| Abstract: | Objective To establish a method for the determination of R-（+） -propranolol and S-（-） -propranolol in the incubation system of liver microsomes by HPLC-MS/MS, and to compare the metabolic characteristics of R-（+） -propranolol and S-（-） -propranolol in rat, dog, monkey and human liver microsomes. Methods R-（+）-propranolol and S-（-）-propranolol were respectively dissolved in NADPH activated liver microsome incubation systems of different species, and acetonitrile was added to terminate the reaction after incubation for different time. The concentrations of R-（+） -propranolol and S-（-） -propranolol in each incubation system were determined by using an electrospray ion source（ESI） with carvedilol as the internal standard and positive ion scanning under multi-reaction monitoring mode（MRM）. The residual percentage, metabolism half-life period in vitro and intrinsic clearance of R-（+） -propranolol and S-（-） -propranolol in incubation systems of different species liver microsomes were calculated based on the mass concentration of propranolol with different configurations at 0 min. Results The linear range of propranolol: 0.05～10.00 μg/mL; and the lower limit of quantitation: 0.05 μg/mL; The RSD% of intra-day and inter-day were less than 13%. In the incubation system of four species liver microsomes, the metabolism of R-（+）-propranolol in rat liver microsomes was fast, followed by monkey liver microsomes, and was slow in dog and human liver microsomes, and the t1/2 values in vitro were as follows: rat < monkey < dog < human. S-（-）-propranolol was metabolized quickly in rat and dog liver microsomes, but slowly in monkey and human liver microsomes, and the t1/2 values in vitro were as follows: rat < dog < monkey < human. In rat, dog and monkey liver microsomes, the t1/2 of R configuration is greater than S configuration, and CLint is smaller than S configuration, but in human liver microsomes, the opposite is true. Using univariate analysis of variance, the in vitro metabolic half-life t1/2 and intrinsic clearance CLint of R -（+） - propranolol and S -（-） - propranolol in four liver microsomes were significantly different（P < 0.05）. Conclusion The established HPLC-MS/MS method is simple, rapid and accurate, and can be used for the determination of propranolol enantiomer mass concentration in liver microsome. The metabolic stability of propranolol with different configurations in rat, dog, monkey and human liver microsomes was significant different. |
