RP-HPLC法测定人血浆中醋酸甲羟孕酮及片剂生物等效性研究

Determination of medroxyprogesterone acetate in human plasma by RP-HPLC and study of bioequivalence

目的 :建立人血浆中醋酸甲羟孕酮的HPLC测定法 ,并研究醋酸甲羟孕酮片剂的生物等效性。方法 :血样用重蒸环己烷提取后 ,进行HPLC分析 ,色谱柱为HypersilODS ( 2 5 0mm× 4.6mm ,5 μm) ,流动相为乙腈 水( 5 3∶47) ,内标为甲地孕酮 ,检测波长为 2 40nm。 2 0例健康志愿者交叉口服供试片和参比片 ,计算主要药代动力学参数及相对生物利用度 ,以判断生物等效性。结果 :在 1～ 16 0ng·mL-1范围内醋酸甲羟孕酮与内标的峰面积比值与浓度线性关系良好 (r =0 .9998) ,最低可定量浓度为 1ng·mL-1,提取回收率为 89.1%～ 92 .9%。以AUC0 -4 8计算的醋酸甲羟孕酮片剂的相对生物利用度为 ( 10 0 .3± 6 .9) %。结论 :本实验建立的分析方法灵敏、准确、简便。统计学结果表明 2种制剂生物等效。

Objective :To develop a RP-HPLC assay for determination of medroxyprogesterone acetate in human plasma and investigate the bioequaivalence in healthy volunteers. Methods :Plasma was extracted by cycle-hexane and separated by a C 18 reversed-phase column using a mobile phase of acetonitrile-water (53∶47).The detection wavelength was 240nm.A randomized crossover design was performed in 20 healthy volunteers.In the two study periods,a single 500mg dose of each tablet was administered to each volunteer. Results :Calibration curves were linear over the range 1～160ng·mL -1 ( r =0.9998).The limit of quantification for medroxyprogesterone acetate in plasma was 1ng·mL -1 .The extraction recovery of medroxyprogesterone acetate from plasma was in the range of 89.1%～ 92.9%. The relative bioavailability of medroxyprogesterone acetate tablets was (100.3±6.9)%. Conclusion :The assay is sensitive, accurate and convenient. The two formulations are bioequivalent.