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目的 建立人肝微粒体中西尼地平及其脱氢代谢物的 HPLC测定法 ,并用本法研究西尼地平在人肝微粒体中代谢的动力学。方法 色谱柱为 Hypersil C1 8柱 (2 5 0 mm× 4.6mm ID,5μm) ,流动相为乙腈 -甲醇 -0 .0 1 mol/ L四丁基溴化铵溶液 (5 5∶ 5∶ 40 ,V/ V/ V) ,柱温 5 0℃ ,检测波长 UV 2 3 8nm,样品用乙醚 -正己烷 (1∶ 1 ,V/ V)提取。用人肝微粒体研究西尼地平的代谢。结果 本法的回收率为 90 .62 %～ 1 0 9.90 % ,西尼地平和其脱氢代谢物的日间、日内 RSD分别为 4.70 %～ 8.3 5 %和 3 .88%～ 8.1 1 % ,西尼地平及其脱氢代谢物的浓度分别在 0 .77μg/ ml～ 98.2 4μg/ ml和 0 .1 3 μg/ ml～ 5 .3 6μg/ ml范围内与峰面积呈良好的线性相关性。在温孵时间为1 0～ 60 min,微粒体蛋白浓度为 1 mg/ ml时 ,西尼地平呈线性消除 ,而其脱氢代谢物呈线性增加。结论 西尼地平在人肝微粒体内被迅速代谢 ,人肝 P45 0酶参与了西尼地平的脱氢氧化
Objective To establish an HPLC method for the determination of cilnidipine and its metabolites in human liver microsomes and to study the kinetics of cilnidipine metabolism in human liver microsomes by this method. Methods The chromatographic column was Hypersil C1 8 column (250 mm × 4.6 mm ID, 5 μm) with a mobile phase of acetonitrile-methanol-0.1 mol / L tetrabutylammonium bromide (5:5:40, V / V / V). The column temperature was 50 ℃ and the detection wavelength was UV 2 38 nm. The sample was extracted with ether-n-hexane (1: 1, V / V). Study of cilnidipine metabolism using human liver microsomes. Results The recoveries of this method ranged from 90.62% to 109.90%. The daytime and day RSD of cilnidipine and its dehydrogenation metabolites were 4.70% ~ 8.35% and 3.88% ~ 8.11%, respectively. The concentration of cilnidipine and its dehydrogenation metabolites showed a good linear correlation with the peak area in the range of 0 .77μg / ml ~ 98.2 4μg / ml and 0.13μg / ml ~ 5.36μg / ml respectively. When incubation temperature was 10-60 min and microsomal protein concentration was 1 mg / ml, cilnidipine was linearly eliminated and its dehydrogenation metabolites increased linearly. Conclusion Cilnidipine is rapidly metabolized in human liver microsomes and human liver P45 0 enzyme is involved in the dehydrogenation of cilnidipine