UPLC-MS/MS法测定人血浆中左乙拉西坦浓度及其药代动力学研究  

Quantification of levetiracetam in human plasma by ultra performance liquid chromatography-tandem mass spectrometry and its application in a pharmacokinetic study

作　　者：胡岚岚[1] 徐颖[1] 周娟[1] 邹欢[1] 张玥[1] 汤建林[1] 

机构地区：[1]第三军医大学新桥医院临床药理基地,重庆400037

出　　处：《药物分析杂志》2016年第4期655-661,共7页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的：建立专属、快速的超高效液相色谱-串联质谱（UPLC-MS/MS）的分析方法测定人血浆中左乙拉西坦的浓度,进行药代动力学研究。方法：以3-氨基-2-萘甲酸为内标,0.2 m L血浆样品加入280·L甲醇除蛋白后,涡旋提取,取上清液3·L进样。色谱柱：Acquity UPLC BEH C18（2.1 mm×50 mm,1.7·m）,流动相：乙腈和0.01 mol·L-1甲酸铵（p H 3.6）,梯度洗脱,流速0.3 m L·min-1,分析时间4 min;采用电喷雾MS检测,正离子检测模式。结果：左乙拉西坦质量浓度在0.5～128.0·g·m L-1范围内呈线性,日内、日间精密度分别小于13.1%和11.7%,基质效应影响较小,相对回收率在95.9%～106.8%之间。口服左乙拉西坦片剂后达峰浓度和达峰时间分别为18.23·g·m L-1和1.31 h。结论：该方法可用于微量血浆测定,并可应用于成人或新生儿癫痫发作,给予左乙拉西坦后对其临床药动学的研究。Objective：To develop a specific and rapid method for the determination of levetiracetam in plasma using ultra performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）,and study pharmacokinetics of levetiracetam.Methods：A plasma aliquot of 0.2 m L was deproteinized by addition of 280 μL methanol which contained 3-amino-2-naphthoic acid as an internal standard.After centrifugation,3 μL of supernatant was injected into the UPLC-system.Compounds were separated on an Acquity UPLC BEH C18（2.1 mm×50 mm,1.7 μm）column using gradient elution with mobile phase acetonitrile and 0.01 mol·L-1 ammonium formate（p H=3.6）with a flow rate of 0.3 m L·min-1 and a total runtime of 4.0 min.Levetiracetam and the internal standard were detected using positive ion electrospray ionization followed by tandem mass spectrometry（ESI-MS/MS）.Results：The assay allowed quantification of levetiracetam plasma concentrations in the range of 0.5-128 μg·m L-1.Intra-assay accuracy was below 13.11% and inter-assay precision was less than 11.7%.Matrix effects were minor,and the recovery of levetiracetam was between 95.9% and 106.8%.The maximum concentration（Cmax）were 18.23 μg·m L-1,and the time to reach this concentration（Tmax）was 1.31 h.Conclusion：The developed method required less sample volume of plasma.The method has been applied in a clinical pharmacokinetic study in which adult/neonates received levetiracetam for the treatment of seizures.

关 键 词：左乙拉西坦 抗癫痫药物 超高效液相色谱-串联质谱 药代动力学 血浆样品 

分 类 号：R917[医药卫生—药物分析学]