使用气相色谱-串联质谱法对总F(2)-异前列腺素进行定量的优化方法。
Optimized method for quantification of total F(2)-isoprostanes using gas chromatography-tandem mass spectrometry.
作者信息
Briskey David R, Wilson Gary R, Fassett Robert G, Coombes Jeff S
机构信息
Exercise and Oxidative Stress Research Group, School of Human Movement Studies, University of Queensland, Brisbane, Australia; School of Medicine, University of Queensland, Brisbane, Australia.
Exercise and Oxidative Stress Research Group, School of Human Movement Studies, University of Queensland, Brisbane, Australia.
出版信息
J Pharm Biomed Anal. 2014 Mar;90:161-6. doi: 10.1016/j.jpba.2013.11.028. Epub 2013 Dec 1.
F2-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/μg protein. This method provides increased ion selectivity and detection capabilities with economical sample through-put.
F2-异前列腺素由花生四烯酸氧化降解产生，被认为是生物样品中脂质过氧化的金标准标志物。我们开发了一种液-液萃取方法，用于在血浆和组织匀浆中使用负化学电离气相色谱-串联质谱法测定总异前列腺素。与目前的方法相比，采用液-液萃取可实现更高的样品通量。在此，我们描述该方案，并给出众多故障排除建议。该方法发现健康个体每毫升血浆中异前列腺素含量为150 - 250皮克，而终末期肾病患者的测量值最高可达1100皮克/毫升。该检测方法的准确工作线性范围为40 - 1000皮克异前列腺素（100 - 2500皮克/毫升），平均变异系数为7%。健康小鼠肝脏的组织值为50 - 70皮克/微克蛋白质。该方法提高了离子选择性和检测能力，且样品通量经济。
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