亲水作用色谱串联质谱法测定人血浆中的拉米夫定

<p><b>目的: </b>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。 <b>方法: </b>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&times;3.0 mm,3 &mu;m)柱分离。流动相为乙腈-5 mmol&middot;L^-1醋酸铵-甲酸(95∶ 5∶ 0.01,<em>v/v/v</em>),进样体积2 &mu;L,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<em>m/z</em> 230 (112(拉米夫定)和<em>m/z</em> 233 (115(内标)。 <b>结果: </b>测定人血浆中拉米夫定的线性范围为8.0~2000 ng&middot;mL^-1,定量下限为8.0 ng&middot;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。 <b>结论: </b>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。</p>

<p><b>Objective: </b>To develop and validate a hydrophilic interaction chromatography-tandem mass spectrometric (HILIC-MS/MS) method for the determination of lamivudine in human plasma. <b>Method: </b>The plasma sample was protein precipitated with acetonitrile using the stable labelled ¹³C₁,¹⁵N₂-lamivudine as the internal standard.After sample preparation,the analyte and the IS were separated on a Luna HILIC(100 mm&times;3.0 mm,3 &mu;m)column using 2 &mu;L injection volume with a run time of 3.0 min.The mobile phase consisited of acetonitrile-5 mmol&middot;L^-1 ammonium acetate-formic acid (95∶ 5∶ 0.01,<em>v/v/v</em>).Detection was performed by tandem mass spectrometry using an electrosprasy source (ESI) in the positive ion mode,operating in the multiple reaction monitoring (MRM) of the transitions of <em>m/z</em> 230&rarr;112 and <em>m/z</em> 233&rarr;115 for lamivudine and the IS,respectively. <b>Results: </b>The linear calibration curves were obtained in the concentration range of 8.0-2000 ng&middot;mL^-1.The lower limit of quantification was 8.0 ng&middot;mL^-1.The intra-day and inter-day relative standard deviation over the entire concentration range was less than 9.0%.The accuracy was in the range of-7.1% to 2.7% in terms of relative error.The method was applied to evaluate the bioequivalence of two lamivudine tablet formulations in healthy volunteers. <b>Conclusion: </b>This method is simple,selective,rapid and suitable for the determination of lamivudine in human plasma.</p>