Optimized method for quantification of total F2-isoprostanes using gas chromatography-tandem mass spectrometry

被引：32
作者：
Briskey, David R. [1 ,2 ]
Wilson, Gary R. [1 ]
Fassett, Robert G. [1 ]
Coombes, Jeff S. [1 ]
机构：
[1] Univ Queensland, Sch Human Movement Studies, Exercise & Oxidat Stress Res Grp, St Lucia, Qld 4072, Australia
[2] Univ Queensland, Sch Med, Brisbane, Qld, Australia
来源：
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS | 2014年 / 90卷
关键词：
Gas chromatography; Tandem mass spectrometry; Oxidative stress; Lipid peroxidation; Liquidp-liquid extraction; VIVO LIPID-PEROXIDATION; OXIDATIVE STRESS; IN-VIVO; HUMAN PLASMA; URINARY; CYCLOOXYGENASE; BIOMARKERS; MARKERS; ISOPROSTANES; F2-ALPHA;
D O I：
10.1016/j.jpba.2013.11.028
中图分类号：
O65 [分析化学];
学科分类号：
070302 ; 081704 ;
摘要：
F-2-isoprostanes are produced from the oxidative degradation of arachidonic acid and are considered the gold standard marker of lipid peroxidation in biological samples. We developed a liquid-liquid extraction method for the determination of total isoprostanes using negative chemical ionization gas chromatography-tandem mass spectrometry in plasma and tissue homogenates. Incorporating liquid-liquid extraction allows for greater sample through-put than current approaches. Here we describe the protocol and include numerous trouble-shooting suggestions. The method found healthy individuals with 150-250 pg of isoprostanes per ml of plasma and end stage kidney disease patients to have the highest measured values of up to 1100 pg/ml. This assay has an accurate working linear range of 40-1000 pg of isoprostanes (100-2500 pg/ml) and an average coefficient of variance of 7%. Tissue values for healthy mice liver were 50-70 pg/mu g protein. This method provides increased ion selectivity and detection capabilities with economical sample through-put. (C) 2013 Elsevier B.V. All rights reserved.
引用
收藏
页码：161 / 166
页数：6
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