UPLC-MS/MS法同时检测人血浆中6种抗结核药的浓度

目的:建立UPLC-MS/MS法用于人血浆中6种抗结核药的同时检测分析,并应用于肺结核患者血药浓度监测。方法:利用蛋白沉淀法将被分析物与生物基质分离,采用Waters ACQUITY UPLC HSS T3(1.8 μm,2.1 mm×100 mm)色谱柱进行色谱分离,以乙腈-15 mmol·L^-1甲酸铵-0.05%甲酸为流动相进行梯度洗脱,流速为0.2 mL·min^-1,柱温40℃,进样量5 μL;采用电喷雾离子源正离子模式,多反应监测进行测定。结果:乙胺丁醇、异烟肼、吡嗪酰胺、利福平、利福喷丁、利福布汀质量浓度分别在0.05~5、0.1~10、0.2~20、0.05~10、0.05~10、0.05~5 μg·mL^-1范围内线性关系良好,相关系数均大于0.99。除吡嗪酰胺在低浓度点的日间精密度小于15.4%外,其他化合物低、中、高3个浓度的日内和日间精密度RSD均小于15%,准确度为88.1%~109.1%,准确度良好。结论:本法经方法学验证,可同时对6种以上抗结核药物进行检测,一步完成,降低了分析成本,为临床调整用药提供了重要的分析手段。

Objective:To develop an ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for determination of six antituberculosis drugs,and to analyze biological samples from clinical trials.Methods:The analytes were isolated from the biological matrix by protein precipitation.The samples were analyzed with a Waters ACQUITY UPLC HSS T3 column(1.8 μm,2.1 mm×100 mm) and eluted with acetonitrile -15 mmol·L^-1 ammonium formate-0.05% formic acid solution as mobile phase in gradient mode at a flow rate of 0.2 mL·min^-1.The column temperature was controlled at 40℃ and the injection volume was 5 μL.Electrospray ionization(ESI) source was applied and operated in the positive multiple reaction monitoring (MRM) mode.Results:There were good linear relationship between peak areas and injection quality in the range of 0.05-5,0.1-10,0.2-20,0.05-10,0.05-10 and 0.05-5 μg·mL^-1 for ethambutol,isoniazid,pyrazinamide,rifampicin,rifapentine and rifabutine,respectively.The inter day precision of pyrazinamide was 15.4% in low concentration,the intra-day and inter-day precision(RSD) other compounds were all below 15%.The accuracy was 88.1%-109.1%.Conclusion:The established method is proved suitable for the simultaneous determination of ethambutol,isoniazid,pyrazinamide,rifampicin,rifapentine and rifabutine in human plasma from clinical plasma samples.By one step,which will significantly reduce the analysis cost,and provide an analysis means for the clinical medication adjustment.

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