液相色谱-串联质谱法测定人血浆中培哚普利及其活性代谢物培哚普利拉

Determination of perindopril and perindoprilat in human plasma by liquid chromatography-tandem mass spectrometry

目的：建立人血浆中培哚普利及其活性代谢物培哚普利拉的高效液相-串联质谱测定法。方法：采用不同的沉淀法对血浆样品进行处理,分别对培哚普利和培哚普利拉进行测定,采用Phenomenex Cu-rosil-PFP柱对样品和内标进行分离,以不同比例的甲醇和醋酸盐缓冲液为流动相,通过电喷雾离子化四级杆串联质谱,以选择反应监测（SRM）正离子方式进行检测。用于定量分析的离子反应分别为m/z369→172.2（培哚普利）和m/z377.0→234.1（内标依那普利）;m/z341→170.2（培哚普利拉）和m/z349.0→206.0（内标依那普利拉）。结果：培哚普利标准曲线范围为0.5～200μg.L-1,定量下限为0.5μg.L-1;培哚普利拉标准曲线范围为0.2～20μg.L-1,定量下限为0.2μg.L-1。原药和代谢物的批内批间精密度（RSD）均〈15%。应用该法测试了18名健康男性受试者口服培哚普利片后血浆中培哚普利和培哚普利拉的浓度。结论：该法灵敏、快速、准确,适用于培哚普利的临床药动学研究。

Objective：To develop a sensitive and specific LC-MS/MS method for the determination of perindopril and its active metabolite-perindoprilat in human plasma.Methods：Perindopril and perindoprilat were assayed respectively.Acetonitrile and perchloric acid were used for protein precipitation in plasma.The samples were subsequently separated using a Phenomenex Curosil-PFP column.The mobile phase consisted of methanol and acetate buffer solution.A tandem mass spectrometer equipped with electrospray ionization source was used as detector and operated in the positive ion mode.Selected reaction monitoring（SRM） using the ion combinations of m/z 369172.2（perindopril） and m/z 377.0234.1（Internal standard：enalapril）;m/z 341170.2（Perindoprilat）and m/z 349.0206.0（Internal standard：enalaprilat） were performed,respectively.Results：The linear calibration curve of perindopril was obtained in the concentration range of 0.5～200μg·L-1.The lower limit of quantification was 0.5μg·L-1.The linear calibration curve for perindoprilat was obtained in the concentration range of 0.2～20μg·L-1 with the lower limit of quantification of 0.2μg·L-1.The inter-and intra-batch precisions（RSD）of three validation runs were both less than 15%.The method was successfully applied to a clinical pharmacokine-tic study for 18 healthy male volunteers.Conclusion：The method was proved to be sensitive,rapid and accurate which can be applied to the clinical pharmacokinetic investigations of perindopril and perindoprilat.