Development of a high-performance liquid chromatographic-mass spectrometric technique, with an ionspray interface, for the determination of platelet-activating factor (PAF) and lyso-PAF in biological samples.

Journal of chromatography Pub Date : 1993-09-24 DOI:10.1016/0021-9673(93)83406-i

L Silvestro, R Da Col, E Scappaticci, D Libertucci, L Biancone, G Camussi

{"title":"Development of a high-performance liquid chromatographic-mass spectrometric technique, with an ionspray interface, for the determination of platelet-activating factor (PAF) and lyso-PAF in biological samples.","authors":"L Silvestro, R Da Col, E Scappaticci, D Libertucci, L Biancone, G Camussi","doi":"10.1016/0021-9673(93)83406-i","DOIUrl":null,"url":null,"abstract":"<p><p>An HPLC-mass spectrometric technique with an ionspray interface was developed for the determination of platelet-activating factor (PAF) and PAF-related compounds in biological samples. HPLC separations were performed using a reversed-phase column. The mass spectra showed intense [M + H]+ ions. Collision-induced dissociation of protonated molecular ions gave characteristic daughter ions corresponding to the phosphorylcholine group. By selective-ion monitoring, a detection limit of 0.3 ng was obtained for all molecules; by multiple reaction monitoring, the same sensitivity was achieved for PAF whereas for lyso-PAF the limit was 3 ng. Finally, PAF was comparatively determined by bioassay and HPLC-MS after extraction from the cell pellets and the supernatants of human polymorphonuclear neutrophils unstimulated or stimulated with opsonized zymosan. The good correlation observed between these techniques indicated the reliability of HPLC-MS for biochemical studies on PAF and PAF-related molecules.</p>","PeriodicalId":15508,"journal":{"name":"Journal of chromatography","volume":"647 2","pages":"261-9"},"PeriodicalIF":0.0000,"publicationDate":"1993-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/0021-9673(93)83406-i","citationCount":"24","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of chromatography","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1016/0021-9673(93)83406-i","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}

引用次数: 24

Abstract

An HPLC-mass spectrometric technique with an ionspray interface was developed for the determination of platelet-activating factor (PAF) and PAF-related compounds in biological samples. HPLC separations were performed using a reversed-phase column. The mass spectra showed intense [M + H]+ ions. Collision-induced dissociation of protonated molecular ions gave characteristic daughter ions corresponding to the phosphorylcholine group. By selective-ion monitoring, a detection limit of 0.3 ng was obtained for all molecules; by multiple reaction monitoring, the same sensitivity was achieved for PAF whereas for lyso-PAF the limit was 3 ng. Finally, PAF was comparatively determined by bioassay and HPLC-MS after extraction from the cell pellets and the supernatants of human polymorphonuclear neutrophils unstimulated or stimulated with opsonized zymosan. The good correlation observed between these techniques indicated the reliability of HPLC-MS for biochemical studies on PAF and PAF-related molecules.

查看原文本刊更多论文

基于离子喷雾界面的高效液相色谱-质谱技术用于测定生物样品中血小板活化因子(PAF)和溶索-PAF。

建立了一种离子喷雾界面高效液相色谱-质谱法测定生物样品中血小板活化因子(PAF)及其相关化合物的方法。采用反相柱进行HPLC分离。质谱显示出强烈的[M + H]+离子。质子化分子离子的碰撞诱导解离产生与磷胆碱基团对应的特征性子离子。通过选择性离子监测，所有分子的检出限均为0.3 ng;通过多反应监测，PAF获得相同的灵敏度，而lyso-PAF的极限为3 ng。最后，从人多形核中性粒细胞的细胞微球和未刺激或经增酶酶解刺激的上清液中提取PAF，采用生物测定法和高效液相色谱-质谱法进行比较。这些技术之间的良好相关性表明了高效液相色谱-质谱法用于PAF及其相关分子生化研究的可靠性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Journal of chromatography

自引率

0.00%

发文量