UPLC-MS/MS法同时检测人血浆中6种抗结核药的浓度  

Simultaneous determination of six antituberculosis drugs in human plasma by UPLC-MS/MS

作　　者：安静[1] 董占军[1] 张笑丹[2] 刘洪涛[1] 

机构地区：[1]河北省人民医院药学部,石家庄050051 [2]河北省胸科医院药学部,石家庄050041

出　　处：《药物分析杂志》2016年第2期208-216,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：河北省医学科学研究重点课题计划"UPLC-MS/MS法检测儿童骨结核患者血中抗结核药的浓度(ZD20140491)"

摘　　要：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(1.8μm,2.1 mm×100 mm)色谱柱进行色谱分离,以乙腈^(-1)5 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 m L·min^(-1),柱温40℃,进样量5μL;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、0.05~5μg·m L^(-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 （ 1.8μm, 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.

关 键 词：抗结核药 乙胺丁醇 异烟肼 吡嗪酰胺 利福平 利福喷丁 利福布汀 超高效液相色谱质谱联用技术 生物样本检测 

分 类 号：R917[医药卫生—药物分析学]