Determination of 8-epi PGF2α concentrations as a biomarker of oxidative stress using triple-stage liquid chromatography/tandem mass spectrometry
Determination of 8-epi PGF2α concentrations as a biomarker of oxidative stress using triple-stage liquid chromatography/tandem mass spectrometry
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DOI：
10.1002/rcm.4197
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发表时间：
2009-09-01
影响因子：
2
通讯作者：
Blomhoff, Rune
中科院分区：
文献类型：
--
作者：
Bastani, Nasser E.;Gundersen, Thomas E.;Blomhoff, Rune
F2-异前列腺素是一类前列腺素F2样化合物，由花生四烯酸的自由基催化过氧化反应形成。几种F2-异前列烷，但特别是8-表PGF（2 α），被广泛用作氧化应激生物标志物。建立了一种基于液相色谱-电喷雾负离子（ESI）串联质谱检测（LC/MS/MS）的分析方法，用于测定人血浆、全血、红细胞和尿液中的8-epi PGF（2 α）浓度。8-表PGF（2 α）-d4，8-表PGF（2 α）的稳定同位素衍生物，用作内标（IS）。将50 μ L样品聚焦在柱上，并在两个串联连接的3 μ m粒径SUPELCOSIL（TM）ABZ+Plus HPLC柱（15 cm × 4.6 mm和7.5 cm × 4.6 mm）上分离。具有ESI的Applied Biosystems 4000 Q TRAP LC/MS/MS系统以多反应监测（MRM）模式操作，其中吸附剂至产物离子跃迁m/z 353.4 -> 193.1（8-差向PGF（2 α））、357.4 -> 197.1（8-差向PGF（2 α）-d4）用于定量。对该试验进行了充分验证，发现其具有足够的准确度、精密度、线性、灵敏度和选择性。检测质量限（mLOD）为1 pg从色谱柱洗脱的分析物。该方法已成功地应用于人血浆、全血、红细胞和尿液样品的分析。版权所有（C）2009约翰威利父子有限公司
F2-isoprostanes are a family of prostaglandin F2-like compounds that are formed by free-radical-catalyzed peroxidation of arachidonic acid. Several F2-isoprostanes, but in particular 8-epi PGF(2 alpha), are widely used as oxidative stress biomarkers. An analytical method based on liquid chromatography with negative electrospray ionization (ESI) coupled to tandem mass spectrometric detection (LC/MS/MS) was developed for the determination of 8-epi PGF(2 alpha) concentrations in human plasma, whole blood, erythrocytes and urine. 8-epi PGF(2 alpha)-d4, a stable isotope derivative of 8-epi PGF(2 alpha), was used as an internal standard (IS). A 50 mu L sample was focused on-column and separated on two 3 mu m particle size SUPELCOSIL (TM) ABZ+Plus HPLC columns (15 cm x 4.6 mm and 7.5 cm x 4.6 mm) connected in series. An Applied Biosystems 4000 Q TRAP LC/MS/MS system with ESI was operated in multiple reaction monitoring (MRM) mode with the precursor-to-product ion transitions m/z 353.4 -> 193.1 (8-epi PGF(2 alpha)), 357.4 -> 197.1 (8-epi PGF(2 alpha)-d4), used for quantification. The assay was fully validated and found to have adequate accuracy, precision, linearity, sensitivity and selectivity. The mass limit of detection (mLOD) was 1 pg of analyte eluting from the column. The assay has been successfully applied to the analysis of human plasma, whole blood, erythrocytes and urine samples. Copyright (C) 2009 John Wiley & Sons, Ltd.