Development and validation of an on-line two-dimensional reversed-phase liquid chromatography-tandem mass spectrometry method for the simultaneous determination of prostaglandins E2 and F2α and 13, 14-dihydro-15-keto prostaglandin F2α levels in human plasma
Development and validation of an on-line two-dimensional reversed-phase liquid chromatography-tandem mass spectrometry method for the simultaneous determination of prostaglandins E2 and F2α and 13, 14-dihydro-15-keto prostaglandin F2α levels in human plasma
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DOI：
10.1002/bmc.1117
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发表时间：
2009-03-01
影响因子：
1.8
通讯作者：
Ogawa, Mikio
中科院分区：
文献类型：
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作者：
Komaba, Junji;Matsuda, Dai;Ogawa, Mikio
我们开发并验证了一种在线反相二维LC/MS/MS（2D-LC/MS/MS）系统，用于同时测定人血浆中前列腺素（PG）E-2以及PGF（2 α）及其代谢产物13，14-二氢-15-酮PGF（2 α）（F-2 α-M）的水平。通过三步固相萃取法提取分析物。然后通过在线2D-LC/MS/MS以负离子模式电喷雾电离分析样品。2D-LC系统由两个反相分析柱和连接两个分析柱的捕集柱组成。虽然酸性缓冲液用于两个分离维度，但每个维度使用不同的有机溶剂：第一个维度使用甲醇，第二个维度使用乙腈，以提高分离能力。2D-LC/MS/MS方法具有高度选择性和灵敏度，定量限显著较低（PGE（2）为0.5 pg/mL，PGF（2 α）和F-2 α-M分别为2.5 pg/mL）。证明了2D-LC/MS/MS系统的线性，PGE（2）的校准范围分别为0.5-50 pg/mL，PGF（2 α）和F-2 α-M的校准范围分别为2.5-500 pg/mL。在整个校准曲线范围内获得了可接受的精密度和准确度。这种高选择性和灵敏度的方法被成功地用于测定6名（4名男性和2名女性）正常志愿者血浆样本中PGE（2）、PGF（2 α）和F-2 α-M的内源性水平。每种分析物的平均浓度分别为0.755 pg/mL（PGE（2））、5.70 pg/mL（PGF（2 α））和9.48 pg/mL（F-2 α-M）。版权所有（C）2008约翰威利父子有限公司
We developed and validated an on-line reverse-phase two-dimensional LC/MS/MS (2D-LC/MS/MS) system for simultaneous determination of the levels of prostaglandin (PG) E-2 as well as PGF(2 alpha) and its metabolite 13,14-dihydro-15-keto PGF(2 alpha) (F-2 alpha-M) in human plasma. Analytes were extracted by a three-step solid-phase extraction. Samples were then analyzed by on-line 2D-LC/MS/MS with electrospray ionization in negative mode. The 2D-LC system is composed of two reverse-phase analytical columns with a trapping column linking the two analytical columns. While an acidic buffer was used for both separation dimensions, differing organic solvents were employed for each dimension: methanol for the first and acetonitrile for the second to increase resolving power. The 2D-LC/MS/MS method was highly selective and sensitive with a significantly lower limit of quantitation (0.5 pg/mL for PGE(2) and 2.5 pg/mL for PGF(2 alpha) and F-2 alpha-M, respectively). Linearity of the 2D-LC/MS/MS system was demonstrated for the calibration ranges of 0.5-50 pg/mL for PGE(2) and 2.5-500 pg/mL for PGF(2 alpha) and F-2 alpha-M, respectively. Acceptable precision and accuracy were obtained throughout the calibration curve ranges. This highly selective and sensitive method was successfully utilized to determine the endogenous levels of PGE(2), PGF(2 alpha), and F-2 alpha-M in plasma samples from six (four male and two female) normal volunteers. The mean concentrations for each analyte were 0.755 pg/mL for PGE(2), 5.70 pg/mL for PGF(2 alpha) and 9.48 pg/mL for F-2 alpha-M. Copyright (C) 2008 John Wiley & Sons, Ltd.