LC-MS/MS法测定人血浆中非洛地平的浓度

目的: 建立人血浆中非洛地平浓度的LC-MS/MS测定方法,并将其应用于临床试验中非洛地平血药浓度的测定。 方法: 以尼莫地平为内标,人血浆样品经乙醚-正己烷(1∶ 1,v/v)萃取处理后,进行LC-MS/MS分析。采用Hypersil BOS-C₁₈(150 mm×2.1 mm,5 μm)色谱柱,流动相为乙腈-0.1%甲酸水溶液(12∶ 88,v/v);使用电喷雾离子源(ESI),正离子多反应监测(MRM)模式检测,用于定量分析的非洛地平和内标尼莫地平的监测离子分别为m/z 384.1→338.0和m/z 419.0→301.1。 结果: 4.5 min完成1次分析,血浆内源性物质对测定无干扰,非洛地平在0.1038～10.38 μg·L^-1浓度范围内线性关系良好(r²>0.99),定量下限为0.1038 μg·L^-1,批内精密度(RSD)不大于9.6%,批间精密度(RSD)不大于10.4%。非洛地平与内标化合物的平均提取回收率分别为99.4%和103.5%,平均基质效应分别为110.1%和108.4%,且均不存在显著浓度依赖性。 结论: 本文所建立的方法准确快速,灵敏度高,专属性强,可用于临床试验中非洛地平制剂低剂量给药后非洛地平血药浓度的测定。

Objective: To develop an LC-MS/MS method for the determination of felodipine in human plasma, and to apply it to a pharmacokinetic study of felodipine in healthy Chinese volunteers. Methods: Nimodipine was used as the internal standard. Human plasma samples were extracted from plasma with diethyl ether-hexane(1∶ 1,v/v), and the extractives were then separated on a Hypersil BOS-C₁₈(150 mm×2.1 mm, 5 μm)column. The mobile phase consisted of acetonitrile-0.1% formic acid(12∶ 88,v/v). An electrospray ionization(ESI)source was applied and operated in the positive multiple reaction monitoring(MRM)mode. The fragment ion for felodipine was m/z 384.1→338.0, and the fragment ion for internal standard was m/z 419.0→301.1. Results: Each analysis could be completed in 4.5 min. Chromatograms showed no endogenous interfering peaks with blank samples. The linear calibration curve was obtained over the concentration range of 0.1038-10.38 μg·L^-1, and the limit of quantification in plasma was established at 0.1038 μg·L^-1. The intra-and inter-day precisions(RSDs)were less than 10.4%. The average extraction recoveries of felodipine and nimodipine were respectively 99.4% and 103.5%, and the average matrix effects were respectively 110.1% and 108.4%. Conclusion: The developed and validated method is rapid, sensitive, selective and reliable for the determination of felodipine in human plasma. The assay can be applied for the determination of felodipine in human plasma and the pharmacokinetic study.