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A rapid ultra‐performance liquid chromatography/tandem mass spectrometric methodology for the in vitro analysis of Pooled and Cocktail cytochrome P450 assays
Author(s) -
Alden Peter G.,
Plumb Robert S.,
Jones Michael D.,
Rainville Paul D.,
Shave Darcy
Publication year - 2009
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.4364
Subject(s) - chemistry , chromatography , analyte , resolution (logic) , tandem , tandem mass spectrometry , cytochrome p450 , liquid chromatography–mass spectrometry , substrate (aquarium) , high performance liquid chromatography , bioanalysis , mass spectrometry , enzyme , biochemistry , materials science , oceanography , artificial intelligence , computer science , composite material , geology
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‐2 µm 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 © 2009 John Wiley & Sons, Ltd.