A rapid ultra-performance liquid chromatography/tandem mass spectrometric methodology for the in vitro analysis of Pooled and Cocktail cytochrome P450 assays

被引：22

作者：

Alden, Peter G. ^{[1
]}

Plumb, Robert S. ^{[1
]}

Jones, Michael D. ^{[1
]}

Rainville, Paul D. ^{[1
]}

Shave, Darcy ^{[1
]}

机构：

[1] Waters Corp, Milford, MA 01757 USA

来源：

RAPID COMMUNICATIONS IN MASS SPECTROMETRY | 2010年 / 24卷 / 01期

关键词：

INHIBITION ASSAYS; GRAPEFRUIT JUICE; QUANTIFICATION;

D O I：

10.1002/rcm.4364

中图分类号：

Q5 [生物化学];

学科分类号：

070307 [化学生物学];

摘要：

Drug-drug interaction evaluations of new pharmaceutical candidates are critical to preventing drug withdrawal and are routinely determined through the use of cytochrome P450 assays. The measurement of the effect of test compounds on the metabolism of known substrates allows for the determination of specific CYP450 isoenzyme inhibition and calculation of IC50 values. A sensitive, high-throughput ultra-performance liquid chromatography/tandem mass spectrometric (UPLC/MS/MS) method is presented for the evaluation of CYP450 inhibition. The assay was performed using a cocktail of probe substrates and the results were compared to those obtained with the more time-consuming methodology utilizing individual substrates. The use of a high-resolution, sub-21 mu m particle, LC system allowed for a high-throughput assay of just 1 min. The extra resolution of the UPLC/MS/MS system allowed for the complete resolution of the analytes, with a fast switching MS for comprehensive data collection. The CYP450 inhibition results obtained using the substrate cocktail approach were found to be essentially identical to those obtained using individual substrates. Copyright (C) 2009 John Wiley & Sons, Ltd.

引用

页码：147 / 154

页数：8

共 18 条

[1]

Ayrton J, 1998, RAPID COMMUN MASS SP, V12, P217, DOI 10.1002/(SICI)1097-0231(19980314)12:5<217::AID-RCM146>3.0.CO

[2]

2-I

[3]

Quantification of a cytochrome P450 3A4 substrate, buspirone, in human plasma by liquid chromatography-tandem mass spectrometry [J].