液相色谱-串联质谱法测定大鼠肝微粒体中咪达普利代谢产物的浓度

Determination of the metabolite of imidapril by LC-MS/MS in the incubation system in vitro

目的建立大鼠肝微粒体酶孵育体系中咪达普利拉的液相色谱-串联质谱(LC-MS/MS)测定方法,并对体外孵育条件进行优化。方法用LC-MS/MS法测定微粒体酶孵育体系中咪达普利活性代谢产物咪达普利拉的浓度,并用单因素法对各孵育条件进行优化,用Lineweaver-Burk双倒数法研究羧酸酯酶(CES)的酶促动力学。结果咪达普利拉在5～1000 ng·mL-1内线性关系良好。咪达普利150μmol.L-1与肝微粒体蛋白含量2.2×10-2mg,孵育时间为15min。结论所建立的LC-MS/MS法快捷、灵敏,适于咪达普利拉浓度的测定。

Objective To establish a LC-MS/MS based method for determining imidaprilat,the metabolite of imidapril and to optimize the incubated conditions for rat liver microsomes.Methods Imidaprilat in the incubation system was determined by LC-MS/MS.Single factor design was used to optimize the incubated conditions,and Lineweaver Burk Plot analysis was conducted to study the kinetics of CES1.Results The linear range of imidaprilat in liver microsome incubation system were 5-1000 ng.mL-1.The optimal incubated conditions were 150 μmol.L-1 imidapril,2.2 × 10-2 mg liver microsomes and 15 min of incubation time.Conclusion This method is accurate and suitable for the determination of Imidapril in rat liver microsomes.