Oh, Hyun-A Lee, Hyunbeom Kim, Donghak Jung, Byung Hwa 2024-01-20T02:03:36Z 2024-01-20T02:03:36Z 2021-09-01 2017-02-15 0003-2697 https://pubs.kist.re.kr/handle/201004/123063 As drug interactions with cytochrome P450 enzymes become increasingly important in the field of drug discovery, a high-throughput screening method for analysing the effects of a drug is needed. We have developed a simple and rapid simultaneous analytical method using a cocktail approach for measuring the activities of seven cytochrome P450 enzymes (CYP1A2, CYP2A6, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and CYP3A4). Human liver microsomes were used as a source for the seven cytochrome P450 enzymes, and a gas chromatography-mass spectrometry (GC-MS) was used for analysing their activities. Kinetic studies and inhibition assays of CYP enzymes were performed using known substrates and inhibitors for validating and comparing the reaction rates and time-dependent activities between methods using each substrate versus a method using a cocktail solution. The optimized cocktail method was successfully applied to evaluate the effects of the decoction of Socheongryong-tang (SCRT) on cytochrome P450 enzymes. Our cocktail method provides a simultaneous high-throughput activity assay using GC-MS for the first time. This method is applicable for analysing the drug interactions of various plant-derived mixtures. (C) 2016 Elsevier Inc. All rights reserved. English ACADEMIC PRESS INC ELSEVIER SCIENCE HUMAN LIVER-MICROSOMES IN-VITRO MASS-SPECTROMETRY DRUG-INTERACTION LIQUID-CHROMATOGRAPHY COCKTAIL APPROACH INHIBITION ASSAY CYP450 ENZYMES LC-MS/MS MEDICINE Development of GC-MS based cytochrome P450 assay for the investigation of multi-herb interaction Article 10.1016/j.ab.2016.12.015 1 ANALYTICAL BIOCHEMISTRY, v.519, pp.71 - 83 ANALYTICAL BIOCHEMISTRY 519 71 83 scie scopus 000392560300011 2-s2.0-85007110960 Biochemical Research Methods Biochemistry & Molecular Biology Chemistry, Analytical Biochemistry & Molecular Biology Chemistry Article HUMAN LIVER-MICROSOMES IN-VITRO MASS-SPECTROMETRY DRUG-INTERACTION LIQUID-CHROMATOGRAPHY COCKTAIL APPROACH INHIBITION ASSAY CYP450 ENZYMES LC-MS/MS MEDICINE GC-MS Human liver microsomes Cytochrome P450 Cocktail approach Socheongryong-tang