HPLC法测定生物样品中来氟米特活性代谢产物A77 1726

Determination of the active leflunomide metabolite A77 1726 in biolgical samples by HPLC

目的 建立用于测定生物样品中来氟米特的活性代谢产物A77172 6的HPLC方法 ,进行药代动力学研究。方法 分析柱为SpherecloneODS柱 (15 0× 4 .6mm ,5 μm) ,流动相为乙酸钠缓冲液 -乙腈 (6 3∶37) ,紫外检测波长 2 90nm。血和肝匀浆样品用乙酸乙酯提取。结果 方法的线性范围为 0 .1～ 10 0 μg·ml-1(血 )和 0 .0 0 1～ 0 .2 5 μg·g-1(肝 ) ,r分别为 0 .9994 5和 0 .99998,检测下限 0 .1μg·ml-1和 0 .0 0 1μg·g-1。A77172 6在血和肝匀浆中的回收率 (% )分别为 98.4 2± 3.4 4和 94 .5 0± 2 .5 3;日间日内RSD 均小于 9.5 7%。结论 本方法各项指标均符合新药报批要求 。

Aim To study the pharmacokinetics of leflunomide,a HPLC method was developed for the Quantitative analysis of A77 1726 in biolgical samples.Methods The method used sphereclone ODS column (150×4.6mm,5μm) as analytical column and acetate buffer-acetonitrile (63∶37) as mobile phase,with 290nm of UV detect wavelegth.The A77 1726 in whole blood and liver homgenate were extracted with ethyl acetate.Results The calibration curves were linear within the rage of 0.1～100μg·ml -1 (whole blood) and 0.001～0.25μg·g -1 (liver homqenate),and the detection limits were 0.1μg·ml -1 and 0.001μg·g -1,respectively.The mean recoveries in whole blood and liver homgenate were (98.42±3.44)% and (94.50±2.53)%,and the RSD for the inter and intra-day were all less than 9.57%.Conclusion This method confirms the reqirement of new drug approval,and has been applied to the pharmacokinetics study of leflunomide in animals.