Quantitative high-performance liquid chromatography/electrospray ionization tandem mass spectrometric analysis of 2-and 3-series prostaglandins in cultured tumor cells
Quantitative high-performance liquid chromatography/electrospray ionization tandem mass spectrometric analysis of 2-and 3-series prostaglandins in cultured tumor cells
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DOI：
10.1016/s0003-2697(02)00218-x
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发表时间：
2002-09-01
影响因子：
2.9
通讯作者：
Newman, RA
中科院分区：
文献类型：
--
作者：
Yang, PY;Felix, E;Newman, RA
本文介绍了一种快速简便的液相色谱/电喷雾串联质谱技术，用于同时定量分析培养细胞中PGE(2)、PGE(3)和其他密切相关的前列腺素。该方法允许从细胞提取物中提取花生四烯酸（AA）或二十碳五烯酸（EPA），而无需繁琐的衍生化，冗长的样品制备和基于GC-MS或hplc - uv的方法所需的分离。验证评价结果表明，PGE(2)和PGE(3)在检测范围（1 ~ 500 ng/ml, 0.0028 ~ 1.4 muM）内均呈线性（r(2) > 0.999）， 3天样品分析的变异系数小于10%。PGE(2)和PGE(3)的检出限均为2.5 pg。细胞悬液中PGE(2)和PGE(3)的萃取率为89.4% ~ 98.2%。作为该方法的应用，测定了EPA在人肺癌A549细胞中形成的前列腺素。当A549细胞被100mum的EPA处理时，PGE(2)的形成减少62%。同时，EPA使A549细胞中PGE(3)的形成增加了10倍。这是第一个明确证明EPA可以在人类癌细胞中通过环加氧酶转化为PGE(3)的报告。(C) 2002 Elsevier Science （USA）。版权所有。
This paper describes a rapid and simple technique for the simultaneous quantitative analysis of PGE(2), PGE(3), and other closely related prostaglandins from cultured cells using liquid chromatography/electrospray ionization tandem mass spectrometry. This method permits quantification of selected individual prostaglandins derived either from arachidonic acid (AA) or eicosapentaenoic acid (EPA) from cell extracts without tedious derivatization, lengthy sample preparation, and separation required by GC-MS- or HPLC-UV-based methods. The validation assessment showed that the quantitative determination is linear (r(2) > 0.999) for both PGE(2) and PGE(3) in the range tested (1-500 ng/ml, 0.0028-1.4 muM) and a coefficient of variation lower than 10% was obtained for samples analyzed on 3 separate days. The detection limit was 2.5 pg for both PGE(2) and PGE(3). Extraction efficiency of PGE(2) and PGE(3) from cell suspensions ranged from 89.4 to 98.2%. As an application of the method, prostaglandins formed by EPA in human lung cancer A549 cells were determined. A 62% reduction of PGE(2) formation was noted when A549 cells were treated with 100 muM of EPA. Concomitantly, EPA increased formation of PGE(3) by 10-fold in A549 cells. This is the first report that unequivocally demonstrates that EPA can be converted to PGE(3) by cyclooxygenase in human cancer cells. (C) 2002 Elsevier Science (USA). All rights reserved.