Simultaneous determination of 9 CYP450 enzymes in human liver microsomes using LC-MS/MS

Autor: SHI Zhongbo, ZHAO Dezhang, JIANG Junhao, XIE Baogang, LI Yan, YU Chao
Jazyk: čínština
Rok vydání: 2021
Předmět:
Zdroj: Di-san junyi daxue xuebao, Vol 43, Iss 3, Pp 211-217 (2021)
Druh dokumentu: article
ISSN: 1000-5404
DOI: 10.16016/j.1000-5404.202008230
Popis: Objective To develop a liquid chromatography with tandem mass spectrometry (LC-MS/MS) to simultaneously detect the activities of 9 CYP450 enzymes in human liver microsomes in order to anticipate the possibility of drug-drug interactions. Methods Nine different enzyme substrates were used as probes to incubate with human liver microsomes, with nifedipine as internal standard. All metabolites were eluted by Agilent 1290 series liquid chromatograph and then analyzed under multiple reaction monitoring. The IC50 values obtained by single substrate method and cocktail mixed substrate method were compared. Results Our established method had good specificity, and all the chromatographic peaks were completely separated without interference. The linear relationship was good. The correlation coefficients of 9 metabolites within the range of the standard curve were all greater than 0.99. The accuracy of the method was 85.34%~113.89%, the intra-day relative standard deviation (RSD) was < 12.49%, and the inter-day RSD was < 14.98%, which complied with the relevant standards. The IC50 value of the inhibitor determined by this cocktail method was highly correlated with that of a single probe substrate (R2> 0.997 8). Conclusion Our LC-MS/MS method can quickly and simultaneously measure the activities of 9 CYP450 enzymes in human liver microsomes.
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