论文部分内容阅读
本研究开发了一种简单可靠的HPLC-UV方法用于美氟尼酮的测定.生物分析步骤包括从500 μL肝微粒体系统中通过甲醇沉淀蛋白质提取美氟尼酮.色谱条件:色谱柱为Agilent TC-C18柱(4.6 mm×250 mm,5μm),流动相为10mM甲酸铵(用10％的甲酸调PH至2.9)-乙腈(70:30,v/v),流速为1.0 mL/min,UV检测波长设定在245 nm.美氟尼酮和吡非尼酮(内标物)分别在6.0和9.7分钟洗脱,总运行时间为12分钟.根据美国食品和药物管理局生物分析指南,进行了方法验证,结果符合验收标准.美氟尼酮在肝微粒体中的标准曲线在0.5-16 μg/mL范围内呈线性关系.美氟尼酮内和外间精确度低于9.0％,偏差在士10.0％以内.美氟尼酮在肝微粒体中孵育后,该方法成功应用于药代动力学研究.“,”In the present study,a simple and reliable HPLC-UV method was developed for the determination of mefunidone.The bioanalytical specific procedure involved extraction of mefunidone from a 500-μL hepatic microsomal system through protein precipitation by methanol.Chromatographic separation was achieved using an Agilent TC-C18 column (4.6 mm×250 mm,5 μm) with an isocratic mobile phase consisting of 10 mM ammonium formate (pH 2.9,later adjusted by using 10％ formic acid)-acetonitrile (70:30,v/v) at a flow rate of 1.0 mL/min.The UV detection wavelength was set at 245 nm.Mefunidone and pirfenidone (PFD,internal standard,IS) were eluted at 6.0 and 9.7 min,separately,with the total running time of 12 min.According to US Food and Drug Administration bioanalytical guidelines,method validation was performed,and the results met the acceptance criteria in details.The calibration curve of mefunidone in liver microsomes was linear over the concentration range of 0.5-16 μg.mL 1.Intra-and inter-run precisions of mefunidone were less than 9.0％,and the biases were within ±10.0％.After incubation of mefunidone in liver microsomes,this method was successfully applied to the pharmacokinetic study.