目的 建立大鼠肝微粒体酶孵育体系中伊立替康（CPT-11）及其活性代谢产物7-乙基-10羟基喜树碱（SN-38）的HPLC检测方法,并对体外孵育条件进行优化。方法 采用HPLC测定微粒体酶孵育体系中伊立替康及其代谢产物的浓度,用单因素法对各孵育条件进行优化,采用Lineweaver-Burk双倒数法研究羧酸酯酶(CES)的酶促动力学。结果 伊立替康、7-乙基-10羟基喜树碱分别在20~4 000、2~400 ng·mL^-1内线性关系良好。体外酶孵育条件为：伊立替康 10 μmol·L^-1,肝微粒体蛋白含量0.02 mg,孵育时间为15 min。结论 本实验建立的HPLC的方法快捷、灵敏,适用于伊立替康及其代谢产物的测定。体外酶孵育条件的优化,同时也为研究其他多种经羧酸酯酶代谢的药物对伊立替康代谢影响及相互作用的体外研究奠定了基础。

Objective To establish an HPLC method for determing irinotecan (CPT-11) and its metabolite 7-ethyl-10 HCPT in rat liver microsome incubation system,and to optimize the incubation conditions. METHODS CPT-11 and SN-38 were determined by HPLC. Single factor design was used to optimize the incubation conditions. RESULTS The linear range of CPT-11 and 7-ethyl-10 HCPT in rat liver microsome incubation system were 20-4 000 ng·mL^-1 and 2-400 ng·mL^-1,respectively. The optimal incubation conditions were as follows: 10 μmol·L^-1 CPT-11,0.02 mg liver microsomes and incubation for 15 min. CONCLUSION The HPLC method is accurate and suitable for the determination of CPT-11 and 7-ethyl-10 HCPT in rat liver microsomes. The incubation condition can be applied in drug interaction studies of irinotecan.