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目的:建立简便、快速的亲水作用色谱串联质谱法(HILIC-MS/MS)测定人血浆中的拉米夫定。方法:以13C1,15N2(拉米夫定为内标,血浆样品经乙腈沉淀蛋白后,采用Luna HILIC(100 mm×3.0 mm,3μm)柱分离。流动相为乙腈-5 mmol·L-1醋酸铵-甲酸(95∶5∶0.01,v/v/v),进样体积2μL,样品分析时间3 min。采用ESI源正离子模式、多反应监测(MRM),用于定量分析的离子反应分别为m/z230(112(拉米夫定)和m/z233(115(内标)。结果:测定人血浆中拉米夫定的线性范围为8.0～2000 ng·mL-1,定量下限为8.0 ng·mL-1,日内、日间精密度(RSD)均小于9.0%,准确度(RE)在-7.1%~2.7%之间。本法成功应用于健康受试者口服2种拉米夫定片后的生物等效性研究。结论:采用稳定同位素内标的HILIC-MS/MS法更为简便、快捷和准确,适用于人血浆样品中拉米夫定的测定。
Objective: To establish a simple and rapid hydrophilic interaction chromatography-tandem mass spectrometry (HILIC-MS / MS) for the determination of lamivudine in human plasma. Methods: 13C1 and 15N2 (lamivudine) were used as the internal standard, and the plasma samples were separated by acetonitrile and then separated by Luna HILIC (100 mm × 3.0 mm, 3 μm). The mobile phase was acetonitrile-5 mmol·L -1 acetic acid Ammonia-formic acid (95: 5: 0.01, v / v / v), injection volume 2 μL, sample analysis time 3 min. ESI source positive ion mode, multiple reaction monitoring (MRM), ion reaction for quantitative analysis (Lamivudine) and m / z 233 (internal standard) of m / z 230.Results: The linear range of determination of lamivudine in human plasma was 8.0-2000 ng · mL -1, and the lower limit of quantitation was 8.0 ng · mL-1, the intra-day and inter-day precision (RSD) were less than 9.0% and the accuracy (RE) was between -7.1% and 2.7% .The method was successfully applied to orally administrate two kinds of Lamiv The bioequivalence study after fixed-dose determination of HILIC-MS / MS with stable internal standard was more convenient, rapid and accurate.It is suitable for the determination of lamivudine in human plasma samples.