Quantification of dinor,dihydro metabolites of F2 isoprostanes in urine by liquid chromatography/tandem mass spectrometry
Quantification of dinor,dihydro metabolites of F2 isoprostanes in urine by liquid chromatography/tandem mass spectrometry
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DOI：
10.1016/j.ab.2005.10.012
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发表时间：
2006-01-15
影响因子：
2.9
通讯作者：
Roberts, LJ
中科院分区：
文献类型：
--
作者：
Davies, SS;Zackert, W;Roberts, LJ
F-2-异前列腺素(F-2-IsoP)是一系列前列腺素(PG)-F-2类化合物，由自由基介导的花生四烯酸氧化而成。一种具有很强生物活性的F-2-IsoP是15-F-2t-IsoP，已在几种疾病中检测到15-F-2t-IsoP水平升高。15-F-2t-IsoP(8-iso-PGF(2α))的主要尿代谢物为2，3-二氢-5.6-二氢-15-F-2t-IsoP(15-F-2t-IsoP-M)。此前，我们建立了稳定的15-F-2T-IsoP-M的同位素稀释气相色谱/负化学电离/质谱仪(MS)分析方法，该方法虽然灵敏度高，但需要耗时的衍生化和薄层纯化。现在，我们报告了一种更快速的高效液相色谱与电喷雾电离-串联质谱仪(LC/MS/MS)联用的方法，用于分析F-2-异构体(F-2-IsoP-M)的所有二氢代谢物。该方法精密度为+/-5.0%，准确度为80%。进样量在1-100 ng范围内线性关系良好。LC/MS/MS法测定的F-2-IsoP-M水平与GC/MS法测定的15-F-2t-IsoP-M水平显著相关(R=0.77y=67.2x-0.5)。40例正常人斑点尿中F-2-IsoP-M水平为38.1+/-19.1 ng/mg肌酐(均值+/-SD)。该方法提供了一种准确、快速的方法来评估体内的氧化状态。(C)2005 Elsevier Inc.保留所有权利。
The F-2-isoprostanes (F-2-IsoP) are a series of prostaglandin (PG)-F-2-like compounds that are produced by free-radical-mediated oxidation of arachidonic acid. One F-2-IsoP with potent biological activity is 15-F-2t-IsoP and increased levels of 15-F-2t-IsoP have been measured in several diseases. The major urinary metabolite of 15-F-2t-IsoP (8-iso-PGF(2 alpha)) is 2,3-dinor-5.6-dihydro-15-F-2t-IsoP (15-F-2t-IsoP-M). Previously, we developed a stable isotope dilution gas chromatography/negative chemical ionization/mass spectrometry (MS) assay for 15-F-2t-IsoP-M, which, while highly sensitive, required time-consuming derivatization and thin-layer chromatography purification. We now report the development of a more rapid high-performance liquid chromatography method Coupled to electrospray ionization-tandem mass spectrometry (LC/MS/MS) to analyze all of the dinor,dihydro metabolites of the F-2-IsoP isomers (F-2-IsoP-M). The precision of this assay was +/-5.0% and the accuracy 80%. The assay remained linear over a range of 1-100 ng injected onto the LC column. Levels of F-2-IsoP-M determined by the LC/MS/MS assay method significantly correlated with levels of 15-F-2t-IsoP-M determined by the GC/MS assay (R = 0.77y = 67.2x - 0.5). The levels of F-2-IsoP-M detected in spot urines from 40 normal subjects were 38.1 +/- 19.1 ng/mg creatinine (mean +/- SD). This method provides an accurate and rapid assay to assess oxidative status in vivo. (C) 2005 Elsevier Inc. All rights reserved.