摘要
目的:建立测定临床前大鼠药物代谢酶2D6亚型活性的 HPLC 方法。方法:HPLC 法测定大鼠尿液及肝微粒体中探针药物右美沙芬(DM)、其代谢物去甲右美沙芬(DX)及内标物丁丙诺啡(BP)的含量。采用 Hypersil-C_6H_5柱(250mm×4.6mm,5μm),以0.01mol-L^(-1)磷酸二氢钾-0.01mol·L^(-1)已烷磺酸钠-乙腈-甲醇(33:33:80:54,v/v,pH4.0)为流动相,流速1.0mL·min^(-1),荧光检测器激发波长280nm,发射波长310nm。结果:本方法稳定性良好,尿样 DM 在0.04~1.20mg·L^(-1)内线性良好(r=0.9981),DX 在3.12~100.0mg·L^(-1)内线性良好(r=0.9971)。肝微粒体重组样品 DM 在30.0~180.0mmol·L^(-1)内线性良好(r=0.9980),DX 在5.0~50.0mg·L^(-1)内线性良好(r=0.9976),日内、日间样品 RSD 均小于10%,样品-20℃保存30d 后测定稳定良好,体外肝微粒体实验 RSD 小于10%。结论:本方法稳定、准确,可用于新药临床前药物代谢酶途径及药物相互作用研究。
Objective:To establish HPLC methods for determination of CYP2D6 activity in rat urine and liver microsomes for preclinical research. Methods: probe drug dextromethorphan ( DM ), its metabolite dextrophan ( DX ) and internal standard buprenorphin(BP) were determined by HPLC in rat urine and liver microsomal reconstituted system to estimate activity of CYP2D6 by using Hypersil - C6 H5 analysis column (250 mm ×4. 6 mm,5 μm ). The mobile phase of 0. 01mol·L^-1 potassium dihydrogen phosphate, sodium hexane sulfonate, acetonitrile, methanol (33: 33: 80: 54, V: V,pH4.0)was pumped at 1.0 mL·min^- 1 and the fluorescence detector was set at λex280 nm,λem310 nm. Results :The methods were stable and linear in range of 0. 04 mg · L^-1 - 1.2 mg · L^-1for DM (r =0. 9981 ) ,3.12 mg· L^-1 - 100. 0 mg · L^-1for DX( r =0. 9971 ) in urine samples;30. 0 mg · L^-1 - 180. 0 mg · L^ - 1 for DM ( r = 0. 9980 ), 5.0 mg·L^- 1 - 50.0 mg·L^-1 for DX ( r = 0. 9976 ) in liver microsomes reconstiction samples respectively. The average recovery of within - day and between day for DM and DX were less than 10%. Samples kept in -20 ℃ for 30 days were stable for HPLC analysis. Conclusion The methods were stable and accurate for study on metabolite ways of new drug preclinical study and drug interactions.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2007年第7期1055-1058,共4页
Chinese Journal of Pharmaceutical Analysis