Journal of chromatography. B, Biomedical sciences and applications最新文献_第2页

Measurement of S-nitrosoalbumin by gas chromatography-mass spectrometry. I. Preparation, purification, isolation, characterization and metabolism of S-[15N]nitrosoalbumin in human blood in vitro. 气相色谱-质谱法测定s -亚硝基白蛋白。1 . S-[15N]亚硝基白蛋白的制备、纯化、分离、表征及体外人血代谢。

Journal of chromatography. B, Biomedical sciences and applications Pub Date : 1999-04-16

D Tsikas, J Sandmann, S Rossa, F M Gutzki, J C Frölich

{"title":"Measurement of S-nitrosoalbumin by gas chromatography-mass spectrometry. I. Preparation, purification, isolation, characterization and metabolism of S-[15N]nitrosoalbumin in human blood in vitro.","authors":"D Tsikas, J Sandmann, S Rossa, F M Gutzki, J C Frölich","doi":"","DOIUrl":"","url":null,"abstract":"S-Nitrosoalbumin (SNALB) and S-[15N]nitrosoalbumin (S[15N]ALB) were prepared by various methods, purified and isolated by a novel selective extraction procedure using HiTrapBlue Sepharose affinity columns and characterized by various techniques including SDS-PAGE electrophoresis, UV-Vis spectroscopy and gas chromatography-mass spectrometry (GC-MS). S-Nitrosylation of albumin in freshly obtained human plasma by unlabeled and 15N-labeled butylnitrite at neutral pH revealed the purest preparations. For GC-MS analysis, SNALB and S[15N]ALB were treated with HgCl2 to obtain nitrite and [15N]nitrite, respectively, which were then analysed as their pentafluorobenzyl derivatives. S[15N]ALB preparations were standardized by GC-MS using nitrite as internal standard. S[15N]ALB was prepared and isolated at concentrations of 188+/-43 microM (mean +/- SD, n = 8) at a final yield of about 45%, an isotopic purity of 98%, and SDS-PAGE electrophoretic purity of 90%. 15N-Labeled SNALB was used to study its metabolism in human blood. The half-life of S[15N]ALB (25 microM) in human heparinized blood in vitro was determined by GC-MS as 5.5 h. The GC-MS method described here could be useful for the quantitative determination of SNALB in human plasma using S[15N]ALB as an internal standard.","PeriodicalId":15425,"journal":{"name":"Journal of chromatography. B, Biomedical sciences and applications","volume":"726 1-2","pages":"1-12"},"PeriodicalIF":0.0,"publicationDate":"1999-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21217176","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Measurement of S-nitrosoalbumin by gas chromatography-mass spectrometry. II. Quantitative determination of S-nitrosoalbumin in human plasma using S-[15N]nitrosoalbumin as internal standard. 气相色谱-质谱法测定s -亚硝基白蛋白。2以S-[15N]亚硝基白蛋白为内标定量测定人血浆中S-亚硝基白蛋白。

Journal of chromatography. B, Biomedical sciences and applications Pub Date : 1999-04-16

D Tsikas, J Sandmann, F M Gutzki, D O Stichtenoth, J C Frölich

{"title":"Measurement of S-nitrosoalbumin by gas chromatography-mass spectrometry. II. Quantitative determination of S-nitrosoalbumin in human plasma using S-[15N]nitrosoalbumin as internal standard.","authors":"D Tsikas, J Sandmann, F M Gutzki, D O Stichtenoth, J C Frölich","doi":"","DOIUrl":"","url":null,"abstract":"A gas chromatographic-mass spectrometric method for the quantitative determination of S-nitrosoalbumin (SNALB) in human plasma is described. The method is based on selective extraction of SNALB and its 15N-labeled SNALB analog (S15NALB) used as internal standard on HiTrapBlue Sepharose affinity columns, Hg2+ -catalysed conversion of the S-nitroso groups to nitrite and [15N]nitrite, respectively, followed by their derivatization to the pentafluorobenzyl derivatives and quantification by GC-MS. Mean recovery of SNALB and S15NALB from plasma was 45%. Mean precision and accuracy within the range 0-10 microM was 95% and 99%, respectively. The limit of quantitation was determined as 100 nM at a precision of 93.8% and an accuracy of 94.8%. Considerable improvement of method sensitivity is possible by eliminating nitrite present in the elution buffer. The limit of detection was 0.2 nM corresponding to 67 amol of S15NALB. In 0.4-ml aliquots of plasma samples from healthy humans, endogenous SNALB was determined at concentrations of 181+/-150 nM (mean +/- SD, n = 23). External addition of SNALB to these plasma samples at 2 microM and 5 microM serving as quality control samples resulted in quantitative recovery of SNALB. Our results show that SNALB occurs in human plasma at concentrations at least one-order of magnitude smaller than those reported in the literature from measurements using chemiluminescence.","PeriodicalId":15425,"journal":{"name":"Journal of chromatography. B, Biomedical sciences and applications","volume":"726 1-2","pages":"13-24"},"PeriodicalIF":0.0,"publicationDate":"1999-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21217177","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

High-throughout screening in combinatorial chemistry for drug discovery. 药物发现组合化学的高通量筛选。

Journal of chromatography. B, Biomedical sciences and applications Pub Date : 1999-04-02

A M Krstulović

引用次数: 0

Proteome analysis. I. Gene products are where the biological action is. 蛋白质组分析。1、基因产物是生物作用所在。

Journal of chromatography. B, Biomedical sciences and applications Pub Date : 1999-02-05

M F Lopez

引用次数: 0

Proteome analysis. II. Protein subcellular redistribution: linking physiology to genomics via the proteome and separation technologies involved. 蛋白质组分析。2蛋白质亚细胞再分配:通过蛋白质组学和分离技术将生理学与基因组学联系起来。

Journal of chromatography. B, Biomedical sciences and applications Pub Date : 1999-02-05

W F Patton

{"title":"Proteome analysis. II. Protein subcellular redistribution: linking physiology to genomics via the proteome and separation technologies involved.","authors":"W F Patton","doi":"","DOIUrl":"","url":null,"abstract":"While annotated two-dimensional (2D) gel electrophoresis databases contain thousands of proteins, they do not represent the entire genome. High-molecular-mass proteins in particular are conspicuously absent from such databases. Filamin is prototypical of this class of proteins since it is a dimer with relative molecular mass (Mr) of 520000 containing at least 240 potential phosphorylation sites. Filamin is not readily separated by current 2D procedures, and is difficult to study with respect to cycles of phosphorylation-dephosphorylation. Novel technologies are needed to identify biochemical pathways impinging upon such targets. The success of immunofluorescence microscopy as a research tool can be attributed in part to the fact that proteins redistribute in response to a variety of physiological stimuli. Comparable quantitative methods are required in proteome analysis. Three components are necessary for development of an approach that is capable of screening for protein redistribution events: (1) subcellular fractionation, (2) protein labeling and (3) data acquisition. An integrated approach is presented that utilizes differential detergent fractionation combined with reversible, luminescent protein stains and analytical imaging for high-throughput analysis of signal transduction events leading to protein subcellular redistribution. The procedure has been successfully implemented to rapidly define key second messenger pathways leading to endothelial cell junctional permeability and to guide in the design of a new family of peptide-based anti-inflammatory drugs.","PeriodicalId":15425,"journal":{"name":"Journal of chromatography. B, Biomedical sciences and applications","volume":"722 1-2","pages":"203-23"},"PeriodicalIF":0.0,"publicationDate":"1999-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20941873","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Use of magnetic techniques for the isolation of cells. 利用磁技术分离细胞。

Journal of chromatography. B, Biomedical sciences and applications Pub Date : 1999-02-05

I Safarík, M Safaríková

引用次数: 0

Chloroformates in gas chromatography as general purpose derivatizing agents. 氯甲酸盐在气相色谱中用作通用衍生试剂。

Journal of chromatography. B, Biomedical sciences and applications Pub Date : 1998-10-09

P Husek

{"title":"Chloroformates in gas chromatography as general purpose derivatizing agents.","authors":"P Husek","doi":"","DOIUrl":"","url":null,"abstract":"Chloroformates with simplest alkyls, i.e. methyl, ethyl or isobutyl, already known as favourable reagents for treating amino groups in gas chromatography for years, were revealed randomly as exceptionally rapid esterification agents. Unlike the rather poor results achieved with chloroformate-mediated ester formation in organic chemistry, the pyridine-catalyzed esterification of carboxylic acids appeared to proceed at the analytical microscale smoothly. Along with the catalyzer, an alcohol should also be present in the medium, accompanied by acetonitrile or water, according to the character of the compounds treated. Reaction conditions were optimized for various classes of carboxylic acids and a uniquely rapid derivatization of amino acids in aqueous ethanol was shown to be possible. Most of the analytes, e.g. acidic metabolites in physiological fluids, could be treated directly in the aqueous matrix. A simultaneous analysis of, e.g., amino and fatty acids or amines and their acidic catabolytes was proven to be possible. Along with the low-molecular-mass reagents, still some others, i.e. the hexyl, menthyl or pentafluorobenzyl ones, found their application fields. Results of optimized reaction conditions and a wide range of applications of chloroformate-mediated derivatization in various disciplines have been summarized in this review.","PeriodicalId":15425,"journal":{"name":"Journal of chromatography. B, Biomedical sciences and applications","volume":"717 1-2","pages":"57-91"},"PeriodicalIF":0.0,"publicationDate":"1998-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20742485","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Measurement of nitrite and nitrate in plasma, serum and urine of humans by high-performance liquid chromatography, the Griess assay, chemiluminescence and gas chromatography-mass spectrometry: interferences by biogenic amines and N(G)-nitro-L-arginine analogs. 用高效液相色谱、Griess测定、化学发光和气相色谱-质谱法测定人血浆、血清和尿液中的亚硝酸盐和硝酸盐:生物胺和N(G)-硝基- l -精氨酸类似物的干扰。

Journal of chromatography. B, Biomedical sciences and applications Pub Date : 1998-09-18

D Tsikas, I Fuchs, F M Gutzki, J C Frölich

引用次数: 0

Cellulose as a (bio)affinity carrier: properties, design and applications. 纤维素作为一种(生物)亲和载体:性质、设计和应用。

Journal of chromatography. B, Biomedical sciences and applications Pub Date : 1998-09-11

P Gemeiner, M Polakovic, D Mislovicová, V Stefuca

引用次数: 0

Studies on neurosteroids. VII. Determination of pregnenolone and its 3-stearate in rat brains using high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. 神经类固醇的研究。7高效液相色谱-常压化学电离质谱法测定大鼠脑中的孕烯醇酮及其3-硬脂酸酯。

Journal of chromatography. B, Biomedical sciences and applications Pub Date : 1998-09-04

K Shimada, Y Mukai

引用次数: 0