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Validation of higher‐throughput high‐performance liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry assays to conduct cytochrome P450s CYP2D6 and CYP3A4 enzyme inhibition studies in human liver microsomes
Author(s) -
Chu Inhou,
Favreau Leonard,
Soares Tony,
Lin Chinchung,
Nomeir Amin A.
Publication year - 2000
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/(sici)1097-0231(20000229)14:4<207::aid-rcm863>3.0.co;2-#
Subject(s) - chemistry , cyp3a4 , chromatography , atmospheric pressure chemical ionization , tandem mass spectrometry , mass spectrometry , microsome , cytochrome p450 , cytochrome , liquid chromatography–mass spectrometry , enzyme , cyp2d6 , chemical ionization , ionization , biochemistry , organic chemistry , ion
In the early stage of drug discovery, thousands of new chemical entities (NCEs) may be screened before a single drug candidate can be identified for development. In order to accelerate the drug discovery process, we have developed higher‐throughput enzyme assays to evaluate the inhibition of cytochrome P450 isoforms 2D6 (CYP2D6) and 3A4 (CYP3A4) in human liver microsomes. The assays are based on high‐performance liquid chromatography/tandem mass spectrometry (LC/MS/MS) techniques. The analysis time for each sample was reduced from ∼20 minutes for the conventional HPLC assay to 30 seconds for the LC/MS/MS assay. For both LC/MS/MS assays, the linearity ( r 2 > 0.99), precision (%CV < 15%) and accuracy (% bias <15%) for both inter‐ and intraday validations were satisfactory. Since the implementation of the LC/MS/MS assays, our sample throughput has increased by over 40‐fold. Copyright © 2000 John Wiley & Sons, Ltd.