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Sub one minute inhibition assays for the major cytochrome P450 enzymes utilizing ultra‐performance liquid chromatography/tandem mass spectrometry
Author(s) -
Rainville Paul D.,
Wheaton Jessalynn P.,
Alden Peter G.,
Plumb Robert S.
Publication year - 2008
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.3465
Subject(s) - chemistry , chromatography , tandem mass spectrometry , mass spectrometry , metabolite , substrate (aquarium) , cytochrome p450 , liquid chromatography–mass spectrometry , tandem , high performance liquid chromatography , enzyme , biochemistry , oceanography , materials science , composite material , geology
Abstract The measurement of cytochrome P450 (CYP450) isoenzyme inhibition is often done during evaluation of new chemical entities in drug discovery. Typical assay protocol consists of multiple CYP450 probe substrates incubated with selected drug candidates and CYP450. Results of the assay, the amount of probe substrate metabolite formed with respect to control, are used to determine the level of interaction. Liquid chromatography utilizing columns packed with sub‐2‐micron particles have been shown to provide up to 8× faster analysis time and 3× increases in sensitivity over traditional high‐performance liquid chromatography (HPLC). The work presented here shows the development of a high‐throughput, sub‐2‐micron particle LC method coupled with tandem quadrupole mass spectrometry for the rapid analysis of six CYP450 probe substrate metabolites in 30 s. Copyright © 2008 John Wiley & Sons, Ltd.