RP-HPLC法体外筛选5α-还原酶抑制剂模型的建立

A model to screen 5α-reductase inhibitors in vitro by RP-HPLC

目的:建立高效液相色谱法体外筛选5α-还原酶抑制剂。方法:5α-还原酶提取液从大鼠肝脏组织中获取。采用反相高效液相色谱法测定酶促反应中底物睾酮浓度在反应前后的变化,以此反映酶活性高低,用于5α-还原酶抑制剂的体外筛选。色谱条件 Diamonsil^(TM) C_(18)柱(250 mm×4.6 mm,5 μm),流动相为甲醇-水-乙酸(70:29.5:0.5),流速1.0 mL·min^(-1),柱温30℃,检测波长230 nm。结果:酶促反应体系中其他组分与睾酮有效分离。5α-还原酶抑制剂爱普列特抑制5α-还原酶活性,并呈现良好的量效关系。结论:方法简便可行,可用于5α-还原酶抑制剂的体外筛选。

Objective：To develop an RP - HPLC model in vitro for screening 5α- reductase inhibitors. Methods： The 5α- reductase was abstracted from the liver of rat. The activity of reductase was determined by the change in testosterone concentration using RP - HPLC. The chromatographic conditions were ： column, DiamonsilTM C18 （250 mm × 4.6 mm,5 μm） ;mobile phase,methanol - water acetic acid（70.29.5： 0.5） ;flow rate,1.0 mL · min^-1 - column temperature ,30 ℃ ;detection wavelength,230 nm. Results ：Testosterone was effectively separated from other component under the chromatographic conditions. Epristeride, one effective 5α-reductase inhibitor, inhibited the activity of 5α- reductase in a dose - dependent manner. Conclusion： The method is simple, accurate and suitable for screening of 5α-reductase inhibitor in vitro.