binding to receptors that belong to the nuclear receptor transcription factor superfamily (Mangelsdorf et al. 1994). It is also a highly potent vertebrate teratogen. To determine the function and effects of endogenous and exogenous RA, it is important to have a highly specific, sensitive, accurate, and precise analytical procedure. Current analyses of RA and other retinoids are labor intensive, of poor sensitivity, have limited specificity, or require compatibility with RA reporter cell lines (Chen et al. 1995. BIOCHEM: Pharmacol. 50: 1257-1264; Creech Kraft et al. 1994. BIOCHEM: J. 301: 111-119; Lanvers et al. 1996. J. Chromatogr. B Biomed. Appl. 685: 233-240; Maden et al. 1998. DEVELOPMENT: 125: 4133-4144; Wagner et al. 1992. DEVELOPMENT: 116: 55-66). This paper describes an HPLC/mass spectrometry/mass spectrometry product ion scan (HPLC/MS(n)) procedure for the analysis of retinoids that employs atmospheric pressure chemical ionization MS. The retinoids are separated by normal-phase column chromatography with a linear hexane-isopropanol-dioxane gradient. Each retinoid is detected by a unique series of MS(n) functions set at optimal collision-induced dissociation energy (30% to 32%) for all MS(n) steps. The scan events are divided into three segments, based on HPLC elution order, to maximize the mass spectrometer duty cycle. The all-trans, 9-cis, and 13-cis RA isomers are separated, if desired, by an isocratic hexane-dioxane-isopropanol mobile phase. This paper describes an HPLC/MS(n) procedure possessing high sensitivity and specificity for retinoids.

  11. Precolumn o-phthalaldehyde-N-acetyl-L-cysteine derivatization followed by RP-HPLC separation and fluorescence detection of sitagliptin enantiomers in rat plasma.

    PubMed

    Nageswara Rao, R; Sravan, B; Ramakrishna, K; Saida, Shaik; Padiya, Raju

    2013-12-01

    An indirect reversed-phase high-performance liquid chromatographic separation and fluorescence detection of sitagliptin enantiomers in rat plasma was developed and validated. Deproteinized rat plasma containing racemic sitagliptin was derivatized with o-phthalaldehyde and N-acetyl-L-cysteine under alkaline conditions, converted to diastereomers, and separated on a Lichrospher 100 RP-18e column using 20 mM phosphate buffer and methanol (45:55 v/v) as a mobile phase under isocratic mode of elution at a flow rate of 1.0 mL/min. Fluorescence detection was performed at 330 and 450 nm as excitation and emission wavelengths, respectively. The method was linear in the range of 50-5000 ng/ mL for both enantiomers. The intra- and interday accuracy and precision were within the predefined limits of ≤15% at all concentrations. The method was successfully applied to a pharmacokinetic study of sitagliptin after 5 mg/kg oral administration to Wistar rats. Robustness of the method was evaluated using design of experiments.

  12. Development and validation of a reversed-phase HPLC method for separation and simultaneous determination of process-related substances of mirtazapine in bulk drugs and formulations.

    PubMed

    Rao, R Nageswara; Raju, A Narasa

    2009-03-01

    A simple and rapid reversed-phase high-performance liquid chromatographic method has been developed for the separation and simultaneous determination of related substances of mirtazapine in bulk drugs and pharmaceutical formulations. Six impurities, including one degradation product of mirtazapine, have been separated on a BDS Hypersil (4.6 x 250 mm; particle size 5 microm) column with a mobile phase consisting of 0.3% triethylamine (pH 3.0)-acetonitrile (78:22 v/v) eluted in an isocratic mode and monitored with a photo diode array detector at 215 nm. The chromatographic behavior of all the analytes was studied under variable compositions of different solvent systems, temperatures, buffer concentrations, and pH values. The method was validated in terms of accuracy, precision, and linearity. The inter- and intra-day assay precision was found to be < 0.98% [relative standard deviation; (RSD)] and the recoveries were in the range 95.54-102.22% with RSD < 2.21%. The correlation coefficients for calibration curves for mirtazapine as well as impurities were in the range of 0.9941-0.9999. The method was successfully applied to the analysis of commercial formulations and the recoveries of mirtazapine were in the range of 99.38-100.73% with < 0.52% RSD. The method is useful not only for rapid evaluation of the purity of mirtazapine, but also for the simultaneous determination of related substances in bulk drugs and pharmaceutical formulations.

  13. Rapid separation and determination of process-related substances of paracetamol using reversed-phase HPLC with photo diode array as a detector.

    PubMed

    Rao, R Nageswara; Narasaraju, A

    2006-02-01

    A simple and rapid gradient reversed-phase high-performance liquid chromatographic method for simultaneous separation and determination of paracetamol and its related compounds in bulk drugs and pharmaceutical formulations has been developed. As many as nine process impurities and one degradation product of paracetamol have been separated on a Symmetry C18 column (4.6 x 250 mm i.d., particle size 5 microm) with gradient elution using 0.01 M potassium dihydrogen phosphate buffer (pH 3.0) and acetonitrile as mobile phase and photo diode array detection at 215 nm. The chromatographic behavior of all the compounds was examined under variable compositions of different solvents, temperatures, buffer concentrations and pH values. The correlation coefficients for calibration curves for paracetamol as well as impurities were in the range of 0.9951 - 0.9994. The proposed RP-LC method was successfully applied to the analysis of commercial formulations; the recoveries of paracetamol were in the range of 99-101%. The method could be of use not only for rapid and routine evaluation of the quality of paracetamol in bulk drug manufacturing units but also for detection of its impurities in pharmaceutical formulations.

  14. Analysis of flavonoids and iridoids in Vitex negundo by HPLC-PDA and method validation.

    PubMed

    Roy, Somendu K; Bairwa, Khemraj; Grover, Jagdeep; Srivastava, Amit; Jachak, Sanjay M

    2013-09-01

    The leaves of Vitex negundo have been reported to contain various bioactive constituents including iridoids and flavonoids. This is the first report on the simultaneous determination of iridoids and flavonoids by HPLC in three different samples of V. negundo leaves collected from three regions of India. Separation of iridoids and flavonoids was accomplished by HPLC and further elaborated for their quantification in V. negundo leaves using a C-18 column with detection at 254 and 330 nm, respectively. The developed HPLC method showed good linearity (r2 > or = 0.999), high precision (RSD < 5%) and a good recovery (99.3-103.0%) of the compounds. All the validation parameters of the developed HPLC were found to be within the permissible limits according to the ICH guidelines. The developed method was robust, accurate and reliable for the quality control of V. negundo leaves.

  15. Comparison of a jet separator and an open splitter as an interface between a multi-capillary gas chromatographic column and a time-of-flight mass spectrometer

    PubMed

    Pongpun; Mlynski; Crisp; Guilhaus

    2000-09-01

    A gas chromatographic/time-of-flight mass spectrometric (GC/TOFMS) interface is being developed for fast on-line analysis utilizing multi-capillary column technology. A variable gap-distance jet separator has been constructed and its performance compared with that of a commercially supplied post-column open splitter recommended for use between the multi-capillary column and a mass spectrometer. Both interfaces were found to be compatible with the GC/TOFMS system at high carrier gas flow-rates, facilitating high-speed and high-resolution separations. The systems were investigated and tested with a mixture of volatile organic compounds (VOCs) with molecular masses from 85 to 166: dichloromethane, toluene, m-dichlorobenzene, o-dichlorobenzene and tetrachloroethylene. The optimum tip-to-tip gap distance corresponding to the highest efficiency of the jet separator was found to be 0.030 mm for each compound at carrier gas flow-rates of 20, 40 and 60 ml min(-1) giving, in the ion source housing, ion gauge pressure readings of 1.6 x 10(-6), 5.0 x 10(-6) and 5.8 x 10(-6) mbar, respectively. The efficiency of the jet separator (10-30% yields) was significantly higher than that of the open splitter (6-9% yields). The observation that the open splitter did not provide a constant flow-rate to the ion source was not in agreement with the manufacturer's specifications. A method for measuring the gas flow-rates in all parts of the equipment is described. The correlation between yield in the jet separator and molecular mass for the heterogeneous set of compounds studied was found to be less linear than usually reported for homologous series of compounds in jet separator studies. The result suggests that the pressure conditions in the jet may be sufficient for the separation process to be partly controlled by diffusion rather than predominately by effusion. Copyright 2000 John Wiley & Sons, Ltd.

  16. Preparation of porous polymer monolithic column using functionalized graphene oxide as a functional crosslinker for high performance liquid chromatography separation of small molecules.

    PubMed

    Li, Yaping; Qi, Li; Ma, Huimin

    2013-09-21

    A newly developed porous polymer monolith was prepared through copolymerization of 3-(trimethoxysilyl)propylmethacrylate modified graphene oxide with glycidyl methacrylate and ethylene dimethacrylate as a functional crosslinker, which was synthesized through silanization reaction of graphene oxide prepared by Hummers method with 3-(trimethoxysilyl)propylmethacrylate. The monolith was characterized by Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, transmission electron microscopy, scanning electron microscopy, mercury intrusion porosimetry and nitrogen adsorption measurement. The monolith column was applied as the stationary phase of high performance liquid chromatography and its chromatographic performance was evaluated by separation of small molecules in the isocratic reversed-phase mode. The chromatograms of hydrophobic steroids and polar aromatic amines on the prepared monolith displayed the enhanced separation performance over those on the parent monolith. The reproducibility of the column was less than 3.5% in terms of relative standard deviation of retention time. The results demonstrate that copolymerization of functionalized graphene oxide into porous polymer monolith was an effective tool for chromatography separation enhancement of small molecules in an isocratic mode. PMID:23884304

  17. A Two-Column Method for the Separation of Kr and Xe from Process Off-Gases

    SciTech Connect

    Liu, Jian; Fernandez, Carlos A.; Martin, Paul F.; Thallapally, Praveen K.; Strachan, Denis M.

    2014-07-29

    Two metal organic framework materials were investigated to determine the removal efficiency and capacity of MOF materials for krypton recovery from air at non-cryogenic temperatures. Our two bed breakthrough measurements on NiDOBDC and a partially fluorinated FMOFCu indicate these materials can capture and separate parts per million levels of Xe and Kr from air and, with a two-bed system, separate Xe from Kr. In a two-bed system, the he removal efficiency and adsorption capacity for Kr on these two MOFs were further increased Xe was removed in the first bed. This shows a promising future for MOFs in a radioactive nuclides separation from spent fuel.

  18. Apparatus and process for the separation of hydrophobic and hydrophilic particles using microbubble column flotation together with a process and apparatus for generation of microbubbles

    DOEpatents

    Yoon, R.H.; Adel, G.T.; Luttrell, G.H.

    1998-09-29

    A method and apparatus are disclosed for the microbubble flotation separation of very fine and coarse particles, especially coal and minerals, so as to produce high purity and high recovery efficiency. This is accomplished through the use of a flotation column, microbubbles, recycling of the flotation pulp, and countercurrent wash water to gently wash the froth. Also disclosed are unique processes and apparatus for generating microbubbles for flotation in a highly efficient and inexpensive manner using either a porous tube or in-line static generators. 14 figs.

  19. Apparatus and process for the separation of hydrophobic and hydrophilic particles using microbubble column flotation together with a process and apparatus for generation of microbubbles

    DOEpatents

    Yoon, Roe-Hoan; Adel, Gregory T.; Luttrell, Gerald H.

    1992-01-01

    A method and apparatus are disclosed for the microbubble flotation separation of very fine and coarse particles, especially coal and minerals, so as to produce high purity and high recovery efficiency. This is accomplished through the use of a flotation column, microbubbles, recycling of the flotation pulp, and countercurrent wash water to gently wash the froth. Also disclosed are unique processes and apparatus for generating microbubbles for flotation in a highly efficient and inexpensive manner using either a porous tube or in-line static generators.

  20. Apparatus and process for the separation of hydrophobic and hydrophilic particles using microbubble column flotation together with a process and apparatus for generation of microbubbles

    DOEpatents

    Yoon, Roe-Hoan; Adel, Gregory T.; Luttrell, Gerald H.

    1998-01-01

    A method and apparatus are disclosed for the microbubble flotation separation of very fine and coarse particles, especially coal and minerals, so as to produce high purity and high recovery efficiency. This is accomplished through the use of a flotation column, microbubbles, recycling of the flotation pulp, and countercurrent wash water to gently wash the froth. Also disclosed are unique processes and apparatus for generating microbubbles for flotation in a highly efficient and inexpensive manner using either a porous tube or in-line static generators.

  1. Apparatus for the separation of hydrophobic and hydrophilic particles using microbubble column flotation together with a process and apparatus for generation of microbubbles

    DOEpatents

    Yoon, Roe-Hoan; Adel, Gregory T.; Luttrell, Gerald H.

    1995-01-01

    An apparatus is disclosed for the microbubble flotation separation of very fine and coarse particles, especially coal, and minerals so as to produce high purity and high recovery efficiency. This is accomplished through the use of a flotation column, microbubbles, recycling of the flotation pulp, and countercurrent wash water to gently wash the froth. Also disclosed are unique processes and apparatus for generating microbubbles for flotation in a highly efficient and inexpensive manner using either a porous tube or in-line static generators.

  2. Apparatus for the separation of hydrophobic and hydrophilic particles using microbubble column flotation together with a process and apparatus for generation of microbubbles

    DOEpatents

    Yoon, R.H.; Adel, G.T.; Luttrell, G.H.

    1995-03-14

    An apparatus is disclosed for the microbubble flotation separation of very fine and coarse particles, especially coal, and minerals so as to produce high purity and high recovery efficiency. This is accomplished through the use of a flotation column, microbubbles, recycling of the flotation pulp, and countercurrent wash water to gently wash the froth. Also disclosed are unique processes and apparatus for generating microbubbles for flotation in a highly efficient and inexpensive manner using either a porous tube or in-line static generators. 14 figs.

  3. Apparatus and process for the separation of hydrophobic and hydrophilic particles using microbubble column flotation together with a process and apparatus for generation of microbubbles

    DOEpatents

    Yoon, R.H.; Adel, G.T.; Luttrell, G.H.

    1992-12-01

    A method and apparatus are disclosed for the microbubble flotation separation of very fine and coarse particles, especially coal and minerals, so as to produce high purity and high recovery efficiency. This is accomplished through the use of a flotation column, microbubbles, recycling of the flotation pulp, and countercurrent wash water to gently wash the froth. Also disclosed are unique processes and apparatus for generating microbubbles for flotation in a highly efficient and inexpensive manner using either a porous tube or in-line static generators. 14 figs.

  4. DETERMINATION OF PESTICIDES IN COMPOSITE DIETARY SAMPLES BY GAS CHROMATOGRAPHY/MASS SPECTROMETRY IN THE SELECTED ION MONITORING MODE USING A TEMPERATURE PROGRAMMABLE LARGE VOLUME INJECTOR WITH PRE-SEPARATION COLUMN

    EPA Science Inventory

    Use of a temperature-programmable pre-separation column in the gas chromatographic injection port permits determination of a wide range of semi-volatile pesticides including organochlorines, organophosphates, triazines, and anilines in fatty composite dietary samples while reduci...

  5. Sensitive determination of 4-(4-bromophenyl)-4-hydroxypiperidine, a metabolite of bromperidol, in rat plasma by HPLC with fluorescence detection after pre-column derivatization using 4-fluoro-7-nitro-2,1,3-benzoxadiazole.

    PubMed

    Higashi, Yasuhiko; Nakamura, Shota; Fujii, Youichi

    2006-12-01

    The purpose of this study was to determine the level of 4-(4-bromophenyl)-4-hydroxypiperidine (BPHP), a bromperidol (BRO) metabolite, in rat plasma by HPLC with fluorescence detection after pre-column derivatization using 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F). After basic extraction of the samples with benzene, derivatization with NBD-F was conducted in borate buffer (pH 8.0) at 60 degrees C for 3 min. Mexiletine was utilized through the procedure as an internal standard (IS). Retention times of the BPHP and IS derivatives were 7.7 and 11.5 min, respectively. The regression equation for BPHP showed good linearity in the range of 0.01-1 mg/ml with the detection limit of 0.003 microg/ml. The coefficient of variation was less than 12.0%. The recovery was satisfactory. This method was applied for a pharmacokinetic study of BPHP in comparison with 4-(4-chlorophenyl)-4-hydroxypiperidine (CPHP), the corresponding haloperidol (HAL) metabolite, in rats. The ratio of the area under the plasma concentration curve (AUC) after p.o. administration of BPHP to the AUC after i.p. administration of BPHP (46%) was lower than that of CPHP (56%), indicating that intestinal absorption of BPHP is lower than that of CPHP. The ratio of BRO metabolism to BPHP (48%) was 1.8-fold higher than that of HAL metabolism to CPHP (27%); the ratio was estimated as (AUCp.o.,A-->B/AUCp.o.,B)x100, where AUCp.o.,A-->B is the AUC value of BPHP or CPHP after p.o. administration of BRO or HAL, and AUCp.o.,B is the AUC of BPHP or CPHP after administration of BPHP or CPHP, respectively. Our method provides a sensitive procedure for determination of BPHP in rat plasma and is suitable for pharmacokinetic studies of BPHP after BRO administration. PMID:17142985

  6. Cationized Magnetoferritin Enables Rapid Labeling and Concentration of Gram-Positive and Gram-Negative Bacteria in Magnetic Cell Separation Columns

    PubMed Central

    Spencer, J.; Schwarzacher, W.

    2016-01-01

    ABSTRACT In order to identify pathogens rapidly and reliably, bacterial capture and concentration from large sample volumes into smaller ones are often required. Magnetic labeling and capture of bacteria using a magnetic field hold great promise for achieving this goal, but the current protocols have poor capture efficiency. Here, we present a rapid and highly efficient approach to magnetic labeling and capture of both Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria using cationized magnetoferritin (cat-MF). Magnetic labeling was achieved within a 1-min incubation period with cat-MF, and 99.97% of the labeled bacteria were immobilized in commercially available magnetic cell separation (MACS) columns. Longer incubation times led to more efficient capture, with S. aureus being immobilized to a greater extent than E. coli. Finally, low numbers of magnetically labeled E. coli bacteria (<100 CFU per ml) were immobilized with 100% efficiency and concentrated 7-fold within 15 min. Therefore, our study provides a novel protocol for rapid and highly efficient magnetic labeling, capture, and concentration of both Gram-positive and Gram-negative bacteria. IMPORTANCE Antimicrobial resistance (AMR) is a significant global challenge. Rapid identification of pathogens will retard the spread of AMR by enabling targeted treatment with suitable agents and by reducing inappropriate antimicrobial use. Rapid detection methods based on microfluidic devices require that bacteria are concentrated from large volumes into much smaller ones. Concentration of bacteria is also important to detect low numbers of pathogens with confidence. Here, we demonstrate that magnetic separation columns capture small amounts of bacteria with 100% efficiency. Rapid magnetization was achieved by exposing bacteria to cationic magnetic nanoparticles, and magnetized bacteria were concentrated 7-fold inside the column. Thus, bacterial capture and concentration were achieved

  7. Separation and on-column preconcentration of some nonsteroidal anti-inflammatory drugs by microemulsion electrokinetic capillary chromatography using high-speed separations.

    PubMed

    Macià, Alba; Borrull, Francesc; Calull, Marta; Aguilar, Carme

    2005-02-01

    Various strategies have been investigated for separating a group of nonsteroidal anti-inflammatory drugs (NSAIDs) by microemulsion electrokinetic capillary chromatography (MEEKC) using high-speed separations. The parameters that of affect the separation, such as the nature of the oil droplet and the buffer, and the surfactant concentration have been studied. In addition, several organic solvents were used to decrease the retention of the analytes in the oil droplet phase and to improve the resolution of the NSAIDs. The optimum microemulsion background electrolyte (BGE) solution made of 0.8% w/w ethyl acetate, 6.6% w/w butan-1-ol, 6.0% w/w acetonitrile, 1.0% w/w sodium dodecyl sulfate (SDS), and 85.6% w/w of 10 mM sodium tetraborate at pH 9.2 resolved the drugs within 8 min. The short-end injection procedure is an alternative for reducing the analysis time. When this procedure was used, the microemulsion BGE solution consisted of 0.8% w/w ethyl acetate, 6.6% w/w butan-1-ol, 17.0% w/w methanol, 1.0% w/w SDS, and 74.6% w/w of 10 mM sodium tetraborate, pH 9.2, and the NSAIDs were separated within 3 min. The reversed electrode polarity stacking mode (REPSM) technique was applied to the on-line concentration of the NSAIDs. In this technique, the sample matrix was pumped out of the capillary using a polarity-switching step. When this technique was applied, the sensitivity was enhanced up to 40-fold and the limits of detection (LODs) were in the low microg.L(-1) levels.

  8. Comprehensive off-line, two-dimensional liquid chromotography. Application to the separation of peptide digest

    SciTech Connect

    Marchetti, Nicola; Guiochon, Georges A

    2008-01-01

    The separation of the peptide digests of myoglobin and bovine serum albumin was performed with an off-line combination of two commercial, conventional HPLC columns. The first column was packed with a strong ion exchanger and eluted with a KCl gradient. The second column was packed with particles of C{sub 18}-bonded silica and eluted with an acetonitrile gradient. The conditional peak capacities of the 2D separations achieved exceed 7000 under the experimental conditions investigated. This performance is achieved at the cost of an analysis time of the order of 28 hours. Possible improvements to the separation method described here are discussed.

  9. Analysis of phthalates in wine using liquid chromatography tandem mass spectrometry combined with a hold-back column: Chromatographic strategy to avoid the influence of pre-existing phthalate contamination in a liquid chromatography system.

    PubMed

    Hayasaka, Y

    2014-11-01

    This paper describes the development and application of a novel method for the analysis of phthalates in wine using HPLC-MS/MS combined with a hold-back column. Phthalates are ubiquitous contaminants in the environment and can be widely found in laboratory materials and equipment. A HPLC system is no exception and can be the source of contamination affecting the accuracy and precision of analytical results. The new method successfully separates phthalates from the different sources, a wine sample and HPLC system by a simple technique using an additional HPLC column (a hold-back column) placed upstream of the injection valve. The hold-back column effectively retains the HPLC-derived contaminants during column equilibrium time and delays their elution times from an analytical column. Consequently, a phthalate from a wine sample can be baseline separated as it elutes sufficiently earlier than the same phthalate from the HPLC system. HPLC-MS/MS analysis combined with the hold-back column demonstrated virtually no influence of the HPLC contaminants on the quantification of phthalates present in wine. Together with a simple and rapid sample preparation and the use of labeled internal standards, the method was confirmed to be robust and reliable to determine concentrations of phthalates in wine. Quantification limits were within the range of 1.6-9.8μgL(-1) for dimethyl, diethyl, dibutyl, benzylbutyl, bis(2-ethylhexyl) and dioctyl phthalates, and 7.5-26.6μgL(-1) for multiple isomeric phthalates, di-iso-nonyl and di-iso-dodecyl phthalates.

  10. Separation and purification of five phenylpropanoid glycosides from Lamiophlomis rotata (Benth.) Kudo by a macroporous resin column combined with high-speed counter-current chromatography.

    PubMed

    Yue, Hui-Lan; Zhao, Xiao-Hui; Mei, Li-Juan; Shao, Yun

    2013-09-01

    Five phenylethanoid glycosides (PhGs), forsythoside B, verbascoside, alyssonoside, isoverbascoside, and leucosceptoside B, were isolated and purified from Lamiophlomis rotata (Benth.) Kudo by high-speed counter-current chromatography (HSCCC) combined with macroporous resin (MR) column separation. In the present study, the two-phase solvent system composed of ethyl acetate/n-butanol/water (13:3:10, v/v/v) was used for HSCCC separation. A total of 27 mg of forsythoside B, 41 mg of verbascoside, 29 mg of alyssonoside, 23 mg of isoverbascoside, and 13 mg of leucosceptoside B with purities of 97.7, 99.2, 99.5, 99.3, and 97.3%, respectively, were obtained in a one-step separation within 4 h from 150 mg of crude extract. The recoveries of the five PhGs after MR-HSCCC separation were 74.5, 76.5, 72.5, 76.4, and 77.0%, respectively. The chemical structures of all five compounds were identified by (1) H and (13) C NMR spectroscopy.

  11. Biosorption and desorption of lanthanum(III) and neodymium(III) in fixed-bed columns with Sargassum sp.: perspectives for separation of rare earth metals.

    PubMed

    Oliveira, Robson C; Guibal, Eric; Garcia, Oswaldo

    2012-01-01

    Rare earth (RE) metals are essentials for the manufacturing of high-technology products. The separation of RE is complex and expensive; biosorption is an alternative to conventional processes. This work focuses on the biosorption of monocomponent and bicomponent solutions of lanthanum(III) and neodymium(III) in fixed-bed columns using Sargassum sp. biomass. The desorption of metals with HCl 0.10 mol L(-1) from loaded biomass is also carried out with the objective of increasing the efficiency of metal separation. Simple models have been successfully used to model breakthrough curves (i.e., Thomas, Bohart-Adams, and Yoon-Nelson equations) for the biosorption of monocomponent solutions. From biosorption and desorption experiments in both monocomponent and bicomponent solutions, a slight selectivity of the biomass for Nd(III) over La(III) is observed. The experiments did not find an effective separation of the RE studied, but their results indicate a possible partition between the metals, which is the fundamental condition for separation perspectives.

  12. Separation of intact proteins on γ-ray-induced polymethacrylate monolithic columns: A highly permeable stationary phase with high peak capacity for capillary high-performance liquid chromatography with high-resolution mass spectrometry.

    PubMed

    Simone, Patrizia; Pierri, Giuseppe; Foglia, Patrizia; Gasparrini, Francesca; Mazzoccanti, Giulia; Capriotti, Anna Laura; Ursini, Ornella; Ciogli, Alessia; Laganà, Aldo

    2016-01-01

    Polymethacrylate-based monolithic capillary columns, prepared by γ-radiation-induced polymerization, were used to optimize the experimental conditions (nature of the organic modifiers, the content of trifluoroacetic acid and the column temperature) in the separation of nine standard proteins with different hydrophobicities and a wide range of molecular weights. Because of the excellent permeability of the monolithic columns, an ion-pair reversed-phase capillary liquid chromatography with high-resolution mass spectrometry method has been developed by coupling the column directly to the mass spectrometer without a flow-split and using a standard electrospray interface. Additionally, the high working flow and concomitant high efficiency of these columns allowed us to employ a longer column (up to 50 cm) and achieve a peak capacity value superior to 1000. This work is motivated by the need to develop new materials for high-resolution chromatographic separation that combine chemical stability at elevated temperatures (up to 75°C) and a broad pH range, with a high peak capacity value. The advantage of the γ-ray-induced monolithic column lies in the batch-to-batch reproducibility and long-term high-temperature stability. Their proven high loading capacity, recovery, good selectivity and high permeability, moreover, compared well with that of a commercially available poly(styrene-divinylbenzene) monolithic column, which confirms that such monolithic supports might facilitate analysis in proteomics.

  13. Preliminary results from a microvolume, dynamically heated analytical column for preconcentration and separation of simple gases prior to stable isotopic analysis

    NASA Astrophysics Data System (ADS)

    Panetta, Robert James; Seed, Mike

    2016-04-01

    Stable isotope applications that call for preconcentration (i.e., greenhouse gas measurements, small carbonate samples, etc.) universally call for cryogenic fluids such as liquid nitrogen, dry ice slurries, or expensive external recirculation chillers. This adds significant complexity, first and foremost in the requirements to store and handle such dangerous materials. A second layer of complexity is the instrument itself - with mechanisms to physically move either coolant around the trap, or move a trap in or out of the coolant. Not to mention design requirements for hardware that can safely isolate the fluid from other sensitive areas. In an effort to simplify the isotopic analysis of gases requiring preconcentration, we have developed a new separation technology, UltiTrapTM (patent pending), which leverage's the proprietary Advanced Purge & Trap (APT) Technology employed in elemental analysers from Elementar Analysensysteme GmbH products. UltiTrapTM has been specially developed as a micro volume, dynamically heated GC separation column. The introduction of solid-state cooling technology enables sub-zero temperatures without cryogenics or refrigerants, eliminates all moving parts, and increases analytical longevity due to no boiling losses of coolant . This new technology makes it possible for the system to be deployed as both a focussing device and as a gas separation device. Initial data on synthetic gas mixtures (CO2/CH4/N2O in air), and real-world applications including long-term room air and a comparison between carbonated waters of different origins show excellent agreement with previous technologies.

  14. Development of HPLC/ESI-MS and HPLC/1H NMR methods for the identification of photocatalytic degradation products of iodosulfuron.

    PubMed

    Sleiman, Mohamad; Ferronato, Corinne; Fenet, Bernard; Baudot, Robert; Jaber, Farouk; Chovelon, Jean-Marc

    2006-05-01

    In the present study, HPLC/ESI-MS and stopped-flow HPLC/1H NMR methods were developed and applied to separate and characterize the byproducts arising from TiO2-catalyzed photodegradation of the herbicide iodosulfuron methyl ester (IOME) in aqueous solution under UV irradiation. Prior to identification, irradiated solutions of IOME (200 and 1000 mg.L(-1)) were concentrated by solid-phase extraction using two cartridges: Isolute C18 and Isolute ENV+. Analytical separation was achieved on a C18 reversed-phase column with ACN/H2O (HPLC/MS) or ACN/D2O (HPLC/NMR) as mobile phase and a linear gradient with a chromatographic run time of 35 min. The combination of UV and MS data allowed the structural elucidation of more than 20 degradation products, whereas 1H NMR data permitted an unequivocal confirmation of the identities of major products and the differentiation of several positional isomers, in particular, the hydroxylation isomers. The obtained results permitted us to propose a possible degradation scheme and to put in evidence the presence of privileged sites for the attack of OH radicals. This work shows, for the first time, the application of combined HPLC with UV, MS, and NMR detection for complete structural elucidation of photocatalytic degradation products, and it will be of particular value in studies on the elimination of pollutants in aqueous solutions by photocatalysis.

  15. Core-Shell Columns in High-Performance Liquid Chromatography: Food Analysis Applications

    PubMed Central

    Preti, Raffaella

    2016-01-01

    The increased separation efficiency provided by the new technology of column packed with core-shell particles in high-performance liquid chromatography (HPLC) has resulted in their widespread diffusion in several analytical fields: from pharmaceutical, biological, environmental, and toxicological. The present paper presents their most recent applications in food analysis. Their use has proved to be particularly advantageous for the determination of compounds at trace levels or when a large amount of samples must be analyzed fast using reliable and solvent-saving apparatus. The literature hereby described shows how the outstanding performances provided by core-shell particles column on a traditional HPLC instruments are comparable to those obtained with a costly UHPLC instrumentation, making this novel column a promising key tool in food analysis. PMID:27143972

  16. Methods and applications of HPLC-AMS (WBio 5)

    SciTech Connect

    Bucholz, B A; Clifford, A J; Duecker, S R; Lin, Y; Vogel, J S

    1999-09-29

    Pharmacokinetics of physiologic doses of nutrients, pesticides, and herbicides can easily be traced in humans using a {sup 14}C-labelled compound. Basic kinetics can be monitored in blood or urine by measuring the elevation in the {sup 14}C content above the control predose tissue and converting to equivalents of the parent compound. High Performance Liquid Chromatography (HPLC) is an excellent method for the chemical separation of complex mixtures whose profiles afford estimation of biochemical pathways of metabolism. Compounds elute from the HPLC systems with characteristic retention times and can be collected in fractions that can then be graphitized for AMS measurement. Unknowns are identified by coelution with known standards and chemical tests that reveal functional groupings. Metabolites are quantified with the {sup 14}C signal. Thoroughly accounting for the carbon inventory in the LC solvents, ion-pairing agents, samples, and carriers adds some complexity to the analysis. In most cases the total carbon inventory is dominated by carrier. Baseline background and stability need to be carefully monitored. Limits of quantitation near 10 amol of {sup 14}C per HPLC fraction are typically achieved. Baselines are maintained by limiting injected {sup 14}C activity <0.17 Bq (4.5 pCi) on the HPLC column.

  17. Comparison of chiral separations of aminophosphonic acids and their aminocarboxylic acid analogs using a crown ether column.

    PubMed

    Barnhart, Wesley W; Xia, Xiaoyang; Jensen, Randy; Gahm, Kyung H

    2013-07-01

    Crown ethers are capable of complexing with primary amines and have been utilized in chromatography to separate amino acid racemates. This application has been extended to resolve (1-amino-1-phenylmethyl)phosphonic acid and (1-aminoethyl)phosphonic acid racemates, along with their aminocarboxylic acid analogs (2-phenylglycine and alanine, respectively), via a ChiroSil RCA crown ether based chiral stationary phase. Effects of the organic modifier, temperature, and acid type and concentration on retention and selectivity were also investigated. Trends in retention and selectivity varied between aminophosponic acids and their aminocarboxylic analogs. Computer modeling and (1)H NMR analyses were performed to potentially gain a better understanding of interactions of the aforementioned molecules with the ChiroSil RCA chiral stationary phase. Theoretical predictions of the most stable conformations for (R)- and (S)-enantiomers were compared to elution order; it was found that the elution order agreed with molecular modeling such that the longest retention correlated with the predicted most stable complex between the enantiomer and crown ether. (1)H NMR demonstrated interactions of aminophosphonic and aminocarboxylic racemates with (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid in solution and was utilized to determine enantiomeric excess of (1-amino-1-phenylmethyl)phosphonic acid after its enantioenrichment via crystallization through diastereomeric salt formation with the crown ether followed by filtration. PMID:23703726

  18. Comparison of chiral separations of aminophosphonic acids and their aminocarboxylic acid analogs using a crown ether column.

    PubMed

    Barnhart, Wesley W; Xia, Xiaoyang; Jensen, Randy; Gahm, Kyung H

    2013-07-01

    Crown ethers are capable of complexing with primary amines and have been utilized in chromatography to separate amino acid racemates. This application has been extended to resolve (1-amino-1-phenylmethyl)phosphonic acid and (1-aminoethyl)phosphonic acid racemates, along with their aminocarboxylic acid analogs (2-phenylglycine and alanine, respectively), via a ChiroSil RCA crown ether based chiral stationary phase. Effects of the organic modifier, temperature, and acid type and concentration on retention and selectivity were also investigated. Trends in retention and selectivity varied between aminophosponic acids and their aminocarboxylic analogs. Computer modeling and (1)H NMR analyses were performed to potentially gain a better understanding of interactions of the aforementioned molecules with the ChiroSil RCA chiral stationary phase. Theoretical predictions of the most stable conformations for (R)- and (S)-enantiomers were compared to elution order; it was found that the elution order agreed with molecular modeling such that the longest retention correlated with the predicted most stable complex between the enantiomer and crown ether. (1)H NMR demonstrated interactions of aminophosphonic and aminocarboxylic racemates with (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid in solution and was utilized to determine enantiomeric excess of (1-amino-1-phenylmethyl)phosphonic acid after its enantioenrichment via crystallization through diastereomeric salt formation with the crown ether followed by filtration.

  19. Determination of citrus limonoid glucosides by high performance liquid chromatography coupled to post-column reaction with Ehrlich’s Reagent

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A method for the identification and quantification of citrus limonoid glucosides in juices based upon high performance liquid chromatography (HPLC) separation coupled to post-column reaction with Ehrlichs’s reagent has been developed. This method utilizes a phenyl stationary phase and an isocratic ...

  20. A confirmatory HPLC-MS/MS method for ten synthetic corticosteroids in bovine urines.

    PubMed

    Savu, S R; Silvestro, L; Haag, A; Sörgel, F

    1996-12-01

    In the present study, an HPLC-MS/MS method to confirm, in bovine urine, the most common synthetic corticosteroids illegally used as growth promoters in livestock breeding will be presented. An API III-Plus (PE-Sciex) triple quadrupole mass spectrometer, interfaced by means of an atmospheric pressure chemical ionization source to the HPLC system, was used. Urine samples were treated with a sulfatase-glucuronidase mixture to cleave the drug-conjugates and then extracted on C18 disposable columns. LC separations were performed on a reversed-phase C18 column with ammonium acetate 0.1 M/acetonitrile (60/40, v/v) as mobile phase. Detection was performed in multiple reaction monitoring mode, negative ions, selecting fragmentations characteristic of 10 corticosteroids used more frequently. Good results, in terms of sensitivity and specificity have been obtained for nine corticosteroids that can be analyzed in the same HPLC run; the limits of sensitivity achieved were 0.05-1.0 ng/ml in urine. Only a more polar corticosteroid, required a different HPLC separation. Practical applications of this technique to real samples proved that it is an effective method to confirm the illegal use of corticosteroids as growth promoter in animal. In comparison with the chemical GC-MS methods the simpler sample preparation and the faster time of analysis permit a considerable increase of sample testing per day without compromising on analytical sensitivity and specificity.

  1. Combination of preparative HPLC and HSCCC methods to separate phosphodiesterase inhibitors from Eucommia ulmoides bark guided by ultrafiltration-based ligand screening.

    PubMed

    Shi, Shu-Yun; Peng, Mi-Jun; Zhang, Yu-Ping; Peng, Sheng

    2013-05-01

    Phosphodiesterase (PDE) inhibitors are widely used because of their various pharmacological properties, and natural products are considered the most productive source of PDE inhibitors. In this work, a new ultrafiltration-high-performance liquid chromatography (HPLC)-diode-array detection-mass spectrometry based ligand screening was developed for the first screening of PDE inhibitors from Eucommia ulmoides bark, and then the target bioactive compounds were prepared by combination of stepwise preparative HPLC and high-speed countercurrent chromatography (HSCCC) methods. Experiments were conducted to optimize the parameters in ultrafiltration, stepwise preparative HPLC, and HSCCC to allow rapid and effective screening and isolation of active compounds from complex mixtures. Seven lignans with purity over 97 % were isolated and identified by their UV, electrospray ionization mass spectrometry, and NMR data as (+)-pinoresinol-4,4'-di-O-β-D-glucopyranoside (1), (+)-pinoresinol-4-O-β-D-glucopyranosyl(1 → 6)-β-D-glucopyranoside (2), (+)-medioresinol-4,4'-di-O-β-D-glucopyranoside (3), (+)-syringaresinol-4,4'-di-O- β-D-glucopyranoside (4), (-)-olivil-4'-O-β-D-glucopyranoside (5), (-)-olivil-4-O-β-D- glucopyranoside (6), and (+)-pinoresinol-4-O-β-D-glucopyranoside (7). Compound 2 was first isolated from the genus Eucommia. Lignan diglucopyranosides (compounds 1-4) shower a greater inhibitory effect than lignan monoglucopyranosides (compounds 5-7). The method developed could be widely applied for high-throughput screening and preparative isolation of PDE inhibitors from natural products.

  2. Acaricidal activity of petroleum ether extract of neem (Azadirachta indica) oil and its four fractions separated by column chromatography against Sarcoptes scabiei var. cuniculi larvae in vitro.

    PubMed

    Deng, Yunxia; Shi, Dongxia; Yin, Zhongqiong; Guo, Jianhong; Jia, Renyong; Xu, Jiao; Song, Xu; Lv, Cheng; Fan, Qiaojia; Liang, Xiaoxia; Shi, Fei; Ye, Gang; Zhang, Wei

    2012-04-01

    The petroleum ether extract of neem oil and its four fractions separated by column chromatography was diluted at different concentrations with liquid paraffin. The acaricidal bioassay was conducted using a dipping method. The results indicated that the median lethal concentration (LC50) of the petroleum ether extract (at the concentration of 500.0ml/l) was 70.9ml/l, 24h after treatment. At concentrations of 500.0, 250.0, 125.0, 62.5 and 31.2ml/l, the median lethal times (LT50) of the petroleum ether extract were 8.7, 8.8, 10.8, 11.5 and 13.1h, respectively. Thin-layer chromatography (TLC) showed that the petroleum ether extract of neem oil separated into four fractions (F1-F4). Acaricidal activity of 68.3% and 100.0% in the F2 and F4 was confirmed. These results suggest that petroleum ether extracts of neem oil and its four fractions possess useful acaricidal activity in vitro. PMID:22349080

  3. Ion-Exclusion High-Performance Liquid Chromatography of Aliphatic Organic Acids Using a Surfactant-Modified C18 Column.

    PubMed

    Fasciano, Jennifer M; Mansour, Fotouh R; Danielson, Neil D

    2016-07-01

    Ion exclusion chromatography (IELC) of short chain aliphatic carboxylic acids is normally done using a cation exchange column under standard HPLC conditions but not in the ultra-HPLC (UHPLC) mode. A novel IELC method for the separation of this class of carboxylic acids by either HPLC or UHPLC utilizing a C18 column dynamically modified with sodium dodecyl sulfate has been developed. The sample capacity is estimated to be near 10 mM for a 20 µL injection or 0.2 µmol using a 150 × 4.6 mm column. The optimum mobile phase determined for three standard mixtures of organic acids is 1.84 mM sulfuric acid at pH 2.43 and a flow rate of 0.6 mL/min. Under optimized conditions, a HPLC separation of four aliphatic carboxylic acids such as tartaric, malonic, lactic and acetic can be achieved in under 4 min and in <2 min in the UHPLC mode at 2.1 mL/min. A variety of fruit juice and soft drink samples are analyzed. Stability of the column as measured by the retention order of maleic and fumaric acid is estimated to be ∼4,000 column volumes using HPLC and 600 by UHPLC. Reproducible chromatograms are achieved over at least a 2-month period. This study shows that the utility of a C18 column can be easily extended when needed to IELC under either standard or UHPLC conditions. PMID:27006111

  4. Validated HPLC method for quantifying permethrin in pharmaceutical formulations.

    PubMed

    García, E; García, A; Barbas, C

    2001-03-01

    An isocratic HPLC method for permethrin determination in raw material and pharmaceutical presentations as lotion and shampoo has been developed and validated following ICH recommendations. Cis and trans- isomers, impurities and degradation products are well separated. The chromatographic analysis were performed on a 4 microm particle C-18 Nova-Pak (Waters, Madrid, Spain) column (15 x 0.39 cm) kept in a Biorad column oven at 35 degrees C. Mobile phase consisted of methanol--water (78:22, v/v) at a flow rate of 1 ml/min. UV detection was performed at 272 nm and peaks were identified with retention times as compared with standards and confirmed with characteristic spectra using the photodiode array detector.

  5. [Determination of amanitatoxins by HPLC].

    PubMed

    Nishizawa, Chiemi; Yamaura, Yoshio

    2003-10-01

    High-performance liquid chromatographic (HPLC) assay has been developed for the simultaneous determination of alpha-amanitin, beta-amanitin and phalloidin in serum. Three toxins were extracted by reflux in a water bath at 80 degrees C for one hour and purified by Sep-Pak Plus tC18 cartridges. The HPLC assay was performed under gradient conditions using Develosil RP AQUEOUS column. The moble phase consisted with a mixture of acetonitorile containing 0.01 M ammonium acetate(pH 5.0). The column effluence was monitored at 295 nm, 302 nm and 230 nm for 35 min. Detection limit of three toxins in serum were 0.2 microgram/ml respectively. High recovery yields in the range of 81.5-88.1% for toxins were obtained by using this method. PMID:14740566

  6. Preparation and evaluation of monolithic poly(N-vinylcarbazole-co-1,4-divinylbenzene) capillary columns for the separation of small molecules.

    PubMed

    Koeck, Rainer; Fischnaller, Martin; Bakry, Rania; Tessadri, Richard; Bonn, Guenther K

    2014-09-01

    Short-term polymerization or the so-called low-conversion polymerization was applied for the preparation of N-vinylcarbazole (NVC) and 1,4-divinylbenzene (DVB) monolithic capillary columns. The synthesis was carried out by thermally initiated free radical copolymerization under the influence of inert micro- (toluene) and macroporogen (1-decanol) and α,α'-azoisobutyronitrile (AIBN) as radical initiator. The morphological and porous properties were studied by scanning electron microscopy (SEM), nitrogen adsorption, and mercury intrusion porosimetry (MIP). The copolymerization process was studied by monomer conversion measurements. This approach led to increased porosity and specific surface area. A specific surface area above 400 m(2)/g of the monolith and a distinct bimodal pore size distribution were obtained. The chromatographic performance was determined in terms of theoretical plate heights and number of theoretical plates. The lowest plate height value was found to be 3.9 μm (corresponding to ≈256,000 plates per meter) applying methylparaben utilizing an 80 mm × 0.2 mm i.d. monolithic capillary. The developed NVC/DVB monolithic supports showed high separation efficiency towards small molecules, which was exemplified applying reversed-phase (RP) separation of alkylbenzenes, beta-blockers, flavanoids, parabens, and phenones. The loading capacity was analyzed for isocratic separation of seven alkylbenzenes and was found to be up to 77 ng total mass of alkylbenzenes. Furthermore, a long-term stability test of 1,000 consecutive runs was performed and resulted in a maximum variance of 0.97, 0.85, and 0.16 % RSD for resolution, peak width at half height, and retention times, respectively. The material was proven to have a high permeability of 1.11E-14 m(2), applying water as a mobile phase. PMID:25056873

  7. Distillation Column Flooding Predictor

    SciTech Connect

    2002-02-01

    This factsheet describes a research project whose goal is to develop the flooding predictor, an advanced process control strategy, into a universally useable tool that will maximize the separation yield of a distillation column.

  8. Optimization of a CUPRAC-Based HPLC Postcolumn Assay and Its Applications for Ginkgo biloba L. Extracts.

    PubMed

    Rimkiene, Laura; Ivanauskas, Liudas; Kubiliene, Asta; Vitkevicius, Konradas; Kiliuviene, Guoda; Jakstas, Valdas

    2015-01-01

    The aim of the present work was to improve and validate the HPLC-CUPRAC postcolumn method for the evaluation of active antioxidant markers from the acetonic extracts of Ginkgo biloba leaves. Improvement of the HPLC online assay was performed by evaluating the suitable loop temperature, the reaction loop length, and the impact of flow rate. Separation of the analytes was performed by the HPLC method on an ACE C18 analytical column using a gradient elution program. The separated antioxidant markers in the extracts reacted with copper(II)-neocuproine (Cu(II)-Nc) reagent in the postcolumn reaction coil. The reagent was reduced by antioxidants to the copper(I)-neocuproine (Cu(I)-Nc) chelate with a maximum absorption at 450 nm. Validation experiments confirmed sufficient precision, sensitivity, and effectiveness of the corresponding method, which could be used for further evaluations of active antioxidant compounds in similar plant materials.

  9. Optimization of a CUPRAC-Based HPLC Postcolumn Assay and Its Applications for Ginkgo biloba L. Extracts

    PubMed Central

    Rimkiene, Laura; Ivanauskas, Liudas; Kubiliene, Asta; Vitkevicius, Konradas; Kiliuviene, Guoda; Jakstas, Valdas

    2015-01-01

    The aim of the present work was to improve and validate the HPLC-CUPRAC postcolumn method for the evaluation of active antioxidant markers from the acetonic extracts of Ginkgo biloba leaves. Improvement of the HPLC online assay was performed by evaluating the suitable loop temperature, the reaction loop length, and the impact of flow rate. Separation of the analytes was performed by the HPLC method on an ACE C18 analytical column using a gradient elution program. The separated antioxidant markers in the extracts reacted with copper(II)-neocuproine (Cu(II)-Nc) reagent in the postcolumn reaction coil. The reagent was reduced by antioxidants to the copper(I)-neocuproine (Cu(I)-Nc) chelate with a maximum absorption at 450 nm. Validation experiments confirmed sufficient precision, sensitivity, and effectiveness of the corresponding method, which could be used for further evaluations of active antioxidant compounds in similar plant materials. PMID:26236538

  10. [HPLC analysis of dehydroepiandrosterone sulfate in serum with use of solid-phase extraction on hyper cross-linked polystyrene (Purosep-200)].

    PubMed

    Dutov, A A; Nikitin, D A; Luk'ianova, Yu L; Sverkunova, A V; Martynova, A V; Ermolina, A V

    2014-01-01

    We have developed a simple HPLC method for analysis of the dehydroepiandrosterone sulfate (DHEA-sulfate) in serum with use a new procedure of solid-phase extraction (SPE) on hyper cross-linked polystyrene (Purosep-200) and fast chromatographic separation on the monolithic column under isocratic elution and UV detection at 200 nm. Complete SPE procedure lasts for about 7 min, chromatographic separation takes less than 6 min. Simplicity and high reproducibility of this method makes it attractive in routine clinical practice.

  11. 32P-postlabeling and HPLC separation of DNA adducts formed by diesel exhaust extracts in vitro and in mouse skin and lung after topical treatment.

    PubMed

    Savela, K; King, L; Gallagher, J; Lewtas, J

    1995-09-01

    Diesel exhaust extracts contain many carcinogenic compounds which have been shown to form polycyclic aromatic hydrocarbon (PAH)- and nitrated PAH-DNA adducts in rodent skin and lung. The aim of this study was to characterize by 32P-postlabeling, TLC and HPLC the primary postlabeled PAH-DNA adduct(s) formed in vitro and in vivo by diesel extracts. The diesel particle extracts had known concentrations of benzo[a]pyrene, benzo[b,j,k]-fluoranthenes (B[b,j,k]F) and chrysene. DNA adducts were analyzed in calf thymus DNA incubated in vitro with PAHs activated by S9 mix and in skin and lung DNA from topically treated mice. The main diesel-derived DNA adduct formed in vitro and in vivo did not co-migrate on HPLC and large TLC plates with (+/-)-r-7,t-8-dihydroxy-t-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (anti BPDE)-, B[b]F-,B[j]F-,B[k]F-or chrysene-DNA adduct standards. By co-chromatography DNA adducts formed by chrysene from both in vitro and in vivo samples were identified. Nissan diesel extract containing higher PAH concentrations than Volkswagen automobile extract formed skin DNA adducts that co-migrated with chrysene- and anti BPDE- DNA-derived adducts. We conclude that the use of a highly sensitive 32P-postlabeling method combined with HPLC improves the identification of PAH adducts formed by complex mixtures such as diesel exhaust extracts.

  12. 32P-postlabeling and HPLC separation of DNA adducts formed by diesel exhaust extracts in vitro and in mouse skin and lung after topical treatment.

    PubMed

    Savela, K; King, L; Gallagher, J; Lewtas, J

    1995-09-01

    Diesel exhaust extracts contain many carcinogenic compounds which have been shown to form polycyclic aromatic hydrocarbon (PAH)- and nitrated PAH-DNA adducts in rodent skin and lung. The aim of this study was to characterize by 32P-postlabeling, TLC and HPLC the primary postlabeled PAH-DNA adduct(s) formed in vitro and in vivo by diesel extracts. The diesel particle extracts had known concentrations of benzo[a]pyrene, benzo[b,j,k]-fluoranthenes (B[b,j,k]F) and chrysene. DNA adducts were analyzed in calf thymus DNA incubated in vitro with PAHs activated by S9 mix and in skin and lung DNA from topically treated mice. The main diesel-derived DNA adduct formed in vitro and in vivo did not co-migrate on HPLC and large TLC plates with (+/-)-r-7,t-8-dihydroxy-t-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (anti BPDE)-, B[b]F-,B[j]F-,B[k]F-or chrysene-DNA adduct standards. By co-chromatography DNA adducts formed by chrysene from both in vitro and in vivo samples were identified. Nissan diesel extract containing higher PAH concentrations than Volkswagen automobile extract formed skin DNA adducts that co-migrated with chrysene- and anti BPDE- DNA-derived adducts. We conclude that the use of a highly sensitive 32P-postlabeling method combined with HPLC improves the identification of PAH adducts formed by complex mixtures such as diesel exhaust extracts. PMID:7554058

  13. Chromatographic Separations Using Solid-Phase Extraction Cartridges: Separation of Wine Phenolics

    NASA Astrophysics Data System (ADS)

    Brenneman, Charles A.; Ebeler, Susan E.

    1999-12-01

    We describe a simple laboratory experiment that demonstrates the principles of chromatographic separation using solid-phase extraction columns and red wine. By adjusting pH and mobile phase composition, the wine is separated into three fractions of differing polarity. The content of each fraction can be monitored by UV-vis spectroscopy. When the experiment is combined with experiments involving HPLC or GC separations, students gain a greater appreciation for and understanding of the highly automated instrumental systems currently available. In addition, they learn about the chemistry of polyphenolic compounds, which are present in many foods and beverages and which are receiving much attention for their potentially beneficial health effects.

  14. Separation of cordycepin from Cordyceps militaris fermentation supernatant using preparative HPLC and evaluation of its antibacterial activity as an NAD+-dependent DNA ligase inhibitor

    PubMed Central

    Zhou, Xiaofeng; Cai, Guoqiang; He, Yi; Tong, Guotong

    2016-01-01

    Cordycepin exhibits various bio-activities, including anticancer, antibacterial, antiviral and immune regulation activities, and is a significant focus of research. However, the preparation of high-purity cordycepin remains challenging. Also, the molecular target with which cordycepin interacts to cause an antibacterial effect remains unknown. In the present study, cordycepin was prepared by preparative high-performance liquid chromatography (prep-HPLC) and the purity obtained was 99.6%, indicating that this technique may be useful for the large-scale isolation of cordycepin in the future. The results of computational molecular docking analysis indicated that the interaction energy between cordycepin and NAD+-dependent DNA ligase (LigA) was lower than that between cordycepin and other common antibacterial targets. The highly pure cordycepin obtained by prep-HPLC demonstrated inhibitory activity against LigA from various bacteria in vitro. In conclusion, cordycepin may be useful as a broad-spectrum antibiotic targeting LigA in various bacteria. PMID:27588098

  15. Numerical investigation with a coupled single-column lake-atmosphere model: using the Alpert-Stein factor separation methodology to assess the sensitivity of surface interactions

    NASA Astrophysics Data System (ADS)

    Goyette, Stéphane

    2016-06-01

    A coupled single-column atmosphere-lake model, along with the Stein-Alpert factor separation methodology, is used to explore some of the non-linear interactions in the vertical dimension between the lower atmosphere and the deep-Lake Geneva, Switzerland, during three selected periods in 1990. The first from the end of April to the end of May when Lake Geneva was building its stratification, the second from mid-August to mid-September during stable stratification, and the third from the end of November to the end of December during destratification. It is recognized that the large thermal inertia of Lake Geneva reduces the surface annual and diurnal temperature variations for neighbouring regions. However, the question of how the open water and the overlying atmosphere interact and which of these "factors" has the most influence needs much attention. The sole presence of the lake is shown to be a major feature with regard to the surface energy budget components whose contributions counteract those of the lower atmosphere, thus supporting the fact that Lake Geneva acts as a damping factor to the regional climate system. It is also shown that not only did the presence of the lake and the overlying atmosphere independently modulate the surface energy budget, but also the synergistic nonlinear interaction among them, either positive or negative, was often found non-negligible. Moreover, some processes may turn out to be important on short time scales while being negligible on the long term.

  16. Distribution and Niche Separation of Planktonic Microbial Communities in the Water Columns from the Surface to the Hadal Waters of the Japan Trench under the Eutrophic Ocean

    PubMed Central

    Nunoura, Takuro; Hirai, Miho; Yoshida-Takashima, Yukari; Nishizawa, Manabu; Kawagucci, Shinsuke; Yokokawa, Taichi; Miyazaki, Junichi; Koide, Osamu; Makita, Hiroko; Takaki, Yoshihiro; Sunamura, Michinari; Takai, Ken

    2016-01-01

    The Japan Trench is located under the eutrophic Northwestern Pacific while the Mariana Trench that harbors the unique hadal planktonic biosphere is located under the oligotrophic Pacific. Water samples from the sea surface to just above the seafloor at a total of 11 stations including a trench axis station, were investigated several months after the Tohoku Earthquake in March 2011. High turbidity zones in deep waters were observed at most of the sampling stations. The small subunit (SSU) rRNA gene community structures in the hadal waters (water depths below 6000 m) at the trench axis station were distinct from those in the overlying meso-, bathy and abyssopelagic waters (water depths between 200 and 1000 m, 1000 and 4000 m, and 4000 and 6000 m, respectively), although the SSU rRNA gene sequences suggested that potential heterotrophic bacteria dominated in all of the waters. Potential niche separation of nitrifiers, including ammonia-oxidizing archaea (AOA), was revealed by quantitative PCR analyses. It seems likely that Nitrosopumilus-like AOAs respond to a high flux of electron donors and dominate in several zones of water columns including shallow and very deep waters. This study highlights the effects of suspended organic matter, as induced by seafloor deformation, on microbial communities in deep waters and confirm the occurrence of the distinctive hadal biosphere in global trench environments hypothesized in the previous study. PMID:27559333

  17. Distribution and Niche Separation of Planktonic Microbial Communities in the Water Columns from the Surface to the Hadal Waters of the Japan Trench under the Eutrophic Ocean.

    PubMed

    Nunoura, Takuro; Hirai, Miho; Yoshida-Takashima, Yukari; Nishizawa, Manabu; Kawagucci, Shinsuke; Yokokawa, Taichi; Miyazaki, Junichi; Koide, Osamu; Makita, Hiroko; Takaki, Yoshihiro; Sunamura, Michinari; Takai, Ken

    2016-01-01

    The Japan Trench is located under the eutrophic Northwestern Pacific while the Mariana Trench that harbors the unique hadal planktonic biosphere is located under the oligotrophic Pacific. Water samples from the sea surface to just above the seafloor at a total of 11 stations including a trench axis station, were investigated several months after the Tohoku Earthquake in March 2011. High turbidity zones in deep waters were observed at most of the sampling stations. The small subunit (SSU) rRNA gene community structures in the hadal waters (water depths below 6000 m) at the trench axis station were distinct from those in the overlying meso-, bathy and abyssopelagic waters (water depths between 200 and 1000 m, 1000 and 4000 m, and 4000 and 6000 m, respectively), although the SSU rRNA gene sequences suggested that potential heterotrophic bacteria dominated in all of the waters. Potential niche separation of nitrifiers, including ammonia-oxidizing archaea (AOA), was revealed by quantitative PCR analyses. It seems likely that Nitrosopumilus-like AOAs respond to a high flux of electron donors and dominate in several zones of water columns including shallow and very deep waters. This study highlights the effects of suspended organic matter, as induced by seafloor deformation, on microbial communities in deep waters and confirm the occurrence of the distinctive hadal biosphere in global trench environments hypothesized in the previous study. PMID:27559333

  18. Distribution and Niche Separation of Planktonic Microbial Communities in the Water Columns from the Surface to the Hadal Waters of the Japan Trench under the Eutrophic Ocean.

    PubMed

    Nunoura, Takuro; Hirai, Miho; Yoshida-Takashima, Yukari; Nishizawa, Manabu; Kawagucci, Shinsuke; Yokokawa, Taichi; Miyazaki, Junichi; Koide, Osamu; Makita, Hiroko; Takaki, Yoshihiro; Sunamura, Michinari; Takai, Ken

    2016-01-01

    The Japan Trench is located under the eutrophic Northwestern Pacific while the Mariana Trench that harbors the unique hadal planktonic biosphere is located under the oligotrophic Pacific. Water samples from the sea surface to just above the seafloor at a total of 11 stations including a trench axis station, were investigated several months after the Tohoku Earthquake in March 2011. High turbidity zones in deep waters were observed at most of the sampling stations. The small subunit (SSU) rRNA gene community structures in the hadal waters (water depths below 6000 m) at the trench axis station were distinct from those in the overlying meso-, bathy and abyssopelagic waters (water depths between 200 and 1000 m, 1000 and 4000 m, and 4000 and 6000 m, respectively), although the SSU rRNA gene sequences suggested that potential heterotrophic bacteria dominated in all of the waters. Potential niche separation of nitrifiers, including ammonia-oxidizing archaea (AOA), was revealed by quantitative PCR analyses. It seems likely that Nitrosopumilus-like AOAs respond to a high flux of electron donors and dominate in several zones of water columns including shallow and very deep waters. This study highlights the effects of suspended organic matter, as induced by seafloor deformation, on microbial communities in deep waters and confirm the occurrence of the distinctive hadal biosphere in global trench environments hypothesized in the previous study.

  19. Determination of the major constituents in fruit of Arctium lappa L. by matrix solid-phase dispersion extraction coupled with HPLC separation and fluorescence detection.

    PubMed

    Liu, He; Zhang, Yupu; Sun, Yantao; Wang, Xue; Zhai, Yujuan; Sun, Ye; Sun, Shuo; Yu, Aimin; Zhang, Hanqi; Wang, Yinghua

    2010-10-15

    The arctiin and arctigenin in the fruit of Arctium lappa L. were extracted by matrix solid-phase dispersion (MSPD) and determined by high-performance liquid chromatography (HPLC) with fluorescence detection. The experimental conditions for the MSPD were optimized. Silica gel was selected as dispersion adsorbent and methanol as elution solvent. The calibration curve showed good relationship (r>0.9998) in the concentration range of 0.010-5.0μgmL(-1) for arctiin and 0.025-7.5μgmL(-1) for arctigenin. The recoveries were between 74.4% and 100%. The proposed method consumed less sample, time and solvent compared with conventional methods, including ultrasonic and Soxhlet extraction.

  20. Synthesis of. beta. -D-glucan in vitro: HPLC separation of oligosaccharides from enzymic digests of glucans synthesized by Golgi apparatus and UDP-glucose

    SciTech Connect

    Gibeaut, D.M.; Carpita, N.C. )

    1990-05-01

    Anion exchange HPLC (Dionex) resolves {beta}-D-glucose oligosaccharides which vary in (1{yields}3) and (1{yields}4) linkage structure. The products from incubation of UDPG and cellular membranes from maize enriched for Golgi apparatus, ER, and plasma membrane are digested with glucan hydrolases specific for either (1{yields}3) or (1{yields}4) linkages and one which requires a specific (1{yields}3){beta}-D-glc-(1{yields}4){beta}-D-glc sequence. Two oligomers released by the latter enzyme are diagnostic of cereal grass mixed-linkage {beta}-D-glucan and are used as standards to examine reaction conditions for synthesis of this polymer with radioactive substrates. Our preliminary data indicate that formation of specific sequences of {beta}-D-glucan is increased by high concentrations of UDPG. Other radioactive oligomers are present in greater quantity than those from authentic mixed-linkage glucan and their linkage structure is now under investigation.

  1. Differentiation of Cirsium japonicum and C. setosum by TLC and HPLC-MS.

    PubMed

    Ganzera, Markus; Pöcher, Astrid; Stuppner, Hermann

    2005-01-01

    The Chinese Pharmacopoeia indicates the use of field thistle (Cirsium setosum) and Japanese field thistle (C. japonicum) in the treatment of bleeding and inflammation. In the absence of an analytical method for the differentiation and analysis of these two species, TLC and HPLC-MS methods have been developed for this purpose. Both species could be readily distinguished by their flavonoid pattern as revealed by TLC on silica gel layers eluted with ethyl acetate:formic acid:acetic acid:water. The quantitative determination of four flavonoids, namely hispidulin-7-neohesperidoside, linarin, pectolinarin and luteolin, was possible using HPLC. Their optimum separation was achieved on a C12 column eluted with water and 0.025% trifluoroacetic acid in acetonitrile. HPLC-MS experiments were performed to confirm peak identity. In samples of C. japonicum, pectolinarin was the major flavonoid (0.32-2.00%), followed by linarin, hispidulin-7-neohesperidoside and luteolin; the total flavonoid content varied from 0.81 to 3.67%. In C. setosum only one flavonoid (linarin; 1.36-2.83%) was assignable. The HPLC method was validated for linearity, limit of detection (< or = 1.7 ng on-column), peak purity, repeatability (< or = 2.3%) and accuracy (recovery rates of spiked samples were between 99.2 and 101.6%).

  2. Analysis of munitions constituents in IMX formulations by HPLC and HPLC-MS.

    PubMed

    Russell, A L; Seiter, J M; Coleman, J G; Winstead, B; Bednar, A J

    2014-10-01

    The use of Insensitive Munitions eXplosives (IMX) is increasing as the Army seeks to replace certain conventional munitions constituents, such as 2,4,6-trinitrotolene (TNT), for improved safety. The IMX formulations are more stable and therefore less prone to accidental detonation while designed to match the performance of legacy materials. Two formulations, IMX 101 and 104 are being investigated as a replacement for TNT in artillery rounds and composition B Army mortars, respectively. The chemical formulations of IMX-101 and 104 are comprised of four constituents;2,4-dinitroanisole (DNAN), 3-nitro-1,2,4-triazol-5-one (NTO), 1-nitroguanidine (NQ), and Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) which are mixed in various ratios to achieve the desired performance. The current work details the analysis of the IMX constituents by single column HPLC-UV-ESI-MS. Detection limits determined are in agreement with similar HPLC analysis of compounds, ranging from 7 to 9μg/L. Gradient mobile phases are used to allow separation of the 4 target compounds in more complex mixture of other concomitant compounds. Mass spectra are used to confirm analyte identity with chromatographic retention time. PMID:25059196

  3. Analysis of munitions constituents in IMX formulations by HPLC and HPLC-MS.

    PubMed

    Russell, A L; Seiter, J M; Coleman, J G; Winstead, B; Bednar, A J

    2014-10-01

    The use of Insensitive Munitions eXplosives (IMX) is increasing as the Army seeks to replace certain conventional munitions constituents, such as 2,4,6-trinitrotolene (TNT), for improved safety. The IMX formulations are more stable and therefore less prone to accidental detonation while designed to match the performance of legacy materials. Two formulations, IMX 101 and 104 are being investigated as a replacement for TNT in artillery rounds and composition B Army mortars, respectively. The chemical formulations of IMX-101 and 104 are comprised of four constituents;2,4-dinitroanisole (DNAN), 3-nitro-1,2,4-triazol-5-one (NTO), 1-nitroguanidine (NQ), and Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) which are mixed in various ratios to achieve the desired performance. The current work details the analysis of the IMX constituents by single column HPLC-UV-ESI-MS. Detection limits determined are in agreement with similar HPLC analysis of compounds, ranging from 7 to 9μg/L. Gradient mobile phases are used to allow separation of the 4 target compounds in more complex mixture of other concomitant compounds. Mass spectra are used to confirm analyte identity with chromatographic retention time.

  4. Fabrication of an ionic liquid-based macroporous polymer monolithic column via atom transfer radical polymerization for the separation of small molecules.

    PubMed

    Zhang, Hang; Bai, Ligai; Wei, Zhen; Liu, Sha; Liu, Haiyan; Yan, Hongyuan

    2016-03-01

    A polymer monolithic column was prepared in a stainless steel column (50×4.6mm i.d.) via atom transfer radical polymerization technique using triallyl isocyanurate and ionic liquid (1-allyl-3-methylimidazolium chloride) as co-monomers, ethylene dimethacrylate as cross linking agent, polyethylene glycol 200, 1,4-butanediol, and N, N- dimethylformamide as porogen system, CCl4 as initiator, and FeCl2 as catalyst. The optimized polymer columns were characterized by scanning electron microscope, nitrogen adsorption-desorption instrument, mercury intrusion porosimetry, infrared spectrometer, and thermogravimetric analysis technique. Respectively, all of these factors above could illustrate that the optimized columns had relative uniform macroporous structure and high thermal stability. A series of basic and acidic small molecules, isomers, and homologues were used to evaluate the performance of these monoliths and enhanced column efficiency was obtained. PMID:26717814

  5. Surface confined ionic liquid as a stationary phase for HPLC

    SciTech Connect

    Wang, Qian; Baker, Gary A; Baker, Sheila N; Colon, Luis

    2006-01-01

    Trimethoxysilane ionosilane derivatives of room temperature ionic liquids based on alkylimidazolium bromides were synthesized for attachment to silica support material. The derivatives 1-methyl-3-(trimethoxysilylpropyl)imidazolium bromide and 1-butyl-3-(trimethoxysilylpropyl)imidazolium bromide were used to modify the surface of 3 {micro}m diameter silica particles to act as the stationary phase for HPLC. The modified particles were characterized by thermogravimetric analysis (TGA) and {sup 13}C and {sup 29}Si NMR spectroscopies. The surface modification procedure rendered particles with a surface coverage of 0.84 {micro}mol m{sup -2} for the alkylimidazolium bromide. The ionic liquid moiety was predominantly attached to the silica surface through two siloxane bonds of the ionosilane derivative (63%). Columns packed with the modified silica material were tested under HPLC conditions. Preliminary evaluation of the stationary phase for HPLC was performed using aromatic carboxylic acids as model compounds. The separation mechanism appears to involve multiple interactions including ion exchange, hydrophobic interaction, and other electrostatic interactions.

  6. Simultaneous Speciation of Arsenic, Selenium, and Chromium by HPLC-ICP-MS

    USGS Publications Warehouse

    Wolf, Ruth E.; Morman, Suzette A.; Morrison, Jean M.; Lamothe, Paul J.

    2008-01-01

    An adaptation of an analytical method developed for chromium speciation has been utilized for the simultaneous determination of As(III), As(V), Se(IV), Se(VI), Cr(III), and Cr(VI) species using high performance liquid chromatography (HPLC) separation with ICP-MS detection. Reduction of interferences for the determination of As, Se, and Cr by ICP-MS is a major consideration for this method. Toward this end, a Dynamic Reaction Cell (DRC) ICP-MS system was used to detect the species eluted from the chromatographic column. A variety of reaction cell gases and conditions may be utilized, and the advantages and limitations of the gases tested to date will be presented and discussed. The separation and detection of the As, Se, and Cr species of interest can be achieved using the same chromatographic conditions in less than 2 minutes by complexing the Cr(III) with EDTA prior to injection on the HPLC column. Practical aspects of simultaneous speciation analysis will be presented and discussed, including issues with HPLC sample vial contamination, standard and sample contamination, species stability, and considerations regarding sample collection and preservation methods. The results of testing to determine the method's robustness to common concomitant element and anion effects will also be discussed. Finally, results will be presented using the method for the analysis of a variety of environmental and geological samples including waters, soil leachates and simulated bio-fluid leachates.

  7. Industrial application of green chromatography--I. Separation and analysis of niacinamide in skincare creams using pure water as the mobile phase.

    PubMed

    Yang, Yu; Strickland, Zackary; Kapalavavi, Brahmam; Marple, Ronita; Gamsky, Chris

    2011-03-15

    In this work, chromatographic separation of niacin and niacinamide using pure water as the sole component in the mobile phase has been investigated. The separation and analysis of niacinamide have been optimized using three columns at different temperatures and various flow rates. Our results clearly demonstrate that separation and analysis of niacinamide from skincare products can be achieved using pure water as the eluent at 60°C on a Waters XTerra MS C18 column, a Waters XBridge C18 column, or at 80°C on a Hamilton PRP-1 column. The separation efficiency, quantification quality, and analysis time of this new method are at least comparable with those of the traditional HPLC methods. Compared with traditional HPLC, the major advantage of this newly developed green chromatography technique is the elimination of organic solvents required in the HPLC mobile phase. In addition, the pure water chromatography separations described in this work can be directly applied in industrial plant settings without further modification of the existing HPLC equipment.

  8. Development and evaluation of a new method for the determination of the carotenoid content in selected vegetables by HPLC and HPLC-MS-MS.

    PubMed

    Huck, C W; Popp, M; Scherz, H; Bonn, G K

    2000-10-01

    Epidemologic studies have shown inverse correlation between the consumption of carotenoid-rich vegetables and the incidence of cancer. Therefore, analytical techniques for the quantitative determination of carotenoids in complex sample matrices are important. The most used method is reversed-phase (RP)-high-performance liquid chromatography (HPLC). In this study, seventeen mobile-phase systems described in the literature and six RP-HPLC columns with differences in particle size and porosity are evaluated. Derived from these results, a new mobile-phase (acetonitrile, methanol, chloroform, and n-heptane) including solvent modifiers is presented, which allows an improved and more efficient separation of carotenoids. From all columns tested, the best chromatographic parameters are found using a silica C18 column (250 x 2 mm, 5 microm, 100 A). As was found, absorbance detection at 450 nm allows the determination of the carotenoids down to the picogram range with good linearity (R2 > 0.98). For the identification and quantitation of carotenoids in complex sample matrices (containing additionally other ultraviolet-absorbing compounds), the optimized RP chromatographic system is coupled to a mass spectrometer (MS) using an atmospheric pressure ionization interface. The calibration plots show high linearity (R2 > 0.99), and the detection limit is found in the lower nanogram range. Furthermore, collision-induced dissociation in the ion source allows for the identification of carotenoids by their characteristic fragmentation pathways. In this study, a total of nine species of vegetables commonly consumed in Central Europe are analyzed for their contents of carotenoids (namely lutein, zeaxanthin, beta-cryptoxanthin, and beta-carotene) by RP-HPLC and RP-HPLC-MS-MS. It is found that good sources for lutein are spinach, kale, and broccoli, and sources for beta-carotene are broccoli, spinach, kale, carrots, and tomatoes. This new method is an improvement for the identification and

  9. Post-synthetic modification of MIL-101(Cr) with pyridine for high-performance liquid chromatographic separation of tocopherols.

    PubMed

    Yang, Fang; Yang, Cheng-Xiong; Yan, Xiu-Ping

    2015-05-01

    Effective separation of tocopherols is challenging and significant due to their structural similarity and important biological role. Here we report the post-synthetic modification of metal-organic framework (MOF) MIL-101(Cr) with pyridine for high-performance liquid chromatographic (HPLC) separation of tocopherols. Baseline separation of four tocopherols was achieved on a pyridine-grafted MIL-101(Cr) packed column within 10 min using hexane/isopropanol (96:4, v/v) as the mobile phase at a flow rate of 0.5 mL min(-1). The pyridine-grafted MIL-101(Cr) packed column gave high column efficiency (85,000 plates m(-1) for δ-tocopherol) and good precision (0.2-0.3% for retention time, 1.8-3.4% for peak area, 2.6-2.7% for peak height), and also offered much better performance than unmodified MIL-101(Cr) and commercial amino-bonded silica packed column for HPLC separation of tocopherols. The results not only show the promising application of pyridine-grafted MIL-101(Cr) as a novel stationary phase for HPLC separation of tocopherols, but also reveal a facile post-modification of MOFs to expand the application of MOFs in separation sciences.

  10. Simultaneous separation and identification of oligomeric procyanidins and anthocyanin-derived pigments in raw red wine by HPLC-UV-ESI-MSn.

    PubMed

    Pati, S; Losito, I; Gambacorta, G; La Notte, E; Palmisano, F; Zambonin, P G

    2006-07-01

    Samples of raw red wine (Primitivo di Manduria, Apulia, Southern Italy) were analysed without any pre-treatment (except 1:2 dilution with water) using HPLC with detection based on UV absorbance and Electrospray Ionisation Sequential Mass Spectrometry (ESI-MSn, with n = 1-3) in a series configuration. In particular, absorbance at 520 nm was monitored for UV detection in order to identify pigments responsible for wine colour. On the other hand, two subsequent stages of MS detection based on positive ions were adopted. The first consisted of an explorative MS acquisition, aimed at the individuation of the m/z ratios for positively charged compounds; the second was based on fragmentation of the detected ions within an ion trap analyser, followed by MS/MS and, if required, MS3 acquisitions. The synergy between UV detection and MSn analysis led to the identification of 41 pigments, which can be classified into five groups: grape anthocyanins, pyranoanthocyanins, vinyl-linked anthocyanin-flavanol pigments, ethyl-bridged anthocyanin-flavanol pigments and flavanol-anthocyanin compounds. Many isomeric and oligomeric structures were found within each group. A further class of compounds, not absorbing in the visible spectrum, could be also characterised by ESI-MSn and corresponded to B-type procyanidins, i.e. proanthocyanidins arising from C4-->C8/C4-->C6 couplings between catechin or epicatechin units. In particular, oligomeric structures (from dimers to pentamers), often present with several isomers, were identified and their fragmentation patterns clarified. PMID:16770836

  11. SEPARATION AND CHARACTERIZATION OF TETROL METABOLITES OF BENZO[A]PYRENE-DNA ADDUCTS USING HPLC AND SOLID-MATRIX ROOM TEMPERATURE LUMINESCENCE. (R824100)

    EPA Science Inventory

    Abstract

    Four tetrols of benzo[a]pyrene-DNA adducts were separated using reversed-phase high performance liquid chromatography. Chromatographic fractions containing a given tetrol were readily characterized with solid-matrix room temperature luminescence techniques. So...

  12. Reverse-phase HPLC of benzylpropionitrile dithiocarbamate complexes for the determination of priority pollutant metals

    SciTech Connect

    Park, Y.J.

    1990-01-01

    A new dithiocarbamate, benzylpropionitrile dithiocarbamate (BPDTC), has been synthesized for use in metal analysis. The HPLC behavior of metal chelates of BPDTC has been investigated for the simultaneous determination of antimony, cadmium, chromium, copper, mercury, nickel, lead, selenium, thallium, and zinc, all of which are on the Environmental Protection Agency's list of priority pollutant metals. Metals are extracted into dichloromethane as BPDTC chelates, and then separated on a C-18 column. Cobalt is added as an internal standard. The effects of pH and of three organic modifiers (methanol, acetonitrile, tetrahydrofuran) of the mobile phase on retention time have been investigated. Addition of dichloromethane to the mobile phase increases solubility and chelate stability, and improves the separation of metal BPDTC complexes. BPDTC is added to the aqueous mobile phase to reduce on-column dissociation of the complexes. Detection limits at 260 nm are in the range of 0.1 to 3 ppb using a 1 liter sample.

  13. [Determination of azoxystrobin in tea by HPLC].

    PubMed

    Chonan, T

    2001-08-01

    A determination method has been developed for azoxystrobin in tea by HPLC. Azoxystrobin was extracted from a sample with acetone, and the extract was passed through an alumina column to remove tannin. The eluate was concentrated to ca. 25 mL and passed through a Sep-Pak Vac tC18 to remove pigments. The eluate was cleaned-up by using liquid-liquid partition, and Florisil and silica-gel columns. The HPLC analysis for azoxystrobin was carried out on a C18 column with acetonitrile-water (9:11) as the mobile phase, with ultraviolet detection at 260 nm. The recovery of azoxystrobin fortified at the level of 0.4 microgram/g was 90.2% and the limit of determination was 0.2 microgram/g.

  14. Application of an efficient strategy based on MAE, HPLC-DAD-MS/MS and HSCCC for the rapid extraction, identification, separation and purification of flavonoids from Fructus Aurantii Immaturus.

    PubMed

    Wang, Chen; Pan, Yaju; Fan, Guorong; Chai, Yifeng; Wu, Yutian

    2010-03-01

    This study presents an efficient strategy based on microwave-assisted extraction (MAE), HPLC-DAD-MS/MS and high-speed counter-current chromatography (HSCCC) for the rapid extraction, identification, separation and purification of active components from the traditional Chinese medicine Fructus Aurantii Immaturus. An LC-DAD-MS/MS method was applied for the screening and structural identification of main components in crude extract, and five components were preliminarily identified as neoeriocitrin, narirutin, naringin, hesperidin and neohesperidin according to their UV and mass spectra. An efficient MAE method for the extraction of the three most abundant components (narirutin, naringin and neohesperidin) was optimized by the combination of univariate and multivariate approaches. The crude extract was then separated and purified by HSCCC and a total of 61.6 mg of narirutin, 207.3 mg of naringin and 159.5 mg of neohesperidin at high purities of 98.1, 97.2 and 99.5%, respectively, were obtained from 1.42 g of crude extract. The recoveries of these compounds were 86, 93 and 89%, respectively.

  15. [A simple preparation method of an electric heating apparatus for heating capillary chromatographic columns and its application in liquid chromatography-mass spectrometry system].

    PubMed

    Jin, Zuyao; Lü, Yayao; Zhou, Shanshan; Hao, Feiran; Fu, Bin; Ying, Wantao; Qian, Xiaohong; Zhang, Yangjun

    2015-06-01

    For deep coverage of proteome, especially in performing qualitative identification and quantitative analysis of low-abundance proteins, the most commonly used method is the application of a longer capillary chromatographic column or a capillary column packed with smaller particle sizes. However, this causes another problem, the very high back pressure which results in liquid leaks in some connection parts in a liquid chromatograph. To solve this problem, an electric heating apparatus was developed to raise the temperature of a capillary column for reducing its back pressure, which was further applied in a capillary high performance liquid chromatography-tandem mass spectrometry system (cHPLC-MS/MS), and evaluated in the terms of chromatographic column back pressure and chromatographic column efficiency using bovine serum albumin (BSA) tryptic digests and yeast tryptic digests, separately. The results showed that at the optimum current, our electric heating apparatus could reduce the column pressure of a capillary column packed with 3 µm packing materials by at least 50% during the separation of BSA tryptic digestion and yeast tryptic digestion, compared with that without electric heating. The column efficiency was also increased slightly. This suggested that the electric heating apparatus can significantly reduce the column pressure, which provides an efficient way to use capillary chromatographic columns packed with smaller sizes of particles at a lower pressure.

  16. Distillation Column Flooding Predictor

    SciTech Connect

    George E. Dzyacky

    2010-11-23

    The Flooding Predictor™ is a patented advanced control technology proven in research at the Separations Research Program, University of Texas at Austin, to increase distillation column throughput by over 6%, while also increasing energy efficiency by 10%. The research was conducted under a U. S. Department of Energy Cooperative Agreement awarded to George Dzyacky of 2ndpoint, LLC. The Flooding Predictor™ works by detecting the incipient flood point and controlling the column closer to its actual hydraulic limit than historical practices have allowed. Further, the technology uses existing column instrumentation, meaning no additional refining infrastructure is required. Refiners often push distillation columns to maximize throughput, improve separation, or simply to achieve day-to-day optimization. Attempting to achieve such operating objectives is a tricky undertaking that can result in flooding. Operators and advanced control strategies alike rely on the conventional use of delta-pressure instrumentation to approximate the column’s approach to flood. But column delta-pressure is more an inference of the column’s approach to flood than it is an actual measurement of it. As a consequence, delta pressure limits are established conservatively in order to operate in a regime where the column is never expected to flood. As a result, there is much “left on the table” when operating in such a regime, i.e. the capacity difference between controlling the column to an upper delta-pressure limit and controlling it to the actual hydraulic limit. The Flooding Predictor™, an innovative pattern recognition technology, controls columns at their actual hydraulic limit, which research shows leads to a throughput increase of over 6%. Controlling closer to the hydraulic limit also permits operation in a sweet spot of increased energy-efficiency. In this region of increased column loading, the Flooding Predictor is able to exploit the benefits of higher liquid

  17. Separations of substituted benzenes and polycyclic aromatic hydrocarbons using normal- and reverse-phase high performance liquid chromatography with UiO-66 as the stationary phase.

    PubMed

    Zhao, Wei-Wei; Zhang, Chao-Yan; Yan, Zeng-Guang; Bai, Li-Ping; Wang, Xiayan; Huang, Hongliang; Zhou, You-Ya; Xie, Yabo; Li, Fa-Sheng; Li, Jian-Rong

    2014-11-28

    Metal-organic frameworks (MOFs) have great potential for applications in chromatography due to their highly tailorable porous structures and unique properties. In this work, the stable MOF UiO-66 was evaluated as both a normal-phase (NP-) and a reverse-phase (RP-) stationary phase in the high performance liquid chromatography (HPLC) to separate substituted benzenes (SBs) and polycyclic aromatic hydrocarbons (PAHs). It was found that the mobile phase composition has a significant effect on the HPLC separation. Baseline RP-HPLC separations of xylene isomers; naphthalene and anthracene; naphthalene and chrysene; and naphthalene, fluorene, and chrysene were achieved using MeOH/H2O ratios of 80:20, 75:25, 85:15, and 75:25, respectively, on the UiO-66 column. Similarly, baseline NP-HPLC separations of xylene isomers and ethylbenzene; ethylbenzene, styrene, o-xylene, and m-xylene; and several PAHs were also obtained on the UiO-66 column with different mobile phase compositions. The relative standard deviations (RSDs) of retention time, peak height, peak area, and half peak width for five replicate separations of the tested analytes were within the ranges 0.2-0.4%, 0.2-1.6%, 0.7-3.9%, 0.4-1.1%, respectively. We also evaluated other critical HPLC parameters, including injected sample mass, column temperature, and the thermodynamic characters of both the RP-HPLC and the NP-HPLC separation processes. It was confirmed that the separation of SBs on a UiO-66 column was an exothermic process, controlled by both enthalpy change (ΔH) and entropy change (ΔS). The reverse shape selectivity, size selectivity, stacking effect, and electrostatic force played vital roles in the separations of these analytes. To the best of our knowledge, this method is one of the very few examples of using MOFs as the stationary phase in both NP-HPLC and RP-HPLC. MOF-based stationary phases may thus be applied in the separations and analyses of SBs and PAHs in environmental samples.

  18. Stability indicating HPLC-method for the determination of econazole nitrate in cream and lotion formulations.

    PubMed

    Christinat, R; Zulliger, H W

    1984-01-01

    A simple, fast HPLC-method for the determination of econazole nitrate in cream (Pevaryl, Pevisone) and lotion formulations (based on polyethylenic oleic glycerides and mono/di-stearic esters of ethylene- and polyethylene glycol) is described. The method is stability indicating as well as linear (range 5-15 mg econazole nitrate/g) and shows a good recovery (98.7-100.2%) and a good reproducibility (cv less than 1%, n = 10). The chromatographic separation is achieved on a RP-18 column using methanol/aqueous ammoniumcarbonate solution/tetrahydrofurane as the mobile phase. Quantification of the chromatograms is done by internal standard method using peak areas.

  19. Analysis of multiple sweeteners and their degradation products in lassi by HPLC and HPTLC plates.

    PubMed

    George, V; Arora, S; Wadhwa, B K; Singh, A K

    2010-08-01

    A solid phase extraction method using C18 cartridges was standardized for the isolation of multiple sweeteners (aspartame, acesulfame-K and saccharin) and their degradation products (diketopiperazine, Lphenylalanine, acetoacetamide and 2-sulfobenzoic acid) from lassi. Analytical conditions for HPLC were standardized over C18 column using UV detector for the simultaneous separation and estimation of multiple sweeteners and their degradation products in lassi sample isolates. A simple cartridge free method was developed for the isolation of sucralose from lassi. Method was also standardized for qualitative detection and quantitative estimation of sucralose over amino and silica gel plates of HPTLC.

  20. Stereoselective metabolism of chrysene by rat liver microsomes. Direct separation of diol enantiomers by chiral stationary phase h.p.l.c.

    PubMed

    Weems, H B; Fu, P P; Yang, S K

    1986-07-01

    The direct enantiomeric resolution of non-K region trans-1,2-dihydrodiol, 1,2,3,4-tetrahydro-trans-1,2-diol, trans-3,4-dihydrodiol and 1,2,3,4-tetrahydro-trans-3,4-diol, K region trans- and cis-5,6-dihydrodiols and their monomethyl ethers of chrysene was studied by chiral stationary phase high-performance liquid chromatography (CSP-h.p.l.c.). The chiral stationary phase columns were packed with gamma-aminopropylsilanized silica to which either (R)-N-(3,5-dinitrobenzoyl)-phenylglycine or (S)-N-(3,5-dinitrobenzoyl)leucine was bonded either ionically or covalently. Enantiomers of all dihydrodiol derivatives were resolved by one or more, but not all, of the chiral stationary phases utilized. Enantiomeric resolutions were substantially improved when the non-K region dihydrodiols were converted to tetrahydrodiols. The absolute configurations of the K region trans- and cis-5,6-dihydrodiols were established by the exciton chirality circular dichroism method. The (R,R):(S,S) enantiomer ratios, determined by CSP-h.p.l.c., of the 1,2-, 3,4- and 5,6-trans-dihydrodiols formed in the metabolism of chrysene by liver microsomes from untreated male rats of the Sprague--Dawley strain were found to be 51:49, 99:1 and 86:14, respectively; from phenobarbital-treated rats, 41:59, 99:1 and 87:13, respectively; from 3-methylcholanthrene-treated rats, 96:4, 99:1 and 92:8, respectively. The absolute configurations of chrysene 5,6-epoxide enantiomers, resolved by CSP-h.p.l.c., were elucidated by the determination of the structures and absolute configurations of their methoxylation products. Both enantiomers of chrysene 5,6-epoxide were hydrated by microsomal epoxide hydrolase to chrysene trans-5,6-dihydrodiol enriched (67-92%) in the 5R,6R enantiomer. Chrysene 5R,6S-epoxide was hydrated to trans-5,6-dihydrodiol at a rate approximately 6-fold faster than chrysene 5S,6R-epoxide.

  1. [Research on the separation of limonoid glucosides by reversed-phase preparative high performance liquid chromatography].

    PubMed

    Tian, Q G; Dai, J; Ding, X L

    2000-03-01

    Obacunone-17-beta-D-glucopyranoside (OG) was isolated from the seeds of Citrus Sinensis Osbeck by using solvent extraction, classical polymer adsorption column separation and weak base anion ion-exchange separation, OG was finally purified by C18 reversed-phase preparative high performance liquid chromatography and was identified by thin-layer chromatography. The purity of OG was analyzed by analytical reversed-phase HPLC. At last the structure of OG was determined by 1H and 13C nuclear magnetic resonance spectrometry (NMR). In this work, the conditions of the reversed-phase preparative HPLC technique to purify limonoid glucosides was optimized. The reversed-phase preparative HPLC on a C18 column with a mobile phase of acidic acetonitrile-water (about 0.2% TFA, V/V) at pH 3 enabled the baseline separation of limonoid glucosides in the extract. The results show that OG is the predominant limonoid glucoside in the seeds of Citrus Sinensis Osbeck and nomilin glucoside is the second one. The results also show that the classical polymer adsorption column separation and weak base anion ion-exchange separation are effective for purifying limonoid glucosides.

  2. Simultaneous separation/enrichment and detection of trace ciprofloxacin and lomefloxacin in food samples using thermosensitive smart polymers aqueous two-phase flotation system combined with HPLC.

    PubMed

    Lu, Yang; Chen, Bo; Yu, Miao; Han, Juan; Wang, Yun; Tan, Zhenjiang; Yan, Yongsheng

    2016-11-01

    Smart polymer aqueous two phase flotation system (SPATPF) is a new separation and enrichment technology that integrated the advantages of the three technologies, i.e., aqueous two phase system, smart polymer and flotation sublation. Ethylene oxide and propylene oxide copolymer (EOPO)-(NH4)2SO4 SPATPF is a pretreatment technique, and it is coupled with high-performance liquid chromatography to analyze the trace ciprofloxacin and lomefloxacin in real food samples. The optimized conditions of experiment were determined in the multi-factor experiment by using response surface methodology. The flotation efficiency of lomefloxacin and ciprofloxacin was 94.50% and 98.23% under the optimized conditions. The recycling experimentsshowed that the smart polymer EOPO could use repeatedly, which will reduce the cost in the future application.

  3. Simultaneous separation/enrichment and detection of trace ciprofloxacin and lomefloxacin in food samples using thermosensitive smart polymers aqueous two-phase flotation system combined with HPLC.

    PubMed

    Lu, Yang; Chen, Bo; Yu, Miao; Han, Juan; Wang, Yun; Tan, Zhenjiang; Yan, Yongsheng

    2016-11-01

    Smart polymer aqueous two phase flotation system (SPATPF) is a new separation and enrichment technology that integrated the advantages of the three technologies, i.e., aqueous two phase system, smart polymer and flotation sublation. Ethylene oxide and propylene oxide copolymer (EOPO)-(NH4)2SO4 SPATPF is a pretreatment technique, and it is coupled with high-performance liquid chromatography to analyze the trace ciprofloxacin and lomefloxacin in real food samples. The optimized conditions of experiment were determined in the multi-factor experiment by using response surface methodology. The flotation efficiency of lomefloxacin and ciprofloxacin was 94.50% and 98.23% under the optimized conditions. The recycling experimentsshowed that the smart polymer EOPO could use repeatedly, which will reduce the cost in the future application. PMID:27211613

  4. Determination of zinc pyrithione in shampoos by HPLC and HPLC-MS/MS.

    PubMed

    Gu, Yu-Xiang; Wang, Qing-He; Zhou, Ze-Lin; Lv, Qing; Mai, Cheng-Hua

    2014-01-01

    Methods have been developed for the determination of zinc pyrithione (ZPT) in shampoos using high-performance liquid chromatography (HPLC) and high-performance liquid chromatography-mass spectrometry/mass spectrometry (HPLC-MS/MS). Samples were washed by water first to remove surfactant and water-soluble impurities, then ultrasonic-extracted by acetonitrile-methanol for 30 min, and finally analyzed by MG C18 column (250 mm x 4.6 mm, 5 μm) or RP-18e (100 mm x 3 mm, 2 μm) plus APCI-MS/MS. Limits of detection were determined as 0.015% (HPLC) and 0.003% (HPLC-MS/MS), with a limit of quantization of 0.05% and 0.01%, respectively. The recoveries were 85.8-104% (HPLC) and 87.6-107% (HPLC-MS/MS). A good linear relationship was obtained from 3.20 μg·ml(-1) to 200 μg·ml(-1) (HPLC) and 1.00 μg·ml(-1) to 200 μg·ml(-1) (HPLC-MS/MS). The proposed methods have been successfully applied to the analysis of ZPT in many shampoos. The established two methods were rapid and reproducible with low interference.

  5. Determination of zinc pyrithione in shampoos by HPLC and HPLC-MS/MS.

    PubMed

    Gu, Yu-Xiang; Wang, Qing-He; Zhou, Ze-Lin; Lv, Qing; Mai, Cheng-Hua

    2014-01-01

    Methods have been developed for the determination of zinc pyrithione (ZPT) in shampoos using high-performance liquid chromatography (HPLC) and high-performance liquid chromatography-mass spectrometry/mass spectrometry (HPLC-MS/MS). Samples were washed by water first to remove surfactant and water-soluble impurities, then ultrasonic-extracted by acetonitrile-methanol for 30 min, and finally analyzed by MG C18 column (250 mm x 4.6 mm, 5 μm) or RP-18e (100 mm x 3 mm, 2 μm) plus APCI-MS/MS. Limits of detection were determined as 0.015% (HPLC) and 0.003% (HPLC-MS/MS), with a limit of quantization of 0.05% and 0.01%, respectively. The recoveries were 85.8-104% (HPLC) and 87.6-107% (HPLC-MS/MS). A good linear relationship was obtained from 3.20 μg·ml(-1) to 200 μg·ml(-1) (HPLC) and 1.00 μg·ml(-1) to 200 μg·ml(-1) (HPLC-MS/MS). The proposed methods have been successfully applied to the analysis of ZPT in many shampoos. The established two methods were rapid and reproducible with low interference. PMID:25682618

  6. Quantitative and qualitative HPLC analysis of thermogenic weight loss products.

    PubMed

    Schaneberg, B T; Khan, I A

    2004-11-01

    An HPLC qualitative and quantitative method of seven analytes (caffeine, ephedrine, forskolin, icariin, pseudoephedrine, synephrine, and yohimbine) in thermogenic weight loss preparations available on the market is described in this paper. After 45 min the seven analytes were separated and detected in the acetonitrile: water (80:20) extract. The method uses a Waters XTerra RP18 (5 microm particle size) column as the stationary phase, a gradient mobile phase of water (5.0 mM SDS) and acetonitrile, and a UV detection of 210 nm. The correlation coefficients for the calibration curves and the recovery rates ranged from 0.994 to 0.999 and from 97.45% to 101.05%, respectively. The qualitative and quantitative results are discussed. PMID:15587578

  7. Characterization of nutraceuticals and functional foods by innovative HPLC methods.

    PubMed

    Corradini, Claudio; Galanti, Roberta; Nicoletti, Isabella

    2002-04-01

    In recent years there is a growing interest in food and food ingredient which may provide health benefits. Food as well as food ingredients containing health-preserving components, are not considered conventional food, but can be defined as functional food. To characterise such foods, as well as nutraceuticals specific, high sensitive and reproducible analytical methodologies are needed. In light of this importance we set out to develop innovative HPLC methods employing reversed phase narrow bore column and high-performance anion-exchange chromatographic methods coupled with pulsed amperometric detection (HPAEC-PAD), which are specific for carbohydrate analysis. The developed methods were applied for the separation and quantification of citrus flavonoids and to characterize fructooligosaccharide (FOS) and fructans added to functional foods and nutraceuticals.

  8. Simultaneous analysis of neuroendocrine tumor markers by HPLC-electrochemical detection.

    PubMed

    Manickum, T

    2009-12-15

    A validated, high pressure liquid chromatographic (HPLC) method for simultaneous quantitation of urinary catecholic acids 4-hydroxy-3-methoxymandelic acid (HMMA) (vanylmandelic acid) (VMA), 4-hydroxy-3-methoxyphenylacetic acid (HVA) and 5-hydroxyindole-3-acetic acid (5-HIAA) was developed. Sample preparation involved liquid-liquid extraction of acidified urine, containing iso-HMMA (IS) as internal standard, with ether, evaporation of the organic extract, followed by reconstitution of the residue in phosphate buffer at pH 3.3. After reversed-phase HPLC at 35 degrees C and separation on a Licrospher 125mmx4mm C(18) column (5microm particle size) with phosphate buffer (pH 3.5)-acetone (950:50, v/v) as eluent, quantitation is achieved by electrochemical detection using coulometric detection at a potential of +350mV. The method was successfully applied to routine diagnosis of neuroblastoma, carcinoid syndrome and pheochromocytoma. PMID:19926540

  9. [Preparation of ferulic acid, senkyunolide I and senkyunolide H from Ligusticum chuanxiong by preparative HPLC].

    PubMed

    Xiong, Yao-Kun; Liang, Shuang; Hong, Yan-Long; Yang, Xiu-Juan; Shen, Lan; Du, Yan; Feng, Yi

    2013-06-01

    Preparative HPLC was used to prepare ferulic acid, senkyunolide I and senkyunolide H from Ligusticum chuanxiong. The separation was conducted on a Shim-Pack Prep-ODS (20.0 mm x 250 mm, 5 microm) column with the mobile phase of methanol-0.2% glacial acetic acid (50:50)at the flow rate of 5 mL x min(-1). The detection wavelength was 278 nm, and the purity of each compound was detected by HPLC analysis. Spectral data analyses including UV, ESI-MS and NMR were used to identify their structures. This method is simple, fast, which is suitable for preparation of standard reference of ferulic acid, senkyunolide I and senkyunolide H from L. chuanxiong and can meet the requirement of new drug research and development. PMID:24066590

  10. An HPLC chromatographic reactor approach for investigating the hydrolytic stability of a pharmaceutical compound.

    PubMed

    Skrdla, Peter J; Abrahim, Ahmed; Wu, Yan

    2006-06-01

    The solution-phase hydrolysis kinetics of the Aprepitant (Emend) prodrug, Fosaprepitant Dimeglumine, were investigated using an HPLC chromatographic reactor approach. The term 'chromatographic reactor' refers to the use of an analytical-scale column as both a flow-through reactor and, simultaneously, as separation medium for the reactant(s) and product(s). Recently, we reported a novel mathematical treatment for the kinetic data obtained from chromatographic reactors, which we believe is superior to other treatments in terms of its accuracy, robustness and ease of implementation. In this work, we demonstrate that our treatment may be applied equally well to HPLC reactors, as previously we studied only GC reactors. It is found that the hydrolysis of Fosaprepitant Dimeglumine (FD) has an apparent activation energy of 107 kJ/mol when the reaction is investigated on-column, using the gradient elution conditions of the validated HPLC impurity profile method for this compound. For comparison, the activation energy determined for the same reaction occurring in a quiescent solution consisting of a fixed ratio of acetonitrile-0.1% v/v aqueous H3PO4 (50:50, v/v) is 91 kJ/mol, calculated using direct application of the Arrhenius equation. The data presented show that, when used as a screening tool, chromatographic reactors may be feasible for use in the pharmaceutical industry to quickly gauge the relative stabilities of various compounds with similar degradation pathways. PMID:16524680

  11. Supercharging Reagent for Enhanced Liquid Chromatographic Separation and Charging of Sialylated and High-Molecular-Weight Glycopeptides for NanoHPLC-ESI-MS/MS Analysis.

    PubMed

    Lin, Chia-Wei; Haeuptle, Micha A; Aebi, Markus

    2016-09-01

    Recent developments in proteomic techniques have led to the development of mass spectrometry (MS)-based methods to characterize site-specific glycosylation of proteins. However, appropriate analytical tools to characterize acidic and high-molecular-weight (hMW) glycopeptides are still lacking. In this study, we demonstrate that the addition of supercharging reagent, m-nitrobenzyl alcohol (m-NBA), into mobile phases greatly facilitates the analysis of acidic and hMW glycopeptides. Using commercial glycoproteins, we demonstrated that in the presence of m-NBA the charge state of sialylated glycopeptides increased and the chromatographic separation of neutral and acidic glycopeptides revealed a remarkable improvement. Next, we applied this system to the characterization of a glycoconjugate vaccine candidate consisting of a genetically detoxified exotoxin A of Pseudomonas aeruginosa covalently linked to Shigella flexneri type 2a O-antigen (Sf2E) produced by engineered Escherichia coli. The addition of m-NBA, allowed us to identify peptides with glycan chains of unprecedented size, up to 20 repeat units (98 monosaccharides). Our results indicated that incorporation of m-NBA into reversed-phase liquid chromatography (LC) solvents improves sensitivity, charging, and chromatographic resolution for acidic and hMW glycopeptides. PMID:27487254

  12. Supercharging Reagent for Enhanced Liquid Chromatographic Separation and Charging of Sialylated and High-Molecular-Weight Glycopeptides for NanoHPLC-ESI-MS/MS Analysis.

    PubMed

    Lin, Chia-Wei; Haeuptle, Micha A; Aebi, Markus

    2016-09-01

    Recent developments in proteomic techniques have led to the development of mass spectrometry (MS)-based methods to characterize site-specific glycosylation of proteins. However, appropriate analytical tools to characterize acidic and high-molecular-weight (hMW) glycopeptides are still lacking. In this study, we demonstrate that the addition of supercharging reagent, m-nitrobenzyl alcohol (m-NBA), into mobile phases greatly facilitates the analysis of acidic and hMW glycopeptides. Using commercial glycoproteins, we demonstrated that in the presence of m-NBA the charge state of sialylated glycopeptides increased and the chromatographic separation of neutral and acidic glycopeptides revealed a remarkable improvement. Next, we applied this system to the characterization of a glycoconjugate vaccine candidate consisting of a genetically detoxified exotoxin A of Pseudomonas aeruginosa covalently linked to Shigella flexneri type 2a O-antigen (Sf2E) produced by engineered Escherichia coli. The addition of m-NBA, allowed us to identify peptides with glycan chains of unprecedented size, up to 20 repeat units (98 monosaccharides). Our results indicated that incorporation of m-NBA into reversed-phase liquid chromatography (LC) solvents improves sensitivity, charging, and chromatographic resolution for acidic and hMW glycopeptides.

  13. HPLC determination of naproxen in plasma.

    PubMed

    Tashtoush, B M; Al-Taani, B M

    2003-09-01

    An assay method using isocratic HPLC with fluorometric detection for the determination of naproxen sodium in plasma is presented. A reverse phase Microbondapack column was used with a mobile phase consisting of 42% acetonitrile and 58% water adjusted to pH 3 using phosphoric acid. The fluorometric detector with an excitation wavelength of 270 nm and emission wavelength of 340 nm provided high sensitivity and no interferences from plasma constituents. Plasma samples were injected to HPLC without any extraction. The method was precise and reproducible as was demonstrated by replicate analysis of pooled plasma sample containing 0.5-80 microg/ml naproxen sodium.

  14. HPLC for quality control of polyimides

    NASA Technical Reports Server (NTRS)

    Young, P. R.; Sykes, G. F.

    1979-01-01

    High Pressure Liquid Chromatography (HPLC) as a quality control tool for polyimide resins and prepregs are presented. A data base to help establish accept/reject criteria for these materials was developed. This work is intended to supplement, not replace, standard quality control tests normally conducted on incoming resins and prepregs. To help achieve these objectives, the HPLC separation of LARC-160 polyimide precursor resin was characterized. Room temperature resin aging effects were studied. Graphite reinforced composites made from fresh and aged resin were fabricated and tested to determine if changes observed by HPLC were significant.

  15. Dual high-resolution α-glucosidase and radical scavenging profiling combined with HPLC-HRMS-SPE-NMR for identification of minor and major constituents directly from the crude extract of Pueraria lobata.

    PubMed

    Liu, Bingrui; Kongstad, Kenneth T; Qinglei, Sun; Nyberg, Nils T; Jäger, Anna K; Staerk, Dan

    2015-02-27

    The crude methanol extract of Pueraria lobata was investigated by dual high-resolution α-glucosidase inhibition and radical scavenging profiling combined with hyphenated HPLC-HRMS-SPE-NMR. Direct analysis of the crude extract without preceding purification was facilitated by combining chromatograms from two analytical-scale HPLC separations of 120 and 600 μg on-column, respectively. High-resolution α-glucosidase and radical scavenging profiles were obtained after microfractionation of the eluate in 96-well microplates. This allowed full bioactivity profiling of individual peaks in the HPLC chromatogram of the crude methanol extract. Subsequent HPLC-HRMS-SPE-NMR analysis allowed identification of 21 known compounds in addition to two new compounds, i.e., 3'-methoxydaidzein 8-C-[α-D-apiofuranosyl-(1→6)]-β-D-glucopyranoside and 6″-O-malonyl-3'-methoxydaidzin, as well as an unstable compound tentatively identified as 3'-de-O-methylpuerariafuran. PMID:25679337

  16. Validated HPLC and Ultra-HPLC Methods for Determination of Dronedarone and Amiodarone Application for Counterfeit Drug Analysis.

    PubMed

    El-Bagary, Ramzia I; Elkady, Ehab F; Mowaka, Shereen; Attallah, Maria

    2015-01-01

    Two simple, accurate, and precise chromatographic methods have been developed and validated for the determination of dronedarone (DRO) HCl and amiodarone (AMI) HCl either alone or in binary mixtures due to the possibility of using AMI as a counterfeit of DRO because of its lower price. First, an RP-HPLC method is described for the simultaneous determination of DRO and AMI. Chromatographic separation was achieved on a BDS Hypersil C18 column (150×4.6 mm, 5 μm). Isocratic elution based on potassium dihydrogen phosphate buffer with 0.1% triethylamine pH 6-methanol (10+90, v/v) at a flow rate of 2 mL/min with UV detection at 254 nm was performed. The second method is RP ultra-HPLC in which the chromatographic separation was achieved on an AcclaimTM RSLC 120 C18 column (100×2.1 mm, 2.2 μm) using isocratic elution with potassium dihydrogen phosphate buffer with 0.1% triethylamine pH 6-methanol (5+95, v/v) at a flow rate of 1 mL/min with UV detection at 254 nm. Linearity, accuracy, and precision of the two methods were found to be acceptable over the concentration ranges of 5-80 μg/mL for both DRO and AMI. The results were statistically compared using one-way analysis of variance. The optimized methods were validated and proved to be specific, robust, precise, and accurate for the QC of the drugs in their pharmaceutical preparations. PMID:26651561

  17. Efficient extraction and preparative separation of four main isoflavonoids from Dalbergia odorifera T. Chen leaves by deep eutectic solvents-based negative pressure cavitation extraction followed by macroporous resin column chromatography.

    PubMed

    Li, Lu; Liu, Ju-Zhao; Luo, Meng; Wang, Wei; Huang, Yu-Yan; Efferth, Thomas; Wang, Hui-Mei; Fu, Yu-Jie

    2016-10-15

    In this study, green and efficient deep eutectic solvent-based negative pressure cavitation-assisted extraction (DES-NPCE) followed by macroporous resin column chromatography was developed to extract and separate four main isoflavonoids, i.e. prunetin, tectorigenin, genistein and biochanin A from Dalbergia odorifera T. Chen leaves. The extraction procedure was optimized systematically by single-factor experiments and a Box-Behnken experimental design combined with response surface methodology. The maximum extraction yields of prunetin, tectorigenin, genistein and biochanin A reached 1.204, 1.057, 0.911 and 2.448mg/g dry weight, respectively. Moreover, the direct enrichment and separation of four isoflavonoids in DES extraction solution was successfully achieved by macroporous resin AB-8 with recovery yields of more than 80%. The present study provides a convenient and efficient method for the green extraction and preparative separation of active compounds from plants. PMID:27517524

  18. Efficient extraction and preparative separation of four main isoflavonoids from Dalbergia odorifera T. Chen leaves by deep eutectic solvents-based negative pressure cavitation extraction followed by macroporous resin column chromatography.

    PubMed

    Li, Lu; Liu, Ju-Zhao; Luo, Meng; Wang, Wei; Huang, Yu-Yan; Efferth, Thomas; Wang, Hui-Mei; Fu, Yu-Jie

    2016-10-15

    In this study, green and efficient deep eutectic solvent-based negative pressure cavitation-assisted extraction (DES-NPCE) followed by macroporous resin column chromatography was developed to extract and separate four main isoflavonoids, i.e. prunetin, tectorigenin, genistein and biochanin A from Dalbergia odorifera T. Chen leaves. The extraction procedure was optimized systematically by single-factor experiments and a Box-Behnken experimental design combined with response surface methodology. The maximum extraction yields of prunetin, tectorigenin, genistein and biochanin A reached 1.204, 1.057, 0.911 and 2.448mg/g dry weight, respectively. Moreover, the direct enrichment and separation of four isoflavonoids in DES extraction solution was successfully achieved by macroporous resin AB-8 with recovery yields of more than 80%. The present study provides a convenient and efficient method for the green extraction and preparative separation of active compounds from plants.

  19. Rapid determination of Papaver somniferum alkaloids in process streams using monolithic column high-performance liquid chromatography with chemiluminescence detection.

    PubMed

    Costin, Jason W; Lewis, Simon W; Purcell, Stuart D; Waddell, Lucy R; Francis, Paul S; Barnett, Neil W

    2007-07-30

    We have combined high-performance liquid chromatography (HPLC) separations using a monolithic column with acidic potassium permanganate and tris(2,2'-bipyridyl)ruthenium(II) chemiluminescence detection in a rapid and highly sensitive method to monitor the process of extracting opiate alkaloids from Papaver somniferum. Due to the high flow rates allowed with the monolithic column and the inherent selectivity of the chemiluminescence reactions, the four predominant alkaloids--morphine, codeine, oripavine and thebaine--were determined in less than 2 min. The results obtained with numerous process samples compared favourable with those of the standard HPLC methodology. Limits of detection were 1x10(-10) M, 5x10(-10) M, 5x10(-10) M and 1x10(-9) M, for morphine, codeine, oripavine and thebaine, respectively.

  20. Qualitative and quantitative analysis of cinobufacini injection using rapid separation liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry and HPLC-photodiode array detection, a feasible strategy for the quality control of Chinese medicine injections.

    PubMed

    Zhao, Haiyu; Wu, Xu; Wang, Hongjie; Gao, Bo; Yang, Jian; Si, Nan; Bian, Baolin

    2013-02-01

    Cinobufacini injection, prepared from the skin of Bufo bufo gargarizans Cantor, has presented its significant effects on the treatment of hepatitis and various cancers in the clinic. However, as an unclear complex chemical system, the optimization of its quality control markers has been a long-term challenge. In present study, a feasible strategy integrated markers screening, determination, and statistical analysis was efficiently proposed, especially for the undefined Chinese medicine injections. First, rapid separation LC-quadrupole-TOF-MS method was applied in the identification of 19 major compounds in the cinobufacini injection for the first time. Further, nine high-level contents active compounds were selected as quality control markers for the quantification analysis. An acceptable and validated determination method was established in 17 batches of cinobufacini injection by HPLC-photodiode array detection method, including linear regression relationship (r(2), 0.9996-1), precisions (RSD, 0.02-1.35%), repeatability (RSD, 0.05-1.97%), stability (RSD, 0.1-3.85%), and recovery (95.88-104.89%). Each analyte was detected at its maximum ultraviolet spectra wavelength. Finally, based on the quantification results, principal component analysis was performed on the quality assessment of cinobufacini injections. This three-step strategy provides a newly feasible solution for the quality control of Chinese medicine injections.

  1. Simultaneous Determination of Matrine and Tinidazole in Compound Lotion by RH-HPLC Method.

    PubMed

    Yin, Zhikui; Ma, Suying; Wang, Jincai; Shang, Xiaojun

    2013-01-01

    A simple, sensitive, and accurate RP-HPLC coupled with UV detector method was developed and validated for simultaneous determination of matrine and tinidazole in compound lotion. The chromatographic separation of the two compounds was carried out with a SinoChoom ODS-BP C18 column (5  μ m, 4.6 mm × 200 mm) analytical column, using a mobile phase consisting of 0.025 mol/L potassium dihydrogen phosphate (containing triethylamine 0.05%, v/v) and acetonitrile (80 : 20, v/v) at a flow rate of 1.0 mL/min. The detection was monitored at 210 and 310 nm for matrine and tinidazole, respectively. Total run time was 12 min, and the column was maintained at 25°C. The excipients in the compound lotion did not interfere with the drug peaks. The calibration curves of matrine and tinidazole were fairly linear over the concentration ranges of 10.0-100.0  μ g/mL (r = 0.9954) and 20.0-200.0  μ g/mL (r = 0.9968), respectively. The RSD of both the intraday and interday variations was below 1.5% for matrine and tinidazole. The proposed HPLC method was validated according to International Conference on Harmonisation and proved to be suitable for the simultaneous determination of matrine and tinidazole in compound lotion.

  2. Production of Molybdenum-99 by (n, ) activation and direct separation of Technetium-99m without column generator fabrication: A viable strategy for enhanced availability of technetium-99m

    SciTech Connect

    Knapp Jr, Russ F; Pillai, M R A

    2012-01-01

    Fission-produced 99Mo (F 99Mo) is traditionally used for fabrication of 99Mo/99mTc adsorption-type column generators. In this paper, several emerging strategies that are being pursued or have been suggested to overcome the continuing shortages of F 99Mo are discussed. To provide an alternative source of 99Mo, the principal focus of this analysis is a detailed discussion of the advantages and strategies for enhanced production of low-specific-activity 99Mo (LSA 99Mo) by direct activation of molybdenum targets in nuclear reactors. In order to enhance the availability of 99Mo, development of an increased network of reactors for production of LSA 99Mo is described, as well as utilization of currently unused reactors. The time spent in manufacturing of 99Mo/99mTc column generators is responsible for the loss of more than 50% of F99Mo produced. Hence, the authors propose a paradigm shift in the use of 99Mo by recovering clinical-grade 99mTc from 99Mo solution as an alternative to use of 99Mo/99mTc column generators, thereby avoiding substantial decreased availability of 99Mo from radioactive decay. Implementation of the suggested strategies would be expected to increase availability of 99mTc to the clinical user community by several folds. Additional important advantages of the use of LSA 99Mo include precluding the need for fission product waste management and phasing out the need for high- and low-enriched uranium as target materials for medical radioisotope production.

  3. Anion-exchange high-performance liquid chromatography with post-column detection for the analysis of phytic acid and other inositol phosphates

    NASA Technical Reports Server (NTRS)

    Rounds, M. A.; Nielsen, S. S.; Mitchell, C. A. (Principal Investigator)

    1993-01-01

    The use of gradient anion-exchange HPLC, with a simple post-column detection system, is described for the separation of myo-inositol phosphates, including "phytic acid" (myo-inositol hexaphosphate). Hexa-, penta-, tetra-, tri- and diphosphate members of this homologous series are clearly resolved within 30 min. This method should facilitate analysis and quantitation of "phytic acid" and other inositol phosphates in plant, food, and soil samples.

  4. STEM imaging of 47-pm-separated atomic columns by a spherical aberration-corrected electron microscope with a 300-kV cold field emission gun.

    PubMed

    Sawada, Hidetaka; Tanishiro, Yasumasa; Ohashi, Nobuhiro; Tomita, Takeshi; Hosokawa, Fumio; Kaneyama, Toshikatsu; Kondo, Yukihito; Takayanagi, Kunio

    2009-12-01

    A spherical aberration-corrected electron microscope has been developed recently, which is equipped with a 300-kV cold field emission gun and an objective lens of a small chromatic aberration coefficient. A dumbbell image of 47 pm spacing, corresponding to a pair of atomic columns of germanium aligned along the [114] direction, is resolved in high-angle annular dark field (HAADF) scanning transmission electron microscopy (STEM) with a 0.4-eV energy spread of the electron beam. The observed image was compared with a simulated image obtained by dynamical calculation.

  5. Boric acid as a mobile phase additive for high performance liquid chromatography separation of ribose, arabinose and ribulose.

    PubMed

    De Muynck, Cassandra; Beauprez, Joeri; Soetaert, Wim; Vandamme, Erick J

    2006-01-01

    A new high performance liquid chromatographic (HPLC) method is described for the analysis of ribose, arabinose and ribulose mixtures obtained from (bio)chemical isomerization processes. These processes gain importance since the molecules can be used for the synthesis of antiviral therapeutics. The HPLC method uses boric acid as a mobile phase additive to enhance the separation on an Aminex HPX-87K column. By complexing with boric acid, the carbohydrates become negatively charged, thus elute faster from the column by means of ion exlusion and are separated because the complexation capacity with boric acid differs from one carbohydrate to another. Excellent separation between ribose, ribulose and arabinose was achieved with concentrations between 0.1 and 10 gL(-1) of discrete sugar.

  6. PULSE COLUMN

    DOEpatents

    Grimmett, E.S.

    1964-01-01

    This patent covers a continuous countercurrent liquidsolids contactor column having a number of contactor states each comprising a perforated plate, a layer of balls, and a downcomer tube; a liquid-pulsing piston; and a solids discharger formed of a conical section at the bottom of the column, and a tubular extension on the lowest downcomer terminating in the conical section. Between the conical section and the downcomer extension is formed a small annular opening, through which solids fall coming through the perforated plate of the lowest contactor stage. This annular opening is small enough that the pressure drop thereacross is greater than the pressure drop upward through the lowest contactor stage. (AEC)

  7. Separating methane emissions from biogenic sources and natural gas by vertical column enhancements of ammonia, ethane, and methane in the Colorado Front Range

    NASA Astrophysics Data System (ADS)

    Chiu, R.; Volkamer, R. M.; Blumenstock, T.; Hase, F.; Hannigan, J. W.; Kille, N.; Frey, M.; Kumar Sha, M.; Orphal, J.

    2015-12-01

    Methane sources in the Colorado Front Range include biogenic sources from cattle feedlots and natural gas operations. Although numerous studies have measured methane emissions, there remains significant uncertainty regarding the relative contributions of these various methane emission sources. Here we present data from a March 2015 field campaign that deployed two Bruker EM27 Sun Fourier Transform Spectrometers (FTS) and the University of Colorado Solar Occultation Flux (CU-SOF) FTS in Eaton, Colorado; the former were used to measure enhancements in the methane vertical column densities (VCD), while the latter was used to measure ethane and ammonia VCDs. A third EM27 FTS was deployed to a background site in Westminster, Colorado which was far removed from cattle and petroleum operations. Northerly winds make possible the determination of methane VCD column enhancement from Westminster to Eaton. All instruments were compared during several background days at the National Center for Atmospheric Research (NCAR) in Boulder, Colorado. This presentation explores the potential of methane source attribution using ammonia as a tracer for feedlot emissions and ethane as a tracer for petroleum emissions.

  8. HPLC determination of calcium pantothenate and two preservatives in topical cream.

    PubMed

    Havlíková, L; Matysová, L; Nováková, L; Solich, P

    2006-05-01

    A RP-HPLC method for simultaneous determination of calcium pantothenate and two preservatives methylparaben and propylparaben present in topical cream was developed. Different analytical columns with various stationary phases were tested. During method development, Supelco Discovery C18 column (125 mmx4.0 mm, 5 microm) and Zorbax SB-CN column (150 mmx4.6 mm, 5 microm) were tested. Both were not convenient for analytical separation because of the co-elution of calcium pantothenate with dead volume, and problems with the peak-shape of all components. Good separation was achieved using Zorbax TSM (250 mmx4.6 mm, 5 microm) and Hypersil ODS column (250 mmx4.6 mm, 5 microm), the latter was finally used for the analysis. The analysis time was 12 min, at flow rate 0.7 ml min-1. Chromatography was performed using binary mobile phase composed of methanol and phosphoric acid, pH 2.5, 65:35 (v/v). UV detection was accomplished at 214 nm. The method was validated according to ICH guideline recommendations. The method is suitable for practical routine analysis of commercially produced topical pharmaceutical preparations. PMID:16473491

  9. Separation of N-derivatized di- and tri-peptide stereoisomers by micro-liquid chromatography using a quinidine-based monolithic column - Analysis of l-carnosine in dietary supplements.

    PubMed

    Wang, Qiqin; Sánchez-López, Elena; Han, Hai; Wu, Huihui; Zhu, Peijie; Crommen, Jacques; Marina, Maria Luisa; Jiang, Zhengjin

    2016-01-01

    In the present study, a new analytical methodology was developed enabling the enantiomeric determination of N-derivatized di- and tri-peptides in dietary supplements using chiral micro-LC on a monolithic column consisting of poly(O-9-[2-(methacryloyloxy)-ethylcarbamoyl]-10,11-dihydroquinidine-co-2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) (poly(MQD-co-HEMA-co-EDMA)). After optimization of the mobile phase conditions, a baseline resolution of the stereoisomers of 24 out of 53 N-derivatized di- and tri-peptides was obtained. 3,5-Dinitrobenzoyl- and 3,5-dichlorobenzoyl-peptide stereoisomers were separated with exceptionally high selectivity and resolution. The monolithic column was then applied to the quantitative analysis of l-carnosine and its enantiomeric impurity in three different commercial dietary supplements. Method validation demonstrated satisfactory results in terms of linearity, precision, selectivity, accuracy and limits of detection and quantification. The determined amounts of l-carnosine in commercial formulations were in agreement with the labeled content for all analyzed samples, and the enantiomeric impurity was found to be below the limit of detection (LOD), showing the potential of the poly(MQD-co-HEMA-co-EDMA) monolithic column as a reliable tool for the quality control of l-carnosine in dietary supplements by micro-LC.

  10. Simple HPLC method for detection of trace ephedrine and pseudoephedrine in high-purity methamphetamine.

    PubMed

    Makino, Yukiko

    2012-03-01

    A simple and sensitive HPLC technique was developed for the qualitative determination of ephedrine and pseudoephedrine (ephedrines), used as precursors of clandestine d-methamphetamine hydrochloride of high purity. Good separation of ephedrines from bulk d-methamphetamine was achieved, without any extraction or derivatization procedure on a CAPCELLPACK C18 MGII (250 × 4.6 mm) column. The mobile phase consisted of 50 mM KH2 PO4-acetonitrile (94:6 v/v %) using an isocratic pump system within 20 min for detecting two analytes. One run took about 50 min as it was necessary to wash out overloaded methamphetamine for column conditioning. The analytes were detected by UV absorbance measurement at 210 nm. A sample (20 mg) was simply dissolved in 1 mL of water, and a 50 μL aliquot of the solution was injected into the HPLC. The detection limits for ephedrine and pseudoephedrine in bulk d-methamphetamine were as low as 3 ppm each. This analytical separation technique made it possible to detect ephedrine and/or pseudoephedrine in seven samples of high-purity d-methamphetamine hydrochloride seized in Japan. The presence of trace ephedrines in illicit methamphetamine may strongly indicate a synthetic route via ephedrine in methamphetamine profiling. This method is simple and sensitive, requiring only commonly available equipment, and should be useful for high-purity methamphetamine profiling.

  11. Analyze distillation columns with thermodynamics

    SciTech Connect

    Ognisty, T.P. )

    1995-02-01

    In a distillation column, heat supplies the work for separating the components of a feed stream into products. Distillation columns consume some 95% of the total energy used in separations. This amounts to roughly 3% of the energy consumed in the US. Since distillation is so energy intensive and requires significant capital outlays, an endless quest to improve the economics has continued since the beginning of the industry. By analyzing the thermodynamics of a distillation column, an engineer can quantify the thermodynamic efficiency of the process, identify the regions where energy can be better utilized, and define the minimum targets for energy consumption. This article reviews the principles of distillation column thermodynamics and outlines the analysis of lost work profiles and column heat profiles. It then illustrates these concepts through three examples.

  12. The detection of radical scavenging compounds in crude extract of borage (Borago officinalis L.) by using an on-line HPLC-DPPH method.

    PubMed

    Bandoniene, Donata; Murkovic, Michael

    2002-01-01

    The rapid evaluation of antioxidant activity of crude borage (Borago officinalis L.) extract was determined by using DPPH free radical method. This borage extract resulted in a rapid decrease of the absorbance and showed very high hydrogen-donating capacity towards the 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical. A new HPLC-DPPH on-line method was applied for a screening of several radical scavenging components in this borage extract as well as for quantitative analysis. This on-line HPLC-DPPH method was developed using a methanolic solution of DPPH-stable radical. The HPLC-separated analytes reacted post-column with the DPPH solution in methanol. The induced bleaching was detected as a negative peak photometrically at 515 nm. The separation of antioxidative components was carried out by gradient HPLC with mobile-phase composition ranging from 2% to 80% acetonitrile with 2% acetic acid in water, UV detection was carried out at 280 nm. The HPLC analysis of borage extract revealed the presence of several radical scavenging components in the borage extract. The results obtained from the chromatograms suggest that some compounds present in the extract possess high radical quenching ability. The dominant antioxidative compound in the crude extract of borage leaves was identified as rosmarinic acid. PMID:12406585

  13. The detection of radical scavenging compounds in crude extract of borage (Borago officinalis L.) by using an on-line HPLC-DPPH method.

    PubMed

    Bandoniene, Donata; Murkovic, Michael

    2002-01-01

    The rapid evaluation of antioxidant activity of crude borage (Borago officinalis L.) extract was determined by using DPPH free radical method. This borage extract resulted in a rapid decrease of the absorbance and showed very high hydrogen-donating capacity towards the 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical. A new HPLC-DPPH on-line method was applied for a screening of several radical scavenging components in this borage extract as well as for quantitative analysis. This on-line HPLC-DPPH method was developed using a methanolic solution of DPPH-stable radical. The HPLC-separated analytes reacted post-column with the DPPH solution in methanol. The induced bleaching was detected as a negative peak photometrically at 515 nm. The separation of antioxidative components was carried out by gradient HPLC with mobile-phase composition ranging from 2% to 80% acetonitrile with 2% acetic acid in water, UV detection was carried out at 280 nm. The HPLC analysis of borage extract revealed the presence of several radical scavenging components in the borage extract. The results obtained from the chromatograms suggest that some compounds present in the extract possess high radical quenching ability. The dominant antioxidative compound in the crude extract of borage leaves was identified as rosmarinic acid.

  14. WATER COLUMN DATA AND SPECTRAL IRRADIANCE MODEL

    EPA Science Inventory

    Water samples collected monthly, for 18 months, from six sites in the Laguna Madre were analyzed to identify and quantify phytopigments using High Performance Liquid Chromatography (HPLC). In addition, water column pigment and nutrient data were acquired at 12 stations in Upper ...

  15. Column chromatography as a useful step in purification of diatom pigments.

    PubMed

    Tokarek, Wiktor; Listwan, Stanisław; Pagacz, Joanna; Leśniak, Piotr; Latowski, Dariusz

    2016-01-01

    Fucoxanthin, diadinoxanthin and diatoxanthin are carotenoids found in brown algae and most other heterokonts. These pigments are involved in photosynthetic and photoprotective reactions, and they have many potential health benefits. They can be extracted from diatom Phaeodactylum tricornutum by sonication, extraction with chloroform : methanol and preparative thin layer chromatography. We assessed the utility of an additional column chromatography step in purification of these pigments. This novel addition to the isolation protocol increased the purity of fucoxanthin and allowed for concentration of diadinoxanthin and diatoxanthin before HPLC separation. The enhanced protocol is useful for obtaining high purity pigments for biochemical studies. PMID:27486920

  16. Normal-Phase Open Column versus Reversed-Phase High Performance Liquid Chromatography: Separation of Chlorophyll a and Chlorophyll b from their Diastereomers.

    ERIC Educational Resources Information Center

    Schaber, Peter M.

    1985-01-01

    Background information, procedures used, and typical results obtained are provided for an experiment involving the separation of chlorophyll a and chlorophyll b from their diastereomers. Reasons why the experiment can be easily integrated into most laboratory curricula where high-performance liquid chromatography capabilities exist are given. (JN)

  17. [Chromatographic separation of aminoglutethimide enantiomers on cellulose tris(3,5-dimethylphenylcarbamate) chiral stationary phase].

    PubMed

    Lin, Xiaoiian; Gong, Rujin; Li, Ping; Yu, Jianguo

    2014-08-01

    Aminoglutethimide (AG) has been used clinically as a drug in the treatment of hormone-dependent metastatic breast cancer. It was reported that S-(-)-AG enantiomer had small activity and sometimes might cause side effects. Therefore, it was of great significance to obtain the high-purity R-(+)-AG by enantioseparation. In this work, aminoglutethimide enantiomers were separated by high performance liquid chromatography (HPLC) using an analytical column which was packed with cellulose tris(3,5-dimethylphenylcarbamate) stationary phase (Chiralcel OD-H). The solubilities of racemic AG in two different solvent compositions, n-hexane/ethanol and n-hexane/isopropanol, were measured, separately. The effects of alcohol content and monoethanolamine additive on the separation performance of racemic AG by HPLC were investigated. According to the experiments, n-hexane-ethanol (30:70, v/v) with 0.1% monoethanolamine additive was selected as the mobile phase. The separation factor, resolution, asymmetry factor, number of theoretical plates and maximum column capacity were measured and analyzed for the chromatographic separation of racemic AG at a flow-rate of 0. 6 mL/min and column temperature of 25-40 °C, with Chiralcel OD-H as stationary phase and n-hexane-ethanol (30:70, v/v) with 0. 1% monoethanolamine as mobile phase. This work provides the basic information of chromatographic separation for the batch and continuous production of aminoglutethimide enantiomers.

  18. Column internals

    SciTech Connect

    Bravo, J.L.

    1998-02-01

    In the fields of distillation, absorption, stripping and extraction, theory and technology go hand in hand. The thermodynamic principles of phase equilibrium and the concepts of mass transfer and fluid flow are of primary importance in all of these operations. The engineer must understand these phenomena to select equipment effectively. This article discusses the latest in commercial technology in column internals for gas-liquid and liquid-liquid contacting. The principles of operation are explained vis-a-vis the characteristics of the applications in which they are used. The focus is on moderate-to-large columns for refining and chemical applications. Guidelines for selecting the most appropriate type of device are presented, and examples of typical applications are described.

  19. Separation and purification of α-glucosidase inhibitors from Polygonatum odoratum by stepwise high-speed counter-current chromatography combined with Sephadex LH-20 chromatography target-guided by ultrafiltration-HPLC screening.

    PubMed

    Zhou, Xiaoling; Liang, Junsheng; Zhang, Yi; Zhao, Huading; Guo, Ying; Shi, Shuyun

    2015-03-15

    Although Polygonatum odoratum has been widely used as medicinal plant and food supplement for treating diabetes, little is known regarding its bioactive components. In this study, ultrafiltration-HPLC based ligand screening was developed to screen α-glucosidase inhibitors from P. odoratum for the first time. Then bioactive components were target-guided separated by combining stepwise high-speed counter-current chromatography (HSCCC) using petroleum ether-ethyl acetate-methanol-water (1:4:0.8:4.2, v/v/v/v), (1:4:1.8:3.2, v/v/v/v) and (1:4:2.3:2.7, v/v/v/v) as solvent systems with Sephadex LH-20 chromatography eluted by MeCN-MeOH (1:1, v/v). Five phenethyl cinnamides, N-cis-feruloyloctopamine (1); N-trans-p-coumaroyloctopamine (2), N-trans-feruloyloctopamine (3), N-trans-p-coumaroyltyramine (4) and N-trans-feruloyltyramine (5), and four homoisoflavanones, (3R)-5,7-dihydroxyl-3-(2',4'-dihydroxylbenzyl)-chroman-4-one (6), (3R)-5,7-dihydroxyl-6-methyl-3-(4'-hydroxylbenzyl)-chroman-4-one (7), (3R)-5,7-dihydroxyl-6-methyl-8-methoxyl-3-(4'-hydroxylbenzyl)-chroman-4-one (8); and (3R)-5,7-dihydroxyl-6,8-dimethyl-3-(4'-hydroxylbenzyl)-chroman-4-one) (9), with purity over 98.5% were purified, and their structures were identified by UV, MS, and (1)H NMR. Notably, compounds 2 and 4 were first reported in genus Polygonatum, while compound 1 was first obtained from family Liliaceae. In addition, α-glucosidase inhibitory activities of compounds 1-9 were evaluated, and compounds 2 and 4 exhibited stronger α-glucosidase inhibitory activity with IC50 values of 2.3 and 2.7μM. The results suggested the potential medicinal use of P. odoratum, and the technology could be widely applied for rapid screening and preparative separation of a group of bioactive compounds from complex matrix.

  20. Blind column selection protocol for two-dimensional high performance liquid chromatography.

    PubMed

    Burns, Niki K; Andrighetto, Luke M; Conlan, Xavier A; Purcell, Stuart D; Barnett, Neil W; Denning, Jacquie; Francis, Paul S; Stevenson, Paul G

    2016-07-01

    The selection of two orthogonal columns for two-dimensional high performance liquid chromatography (LC×LC) separation of natural product extracts can be a labour intensive and time consuming process and in many cases is an entirely trial-and-error approach. This paper introduces a blind optimisation method for column selection of a black box of constituent components. A data processing pipeline, created in the open source application OpenMS®, was developed to map the components within the mixture of equal mass across a library of HPLC columns; LC×LC separation space utilisation was compared by measuring the fractional surface coverage, fcoverage. It was found that for a test mixture from an opium poppy (Papaver somniferum) extract, the combination of diphenyl and C18 stationary phases provided a predicted fcoverage of 0.48 and was matched with an actual usage of 0.43. OpenMS®, in conjunction with algorithms designed in house, have allowed for a significantly quicker selection of two orthogonal columns, which have been optimised for a LC×LC separation of crude extractions of plant material. PMID:27154652

  1. High-performance liquid chromatographic separation and indirect fluorescence detection of thiols.

    PubMed

    Pelletier, Sarah; Lucy, Charles A

    2002-10-01

    A fluorescent post-column reaction detection scheme has been devised for selective determination of thiols. The post-column reagent is 40 microM Cd2+ and 100 microM 8-hydroxyquinoline-5-sulfonic acid (HQS) in non-complexing buffer at pH 10. HQS complexes Cd2+ to form a fluorescent product. Thiols in the HPLC effluent compete for complexation of the Cd2+, resulting in a decrease in the fluorescence response. Detection limits of 0.2 microM (0.04 ppm) are achieved for cysteine, homocysteine and glutathione in a 5 min separation. Recoveries from spiked synthetic urine samples are 87-120%.

  2. Effect of basic and acidic additives on the separation of some basic drug enantiomers on polysaccharide-based chiral columns with acetonitrile as mobile phase.

    PubMed

    Gogaladze, Khatuna; Chankvetadze, Lali; Tsintsadze, Maia; Farkas, Tivadar; Chankvetadze, Bezhan

    2015-03-01

    The separation of enantiomers of 16 basic drugs was studied using polysaccharide-based chiral selectors and acetonitrile as mobile phase with emphasis on the role of basic and acidic additives on the separation and elution order of enantiomers. Out of the studied chiral selectors, amylose phenylcarbamate-based ones more often showed a chiral recognition ability compared to cellulose phenylcarbamate derivatives. An interesting effect was observed with formic acid as additive on enantiomer resolution and enantiomer elution order for some basic drugs. Thus, for instance, the enantioseparation of several β-blockers (atenolol, sotalol, toliprolol) improved not only by the addition of a more conventional basic additive to the mobile phase, but also by the addition of an acidic additive. Moreover, an opposite elution order of enantiomers was observed depending on the nature of the additive (basic or acidic) in the mobile phase.

  3. RACORO continental boundary layer cloud investigations. 3. Separation of parameterization biases in single-column model CAM5 simulations of shallow cumulus

    SciTech Connect

    Lin, Wuyin; Liu, Yangang; Vogelmann, Andrew M.; Fridlind, Ann; Endo, Satoshi; Song, Hua; Feng, Sha; Toto, Tami; Li, Zhijin; Zhang, Minghua

    2015-06-19

    Climatically important low-level clouds are commonly misrepresented in climate models. The FAst-physics System TEstbed and Research (FASTER) project has constructed case studies from the Atmospheric Radiation Measurement (ARM) Climate Research Facility's Southern Great Plain site during the RACORO aircraft campaign to facilitate research on model representation of boundary-layer clouds. This paper focuses on using the single-column Community Atmosphere Model version 5 (SCAM5) simulations of a multi-day continental shallow cumulus case to identify specific parameterization causes of low-cloud biases. Consistent model biases among the simulations driven by a set of alternative forcings suggest that uncertainty in the forcing plays only a relatively minor role. In-depth analysis reveals that the model's shallow cumulus convection scheme tends to significantly under-produce clouds during the times when shallow cumuli exist in the observations, while the deep convective and stratiform cloud schemes significantly over-produce low-level clouds throughout the day. The links between model biases and the underlying assumptions of the shallow cumulus scheme are further diagnosed with the aid of large-eddy simulations and aircraft measurements, and by suppressing the triggering of the deep convection scheme. It is found that the weak boundary layer turbulence simulated is directly responsible for the weak cumulus activity and the simulated boundary layer stratiform clouds. Increased vertical and temporal resolutions are shown to lead to stronger boundary layer turbulence and reduction of low-cloud biases.

  4. RACORO continental boundary layer cloud investigations. 3. Separation of parameterization biases in single-column model CAM5 simulations of shallow cumulus

    DOE PAGES

    Lin, Wuyin; Liu, Yangang; Vogelmann, Andrew M.; Fridlind, Ann; Endo, Satoshi; Song, Hua; Feng, Sha; Toto, Tami; Li, Zhijin; Zhang, Minghua

    2015-06-19

    Climatically important low-level clouds are commonly misrepresented in climate models. The FAst-physics System TEstbed and Research (FASTER) project has constructed case studies from the Atmospheric Radiation Measurement (ARM) Climate Research Facility's Southern Great Plain site during the RACORO aircraft campaign to facilitate research on model representation of boundary-layer clouds. This paper focuses on using the single-column Community Atmosphere Model version 5 (SCAM5) simulations of a multi-day continental shallow cumulus case to identify specific parameterization causes of low-cloud biases. Consistent model biases among the simulations driven by a set of alternative forcings suggest that uncertainty in the forcing plays only amore » relatively minor role. In-depth analysis reveals that the model's shallow cumulus convection scheme tends to significantly under-produce clouds during the times when shallow cumuli exist in the observations, while the deep convective and stratiform cloud schemes significantly over-produce low-level clouds throughout the day. The links between model biases and the underlying assumptions of the shallow cumulus scheme are further diagnosed with the aid of large-eddy simulations and aircraft measurements, and by suppressing the triggering of the deep convection scheme. It is found that the weak boundary layer turbulence simulated is directly responsible for the weak cumulus activity and the simulated boundary layer stratiform clouds. Increased vertical and temporal resolutions are shown to lead to stronger boundary layer turbulence and reduction of low-cloud biases.« less

  5. RACORO Continental Boundary Layer Cloud Investigations: 3. Separation of Parameterization Biases in Single-Column Model CAM5 Simulations of Shallow Cumulus

    NASA Technical Reports Server (NTRS)

    Lin, Wuyin; Liu, Yangang; Vogelmann, Andrew M.; Fridlind, Ann; Endo, Satoshi; Song, Hua; Feng, Sha; Toto, Tami; Li, Zhijin; Zhang, Minghua

    2015-01-01

    Climatically important low-level clouds are commonly misrepresented in climate models. The FAst-physics System TEstbed and Research (FASTER) Project has constructed case studies from the Atmospheric Radiation Measurement Climate Research Facility's Southern Great Plain site during the RACORO aircraft campaign to facilitate research on model representation of boundary-layer clouds. This paper focuses on using the single-column Community Atmosphere Model version 5 (SCAM5) simulations of a multi-day continental shallow cumulus case to identify specific parameterization causes of low-cloud biases. Consistent model biases among the simulations driven by a set of alternative forcings suggest that uncertainty in the forcing plays only a relatively minor role. In-depth analysis reveals that the model's shallow cumulus convection scheme tends to significantly under-produce clouds during the times when shallow cumuli exist in the observations, while the deep convective and stratiform cloud schemes significantly over-produce low-level clouds throughout the day. The links between model biases and the underlying assumptions of the shallow cumulus scheme are further diagnosed with the aid of large-eddy simulations and aircraft measurements, and by suppressing the triggering of the deep convection scheme. It is found that the weak boundary layer turbulence simulated is directly responsible for the weak cumulus activity and the simulated boundary layer stratiform clouds. Increased vertical and temporal resolutions are shown to lead to stronger boundary layer turbulence and reduction of low-cloud biases.

  6. Determination of CMPO using HPLC -UV

    SciTech Connect

    Gracy Elias; Gary S. Groenewold; Bruce J. Mincher; Stephen P. Mezyk

    2012-06-01

    Octyl(phenyl)-N,N-diisobutylcarbamoylmethylphosphine oxide (CMPO) is an extractant proposed for selective separation of radionuclide metals from used nuclear fuel solutions using solvent extraction. Radiolysis reactions can degrade CMPO and reduce separation performance and hence methods for measuring concentration of CMPO and identifying degradation products are needed. A novel high performance liquid chromatography (HPLC) method employing ultraviolet detection (UV) was developed to detect and quantitate CMPO in dodecane. Some radiolysis products in gamma and alpha irradiated CMPO solutions were identified using HPLC/electrospray ionization-mass spectrometry (ESI-MS). Validation data indicated that the HPLC-UV method for CMPO determination provided good linearity, sensitivity, procedure accuracy and system precision. CMPO-nitric acid complexes were also identified, that account for the apparent loss of CMPO in acidic environment, independent of irradiation.

  7. Quantitative analysis combined with chromatographic fingerprint for comprehensive evaluation of Xiaoer Chaigui Tuire granules by HPLC-DAD.

    PubMed

    Liu, Hong-Ming; Nie, Lei

    2015-01-01

    Quantitative analysis of eight major components combined with chromatographic fingerprint based on high performance liquid chromatography coupled with diode array detector (HPLC-DAD) was developed for the quality evaluation of Xiaoer Chaigui Tuire granules (XCTG), a traditional Chinese medicine (TCM) preparation. Each compound was analyzed by comparing its retention time and UV spectrum of each chromatographic peak with the corresponding retention time and UV spectrum of each standard compound. Baseline separation was achieved on an Agilent Zorbax SB-C18 column with gradient elution of acetonitrile and 0.1% (v/v) phosphoric acid. The developed method was validated by linearity, precision, repeatability, stability and recovery and was subsequently applied to quality evaluation of 12 batches of XCTG with similarity analysis, principal component analysis and cluster analysis. Quantitative analysis combined with HPLC fingerprint could offer an efficient, reliable and practical approach for quality evaluation of XCTG.

  8. Characterization of Jamaican agro-industrial wastes. Part II, fatty acid profiling using HPLC: precolumn derivatization with phenacyl bromide.

    PubMed

    Bailey-Shaw, Y A; Golden, K D; Pearson, A G M; Porter, R B R

    2012-09-01

    This paper describes the determination of fatty acid composition of coffee, citrus and rum distillery wastes using reversed-phase high-performance liquid chromatography (RP-HPLC). Lipid extracts of the waste samples are derivatized with phenacyl bromide and their phenacyl esters are separated on a C8 reversed-phase column by using continuous gradient elution with water and acetonitrile. The presence of saturated and unsaturated fatty acids in quantifiable amounts in the examined wastes, as well as the high percentage recoveries, are clear indications that these wastes have potential value as inexpensive sources of lipids. The HPLC procedures described here could be adopted for further analysis of materials of this nature. PMID:22595260

  9. New "hyphenated" CPC-HPLC-DAD-MS strategy for simultaneous isolation, analysis and identification of phytochemicals: application to xanthones from Garcinia mangostana.

    PubMed

    Michel, Thomas; Destandau, Emilie; Fougère, Laëtitia; Elfakir, Claire

    2012-12-01

    Centrifugal partition chromatography (CPC) coupled online with high-performance liquid chromatography (HPLC) with diode-array detection (DAD) and mass spectrometry (MS) is presented in this work. This strategy offers the possibility to obtain simultaneously CPC fractionation of natural extracts, the HPLC fingerprint of separated fractions and structural information on molecules contained in each fraction. This new approach was applied to the fractionation and purification of xanthones from Garcinia mangostana (Clusiaceae) pericarp. A biphasic solvent system of heptane/ethyl acetate/methanol/water (2:1:2:1, v/v) was used for the CPC separation of 175 mg crude ethanolic extract. The HPLC analysis was conducted with a reversed-phase monolithic column allowing fast and repeatable separation. This combined CPC-HPLC-DAD-MS method led to isolation of 33 mg α-mangostin and 6 mg γ-mangostin at 98% and 98.5% purity, respectively, in 140 min. Furthermore, in the same time a total of 16 other xanthones were detected in the extract, and ten of them were identified on the basis of their UV and MS spectra.

  10. Solvent viscosity mismatch between the solute plug and the mobile phase: Considerations in the applications of two-dimensional HPLC

    SciTech Connect

    Shalliker, R. Andrew; Guiochon, Georges A

    2010-01-01

    Understanding the nature of viscosity contrast induced flow instabilities is an important aspect in the design of two-dimensional HPLC separations. When the viscosity contrast between the sample plug and the mobile phase is sufficiently large, the phenomenon known as viscous fingering can be induced. Viscous fingering is a flow instability phenomenon that occurs at the interface between two fluids with different viscosities. In liquid chromatography, viscous fingering results in the solute band undergoing a change in form as it enters into the chromatography column. Moreover, even in the absence of viscous fingering, band shapes change shape at low viscosity contrasts. These changes can result in a noticeable change in separation performance, with the result depending on whether the solvent pushing the solute plug has a higher or lower viscosity than the solute plug. These viscosity induced changes become more important as the solute injection volume increases and hence understanding the process becomes critical in the implementation of multidimensional HPLC techniques, since in these techniques the sample injection plug into the second dimension is an order of magnitude greater than in one-dimensional HPLC. This review article assesses the current understanding of the viscosity contrast induced processes as they relate to liquid chromatographic separation behaviour.

  11. Development and Validation of HPLC and HPTLC Methods for Estimation of Glabridin in Extracts of Glycyrrhiza glabra.

    PubMed

    Viswanathan, Vivek; Mukne, Alka P

    2016-01-01

    Glabridin is a major bioactive phytoconstituent of licorice. This work discusses the development and validation of HPLC and HPTLC methods for analysis of glabridin in licorice. The HPLC separation was performed using a Purospher STAR RP-18e column (5 μm silica particle size, 250 mm × 4.6 mm inner diameter) with gradient elution of 0.2% acetic acid in water-acetonitrile. The flow rate was 1 mL/min. Quantification was performed at a detection wavelength of 280 nm. HTPLC separation was performed on precoated silica gel 60 F254 aluminum plate (10 × 10 cm, 250 μm thickness). A linear ascending development was done using a mobile phase of hexane-ethyl acetate-chloroform (5 + 4 + 3, v/v/v). After development, the plates were scanned at 285 nm. Both of the methods provided good separation of glabridin from other constituents of licorice extract. The methods were validated as per ICH guidelines. Comparison by Student t-test showed that there was a statistically insignificant difference between the mean glabridin content estimated by both methods at 95% confidence interval. The glabridin content in licorice extract was 3.90% by HPLC and 3.79% by HPTLC. PMID:27103104

  12. Quality by design (QbD) based development of a stability indicating HPLC method for drug and impurities.

    PubMed

    Karmarkar, S; Garber, R; Genchanok, Y; George, S; Yang, X; Hammond, R

    2011-01-01

    In this paper, an application of Quality by Design (QbD) concepts to the development of a stability indicating HPLC method for a complex pain management drug product containing drug substance, two preservatives, and their degradants is described. The QbD approach consisted of (i) developing a full understanding of the intended purpose, (ii) developing predictive solutions, (iii) designing a meaningful system suitability solution that helps to identify failure modes, and (iv) following design of experiments (DOE) approach. The starting method lacked any resolution among drug degradant and preservative oxidative degradant peaks, and peaks for preservative and another drug degradant. The method optimization was accomplished using Fusion AE™ software (S-Matrix Corporation, Eureka, CA) that follows a DOE approach. Column temperature (50 ± 5°C), mobile phase buffer pH (2.9 ± 0.2), initial % acetonitrile (ACN, 2 ± 1%), and initial hold time (2.5, 5, or 10 min) of the HPLC method were simultaneously studied to optimize separation of the unresolved peaks. The optimized HPLC conditions (column temperature of 50°C, buffer pH of 3.1, 3% initial ACN with 2.5 min initial hold) resulted in fully resolved peaks in the two critical pairs. The QbD based method development helped in generating a design space and operating space with knowledge of all method performance characteristics and limitations and successful method robustness within the operating space. PMID:21682993

  13. Temperature programmable microfabricated gas chromatography column

    DOEpatents

    Manginell, Ronald P.; Frye-Mason, Gregory C.

    2003-12-23

    A temperature programmable microfabricated gas chromatography column enables more efficient chemical separation of chemical analytes in a gas mixture by the integration of a resistive heating element and temperature sensing on the microfabricated column. Additionally, means are provided to thermally isolate the heated column from their surroundings. The small heat capacity and thermal isolation of the microfabricated column improves the thermal time response and power consumption, both important factors for portable microanalytical systems.

  14. Effect of physicochemical parameters on the retention of some monoamine oxidase inhibitory drugs on a porous graphitized carbon column.

    PubMed

    Forgács, E; Cserháti, T

    1996-05-31

    The retention of sixteen monoamine oxidase inhibitory drugs (proparlgylamine derivatives) was determined on a porous graphitized carbon (PGC) column using ethanol-water mixtures as eluents. The HPLC retention characteristics of drugs were correlated with their physicochemical properties using stepwise regression analysis and principal component (PC) analysis. The dimensions of the matrices for PC loadings and variables was reduced using a non-linear mapping technique, varimax rotation and cluster analysis. It has been established that the drugs can be well separated on the PGC column in ethanol-water eluents. Calculations proved that the retention behavior of monoamine oxidase drugs on PGC column is of mixed character: both steric and electronic parameters influence the retention.

  15. An Inexpensive Digital Gradient Controller for HPLC.

    ERIC Educational Resources Information Center

    Brady, James E.; Carr, Peter W.

    1983-01-01

    Use of gradient elution techniques in high performance liquid chromatography (HPLC) is often essential for direct separation of complex mixtures. Since most commercial controllers have features that are of marginal value for instructional purposes, a low-cost controller capable of illustrating essential features of gradient elution was developed.…

  16. Determination of the Enantiomerization Barrier of the Residual Enantiomers of C3 -Symmetric Tris[3-(1-Methyl-2-Alkyl)Indolyl]Phosphane Oxides: Case Study of a Multitasking HPLC Investigation Based on an Immobilized Polysaccharide Stationary Phase.

    PubMed

    Rizzo, Simona; Menta, Sergio; Benincori, Tiziana; Ferretti, Rosella; Pierini, Marco; Cirilli, Roberto; Sannicolò, Francesco

    2015-12-01

    The residual enantiomers of three tris-(3-indolyl)-phosphane oxides bearing different alkyl groups (methyl, ethyl or i-propyl) in position 2 of the indole rings constituting the blades were separated on the immobilized type Chiralpak IC column in polar organic and reversed-phase modes. The good enantioselectivity and versatility of the IC CSP allowed easy isolation of the enantiomerically highly enriched samples suitable for configurational stability studies. The enantiomerization barriers of residual phosphane oxides were evaluated both by off-column techniques (CD signal and enantiomeric purity decay kinetics) and by dynamic enantioselective high-performance liquid chromatography (HPLC). PMID:26402152

  17. Determination of the Enantiomerization Barrier of the Residual Enantiomers of C3 -Symmetric Tris[3-(1-Methyl-2-Alkyl)Indolyl]Phosphane Oxides: Case Study of a Multitasking HPLC Investigation Based on an Immobilized Polysaccharide Stationary Phase.

    PubMed

    Rizzo, Simona; Menta, Sergio; Benincori, Tiziana; Ferretti, Rosella; Pierini, Marco; Cirilli, Roberto; Sannicolò, Francesco

    2015-12-01

    The residual enantiomers of three tris-(3-indolyl)-phosphane oxides bearing different alkyl groups (methyl, ethyl or i-propyl) in position 2 of the indole rings constituting the blades were separated on the immobilized type Chiralpak IC column in polar organic and reversed-phase modes. The good enantioselectivity and versatility of the IC CSP allowed easy isolation of the enantiomerically highly enriched samples suitable for configurational stability studies. The enantiomerization barriers of residual phosphane oxides were evaluated both by off-column techniques (CD signal and enantiomeric purity decay kinetics) and by dynamic enantioselective high-performance liquid chromatography (HPLC).

  18. Extraction and identification of flavonoids from parsley extracts by HPLC analysis

    NASA Astrophysics Data System (ADS)

    Stan, M.; Soran, M. L.; Varodi, C.; Lung, I.

    2012-02-01

    Flavonoids are phenolic compounds isolated from a wide variety of plants, and are valuable for their multiple properties, including antioxidant and antimicrobial activities. In the present work, parsley (Petroselinum crispum L.) extracts were obtained by three different extraction techniques: maceration, ultrasonic-assisted and microwave-assisted solvent extractions. The extractions were performed with ethanol-water mixtures in various ratios. From these extracts, flavonoids like the flavones apigenin and luteolin, and the flavonols quercetin and kaempferol were identified using an HPLC Shimadzu apparatus equipped with PDA and MS detectors. The separation method involved a gradient step. The mobile phase consisted of two solvents: acetonitrile and distilled water with 0.1% formic acid. The separation was performed on a RP-C18 column.

  19. Determination of 2-propanol in surface cleaning solutions used for copper continuous casting process by flow injection-spectrophotometric detection with on-line column separation.

    PubMed

    Hayashibe, Yutaka; Tokuda, Masahiro; Takeya, Minoru

    2003-09-01

    A flow-injection system has been developed for the determination of 2-propanol in the surface cleaning solutions used in the copper continuous cast rod making system. Adsorption chromatography in nitric acid medium was used for the on-line separation of oily substances in the sample solution. Cerium(IV) diammonium nitrate was utilized as the chromogenic reagent for the spectrophotometric detection of 2-propanol. The system permits a throughput of one sample per hour for the oily sample, and of 12 samples per hour for the none-oily sample. The reproducibility has been proven to be satisfactory with a relative standard deviation of less than 6.0% (2.2%(V/V) 2-propanol level, n = 23). The detection limit is 0.01% (V/V).

  20. [Establishment and application of HPLC-QAMS for quality evaluation of Chuanxiong Rhizoma].

    PubMed

    Qiao, Feng-xian; Cai, Hao; Tu, Peng-fei; Pei, Ke; Song, Xiao-qing

    2015-06-01

    A quantitative analysis method of multi-components with a single marker (QAMS) for simultaneous determination of six marker compounds (one from phenolic acids and five from phthalides) in Chuanxiong Rhizoma was established by applying HPLC and using butylidenephthalide as the internal reference substance. And also the feasibility and accuracy of the established method for quality evaluation and application of Chuanxiong Rhizoma were investigated and validated. The analysis was performed with the mobile phase consisting of acetonitrile - 0.2% aqueous formic acid. The flow rate was 1.0 mL . min-1 and the column temperature was maintained at 30 °C. The detection wavelengths were set at 252 nm (for ferulic acid, Z-ligustilide, and butylidenephthalide) and 266 nm (for senkyunolide I, senkyunolide A, and coniferyl ferulate), separately, and 20 µL was injected for analysis with gradient elution. The results showed that there were no significant differences observed between the HPLC-QAMS method and the external standard method (RSD <5%). The relative correction factors were credible (RSD < 5%) in changed chromatographic conditions. The established HPLC-QAMS method can be accurately used for simultaneously evaluating and controlling the quality of Chuanxiong Rhizoma with multi-components. PMID:26521448

  1. Quantitative analysis of eugenol in clove extract by a validated HPLC method.

    PubMed

    Yun, So-Mi; Lee, Myoung-Heon; Lee, Kwang-Jick; Ku, Hyun-Ok; Son, Seong-Wan; Joo, Yi-Seok

    2010-01-01

    Clove (Eugenia caryophyllata) is a well-known medicinal plant used for diarrhea, digestive disorders, or in antiseptics in Korea. Eugenol is the main active ingredient of clove and has been chosen as a marker compound for the chemical evaluation or QC of clove. This paper reports the development and validation of an HPLC-diode array detection (DAD) method for the determination of eugenol in clove. HPLC separation was accomplished on an XTerra RP18 column (250 x 4.6 mm id, 5 microm) with an isocratic mobile phase of 60% methanol and DAD at 280 nm. Calibration graphs were linear with very good correlation coefficients (r2 > 0.9999) from 12.5 to 1000 ng/mL. The LOD was 0.81 and the LOQ was 2.47 ng/mL. The method showed good intraday precision (%RSD 0.08-0.27%) and interday precision (%RSD 0.32-1.19%). The method was applied to the analysis of eugenol from clove cultivated in various countries (Indonesia, Singapore, and China). Quantitative analysis of the 15 clove samples showed that the content of eugenol varied significantly, ranging from 163 to 1049 ppb. The method of determination of eugenol by HPLC is accurate to evaluate the quality and safety assurance of clove, based on the results of this study.

  2. Validation of a HPLC method for simultaneous determination of five sunscreens in lotion preparation.

    PubMed

    Kedor-Hackmann, E R M; De Lourdes Pérez González, M L; Singh, A K; Santoro, M I R M

    2006-06-01

    The aim of this research was to develop and validate a high-performance liquid chromatographic (HPLC) method for simultaneous determination of five sunscreens, namely benzophenone-3 (B-3), butyl methoxydibenzoylmethane (BM), octyl methoxycinnamate (OM), octyl salicylate (OS) and homosalate (HS). The separation and quantitative determination was made by HPLC at 40 +/-1 degrees C with a gradient elution from 10% to 100% mobile phase B in mobile phase A. The gradient liquid chromatographic system constituted of mobile phase A [acetonitrile : water (10 : 90 v/v)] and mobile phase B [acetonitrile : water (90 : 10 v/v)], at a flow rate of 1.0 mL min(-1) and ultraviolet detection at 310 nm. The separation was obtained with two Waters reversed phase columns: Novapack C-18 and Symmetry((R)) C-18 connected in series. All sunscreens were efficiently separated within 17 min. The coefficient of correlation and average recovery for B-3, BM, OM, OS and HS were 0.9798 and 98.5%, 0.9672 and 98.8%, 0.9922 and 99.1%, 0.9961 and 98.9% and 0.9909 and 99.4% respectively. The relative standard deviations obtained were between 1.07% and 2.44%. The excipients did not interfere in the analysis. The results showed that the proposed method could be used for rapid and simultaneous determination of B-3, BM, OM, OS and HS in sunscreen lotions with precision, accuracy and specificity.

  3. Quantitative HPLC Analysis of an Analgesic/Caffeine Formulation: Determination of Caffeine

    NASA Astrophysics Data System (ADS)

    Ferguson, Glenda K.

    1998-04-01

    A modern high performance liquid chromatography (HPLC) laboratory experiment which entails the separation of acetaminophen, aspirin, and caffeine and the quantitative assay of caffeine in commercial mixtures of these compounds has been developed. Our HPLC protocol resolves these compounds in only three minutes with a straightforward chromatographic apparatus which consists of a C-18 column, an isocratic mobile phase, UV detection at 254 nm, and an integrator; an expensive, sophisticated system is not required. The separation is both repeatable and rapid. Moreover, the experiment can be completed in a single three-hour period. The experiment is appropriate for any chemistry student who has completed a minimum of one year of general chemistry and is ideal for an analytical or instrumental analysis course. The experiment detailed herein involves the determination of caffeine in Goody's Extra Strength Headache Powders, a commercially available medication which contains acetaminophen, aspirin, and caffeine as active ingredients. However, the separation scheme is not limited to this brand of medication nor is it limited to caffeine as the analyte. With only minor procedural modifications, students can simultaneously quantitate all of these compounds in a commercial mixture. In our procedure, students prepare a series of four caffeine standard solutions as well as a solution from a pharmaceutical analgesic/caffeine mixture, chromatographically analyze each solution in quadruplicate, and plot relative average caffeine standard peak area versus concentration. From the mathematical relationship that results, the concentration of caffeine in the commercial formulation is obtained. Finally, the absolute standard deviation of the mean concentration is calculated.

  4. Rapid method for the simultaneous determination of flavonol aglycones in food using u-HPLC coupled with heating block acidic hydrolysis.

    PubMed

    Shim, You-Shin; Kim, Seunghee; Seo, Dongwon; Ito, Masahito; Nakagawa, Hiroaki; Park, Hyun-Jin; Ha, Jaeho

    2013-01-01

    A rapid method for the simultaneous determination of flavonol aglycones in food using ultra-high-performance LC (u-HPLC) coupled with a heating-block acidic hydrolysis method was validated in terms of precision, accuracy, and linearity. The u-HPLC separation was performed on an RP C18 column (particle size 2 micro m id, 2 mm, length 100 mm) with a photodiode array detector. The LOD and LOQ of the u-HPLC analyses were 0.15 and 0.47 mg/kg for myricetin, 0.09 and 0.28 mg/kg for quercetin, 0.16 and 0.49 mg/kg for kaempferol, and 0.08 and 0.25 mg/kg for isorhamnetin. The intraday and interday precisions of the individual flavonol aglycones were less than 9.31%. All calibration curves exhibited good linearity (r2 = 0.99) within the tested ranges. Total run time of u-HPLC was 13 min. The rapid u-HPLC method presented herein significantly improved the speed, sensitivity, and resolution of the analyses of myricetin, quercetin, kaempferol, and isorhamnetin in food.

  5. Determination of neotame in beverages, cakes and preserved fruits by column-switching high-performance liquid chromatography.

    PubMed

    Yang, Dajing; Chen, Bo

    2010-09-01

    A column-switching HPLC method for the determination of neotame in beverages, cakes and preserved fruits was developed. After pre-treatment using a Waters Oasis HLB cartridge, the sample solution was separated on two C(18) columns using a column-switching technique with a six-port valve. UV detection was performed at 210 nm. The effects of eluent composition and eluate volume on the retention and elution of neotame on SPE cartridge were investigated. The method is simple, rapid, sensitive and has good reproducibility. RSD was lower than 5% (n = 5). The calibration curve of neotame was in the range 5-100 microg/ml with good linearity (r(2) = 0.999). Because neotame was concentrated 10 times from an original sample to a sample solution by solid-phase extraction (SPE), the limit of quantification (LOQ) of the method was 0.5 mg/kg. The recovery yields of neotame spiked in foods was >92% with a coefficient of variation <3.2%. The proposed column-switching HPLC method can be successfully used to determine neotame in routine inspection work.

  6. Poly(cyclooctene)-based monolithic columns for capillary high performance liquid chromatography prepared via ring-opening metathesis polymerization.

    PubMed

    Schlemmer, Bettina; Gatschelhofer, Christina; Pieber, Thomas R; Sinner, Frank M; Buchmeiser, Michael R

    2006-11-01

    Monolithic columns for capillary HPLC were prepared via ring-opening metathesis polymerization (ROMP) from cis-cyclooctene (COE), tris(cyclooct-4-enyl-1-oxy)methylsilane (CL) as monomers, 2-propanol and toluene as porogens and RuCl(2)(Py)(2)(IMesH(2))(CHC(6)H(5)) (Py=pyridine, IMesH(2)=1,3-dimesityl-4,5-dihydroimidazolin-2-ylidene) as initiator within the confines of 200 microm i.d. fused silica columns. For evaluation of the novel monolithic capillary HPLC columns, a protein standard consisting of six proteins in the molecular weight range of 5800-66000 g/mol, i.e. ribonuclease A, insulin, albumin, lysozyme, myoglobin and beta-lactoglobulin, was used. Reproducibility of synthesis was checked by determining the relative standard deviation (RSD) in retention times (t(R)), which was found to be in the range of 2.9-3.9% for all analytes. Variations in polymer kinetics were realized by adding different amounts of free pyridine and had a significant influence on the monolith's morphology, the backpressure and retention times. On the contrary, variations in monomer content and COE to CL ratio showed only minor changes on these parameters. Long-term stability of 1000 runs at 50 degrees C showed excellent stability of the columns and no significant alteration in separation performance was observed in combination with slightly decreased retention times (approx. 1.6-7.2% for all analytes).

  7. Pressure programmed microbore column supercritical fluid chromatography-mass spectrometry for the determination of organophosphorus insecticides

    SciTech Connect

    Kalinoski, H.T.; Smith, R.D.

    1988-03-15

    The use of the high flow rate (HFR) interface for supercritical fluid chromatography-mass spectrometry (SFC-MS) is shown to allow operation under conditions which provide efficient pressure programmed separations with appropriate microbore (packed) HPLC columns. The combined advantages of selectivity offered by the microparticle-packed stationary phase and variable solvating power of the supercritical fluid are fully utilized in this approach. The greater sample loadings and lower detection limits possible using packed columns (based on concentration of sample in the injection solvent) compared with commercially available capillary columns are demonstrated for the determination of a series of organophosphorus insecticides. Low concentrations of polar fluid modifiers, generally required for high-quality separations in packed-column SFC, also function as mild chemical ionization reagents. Broad classes of thermally labile, higher molecular weight, moderately polar pesticides are amenable to identification by SFC-MS, which provides a sensitive, selective, and broadly applicable technique for the identification of pesticide compounds with detection limits in the part-per-billion range.

  8. The current revolution in column technology: how it began, where is it going?

    PubMed

    Gritti, Fabrice; Guiochon, Georges

    2012-03-01

    This work revisits the exceptionally rapid evolution of the technology of chromatographic columns and the important progress in speed of analysis and resolution power that was achieved over the last ten years. Whereas columns packed with 10 and 5 μm fully porous particles dominated the field for nearly thirty years (1975-2000), it took barely six years to see the commercialization of monolithic silica rods (2000), their raise to fame and decay to oblivion, the development of finer fully porous particles with size down to 1.7 μm (2006), and of sub-3 μm superficially porous particles (2006). Analysis times and plate heights delivered by columns packed with these recent packing materials have then been improved by more than one order of magnitude in this short period of time. This progress has rendered practically obsolete the age-old design of LC instruments. For low molecular weight compounds, analysts can now achieve peak capacities of 40 peaks in about 15s with a hold-up time of the order of 1.5s , in gradient elution, by operating columns packed with sub-3 μm shell particles at elevated temperatures, provided that they use optimized high pressure liquid chromatographs. This is the ultimate limit allowed by modern instruments, which have an extra-column band broadening contribution of 7 μL² at 4.0 mL/min and data acquisition rate of 160 Hz. The best 2.1 mm × 50 mm narrow-bore columns packed with 1.7 μm silica core-shell particles provide peaks that have a variance of 2.1 μL² for k=1. Finally, this work discusses possible ways to accelerate separations and, in the same time perform these separations at the same level of efficiency as they have today. It seems possible to pack columns with smaller particles, probably down to 1 μm and operate them with current vHPLC equipments for separations of biochemicals. Analyses of low molecular weight compounds will require new micro-HPLC systems able to operate 1mm I.D. columns at pressures up to 5 kbar, which

  9. Enantioselective and diastereoselective separation of synthetic pyrethroid insecticides on a novel chiral stationary phase by high-performance liquid chromatography.

    PubMed

    Tan, Xulin; Hou, Shicong; Wang, Min

    2007-07-01

    A novel chiral packing material for high-performance liquid chromatography (HPLC) was prepared by connecting (R)-1-phenyl-2-(4-methylphenyl) ethylamine (PTE) amide derivative of (S)-isoleucine to aminopropyl silica gel through 2-amino-3,5-dinitro-1-carboxamido-benzene unit. This chiral stationary phase was applied to the enantioselective and diastereoselective separation of five pyrethroid insecticides by HPLC under normal phase condition. To achieve satisfactory baseline separation an optimization of the variables of mobile phase composition was required. The two enantiomers of fenpropathrin and four stereoisomers of fenvalerate were baseline separated using hexane-1,2-dichloroethane-2-propanol as mobile phase. The results show that the enantioselectivity of CSP is better than Pirkle type 1-A column for these compounds. Only partial separations for the cypermethrin and cyfluthrin stereoisomers were observed. Seven peaks and eight peaks were observed for cypermethrin and cyfluthrin, respectively. The elution orders were assigned by using different stereoisomer-enriched products.

  10. Online coupling of molecularly imprinted solid-phase extraction to HPLC for determination of trace tetracycline antibiotic residues in egg samples.

    PubMed

    Jing, Tao; Niu, Jiwei; Xia, Huan; Dai, Qing; Zheng, Hongyan; Hao, Qiaolin; Mei, Surong; Zhou, Yikai

    2011-06-01

    An automated system has been developed for the determination of trace tetracycline antibiotics (TCs) in egg samples, based on online molecularly imprinted solid-phase extraction (MISPE) coupling with high-performance liquid chromatography (HPLC). Oxytetracycline and chlortetracycline were chosen as mixed templates to synthesize highly selective molecularly imprinted polymers for online extraction. Under the optimal online MISPE-HPLC condition, 10 mL egg samples were injected into the MISPE column and then the matrix was washed out. By rotating the switching valve, TCs were transferred to the analytical column and then separated by HPLC. Because sample pretreatment and chromatographic separation were carried out simultaneously, the whole analytical time (18 min) was significantly shortened compared with conventional offline techniques. The detection limits ranged from 0.8 to 1.3 ng/g. The enhancement factors were in the range of 159-410. The spiked recoveries of TCs in real egg samples ranged from 91.6 to 107.6% and the relative standard deviations (RSDs) were not higher than 4.0%.

  11. Preparation of a new chiral stationary phase for HPLC based on the (R)- 1-phenyl-2-(4-methylphenyl)ethylamine amide derivative of (S)-valine and 2-chloro-3,5-dinitrobenzoic acid: enantioseparation of amino acid derivatives and pyrethroid insecticides.

    PubMed

    Tan, Xulin; Hou, Shicong; Jiang, Jingli; Wang, Min

    2007-08-01

    A novel chiral stationary phase (CSP) for HPLC was prepared by bonding (R)-1-phenyl-2-(4-methylphenyl)ethylamine amide derivative of (S)-valine to aminopropyl silica gel through a 2-amino-3,5-dinitro-1-carboxamido-benzene unit. The CSP was used for the separation of some amino acid derivatives and pyrethroid insecticides by chiral HPLC. Satisfactory baseline separation required optimization of the variables of mobile phase composition. Use of dichloromethane as modifier in the mobile phase gave baseline separations of amino acid derivatives. The two enantiomers of fenpropathrin and four stereoisomers of fenvalerate were baseline separated using hexane-dichloromethane-ethanol as mobile phase. The results show that the enantioselectivity of the new CSP is better than Pirkle type 1-A column for these compounds. Only partial separations were observed for the stereoisomers of cypermethrin and cyfluthrin, which gave even and eight peaks, respectively.

  12. Silica-based polypeptide-monolithic stationary phase for hydrophilic chromatography and chiral separation.

    PubMed

    Zhao, Licong; Yang, Limin; Wang, Qiuquan

    2016-05-13

    Glutathione (GSH)-, somatostatin acetate (ST)- and ovomucoid (OV)-functionalized silica-monolithic stationary phases were designed and synthesized for HILIC and chiral separation using capillary electrochromatography (CEC). GSH, ST and OV were covalently incorporated into the silica skeleton via the epoxy ring-opening reaction between their amino groups and the glycidyl moiety in γ-glycidoxypropyltrimethoxysilane (GPTMS) together with polycondensation and copolymerization of tetramethyloxysilane and GPTMS. Not only could the direction and electroosmotic flow magnitude on the prepared GSH-, ST- and OV-silica hybrid monolithic stationary phases be controlled by the pH of the mobile phase, but also a typical HILIC behavior was observed so that the nucleotides and HPLC peptide standard mixture could be baseline separated using an aqueous mobile phase without any acetonitrile during CEC. Moreover, the prepared monolithic columns had a chiral separation ability to separate dl-amino acids. The OV-silica hybrid monolithic column was most effective in chiral separation and could separate dl-glutamic acid (Glu) (the resolution R=1.07), dl-tyrosine (Tyr) (1.57) and dl-histidine (His) (1.06). Importantly, the chiral separation ability of the GSH-silica hybrid monolithic column could be remarkably enhanced when using gold nanoparticles (AuNPs) to fabricate an AuNP-mediated GSH-AuNP-GSH-silica hybrid monolithic column. The R of dl-Glu, dl-Tyr and dl-His reached 1.19, 1.60 and 2.03. This monolithic column was thus applied to separate drug enantiomers, and quantitative separation of all four R/S drug enantiomers were achieved with R ranging from 4.36 to 5.64. These peptide- and protein-silica monolithic stationary phases with typical HILIC separation behavior and chiral separation ability implied their promise for the analysis of not only the future metabolic studies, but also drug enantiomers recognition.

  13. Isolation and monitoring of the endohedral metallofullerenes YC{sub 82} and Sc{sub 3}C{sub 82}. On-line chromatographic separation with EPR detection

    SciTech Connect

    Stevenson, S.; Dorn, H.C.; Burbank, P.; Harich, K.; Sun, Z.; Kiang, C.H.; Salem, J.R.; DeVries, M.S.; Loosdrecht, P.H.M. van; Johnson, R.D.; Yannoni, C.S.; Bethune, D.S.

    1994-09-01

    The direct coupling of high-performance liquid chromatography (HPLC) with on-line electron paramagnetic resonance (EPR) detection is demonstrated for monitoring separations of endohedral metallofullerenes (MC{sub 2a}). The HPLC-EPR approach readily permits detection of the paramagnetic species, such as YC{sub 82} and Sc{sub 3}C{sub 82}, in the presence of the dominant empty-cage fullerenes (C{sub 60}, C{sub 70}) and diamagnetic metallofullerenes (e.g., M{sub 2}C{sub 2n}). The results indicate that on-line EPR provides a noninvasive, selective detector for HPLC metallofullerene separations that is readily adaptable to air-sensitive and/or labile compounds. Specifically, the `EPR-active` metallofullerenes, YC{sub 82} and Sc{sub 3}C{sub 82}, are selectively monitored on-line for an initial separation of the metallofullerene fraction from the dominant empty-cage fullerenes utilizing a combination of polystyrene columns. This preparative `cleanup` procedure is followed by HPLC-EPR separation and monitoring of YC{sub 82} and Sc{sub 3}C{sub 82} species using a selective tripodal {pi}-acidic-phase column (Trident-Tri-DNP) for the final stages of isolation. 30 refs., 8 figs.

  14. Simultaneous Determination of Eight Hypotensive Drugs of Various Chemical Groups in Pharmaceutical Preparations by HPLC-DAD.

    PubMed

    Stolarczyk, Mariusz; Hubicka, Urszula; Żuromska-Witek, Barbara; Krzek, Jan

    2015-01-01

    A new sensitive, simple, rapid, and precise HPLC method with diode array detection has been developed for separation and simultaneous determination of hydrochlorothiazide, furosemide, torasemide, losartane, quinapril, valsartan, spironolactone, and canrenone in combined pharmaceutical dosage forms. The chromatographic analysis of the tested drugs was performed on an ACE C18, 100 Å, 250×4.6 mm, 5 μm particle size column with 0.0.05 M phosphate buffer (pH=3.00)-acetonitrile-methanol (30+20+50 v/v/v) mobile phase at a flow rate of 1.0 mL/min. The column was thermostatted at 25°C. UV detection was performed at 230 nm. Analysis time was 10 min. The elaborated method meets the acceptance criteria for specificity, linearity, sensitivity, accuracy, and precision. The proposed method was successfully applied for the determination of the studied drugs in the selected combined dosage forms. PMID:26651566

  15. Simultaneous Determination of Eight Hypotensive Drugs of Various Chemical Groups in Pharmaceutical Preparations by HPLC-DAD.

    PubMed

    Stolarczyk, Mariusz; Hubicka, Urszula; Żuromska-Witek, Barbara; Krzek, Jan

    2015-01-01

    A new sensitive, simple, rapid, and precise HPLC method with diode array detection has been developed for separation and simultaneous determination of hydrochlorothiazide, furosemide, torasemide, losartane, quinapril, valsartan, spironolactone, and canrenone in combined pharmaceutical dosage forms. The chromatographic analysis of the tested drugs was performed on an ACE C18, 100 Å, 250×4.6 mm, 5 μm particle size column with 0.0.05 M phosphate buffer (pH=3.00)-acetonitrile-methanol (30+20+50 v/v/v) mobile phase at a flow rate of 1.0 mL/min. The column was thermostatted at 25°C. UV detection was performed at 230 nm. Analysis time was 10 min. The elaborated method meets the acceptance criteria for specificity, linearity, sensitivity, accuracy, and precision. The proposed method was successfully applied for the determination of the studied drugs in the selected combined dosage forms.

  16. FRACTIONATING COLUMN PRODUCT COLLECTOR CONTROL

    DOEpatents

    Paxson, G.D. Jr.

    1964-03-10

    Means for detecting minute fluid products from a chemical separation column and for advancing a collector tube rack in order to automatically separate and collect successive fractionated products are described. A charge is imposed on the forming drops at the column orifice to create an electric field as the drop falls in the vicinity of a sensing plate. The field is detected by an electrometer tube coupled to the plate causing an output signal to actuate rotation of a collector turntable rack, thereby positioning new collectors under the orifice. The invention provides reliable automatic collection independent of drop size, rate of fall, or chemical composition. (AEC)

  17. Development of a chiral HPLC method to evaluate in vivo enantiomeric inversion of an unstable, polar radiosensitizer in plasma.

    PubMed

    Kagel, J R; Rossi, D T; Hoffman, K L; Leja, B; Lathia, C D

    1999-11-01

    A chiral HPLC method to quantify in vivo enantiomeric inversion of prodrug CI-1010 (IR) or its drug IIR (PD 146923), a radiosensitizer, upon X-irradiation of dosed rats was developed. These polar enantiomers were separated only by using normal-phase chiral HPLC. A Chiralpak AS column provided the best separation. Isolation of analytes from plasma employed solid-phase extraction (SPE), and required conditions that were compatible with normal-phase HPLC. Options for SPE were restricted by the chemically reactive nature of both prodrug and drug, which produced analyte losses as high as 100%. Acceptable recoveries using SPE required evaluation of conditions for analyte chemical stability. The validated method gave a lower-limit of quantitation (LLOQ) of 200 ng/ml for each enantiomer extracted from 0.15 ml of plasma. The LLOQ of the inverted enantiomer could be detected in the presence of 10,000 ng/ml of the dosed enantiomer. Precision (RSD) ranged from 14.2 to 4.4%, and from 24.2 to 5.1% for IIS and IIR, respectively. Accuracy (RE) was +/- 13.1 and +/- 13.2%, respectively. Recoveries ranged from 44.3 to 71.4%, and from 40.7 to 67.9%, for IIS and IIR, respectively.

  18. Speciation of small molecules and inorganic ions in salmon egg cell cytoplasm by surfactant-mediated HPLC/ICP-MS.

    PubMed

    Matsuura, Hirotaka; Hasegawa, Takuya; Nagata, Hitomi; Takatani, Kohei; Asano, Motoki; Itoh, Akihide; Haraguchi, Hiroki

    2003-01-01

    The speciation of diverse elements in salmon egg cell cytoplasm was performed by a surfactant-mediated HPLC/ICP-MS hyphenated system. In the present experiment, an ODS column coated with CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate), which is a zwitterionic bile acid derivative, was employed as a surfactant-mediated separation column, and ICP-MS was used as an element-selective detector. The present surfactant-mediated HPLC allowed us to separate large and small molecules within 10 min; large molecules, such as proteins, were eluted within 2.5 min, while small molecules were eluted after 2.5 min, but within 10 min. In the present experiment, Fe, Cu, and Zn in egg cell cytoplasm were observed mostly in species with large molecular weights, indicating that these elements are contained as metalloproteins or metalloenzymes in egg cell cytoplasm. On the contrary, it was found that P, S, Mo, and halogens in egg cell cytoplasm were contained as small molecules or inorganic ions. The major species of P in egg cell cytoplasm was identified as the phosphate ion (PO4(3-)). Molybdenum, Cl, and Br in egg cell cytoplasm were molybdate (MoO4(2-), chloride (Cl-), and bromide (Br-) ions, respectively.

  19. A lectin HPLC method to enrich selectively-glycosylated peptides from complex biological samples.

    PubMed

    Johansen, Eric; Schilling, Birgit; Lerch, Michael; Niles, Richard K; Liu, Haichuan; Li, Bensheng; Allen, Simon; Hall, Steven C; Witkowska, H Ewa; Regnier, Fred E; Gibson, Bradford W; Fisher, Susan J; Drake, Penelope M

    2009-01-01

    Glycans are an important class of post-translational modifications. Typically found on secreted and extracellular molecules, glycan structures signal the internal status of the cell. Glycans on tumor cells tend to have abundant sialic acid and fucose moieties. We propose that these cancer-associated glycan variants be exploited for biomarker development aimed at diagnosing early-stage disease. Accordingly, we developed a mass spectrometry-based workflow that incorporates chromatography on affinity matrices formed from lectins, proteins that bind specific glycan structures. The lectins Sambucus nigra (SNA) and Aleuria aurantia (AAL), which bind sialic acid and fucose, respectively, were covalently coupled to POROS beads (Applied Biosystems) and packed into PEEK columns for high pressure liquid chromatography (HPLC). Briefly, plasma was depleted of the fourteen most abundant proteins using a multiple affinity removal system (MARS-14; Agilent). Depleted plasma was trypsin-digested and separated into flow-through and bound fractions by SNA or AAL HPLC. The fractions were treated with PNGaseF to remove N-linked glycans, and analyzed by LC-MS/MS on a QStar Elite. Data were analyzed using Mascot software. The experimental design included positive controls-fucosylated and sialylated human lactoferrin glycopeptides-and negative controls-high mannose glycopeptides from Saccharomyces cerevisiae-that were used to monitor the specificity of lectin capture. Key features of this workflow include the reproducibility derived from the HPLC format, the positive identification of the captured and PNGaseF-treated glycopeptides from their deamidated Asn-Xxx-Ser/Thr motifs, and quality assessment using glycoprotein standards. Protocol optimization also included determining the appropriate ratio of starting material to column capacity, identifying the most efficient capture and elution buffers, and monitoring the PNGaseF-treatment to ensure full deglycosylation. Future directions include

  20. Continuous Flow Liquid Microjunction Surface Sampling Probe Connected On-line with HPLC/MS for Spatially Resolved Analysis of Small Molecules and Proteins

    SciTech Connect

    Van Berkel, Gary J; Kertesz, Vilmos

    2013-01-01

    RATIONALE: A continuous flow liquid microjunction surface sampling probe extracts soluble material from surfaces for direct ionization and detection by MS. Demonstrated here is the on-line coupling of such a probe with HPLC/MS enabling extraction, separation and detection of small molecules and proteins from surfaces in a spatially resolved (~0.5 mm diameter spots) manner. Methods: A continuous flow liquid microjunction surface sampling probe was connected to a 6-port, 2-position valve for extract collection and injection to an HPLC column. A QTRAP 5500 hybrid triple quadrupole linear ion trap equipped with a Turbo V ion source operated in positive ESI mode was used for all experiments. System operation was tested with extraction, separation and detection of propranolol and associated metabolites from drug dosed tissues and proteins from dried sheep blood spots on paper. Results: Confirmed in the tissue were the parent drug and two different hydroxypropranolol glucuronides. The mass spectrometric response for these compounds from different locations in the liver showed an increase with increasing extraction time (5, 20 and 40 s extractions). For on-line separation and detection/identification of extracted proteins from dried sheep blood spots, two major protein peaks dominated the chromatogram and could be correlated with the expected masses for the hemoglobin and chains. Conclusions: Spatially resolved sampling, separation, and detection of small molecules and proteins from surfaces can be accomplished using a continuous flow liquid microjunction surface sampling probe coupled on-line with HPLC/MS detection.

  1. Development of different comprehensive two dimensional systems for the separation of phenolic antioxidants.

    PubMed

    Cacciola, Francesco; Jandera, Pavel; Blahová, Eva; Mondello, Luigi

    2006-11-01

    Three different comprehensive 2-D HPLC systems for the separation of phenolic antioxidants have been developed on the basis of different selectivities of a PEG-silica column in the first dimension and a packed or monolithic C18 or a ZR-CARBON column, respectively, in the second dimension. Two-dimensional comprehensive liquid chromatography using a serially connected short PEG-silica column and a conventional C18-silica or a ZR-CARBON column in the second dimension was tested to improve the resolution of the earlier eluting compounds in the first dimension. Various types of interface were used to connect the columns in the first and in the second dimension: i) two injection sampling loops of 100 microL in conventional arrangement; ii) a 10-port 2-position valve equipped with two trapping X-Terra columns instead of loops; and iii) two analytical D2 columns in parallel. The mobile phase in the first dimension has a lower elution strength than in the second dimension, allowing band compression of the solutes transferred from the first to the second dimension. This effect was enhanced using trapping columns instead of sampling loops as the interface between the two dimensions, thus allowing a decrease in the time of analysis. These systems were used for the analysis of beer samples. The relative location of the components in the 2-D retention plane varied in relation to their chemical structure in each instrumental set-up and allowed positive peak identification. PMID:17154131

  2. Purification of patulin from Penicillium expansum culture: high-speed counter-current chromatography (HSCCC) versus preparative high-performance liquid chromatography (prep-HPLC).

    PubMed

    He, J; Tsao, R; Yang, R; Zhou, T

    2009-01-01

    Patulin is a mycotoxin produced by species of Penicillium and Aspergillus and is toxic to a wide range of organisms, including humans and livestock. To produce large amount of pure patulin for research purposes, high-speed counter-current chromatography (HSCCC) and preparative high-performance liquid chromatography (prep-HPLC) were applied to the purification of patulin. Apple juice was inoculated with P. expansum and containing 0.5 mg patulin per ml was used as a starting material for separation. For HSCCC, a biphasic solvent system consisted of ethyl acetate-hexane-pH 4 acetic acid (7.5:2.5:10, v/v/v) was used. For prep-HPLC, the separation was carried out in a C18 reversed-phase preparative column with a mobile phase containing acetonitrile-pH 4 acetic acid (5:95, v/v). Fractions containing patulin were collected and analysed by analytical HPLC and identified by congruent retention time and ultraviolet/visible (UV-VIS) spectrum of the standard. The structure of the purified patulin was confirmed by mass spectrometry and nuclear magnetic resonance. HSCCC produced 21.9 mg of patulin from 50 ml apple juice culture whereas the prep-HPLC yielded 18.1 mg. HSCCC also produced purer patulin than the prep-HPLC (98.6 versus 96.3%) and higher recovery (86.2 versus 71.3%). In addition, the HSCCC method is advantageous for its lower cost and a simpler procedure compared with the prep-HPLC. This one-step HSCCC method can potentially provide a simple, effective and environmentally friendly tool for obtaining gram-level pure patulin for toxicology, detoxification and many other patulin-related studies.

  3. One-pot preparation of a sulfamethoxazole functionalized affinity monolithic column for selective isolation and purification of trypsin.

    PubMed

    Xiao, Yuan; Guo, Jialiang; Ran, Danni; Duan, Qianqian; Crommen, Jacques; Jiang, Zhengjin

    2015-06-26

    A facile and efficient "one-pot" copolymerization strategy was used for the preparation of sulfonamide drug (SA) functionalized monolithic columns. Two novel SA-immobilized methacrylate monolithic columns, i.e. poly(GMA-SMX-co-EDMA) and poly(GMA-SAA-co-EDMA) were prepared by one-pot in situ copolymerization of the drug ligand (sulfamethoxazole (SMX) or sulfanilamide (SAA)), the monomer (glycidyl methacrylate, GMA) and the cross-linker (ethylene dimethacrylate, EDMA) within 100 μm i.d. capillaries under optimized polymerization conditions. The physicochemical properties and column performance of the fabricated monolithic columns were characterized by elemental analysis, scanning electron microscopy and micro-HPLC. Satisfactory column permeability, efficiency and separation performance were obtained on the optimized poly(GMA-SMX-co-EDMA) monolithic column for small molecules, such as a standard test mixture and eight aromatic ketones. Notably, it was found that the poly(GMA-SMX-co-EDMA) monolith showed a selective affinity to trypsin, while the poly(GMA-SAA-co-EDMA) monolith containing sulfanilamide did not exhibit such affinity at all. This research not only provides a novel monolith for the selective isolation and purification of trypsin, but it also offers the possibility to easily prepare novel drug functionalized methacrylate monoliths through a one-pot copolymerization strategy.

  4. Arsenic speciation in chinese seaweeds using HPLC-ICP-MS and HPLC-ES-MS.

    PubMed

    Van Hulle, Marijn; Zhang, Chao; Zhang, Xinrong; Cornelis, Rita

    2002-05-01

    Three common Chinese edible seaweeds, one brown (Laminaria japonica) and two red (Porphyra crispata and Eucheuma denticulatum), were examined for their total arsenic content. The As species were extracted with yields of 76.4, 69.8 and 25.0%, respectively. Anion-exchange and cation-exchange high-performance liquid chromatography (HPLC) in combination with inductively coupled plasma mass spectrometry (ICP-MS) were used for the separation of the different arsenic species in two of the three seaweed extracts (Laminaria and Porphyra). The main arsenic species in the algal extracts are arseno sugars, although it has been shown that the Laminaria seaweed contains significant amounts of dimethylarsinic acid (DMA). HPLC was coupled with electrospray mass spectrometry (ES-MS) for structural confirmation of the arsenic species. The mass spectrometer settings for the arseno sugars were optimised using standards. The conclusions drawn on the basis of HPLC-ICP-MS were confirmed by the HPLC-ES-MS data. The HPLC-ES-MS method is capable of determining both arseno sugars and DMA in the seaweeds. The unknown compounds seen in the HPLC-ICP-MS chromatogram of Laminaria could not be ascribed to trimethylarsenic oxide or tetramethylarsonium ion. PMID:12081041

  5. Self-regenerating column chromatography

    DOEpatents

    Park, Woo K.

    1995-05-30

    The present invention provides a process for treating both cations and anions by using a self-regenerating, multi-ionic exchange resin column system which requires no separate regeneration steps. The process involves alternating ion-exchange chromatography for cations and anions in a multi-ionic exchange column packed with a mixture of cation and anion exchange resins. The multi-ionic mixed-charge resin column works as a multi-function column, capable of independently processing either cationic or anionic exchange, or simultaneously processing both cationic and anionic exchanges. The major advantage offered by the alternating multi-function ion exchange process is the self-regeneration of the resins.

  6. HPLC and chemometric methods for the simultaneous determination of cyproheptadine hydrochloride, multivitamins, and sorbic acid.

    PubMed

    el-Gindy, Alaa; el-Yazby, Fawzy; Mostafa, Ahmed; Maher, Moustafa M

    2004-06-29

    Three methods are presented for the simultaneous determination of cyproheptadine hydrochloride (CP), thiamine hydrochloride (B1), riboflavin-5-phosphate sodium dihydrate (B2), nicotinamide (B3), pyridoxine hydrochloride (B6), and sorbic acid (SO). The chromatographic method depends on a high performance liquid chromatographic (HPLC) separation on a reversed-phase, RP 18 column. Elution was carried out with 0.1% methanolic hexane sulphonic acid sodium salt (solvent A) and 0.01 M phosphate buffer containing 0.1% hexane sulphonic acid sodium salt, adjusted to an apparent pH of 2.7 (solvent B). Gradient HPLC was used with the solvent ratio changed from 20:80 to 70:30 (over 9 min), then to 80:20 (over 11 min) for solvent A:B, respectively. Quantitation was achieved with UV detection at 220 and 288 nm based on peak area. The other two chemometric methods applied were principal component regression (PCR) and partial least squares (PLS). These approaches were successfully applied to quantify each drug in the mixture using the information included in the UV absorption spectra of appropriate solutions in the range 250-290 nm with the intervals Deltalambda = 0.4 nm at 100 wavelengths. The chemometric methods do not require any separation step. The three methods were successfully applied to a pharmaceutical formulation and the results were compared with each other.

  7. Quantitative analysis of the major constituents of St John's wort with HPLC-ESI-MS.

    PubMed

    Chandrasekera, Dhammitha H; Welham, Kevin J; Ashton, David; Middleton, Richard; Heinrich, Michael

    2005-12-01

    A method was developed to profile the major constituents of St John's wort extracts using high-performance liquid chromatography-electrospray mass spectrometry (HPLC-ESI-MS). The objective was to simultaneously separate, identify and quantify hyperforin, hypericin, pseudohypericin, rutin, hyperoside, isoquercetrin, quercitrin and chlorogenic acid using HPLC-MS. Quantification was performed using an external standardisation method with reference standards. The method consisted of two protocols: one for the analysis of flavonoids and glycosides and the other for the analysis of the more lipophilic hypericins and hyperforin. Both protocols used a reverse phase Luna phenyl hexyl column. The separation of the flavonoids and glycosides was achieved within 35 min and that of the hypericins and hyperforin within 9 min. The linear response range in ESI-MS was established for each compound and all had linear regression coefficient values greater than 0.97. Both protocols proved to be very specific for the constituents analysed. MS analysis showed no other signals within the analyte peaks. The method was robust and applicable to alcoholic tinctures, tablet/capsule extracts in various solvents and herb extracts. The method was applied to evaluate the phytopharmaceutical quality of St John's wort preparations available in the UK in order to test the method and investigate if they contain at least the main constituents and at what concentrations.

  8. Advances in the development of organic polymer monolithic columns and their applications in food analysis--a review.

    PubMed

    Jandera, Pavel

    2013-10-25

    Monolithic continuous separation media are gradually finding their way to sample pre-treatment, isolation, enrichment and final analytical separations of a plethora of compounds, occurring as food components, additives or contaminants, including pharmaceuticals, pesticides and toxins, which have traditionally been the domain of particulate chromatographic materials. In the present review, recent advances in the technology of monolithic columns and the applications in food analysis are addressed. Silica-based monoliths are excellent substitutes to conventional particle-packed columns, improving the speed of analysis for low-molecular weight compounds, due to their excellent efficiency and high permeability. These properties have been recently appreciated in two-dimensional HPLC, where the performance in the second dimension is of crucial importance. Organic-polymer monoliths in various formats provide excellent separations of biopolymers. Thin monolithic disks or rod columns are widely employed in isolation, purification and pre-treatment of sample containing proteins, peptides or nucleic acid fragments. Monolithic capillaries were originally intended for use in electrochromatography, but are becoming more frequently used for capillary and micro-HPLC. Monoliths are ideal highly porous support media for immobilization or imprinting template molecules, to provide sorbents for shape-selective isolation of target molecules from various matrices occurring in food analysis. The separation efficiency of organic polymer monoliths for small molecules can be significantly improved by optimization of polymerization approach, or by post-polymerization modification. This will enable full utilization of a large variety of available monomers to prepare monoliths with chemistry matching the needs of selectivity of separations of various food samples containing even very polar or ionized compounds.

  9. HPLC-DAD-MS identification of bioactive secondary metabolites from Ferula communis roots.

    PubMed

    Arnoldi, Lolita; Ballero, Mauro; Fuzzati, Nicola; Maxia, Andrea; Mercalli, Enrico; Pagni, Luca

    2004-06-01

    A simple HPLC method was developed to distinguish between 'poisonous' and 'non-poisonous' chemotypes of Ferula communis. The method was performed on a C8 reverse phase analytical column using a binary eluent (aqueous TFA 0.01%-TFA 0.01% in acetonitrile) under gradient condition. The two chemotypes showed different fingerprints. The identification of five coumarins and eleven daucane derivatives by HPLC-diode array detection (HPLC-DAD) and HPLC-MS is described. A coumarin, not yet described, was detected.

  10. HPLC-DAD-MS identification of bioactive secondary metabolites from Ferula communis roots.

    PubMed

    Arnoldi, Lolita; Ballero, Mauro; Fuzzati, Nicola; Maxia, Andrea; Mercalli, Enrico; Pagni, Luca

    2004-06-01

    A simple HPLC method was developed to distinguish between 'poisonous' and 'non-poisonous' chemotypes of Ferula communis. The method was performed on a C8 reverse phase analytical column using a binary eluent (aqueous TFA 0.01%-TFA 0.01% in acetonitrile) under gradient condition. The two chemotypes showed different fingerprints. The identification of five coumarins and eleven daucane derivatives by HPLC-diode array detection (HPLC-DAD) and HPLC-MS is described. A coumarin, not yet described, was detected. PMID:15158993

  11. High-performance liquid chromatographic separation of the enantiomers of organophosphorus pesticides on polysaccharide chiral stationary phases.

    PubMed

    Ellington, J J; Evans, J J; Prickett, K B; Champion, W L

    2001-09-14

    High-performance liquid chromatographic separation of the individual enantiomers of 12 organophosphorus pesticides (OPs) was obtained on polysaccharide enantioselective HPLC columns using alkane-alcohol mobile phase. The OP pesticides were crotoxyphos, dialifor, fonofos, fenamiphos, fensulfothion, isofenphos, malathion, methamidophos, profenofos, crufomate, prothiophos and trichloronate. The enantiomers of fenamiphos, fensulfothion, profenofos and crufomate were separated on CHIRALPAK AD; the enantiomers of fenamiphos were also separated on CHIRALPAK AS; the enantiomers of methamidophos, crufomate and trichloronate were separated on CHIRALCEL OD; the enantiomers of crotoxyphos, dialifor, fonofos, malathion, prothiophos and trichloronate were separated on CHIRALCEL OJ; and the enantiomers of isofenphos were separated on CHIRALCEL OG. Baseline or partial separation of the enantiomers of six of these OP pesticides was obtained on CHIRALCEL OJ. In continued method development, the separation of the enantiomers of the 12 OPs was investigated more extensively on CHIRALCEL OJ to determine whether the mobile phase composition, flow-rate and column temperature could be optimized to yield at least partial separation of the enantiomers. Chromatographic conditions were found that gave either baseline or near baseline separations of the enantiomers of the 12 OPs on the CHIRALCEL OJ column. PMID:11587332

  12. Method of filling a microchannel separation column

    DOEpatents

    Arnold, Don W.

    2002-01-01

    A method for packing a stationary phase into a small diameter fluid passageway or flow channel. Capillary action is employed to distribute a stationary phase uniformly along both the length and diameter of the flow channel. The method disclosed here: 1) eliminates the need for high pressure pumps and fittings and the safety hazards associated therewith; 2) allows the use of readily available commercial microparticles, either coated or uncoated, as the stationary phase; 3) provides for different types of particles, different particle sizes, and different particle size distributions to be packed in sequence, or simultaneously; 4) eliminates the need for plugging the flow channel prior to adding the stationary phase to retain the packing particles; and 5) many capillaries can be filled simultaneously.

  13. Gas Chromatograph Method Optimization Trade Study for RESOLVE: 20-meter Column v. 8-meter Column

    NASA Technical Reports Server (NTRS)

    Huz, Kateryna

    2014-01-01

    RESOLVE is the payload on a Class D mission, Resource Prospector, which will prospect for water and other volatile resources at a lunar pole. The RESOLVE payload's primary scientific purpose includes determining the presence of water on the moon in the lunar regolith. In order to detect the water, a gas chromatograph (GC) will be used in conjunction with a mass spectrometer (MS). The goal of the experiment was to compare two GC column lengths and recommend which would be best for RESOLVE's purposes. Throughout the experiment, an Inficon Fusion GC and an Inficon Micro GC 3000 were used. The Fusion had a 20m long column with 0.25mm internal diameter (Id). The Micro GC 3000 had an 8m long column with a 0.32mm Id. By varying the column temperature and column pressure while holding all other parameters constant, the ideal conditions for testing with each column length in their individual instrument configurations were determined. The criteria used for determining the optimal method parameters included (in no particular order) (1) quickest run time, (2) peak sharpness, and (3) peak separation. After testing numerous combinations of temperature and pressure, the parameters for each column length that resulted in the most optimal data given my three criteria were selected. The ideal temperature and pressure for the 20m column were 95 C and 50psig. At this temperature and pressure, the peaks were separated and the retention times were shorter compared to other combinations. The Inficon Micro GC 3000 operated better at lower temperature mainly due to the shorter 8m column. The optimal column temperature and pressure were 70 C and 30psig. The Inficon Micro GC 3000 8m column had worse separation than the Inficon Fusion 20m column, but was able to separate water within a shorter run time. Therefore, the most significant tradeoff between the two column lengths was peak separation of the sample versus run time. After performing several tests, it was concluded that better

  14. Effect of pressure pulses at the interface valve on the stability of second dimension columns in online comprehensive two-dimensional liquid chromatography.

    PubMed

    Talus, Eric S; Witt, Klaus E; Stoll, Dwight R

    2015-01-23

    Users of online comprehensive two-dimensional liquid chromatography (LCxLC) frequently acknowledge that the mechanical instability of HPLC columns installed in these systems, particularly in the second dimension, is a significant impediment to its use. Such instability is not surprising given the strenuous operating environment to which these columns are subjected, including the large number (thousands per day) of fast and large pressure pulses resulting from interface valve switches (on the timescale of tens of milliseconds) associated with very fast second dimension separations. There appear to be no published reports of systematic studies of the relationship between second dimension column lifetime and any of these variables. In this study we focused on the relationship between the lifetimes of commercially available columns and the pressure pulses observed at the inlet of the second dimension column that occur during the switching of the valve that interfaces the two dimensions of a LCxLC system. We find that the magnitude of the pressure drop at the inlet of the second dimension column during the valve switch, which may vary between 10 and 95% of the column inlet pressure, is dependent on valve switching speed and design, and has a dramatic impact on column lifetime. In the worst case, columns fail within the first few hours of use in an LCxLC system. In the best case, using a valve that exhibits much smaller pressure pulses, the same columns exhibit much improved lifetimes and have been used continuously under LCxLC conditions for several days with no degradation in performance. This result represents a first step in understanding the factors that affect second dimension column lifetime, and will significantly improve the usability of the LCxLC technique in general.

  15. [Study on limit detection of flavones in diterpene ginkgolides meglumine injection materials by LC-MS and HPLC-DAD].

    PubMed

    Bi, Sen; Li, Yan-jing; Huang, Wen-zhe; Kang, Dan-yu; Ding, Gang; Xiao, Wei

    2015-08-01

    Limit test of flavones in diterpene ginkgolides meglumine injection materials by UV-Vis and HPLC-DAD method was studied in this essay. The HPLC-DAD method has lower LOD (about 1% of the UV-Vis), that is, the sensitivity is higher than UV-Vis method. Through the analysis of the kinds of flavonoids ingredients in the samples by LC-MS, the three compounds with highest contents are kaempferol, quercetin and isorhamnetin. Kaempferol, quercetin and isorhamnetin were chosen as reference compounds for HPLC analysis, and the HPLC separation analysis was carried on an Agilent Eclipse plus C18 column (4.6 mm x 250 mm, 5 μm) with methanol and water containing 0.4% phosphoric acid (50: 50) as mobile phase, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 360 nm. This method has good specificity, precision and reproducibility. The LODs of quercetin, kaempferide and isorhamnetin were 27.6, 22.3, 29.5 μg x L(-1). The average recovery was 87.9% (RSD 3.3%), 91.7% (RSD 3.1%), 88.3 (RSD 1.3%) for quercetin, kaempferide and isorhamnetin, respectively. Based on the 10 batches of sample results and sensitivity of different HPLC, the content of total flavonoids ingredients of diterpene ginkgolides meglumine injection materials was limited no more than 2 x 10(-5). This method is simple, quick and has good maneuverability, and could be used to the limit test of flavonoids in the diterpene ginkgolides meglumine injection materials.

  16. [Study on limit detection of flavones in diterpene ginkgolides meglumine injection materials by LC-MS and HPLC-DAD].

    PubMed

    Bi, Sen; Li, Yan-jing; Huang, Wen-zhe; Kang, Dan-yu; Ding, Gang; Xiao, Wei

    2015-08-01

    Limit test of flavones in diterpene ginkgolides meglumine injection materials by UV-Vis and HPLC-DAD method was studied in this essay. The HPLC-DAD method has lower LOD (about 1% of the UV-Vis), that is, the sensitivity is higher than UV-Vis method. Through the analysis of the kinds of flavonoids ingredients in the samples by LC-MS, the three compounds with highest contents are kaempferol, quercetin and isorhamnetin. Kaempferol, quercetin and isorhamnetin were chosen as reference compounds for HPLC analysis, and the HPLC separation analysis was carried on an Agilent Eclipse plus C18 column (4.6 mm x 250 mm, 5 μm) with methanol and water containing 0.4% phosphoric acid (50: 50) as mobile phase, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 360 nm. This method has good specificity, precision and reproducibility. The LODs of quercetin, kaempferide and isorhamnetin were 27.6, 22.3, 29.5 μg x L(-1). The average recovery was 87.9% (RSD 3.3%), 91.7% (RSD 3.1%), 88.3 (RSD 1.3%) for quercetin, kaempferide and isorhamnetin, respectively. Based on the 10 batches of sample results and sensitivity of different HPLC, the content of total flavonoids ingredients of diterpene ginkgolides meglumine injection materials was limited no more than 2 x 10(-5). This method is simple, quick and has good maneuverability, and could be used to the limit test of flavonoids in the diterpene ginkgolides meglumine injection materials. PMID:26790294

  17. Development and application of a HPLC method for eight sunscreen agents in suncare products.

    PubMed

    Peruchi, L M; Rath, S

    2012-06-01

    This work describes the development, validation and application of a simple and fast high-performance liquid chromatography-with diode array dectection (HPLC-DAD) method for the determination of eight sunscreen agents: benzophenone-3, octocrylene, ethylhexyl methoxycinnamate, ethylhexyl salicylate, homosalate (used in two isomeric forms), butyl methoxydibenzoylmethane, 4-methylbenzylidene camphor and ethylhexyl dimethyl PABA in sunscreen formulations. The separation of the eight sunscreen compounds was achieved using an ACE C18 column (250 × 4.6 mm, 5 μm), with a column temperature 20°C, and a mobile phase of 88 : 12 (v/v) methanol-water with isocratic elution. Column temperature strongly influences the retention time and resolution of the compounds. The flow rate was 1.0 mL min(-1) and quantitation was performed by external calibration at the maximum wavelength of each compound. The sample preparation was simple and consisted basically of sample dilution with methanol, centrifugation and filtration in syringe filters before quantitation. Total run time was 18 min. The method was validated according to the parameters: linear range, linearity, selectivity, intra-day and inter-day precision and accuracy. Ten samples of sunscreen emulsions commercially available in Brazil (SPF 30) from different manufacturers were analysed using the proposed method. The number of the sunscreen agents varied between one and five in a single sample. The concentrations of all compounds were in the range of 0.9-10% (w/w) and were in accordance with the current Brazilian legislation.

  18. REDISTRIBUTOR FOR LIQUID-LIQUID EXTRACTION COLUMNS

    DOEpatents

    Bradley, J.G.

    1957-10-29

    An improved baffle plate construction to intimately mix immiscible liquid solvents for solvent extraction processes in a liquid-liquid pulse column is described. To prevent the light and heavy liquids from forming separate continuous homogeneous vertical channels through sections of the column, a baffle having radially placed rectangular louvers with deflection plates opening upon alternate sides of the baffle is placed in the column, normal to the axis. This improvement substantially completely reduces strippiig losses due to poor mixing.

  19. Identification and quantification of constituents of Gardenia jasminoides Ellis (Zhizi) by HPLC-DAD-ESI-MS.

    PubMed

    Bergonzi, M C; Righeschi, C; Isacchi, B; Bilia, A R

    2012-09-15

    A simple, rapid and specific HPLC method was carried out for the analysis of characteristic constituents in Gardenia jasminoides Ellis (Zhizi), namely iridoids, caffeoyl quinic acid derivatives and crocins. The separation was successfully obtained using a C(18) column by gradient elution with mixtures of methanol and water as mobile phases; detection wavelength was set at 240 nm for iridoid glycosides, 315 nm for quinic acid derivatives and 438 nm for crocins. The analytical method was validated and the quantification of active compounds, namely iridoids, was performed. Linearity, precision, repeatability, stability, accuracy, limit of detection (LOD) and limit of quantification (LOQ) were also reported. This assay was successfully applied for qualitative and quantitative analysis of five commercial samples of G. jasminoides Ellis. PMID:23107748

  20. Determination of metrafenone in vegetables by matrix solid-phase dispersion and HPLC-UV method.

    PubMed

    Li, Jianjun; Li, Yangyang; Xu, Dongliang; Zhang, Jingyu; Wang, Yuxi; Luo, Chao

    2017-01-01

    A simple method for determination of metrafenone in vegetables by matrix solid-phase dispersion (MSPD) and HPLC was developed. All vegetable samples were extracted with dichloromethane, and then the extracts were directly separated on a reversed-phase column with isocratic elution without a cleanup step. The linearity of metrafenone was good with the concentration between 0.005 and 5mg/kg, and the limit of detection (LOD) of the metrafenone was 0.002mg/kg. The recoveries ranged from 86.5% to 104.8% with the relative standard deviations (RSDs) in the range of 2.1-7.9% (n=6). The results indicated that the method was simple, rapid, highly sensitive and suitable for the determination of metrafenone in vegetables. PMID:27507450