김민규 (제주대학교 약학대학 의약품분석학 연구실)

유준상 (제주대학교 약학대학 의약품분석학 연구실)

김민규 (제주대학교 약학대학 의약품분석학 연구실)

박현민 (제주대학교 약학대학 의약품분석학 연구실)

유혜현 (한양대학교 약학대학)

유준상 (제주대학교 약학대학 의약품분석학 연구실)

Cytochrome P450 (CYP) enzymes play a central role in drug metabolism, and natural products or plant extracts may inhibit or induce CYP activities, potentially causing herb–drug interactions. Therefore, establishing a rapid and reliable system for evaluating CYP inhibition is essential for assessing drug safety and efficacy. In this study, a novel LC-MS/MS-based simultaneous analytical method for nine CYP marker metabolites was established to evaluate the inhibitory effects of natural products and plant extracts on CYP enzymes. The method targeted eight major CYP isoforms in human liver microsomes (HLMs): CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, and CYP3A4. Metoprolol and nifedipine were employed as the marker substrates for CYP2D6 and CYP3A4, respectively, instead of the conventional substrates. In addition, multiple reaction monitoring (MRM) conditions were optimized to achieve high selectivity and sensitivity. The established method enabled the simultaneous evaluation of eight CYP enzyme activities in a single analysis and successfully detected all nine CYP marker metabolites with high selectivity and sensitivity. The established LC-MS/MS-based CYP cocktail assay provides a useful platform for the efficient evaluation of the CYP inhibitory effects of various natural products and plant extracts.​