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

Abstract

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 0.997 8). Conclusion Our LC-MS/MS method can quickly and simultaneously measure the activities of 9 CYP450 enzymes in human liver microsomes.

Keywords

• drug-drug interactions
• cyp450 enzyme
• lc-ms/ms
• cocktail