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Chromatograms
Kinds of Chromatograms Selected AbstractsExtraction and Removal of Caffeine from Green Tea by Ultrasonic-Enhanced Supercritical FluidJOURNAL OF FOOD SCIENCE, Issue 4 2010Wei-Qiang Tang ABSTRACT:, Low-caffeine or caffeine-removed tea and its products are widely welcomed on market in recent years. In the present study, we adopt ultrasonic-enhanced supercritical fluid extraction process to remove caffeine from green tea. An orthogonal experiment (L16 (45)) was applied to optimize the best removal conditions. Extraction pressure, extraction time, power of ultrasound, moisture content, and temperature were the main factors to influence the removal rate of caffeine from green tea. The 5 factors chosen for the present investigation were based on the results of a single-factor test. The optimum removal conditions were determined as follows: extraction pressure of 30 MPa, temperature at 55 °C, time of 4 h, 30% moisture content, and ultrasound power of 100 W. Chromatogram and ultraviolet analysis of raw material and decaffeinates suggests that under optimized conditions, the caffeine of green tea was effectively removed and minished without damaging the structure of active ingredients in green tea. [source] Fast Liquid Chromatography for High-Throughput Screening of PolymersMACROMOLECULAR RAPID COMMUNICATIONS, Issue 1 2003Harald Pasch Abstract Liquid chromatography of polymers is traditionally a slow technique with analysis times of typically 30 min per sample. For the application of liquid chromatographic techniques to combinatorial materials research the analysis time per sample must be reduced considerably. Analysis time in SEC can be reduced to about 2 min per sample when high-throughput columns are used. For HPLC small columns with improved separation efficiencies can be used. As compared to conventional technology, time savings of more than 80% are achieved. Chromatogram from conventional SEC column compared to high-speed SEC column tested on an identical instrument with polystyrene standards in THF. [source] Detection of loci controlling seed glucosinolate content and their association with Sclerotinia resistance in Brassica napusPLANT BREEDING, Issue 1 2003J. Zhao Abstract A genetic linkage map of Brassica napus constructed from a cross between a low glucosinolate cultivar ,H5200' and a high glucosinolate line ,NingRS-1' was used to identify loci associated with seed glucosinolate content and to understand the association between specific glucosinolate components and Sclerotinia resistance. Seed glucosinolate content was assessed by standard High pressure Liquid Chromatogram (HPLC) protocol. Seven components of seed glucosinolate, including four types of aliphatic glucosinolate, two types of indolyl glucosinolates and one aromatic glucosinolate were detected in the seeds. Three quantitative trait loci (QTLs) were identified for seed total glucosinolate content. From three to 15 loci were found to be responsible for different types of glucosinolates, and by comparing the overlapped intervals, eight genomic regions were defined. One of the nine loci associated with aliphatic glucosinolate content was found to be associated with Sclerotinia resistance on the leaf at the seedling stage, and one locus, responsible for 3-indolyl-methyl glucosinolate content, was probably linked with Sclerotinia resistance on the stem of the maturing plant. The association between seed glucosinolate content and Sclerotinia resistance is discussed. [source] Tumor prevalence and biomarkers of exposure and response in brown bullhead (Ameiurus nebulosus) from the Anacostia River, Washington, DC and Tuckahoe River, Maryland, USAENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 3 2004Alfred E. Pinkney Abstract We valuated liver and skin tumor prevalence and biomarkers of exposure and response in brown bullhead (Ameiurus nebulosus) from three locations in the Anacostia River (Washington, DC, USA), a Chesapeake Bay region of concern. The Tuckahoe River (Maryland, USA) served as a reference. Each river was sampled in fall 2000 and spring 2001. In the Anacostia, prevalence of liver tumors was 50 to 68%, and prevalence of skin tumors was 13 to 23% in large (,260 mm, age ,3 years) bullheads. Liver and skin tumor prevalence was 10 to 17% and 0%, respectively, in small (150,225 mm, age 1,2 years) bullheads. Tuckahoe bullhead liver tumor prevalence was 0 to 3% (large) and 0% (small); none had skin tumors. Biliary polynuclear aromatic hydrocarbon (PAH)-like fluorescent metabolites and liver DNA adduct concentrations were elevated in large and small Anacostia bullheads. Mean adduct concentrations were 16 to 28 times higher than those in Tuckahoe fish. Chromatograms revealed a diagonal radioactive zone, indicating polycyclic aromatic compound (PAC)-DNA adducts. The biomarker data and the 10 to 17% liver tumor prevalence at ages 1 to 2 suggest that these year classes are likely to have a high prevalence as they reach age 3 and older. This study provides the strongest evidence to date of the role of PAHs in tumor development in Anacostia bullheads. [source] Vectorization of Harungana madagascariensis Lam. ex Poir. (Hypericaceae) ethanolic leaf extract by using PLG-nanoparticles: antibacterial activity assessmentDRUG DEVELOPMENT RESEARCH, Issue 1 2005B. Moulari Abstract This study was undertaken to compare the in vitro and ex vivo antibacterial activity of an ethanolic Harungana madagascariensis leaf extract (HLE) incorporated into poly (D,L -lactide-co,glycolide) nanoparticles (HLE -PLG-NP). Two concentrations of HLE (500 and 1,000,µg/mL) for the in vitro study and one concentration (500 µg/mL) for the ex vivo study were compared using two gram-positive bacterial strains (Micrococcus luteus and Staphylococcus epidermidis), and one gram-negative bacterial strain (Moraxella sp.). The ex vivo antibacterial activity was evaluated on S. epidermidis CIP 55109 (SE) using an artificial contamination method. SE was inoculated for 12 h onto human skin fragment surfaces treated for 5,min either with HLE loaded, unloaded PLG-NP, or HLE solution. In vitro, the two preparations inhibited completely the growth of all bacterial strains at 1,000,µg/mL. However, the HLE -PLG-NP had a significant antibacterial activity against SE (18.4±1.8,0.4±0.2 CFU/mL, P<0.05), and a marked antibacterial effect against M. luteus (ML) and Moraxella sp. (Msp) compared to HLE solution at 500 µg/mL. Ex vivo, HLE -PLG-NP at 500,µg/mL reduced viable bacteria (6.3,4.8 log10), compared to the HLE solution (6.3,5.5 log10) after 4 h artificial contamination (P<0.05). A thin layer chromatography study of both HLE solution and HLE -PLG-NP showed that among the seven components found on the chromatogram of the HLE solution, only two were present on the nanoparticles, one including a flavonoid heteroside fraction responsible for the antibacterial properties. The incorporation of the HLE into a colloidal carrier improved antibacterial performance. Drug Dev. Res. 65:26,33, 2005. © 2005 Wiley-Liss, Inc. [source] Solid-phase aroma concentrate extraction (SPACEÔ ): a new headspace technique for more sensitive analysis of volatilesFLAVOUR AND FRAGRANCE JOURNAL, Issue 3 2004Masashi Ishikawa Abstract The SPACEÔ (solid-phase aroma concentrate extraction) method is a modi,ed version of the SPME (solid-phase micro extraction) technique for headspace analysis, with increased area of the adsorbent to enable more sensitive analysis of volatiles. The SPACEÔ rod used in the technique is fabricated from stainless steel coated with an adsorbent mixture, consisting mainly of a graphite carbon. Initially, the SPACEÔ rod is ,xed in the head of a closed ,ask, where it adsorbs the aroma. Next, the rod is thermally desorbed on-line with a high-resolution gas chromatography,mass spectrometer (HRGC,MS). In the present experiments, SPACEÔ sampling reproducibility was determined by analysing a standard mixture and roasted coffee beans. The SPACEÔ rod collected the analytes with good reproducibility, with the exception of high polar compounds. Similar analyses of coffee powder were performed by SPME and other methods for comparison with the SPACEÔ method. The SPACEÔ method proved to have superior capabilities with high concentrations, and it produced a well-balanced chromatogram. Copyright © 2004 John Wiley & Sons, Ltd. [source] Supercritical carbon dioxide extraction of 2-acetyl-1-pyrroline and volatile components from pandan leavesFLAVOUR AND FRAGRANCE JOURNAL, Issue 3 2004Natta Laohakunjit Abstract The ,avour of pandan (Pandanus amaryllifolius Roxb.) leaves was extracted by supercritical ,uid with CO2 (SC-CO2) under different conditions of pressure, temperature and contact time to determine the yield of 2-acetyl-1-pyrroline (ACPY) and various other components; 14 volatile compounds on the gas chromatogram were identi,ed, and the predominant constituents were ACPY and 3-methyl-2(5H)-furanone. The interaction of different conditions signi,cantly in,uenced the yield of ACPY and various volatile compounds. There is a potential for high yield of ACPY by SC-CO2 at 200 bar, 500 °C and 20 min. The SDE,ether extract was found to have a very small amount of ACPY and an undesirable odour, as compared to the dark green ethanol extract, which contains a relatively larger quantity of ACPY and even more 3-methyl-2(5H)-furanone. Although at least 34 new components were uncovered from SC-CO2, SDE, and ethanol extraction, both ACPY and 3-methyl-2(5H)-furanone were the components tentatively obtained by all three methods. Copyright © 2004 John Wiley & Sons, Ltd. [source] Opportunities for ultra-high resolution analysis of essential oils using comprehensive two-dimensional gas chromatography: a reviewFLAVOUR AND FRAGRANCE JOURNAL, Issue 3 2003Robert Shellie Abstract In comprehensive 2D gas chromatography, the entire sample is simultaneously subjected to analysis on two capillary columns. By using a suitable modulation interface between the primary and secondary columns, hundreds of fast, second-dimension chromatograms are produced. The data from these chromatograms are treated such that a 3D surface plot or a 2D contour plot of the components' individual retention times, on each column, as well as peak responses, are represented. In a properly tuned comprehensive 2D chromatogram, the individual sample components are spread throughout a 2D separation space, providing a signi,cant increase in the probability of resolving a greater number of sample components without increasing the analysis time. Comprehensive 2D,GC has proved useful for high-resolution conventional essential oil analysis as well as high-resolution enantioselective essential oil analysis. Combining comprehensive 2D,GC with either a quadrupole or time-of-,ight mass spectrometer gives a powerful 3D analysis technique, which is extremely effective for complex sample analysis. The present status and opportunities arising from these ultra-high resolution approaches are discussed herein. Copyright © 2003 John Wiley & Sons, Ltd. [source] Determination of phthalate esters in cosmetics by gas chromatography with flame ionization detection and mass spectrometric detectionINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 4 2005Huiming Chen GC-FID; GC-MS; Produits cosmétiques; Esters Phtaliques Synopsis A gas chromatography coupled with flame ionization detection (GC-FID) and mass spectrometric detection (MSD) method was developed to determine the six kinds of phthalate esters [dimethyl phthalate (DMP), diethyl phthalate (DEP), di- n -butyl phthalate (DBP), benzyl butyl phthalate (BBP), di(2-ethylhexyl) phthalate (DEHP) and di- n -octyl phthalate (DOP)] in cosmetics (solid, cream and liquid cosmetics). The cosmetics were extracted with methanol by ultrasonic and then separated with high-speed centrifugation. The upper clear layer was dried and filtered through a 0.45 ,m pore diameter filter. The filtrate was injected into GC-FID/GC-MS for detection. GC-FID chromatogram was applied for qualitative analysis, external standard method was used for quantitative analysis. Confirmation of phthalate presence was undertaken by GC-EI-MS. The recovery range of all phthalates were between 92.0 and 110.0% with relative standard deviations between 1.95 and 5.92%. The low detection limits of the method were: 0.1 ng for DMP, DEP, DBP and BBP, 0.5 ng for DEHP and DOP. The method had advantages of high precision and sensitivity, simplicity of pretreatment. The method can be used to test the six kinds of phthalate esters in cosmetics. Resume Une méthode d'analyse par chromatographie gazeuse couplée à une détection par ionization de flamme (GC - FID) et une détection spectrométrique de masse (MSD) a été développée pour analyser 6 sortes d'esters phtaliques (phtalate de diméthyle (DMP), phtalate de diéthyle (DEP), phtalate de di- n -butyle (DBP), phtalate de benzylbutyle (BBP), phtalate de di-2-éthylhexyle (DEHP) et phtalate de di- n -octyle (DOP)) dans des produits cosmétiques (solides, crèmes et liquides). Les produits cosmétiques sont extraits au méthanol sous ultrason, puis séparés par ultracentrifugation. La phase supérieure limpide est déshydratée et filtrée sur un filtre de diamètre de pore moyen égal à 0,45 ,m. Le filtrat est injecté dans le système GC - FID/GC-MS pour analyse. Les chromatogrammes GC-FID sont utilisés pour l'analyse qualitative, des standards externes ont été utilisés pour l'analyse quantitative. La GC-EI-MS permet de confirmer la présence des esters phtaliques. Le taux de récupération de tous les esters est compris entre 92 et 110% avec une déviation standard allant de 1,95%à 5,92%. La limite de détection par cette méthode est de 0,1 ng pour DMP, DEP, DBP et BBP, 0,5 ng pour DEHP et DOP. Les avantages de cette méthode sont sa haute précision, sa sensibilité et la simplicité du prétraitement. Cette méthode peut être utilisée pour doser la présence des six sortes d'esters phtaliques dans des produits cosmétiques. [source] Enhanced photodegradation efficiency of polyethylene-TiO2 nanocomposite film with oxidized polyethylene waxJOURNAL OF APPLIED POLYMER SCIENCE, Issue 1 2010Wenjun Fa Abstract A novel photodegradable polyethylene-oxidized polyethylene wax-TiO2 (PE-OPW-TiO2) nanocomposite film was prepared by embedding the organically modified TiO2 nanoparticles into commercial PE in the presence of OPW. The photocatalytic degradation behavior under ultraviolet light or solar light was investigated by examining the weight loss of the composite films, UV,vis transmittance spectrum, scanning electron microscope (SEM), and gel permeation chromatogram (GPC). The results show that OPW, as a dispersant and a compatibiliser, markedly improves the dispersion and compatibility of TiO2 nanoparticles in PE resins. The PE-OPW-TiO2 composite film demonstrates much higher photodegradation efficiency and much better mechanical property than either the PE-TiO2 composite film or the pure PE film. The weight-average molecular weight (Mw) of the PE-OPW-TiO2 composite film decreased 94.3% and the number-average molecular weight (Mn) decreased 84.5% after 38 days solar light irradiation. The photocatalytic degradation mechanism of the film is briefly discussed. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010 [source] Chemical Structure and Physical Properties of Mung Bean Starches Isolated from 5 Domestic CultivarsJOURNAL OF FOOD SCIENCE, Issue 9 2007S.-H. Kim ABSTRACT:, Chemical structure and physical properties of starches isolated from 5 domestic mung bean cultivars (Gyungsun, Geumsung, Sunhwa, Eohul, and Jangan) were examined. The granules were jelly bean like in shape and smooth on surface, and the size was within 10 to 30 ,m. Mung bean starches displayed a CA -type crystalline structure when judged by the X-ray diffraction patterns. Branch chain-length distribution patterns of amylopectin (AP) revealed that peak chain length of APs was at either DP (degree of polymerization) 12 or DP13. Apparent amylose contents of 5 cultivars by iodine affinity test were 31.7% to 33.8%. Mung bean APs showed a unique molecular size distribution that has not been observed from other plant-derived starches. Two distinct peaks of AP fraction were identified on the size-exclusion chromatogram, and the ratios of these 2 peaks were different depending on the mung bean cultivars. Weight-average chain length (CLavg) of APs was in the range of 16.9 (Eohul) and 17.5 (Geumsung). The onset temperature (To) and enthalpy change (,Hgel) of starch gelatinization were 54.6 to 60.2 °C and 11.6 to 13.2 J/g. The ,H of the retrograded mung bean starches was 5.5 to 6.6 J/g, which indicated 44.5% to 52.7% of recrystallization. The pasting temperature, peak viscosity, and setback were 66.1 to 69.2 °C, 510 to 579 Rapid Visco Unit (RVU), and 66 to 90 RVU, respectively. [source] Isomerization of delta-9-THC to delta-8-THC when tested as trifluoroacetyl-, pentafluoropropionyl-, or heptafluorobutyryl- derivatives,,JOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 5 2008Justin M. Holler Abstract For GC,MS analysis of delta-9-tetrahydrocannabinol (delta-9-THC), perfluoroacid anhydrides in combination with perfluoroalcohols are commonly used for derivatization. This reagent mixture is preferred because it allows simultaneous derivatization of delta-9-THC and its acid metabolite, 11-nor-delta-9-THC-9-carboxylic acid present in biological samples. When delta-9-THC was derivatized by trifluoroacetic anhydride/hexafluoroisopropanol (TFAA/HFIPOH) and analyzed by GC,MS using full scan mode (50,550 amu), two peaks (P1 and P2) with an identical molecular mass of 410 amu were observed. On the basis of the total ion chromatogram (TIC), P1 with a shorter retention time (RT) was the major peak (TIC 84%). To identify the peaks, delta-8-THC was also tested under the same conditions. The RT and spectra of the major peak (TIC 95%) were identical with that of P1 for delta-9-THC. A minor peak (5%) present also correlated well with the latter peak (P2) for the delta-9-THC derivative. The fragmentation pathway of P1 was primarily demethylation followed by retro Diels-Alder fragmentation (M , 15,68, base peak 100%) indicating P1 as a delta-8-THC-trifluoroacetyl compound. This indicated that delta-9-THC isomerized to delta-8-THC during derivatization with TFAA/HFIPOH. Similar results were also observed when delta-9-THC was derivatized with pentafluoropropionic anhydride/pentafluoropropanol or heptafluorobutyric anhydride/heptafluorobutanol. No isomerization was observed when chloroform was used in derivatization with TFAA. In this reaction, the peaks of delta-8-THC-TFA and delta-9-THC-TFA had retention times and mass spectra matching with P1 and P2, respectively. Because of isomerization, perfluoroacid anhydrides/perfluoroalcohols are not suitable derivatizing agents for analysis of delta-9-THC; whereas the TFAA in chloroform is suitable for the analysis. Published in 2008 by John Wiley & Sons, Ltd. [source] Analysis of intracellular short organic acid-coenzyme A esters from actinomycetes using liquid chromatography-electrospray ionization-mass spectrometryJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 9 2007Je Won Park Abstract A method employing silicone oil density centrifugation, solid-phase extraction (SPE) cleanup, and LC-ESI-MS/MS analysis was developed for the rapid, selective, sensitive, and quantitative detection of an intracellular pool of short organic acid-CoA esters in actinomycetes. The detection limit was determined to be approximately 0.8 pmol (1.2 ng/ml) for each standard CoA-ester analyzed by the present LC-ESI-MS/MS method. A selected ion chromatogram for a typical fragment ion (m/z 428) specific to CoA-esters enabled the detection of eight intracellular CoA-esters involved in both primary and secondary metabolisms. The application of this method to bacterial metabolomic study is demonstrated by the profiling of the intracellular CoA-ester pools in the wild-type Streptomyces venezuelae strain producing polyketide antibiotics (methymycin and pikromycin), a polyketide synthase (PKS)-deleted S. venezuelae mutant, and a S. venezuelae mutant expressing the heterologous PKS genes. By quantifying the individual CoA-esterlevel in three different genotypes of the S. venezuela e strain, further insight could be gained into the role of CoA-estersin polyketide biosynthesis. This analytical approach can be extended to the quantification of the size and composition of in vivo CoA-ester pools in various microbes, and can provide a detailed understanding of the relationship between the in vivo CoA-ester pool and the production of pharmaceutically important polyketides. Copyright © 2007 John Wiley & Sons, Ltd. [source] Investigation of cis/trans ratios of peptide bonds in AVP analogues containing N -methylphenylalanine enantiomersJOURNAL OF PEPTIDE SCIENCE, Issue 1 2006Emilia Sikorska Abstract The solution conformation of vasopressin analogues, modified at positions 2 and 3 with N -methylphenylalanine or its enantiomer, [D -MePhe2,MePhe3]AVP and [MePhe2,D -MePhe3]AVP, were studied by 2D NMR spectroscopy in H2O/D2O and theoretical calculations (EDMC/ANALYZE). In the case of [MePhe2,D -MePhe3]AVP, the synthesis afforded two products, A and B, which had identical molecular ions and similar retention times on HPLC. This finding was explained by racemization of Cys1, which gave an additional analogue, [D -Cys1,MePhe2,D -MePhe3]AVP (B). The possibility is not excluded of racemization of Cys1 in the remaining analogues of this series. However, only in the case of [MePhe2,D -MePhe3]AVP was this process so distinct that two strong peaks in the HPLC chromatogram were observed. The NMR spectra of all the analogues showed several distinct sets of residual proton resonances. This suggests that the peptides adopt more than two groups of conformations in H2O/D2O. This fact is due to cis/trans isomerization. Two more populated isomers arise from the cis/trans isomerization across the 2,3 peptide bond in [D -MePhe2,MePhe3]AVP and [MePhe2,D -MePhe3]AVP (A) and across the 1,2 peptide bond in [D -Cys1,MePhe2,D -MePhe3]AVP (B). Copyright © 2005 European Peptide Society and John Wiley & Sons, Ltd. [source] Quantitative analysis and chromatographic fingerprinting for the quality evaluation of Forsythia suspensa extract by HPLC coupled with photodiode array detectorJOURNAL OF SEPARATION SCIENCE, JSS, Issue 23-24 2009Yonggang Xia Abstract A simple and reproducible HPLC-photodiode array detector method has been described for evaluating and controlling quality of Forsythia suspensa extract (FSE). First, by analysis of chromatographic fingerprints, the similarities of chromatograms of FSE samples from the same pharmaceutical company exceeded 0.999, 0.997 and 0.960, respectively, although they were much lower from different pharmaceutical companies. Second, by further comparing many batches of extract chromatograph charts with the corresponding reference herb materials, the "common peaks" 3, 5, 7 and 10 were defined as "marker peaks", which were identified as (+)-pinoresinol-,- D -glucoside, forsythiaside, phillyrin and phillygenin, respectively. Third, four "marker peaks" were simultaneously determined based on fingerprint chromatogram for further controlling the quality of FSE quantitatively. Namely, the newly developed method was successfully applied to analyze 38 batches of FSE samples supplied by three pharmaceutical factories, which showed acceptable linearity, intra-day precision (RSD<2.76%), inter-day precision (RSD<3.43%) and the average recovery rates in the range of (95.38±2.96)% to (101.60±3.08)%. At last, hierarchical clustering analysis and Bayes discriminant analysis statistical methods were used to classify and differentiate the 38 FSE samples to provide the basis for guiding reasonable use of FSE and controlling its quality better. [source] Combination of GC-MS with local resolution for determining volatile components in si-wu decoctionJOURNAL OF SEPARATION SCIENCE, JSS, Issue 1-2 2003Fan Gong Abstract In this paper, the combination of gas chromatography-mass spectrometry with chemometric local resolution techniques such as subwindow factor analysis (SFA) and orthogonal projection resolution (OPR) is investigated as a method of determining volatile components present in a traditional Chinese medicinal formulation known as si-wu decoction and its two individual herbs Rhizoma chuanxiong and Radix angelicae sinensis. In order to validate the reliability of the results obtained, the volatile components of interest were further separated on open glass columns and then analyzed in the same way as above. With the help of SFA and OPR approaches, the purity of chromatographic peaks can first be identified. Then, the pure chromatogram and mass spectrum of each component involved in a target peak cluster can be easily resolved and subsequently subjected to similarity searches in the NIST MS database to qualitatively and quantitatively determine the volatile components. Our results showed that about 127, 80, and 97 chemical components could be separated and 81, 49, and 55 of them identified, representing 83.95%, 91.86%, and 85.11% of the total relative content of volatile components from Rhizoma chuanxiong, Radix angelicae sinensis, and si-wu decoction, respectively. The results obtained in this work strongly indicate that the combination of GC-MS with chemometric local resolution methods could greatly improve the chromatographic separation ability by means of mathematical approaches. Moreover, they indicated the reliability and practicability of this combined technique. [source] GC,MS characterization of oligomers in polyadipates used as plasticizers for PVC in food contactPACKAGING TECHNOLOGY AND SCIENCE, Issue 3 2006Maurus Biedermann Abstract Fourteen commercial polyadipates and a polysebacate were analysed for their components of a molecular mass below 1000,Da, primarily with the aim of generating the background data for measuring the migration of this type of polymeric additives from plasticized PVC (e.g. cling films and gaskets of lids) into foods or food simulants. Since the composition of the material <1000,Da varies between the polyadipates, the main components must be identified to enable a correct quantification. Polyadipates differ in the diol used as linker, their termination (acid or alcohol) and in the end-capping (free alcohols, acetylation, acylation with fatty acids, esterification with octanol/decanol). Gas chromatography (GC) provides good separation, but the material remaining in the column up to high temperatures decomposes and forms a hump in the rear part of the chromatogram. Examples of mass spectra are shown, the most indicative fragments pointed out and spectra of 159 components listed. The polyadipates and the sebacate are characterized by their structure, the main components <1000,Da and the fraction of material <1000,Da. Copyright © 2006 John Wiley & Sons, Ltd. [source] Application of Scion image software to the simultaneous determination of curcuminoids in turmeric (Curcuma longa)PHYTOCHEMICAL ANALYSIS, Issue 1 2009Uthai Sotanaphun Abstract Introduction Curcumin, desmethoxycurcumin and bisdesmethoxycurcumin are bioactive constituents of turmeric (Curcuma longa). Owing to their different potency, quality control of turmeric based on the content of each curcuminoid is more reliable than that based on total curcuminoids. However, to perform such an assay, high-cost instrument is needed. Objective To develop a simple and low-cost method for the simultaneous quantification of three curcuminoids in turmeric using TLC and the public-domain software Scion Image. Methodology The image of a TLC chromatogram of turmeric extract was recorded using a digital scanner. The density of the TLC spot of each curcuminoid was analysed by the Scion Image software. The density value was transformed to concentration by comparison with the calibration curve of standard curcuminoids developed on the same TLC plate. Results The polynomial regression data for all curcuminoids showed good linear relationship with R2 > 0.99 in the concentration range of 0.375,6 µg/spot. The limits of detection and quantitation were 43,73 and 143,242 ng/spot, respectively. The method gave adequate precision, accuracy and recovery. The contents of each curcuminoid determined using this method were not significantly different from those determined using the TLC densitometric method. Conclusion TLC image analysis using Scion Image is shown to be a reliable method for the simultaneous analysis of the content of each curcuminoid in turmeric. Copyright © 2008 John Wiley & Sons, Ltd. [source] Quantitative analysis of polymeric proanthocyanidins in birch leaves with normal-phase HPLCPHYTOCHEMICAL ANALYSIS, Issue 3 2006Maarit Karonen Abstract The proanthocyanidin composition and content in the leaves of nine birch species (Betula albosinensis, B. ermanii, B. maximowicziana, B. nana, B. papyrifera, B. pendula, B. platyphylla, B. pubescens, and B. pubescens ssp. czerepanovii) were studied with different methods including colorimetric assay, HPLC coupled with PAD or ESI/MS and NMR. Total proanthocyanidin content was determined using the acid butanol assay. A normal phase-HPLC method was applied for the analysis of polymeric proanthocyanidins. The content of polymeric proanthocyanidins was estimated from a late eluting peak in the chromatogram. With this HPLC method, quantitative analysis of polymeric proanthocyanidins could be performed directly from leaf extracts: no additional purification or preparation steps were required. It was shown that birch leaves contained mainly polymeric proanthocyanidins with a degree of polymerisation greater than 10. Total proanthocyanidin content (expressed as dry weight) was found to vary from 44 mg/g (B. papyrifera) to 145 mg/g (B. nana), and polymeric proanthocyanidin content from 39 mg/g (B. pendula) to 119 mg/g (B. nana). Copyright © 2006 John Wiley & Sons, Ltd. [source] Application of thin-layer chromatography/infrared matrix-assisted laser desorption/ionization orthogonal time-of-flight mass spectrometry to structural analysis of bacteria-binding glycosphingolipids selected by affinity detectionRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 7 2010Anne Müsken Glycosphingolipids (GSLs) play key roles in the manifestation of infectious diseases as attachment sites for pathogens. The thin-layer chromatography (TLC) overlay assay represents one of the most powerful approaches for the detection of GSL receptors of microorganisms. Here we report on the direct structural characterization of microbial GSL receptors by employment of the TLC overlay assay combined with infrared matrix-assisted laser desorption/ionization orthogonal time-of-flight mass spectrometry (IR-MALDI-o-TOF-MS). The procedure includes TLC separation of GSL mixtures, overlay of the chromatogram with GSL-specific bacteria, detection of bound microbes with primary antibodies against bacterial surface proteins and appropriate alkaline phosphatase labeled secondary antibodies, and in situ MS analysis of bacteria-specific GSL receptors. The combined method works on microgram scale of GSL mixtures and is advantageous in that it omits laborious and time-consuming GSL extraction from the silica gel layer. This technique was successfully applied to the compositional analysis of globo-series neutral GSLs recognized by P-fimbriated Escherichia coli bacteria, which were used as model microorganisms for infection of the human urinary tract. Thus, direct TLC/IR-MALDI-o-TOF-MS adds a novel facet to this fast and sensitive method offering a wide range of applications for the investigation of carbohydrate-specific pathogens involved in human infectious diseases. Copyright © 2010 John Wiley & Sons, Ltd. [source] Identification of 1-hydroxypyrene glucuronide in tissue of marine polychaete Nereis diversicolor by liquid chromatography/ion trap multiple mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 16 2002Anders M. B. Giessing 1-Hydroxypyrene glucuronide is identified as the single major aqueous metabolite of the tetracyclic aromatic hydrocarbon pyrene, in tissue from a deposit-feeding marine polychaete, Nereis diversicolor. Identification was performed using an ion trap mass spectrometer fitted with an atmospheric pressure chemical ionization (APCI) probe and connected to a high-performance liquid chromatography/diode array detector (HPLC/DAD) system. Besides 1-hydroxypyrene, the 339-nm UV trace of tissue samples from pyrene-exposed worms showed only one dominant peak that could be related to pyrene metabolism. Negative APCI-MS of this supposed 1- hydroxypyrene conjugate gave a characteristic signal at m/z 429 corresponding to the molecular ion of 1-hydroxypyrene glucuronide plus eluent adducts ([M,,,H,+,2H2O],). Fragmentation pathways were studied by isolating the abundant ion at m/z 429 in the ion trap and performing multiple mass spectrometric experiments (MSn). The fragmentations observed were consistent with the proposed identification. Two low intensity LC peaks that could be related to pyrene metabolism by their DAD absorption spectra were also present in the 339-nm UV chromatogram of tissue samples. However, these peaks could not be identified by their mass spectra in negative ion mode due to ion suppression by very abundant co-eluting impurities. The present method shows that LC/MSn is a fast and useful analytical tool for identification of aqueous polycyclic aromatic hydrocarbon biotransformation products in samples from relatively small marine invertebrates with limited sample preparation. Copyright © 2002 John Wiley & Sons, Ltd. [source] Clinical evaluation of spermatogenic activity of processed Shilajit in oligospermiaANDROLOGIA, Issue 1 2010T. K. Biswas Summary The safety and spermatogenic activity of processed Shilajit (PS) were evaluated in oligospermic patients. Initially, 60 infertile male patients were assessed and those having total sperm counts below 20 million ml,1 semen were considered oligospermic and enrolled in the study (n = 35). PS capsule (100 mg) was administered twice daily after major meals for 90 days. Total semenogram and serum testosterone, luteinising hormone and follicle-stimulating hormone were estimated before and at the end of the treatment. Malondialdehyde (MDA), a marker for oxidative stress, content of semen and biochemical parameters for safety were also evaluated. Twenty-eight patients who completed the treatment showed significant (P < 0.001) improvement in spermia (+37.6%), total sperm count (+61.4%), motility (12.4,17.4% after different time intervals), normal sperm count (+18.9%) with concomitant decrease in pus and epithelial cell count compared with baseline value. Significant decrease of semen MDA content (,18.7%) was observed. Moreover, serum testosterone (+23.5%; P < 0.001) and FSH (+9.4%; P < 0.05) levels significantly increased. HPLC chromatogram revealed inclusion of PS constituents in semen. Unaltered hepatic and renal profiles of patients indicated that PS was safe at the given dose. The present findings provide further evidence of the spermatogenic nature of Shilajit, as attributed in Ayurvedic medicine, particularly when administered as PS. [source] Development of a 25-plex SNP assay for traceability in cattleANIMAL GENETICS, Issue 3 2009B. Karniol Summary Single nucleotide polymorphisms (SNPs) are amenable to automation and therefore have become the marker of choice for DNA profiling. SNaPshot, a primer extension-based method, was used to multiplex 25 SNPs that have been previously validated as useful for identity control. Detection of extended products was based on four different fluorochromes and extension primers with oligonucleotide tails of differing lengths, thus controlling the concise length of the entire chromatogram to 81 bases. Allele frequencies for Holstein, Simmental, Limousin, Angus, Charolais and Tux Cattle were estimated and significant positive Pearson-correlation coefficients were obtained among the analysed breeds. The probability that two randomly unrelated individuals would share identical genotypes for all 25 loci varied from 10,8 to 10,10 for these breeds. For parentage control, the exclusion power was found to be 99.9% when the genotypes of both putative parents are known. A traceability test of duplicated samples indicated a high genotyping precision of >0.998. This was further corroborated by analysis of 60 cases of parent,sib pairs and trio families. The 25-plex SNaPshot assay is adapted for low- and high-throughput capacity and thus presents an alternative for DNA-based traceability in the major commercial cattle breeds. [source] Identification of N -linked oligosaccharide labeled with 1-pyrenesulfonyl chloride by quadrupole time-of-flight tandem mass spectrometry after separation by micro- and nanoflow liquid chromatographyBIOMEDICAL CHROMATOGRAPHY, Issue 9 2009Jun Zhe Min Abstract The development of a qualitative determination method for the N -linked oligosaccharides in glycoproteins was attempted by the combination of micro- or nanoflow LC with Q-TOF-MS/MS. The asparaginyl-oligosaccharides in glycoproteins, liberated from the treatment of Pronase E, were separated, purified and labeled with 1-pyrenesulfonyl chloride (PSC). The resulting derivatives were separated by the microflow LC system utilizing a 0.5 mm diameter microcolumn or nanoflow LC system utilizing a 75 µm diameter chip column. The eluted N -linked oligosaccharide derivatives were then introduced into the Q-TOF-MS instrument and sensitively detected in the ESI+ mode. Several factors (i.e. fragmentor, skimmer, Vcap voltages and collision energy) affecting the sensitivity of Q-TOF-MS/MS detection were optimized in both the micro- and nanoflow LC systems. Various fragment ions based on the carbohydrate units appeared on the MS/MS spectra. Among the peaks, m/z 600.16 corresponding to PSC-labeled Asp-HexNAc is the most important one to identify the asparaginyl-oligosaccharide. The N -linked oligosaccharides in the ovalbumin were easily identified by the selected-ion chromatogram at m/z 600.16 by the MS/MS detection. Therefore, the identification of N -linked oligosaccharides in glycoproteins seems to be possible by the proposed micro- and nanoflow LC separations followed by the Q-TOF-MS/MS detection. Copyright © 2009 John Wiley & Sons, Ltd. [source] Identification of flurbiprofen and its photoproducts in methanol by gas chromatography,mass spectrometryBIOMEDICAL CHROMATOGRAPHY, Issue 5 2007Su-Hui Chao Abstract A sample of 10 mm flurbiprofen in methanol (or ethanol) was photoirradiated with sixteen 8 W low-pressure quartz mercury lamps irradiated at 306 nm in a Panchum PR-2000 photochemical reactor. In total, four major photoproducts derived from each sample were observed from the HPLC chromatogram. The photoproducts were separated and their structures elucidated by various spectroscopic methods. Alternatively, using GC-MS, 11 major photoproducts were observed. A reaction scheme of flurbiprofen in methanol is proposed: the photochemical reaction routes occur mainly via esterification and decarboxylation, followed by oxidation with singlet oxygen to produce a ketone, alcohols and other derivatives. Copyright © 2007 John Wiley & Sons, Ltd. [source] An automated fluorescence protein sequencer using 7-methylthio-4-(2,1,3-benzoxadiazolyl) isothiocyanate (MTBD-NCS) as an Edman reagentBIOMEDICAL CHROMATOGRAPHY, Issue 3 2002Natsuko Okiyama An automated fluorescence protein sequencer using 7-methylthio-5-(2,1,3-benzoxadiazolyl) isithiocyanate (MTBD-NCS), a fluorescent Edman reagent, is developed by the modification of a commercial protein sequencer. The generated MTBD-thiohydantoin amino acids fluoresced strongly, whereas the by-products such as MTBD-thiocarbamoyl amino acids and MTBD-carbamoly amino acids did not fluoresce. A few interfering peaks were observed in the chromatogram and amino acid sequence was easily determined. The coupling and cyclization/cleavage reaction conditions and extraction conditions of generated MTBD-thiazolinone amino acids were optimized using an autonalyzer. Finally, the sequence of a synthetic peptide (25,pmol), leucine-enkephalin-Thr-amide, was determined and up to six residues were successively analyzed. Copyright © 2002 John Wiley & Sons, Ltd. [source] Ultra scale-down approach to correct dispersive and retentive effects in small-scale columns when predicting larger scale elution profilesBIOTECHNOLOGY PROGRESS, Issue 4 2009N. Hutchinson Abstract Ultra scale-down approaches represent valuable methods for chromatography development work in the biopharmaceutical sector, but for them to be of value, scale-down mimics must predict large-scale process performance accurately. For example, one application of a scale-down model involves using it to predict large-scale elution profiles correctly with respect to the size of a product peak and its position in a chromatogram relative to contaminants. Predicting large-scale profiles from data generated by small laboratory columns is complicated, however, by differences in dispersion and retention volumes between the two scales of operation. Correcting for these effects would improve the accuracy of the scale-down models when predicting outputs such as eluate volumes at larger scale and thus enable the efficient design and operation of subsequent steps. This paper describes a novel ultra scale-down approach which uses empirical correlations derived from conductivity changes during operation of laboratory and pilot columns to correct chromatographic profiles for the differences in dispersion and retention. The methodology was tested by using 1 mL column data to predict elution profiles of a chimeric monoclonal antibody obtained from Protein A chromatography columns at 3 mL laboratory- and 18.3 L pilot-scale. The predictions were then verified experimentally. Results showed that the empirical corrections enabled accurate estimations of the characteristics of larger-scale elution profiles. These data then provide the justification to adjust small-scale conditions to achieve an eluate volume and product concentration which is consistent with that obtained at large-scale and which can then be used for subsequent ultra scale-down operations. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 [source] Origin of the Silurian Crude Oils and Reservoir Formation Characteristics in the Tazhong UpliftACTA GEOLOGICA SINICA (ENGLISH EDITION), Issue 5 2010YANG Haijun Abstract: The Silurian stratum in the Tazhong uplift is an important horizon for exploration because it preserves some features of the hydrocarbons produced from multi-stage tectonic evolution. For this reason, the study of the origin of the Silurian oils and their formation characteristics constitutes a major part in revealing the mechanisms for the composite hydrocarbon accumulation zone in the Tazhong area. Geochemical investigations indicate that the physical properties of the Silurian oils in Tazhong vary with belts and blocks, i.e., heavy oils are distributed in the TZ47,15 well-block in the North Slope while normal and light oils in the No. I fault belt and the TZ16 well-block, which means that the oil properties are controlled by structural patterns. Most biomarkers in the Silurian oils are similar to that of the Mid-Upper Ordovician source rocks, suggesting a good genetic relationship. However, the compound specific isotope of n -alkanes in the oils and the chemical components of the hydrocarbons in fluid inclusions indicate that these oils are mixed oils derived from both the Mid-Upper Ordovician and the Cambrian,Lower Ordovician source rocks. Most Silurian oils have a record of secondary alterations like earlier biodegradation, including the occurrence of "UCM" humps in the total ion current (TIC) chromatogram of saturated and aromatic hydrocarbons and 25-norhopane in saturated hydrocarbons of the crude oils, and regular changes in the abundances of light and heavy components from the structural low to the structural high. The fact that the Silurian oils are enriched in chain alkanes, e.g., n -alkanes and 25-norhopane, suggests that they were mixed oils of the earlier degraded oils with the later normal oils. It is suggested that the Silurian oils experienced at least three episodes of petroleum charging according to the composition and distribution as well as the maturity of reservoir crude oils and the oils in fluid inclusions. The migration and accumulation models of these oils in the TZ47,15 well-blocks, the No. I fault belt and the TZ16 well-block are different from but related to each other. The investigation of the origin of the mixed oils and the hydrocarbon migration and accumulation mechanisms in different charging periods is of great significance to petroleum exploration in this area. [source] Comparative analysis of triacylglycerols from Olea europaea L. fruits using HPLC and MALDI-TOFMSEUROPEAN JOURNAL OF LIPID SCIENCE AND TECHNOLOGY, Issue 5 2010Faouzi Sakouhi Abstract MALDI-TOFMS and HPLC are two analytical methods that were used to characterize triacylglycerols (TAG) of the Meski, Sayali, and Picholine Tunisian olive varieties. The HPLC chromatograms of the oils showed the presence of 15 TAG species, among which triolein (OOO) was the most abundant (21,48%). In the Sayali cultivar, OOO was the predominant TAG species followed by POO and LOO. However, the minor TAG molecules were represented by LnLO and LnLP. MALDI mass spectra produced sodiated ([M,+,Na]+) and potassiated ([M,+,K]+) TAG molecules; only the major TAG were potassiated [OOO,+,K] ([OOO,+,K]+, [POO,+,K]+, and [LOO,+,K]+). In contrast to the HPLC chromatograms, the MALDI mass spectra showed 13 peaks of TAG. The major peak was detected at m/z,907, which corresponds to OOO with an Na+ adduct. The results from both HPLC and MALDI techniques predict the fatty acid composition and their percentages for each olive variety. Practical applications: TAG are the main components in vegetable oils. These biomolecules determine the physical, chemical, and nutritional properties of the oils. The nutritional benefits of TAG are related to DAG (moderate plasma lipid level) and esterified FA, which are intermediate biosynthetic molecules of TAG. TAG analysis is necessary to discriminate between oils of different origin, since some oils have similar FA profiles. Olive products, oils, and table olives, are the main diet sources of TAG in the Mediterranean countries. In this work, chromatographic and spectrometric methods were used for TAG analysis and characterization of Tunisian olive varieties. [source] Identification of Trichoderma strains by image analysis of HPLC chromatogramsFEMS MICROBIOLOGY LETTERS, Issue 2 2001Ulf Thrane Abstract Forty-four Trichoderma strains from water-damaged building materials or indoor dust were classified with chromatographic image analysis on full chromatographic matrices obtained by high performance liquid chromatography with UV detection of culture extracts. The classes were compared with morphological identification and rDNA sequence data, and for each class all strains were of the same identity. With all three techniques each strain , except one , was identified as the same species. These strains belonged to Trichoderma atroviride (nine strains), Trichoderma viride (three strains), Trichoderma harzianum (10 strains), Trichoderma citrinoviride (12 strains), and Trichoderma longibrachiatum (nine strains). The odd strain was identified as Trichoderma hamatum by morphology and rDNA sequencing, but not by image analysis as no reference strains of this species were included. It is concluded that the secondary metabolite profile contains sufficient information for classification and species identification. [source] | ||||||||||||||||||||||||||||||||||