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目的:建立测定人全血中他克莫司液相色谱-串联质谱联用(LC-MS/MS)方法,并与微粒子免疫分析(MEIA)方法测定的结果进行了比较。方法:运用LC-MS/MS方法,血样经乙腈沉淀蛋白,上清液氯气吹干流动相复溶进样。色谱柱:ZorbaxExtend-C_(18)柱(150 mm×4.6 mm,5μm);流动相:甲醇-乙腈-20 mmol·L~(-1)醋酸胺溶液(80:15:5);流速:0.4 ml·m~(-1)。采用电喷雾离子化四极杆串联质谱,多反应检测方式测定样品的浓度。检测离子对分别为m/z 821.7→m/z 768.4和m/z821.7→m/z 786.5。结果:他克莫司在0.5～128 ng·ml~(-1)浓度范围内,峰面积与浓度线性关系良好(r=0.999 1,n=5),最低定量限为0.5 ng·ml~(-1),平均回收率为99.0%,日内精密度、日间精密度的RSD分别为3.38%和4.16%。MEIA分析采用标准IMX测定方法两种方法比较,其标准差小于3.0,表明相关性好。结论:本方法具有良好的灵敏度、准确度、精确度及专属性,可用于他克莫司血药浓度监测和人体药药动力学研究。
OBJECTIVE: To establish a method for the determination of tacrolimus in human whole blood by liquid chromatography-tandem mass spectrometry (LC-MS / MS) and to compare the results with those obtained by the method of microparticle immunoassay (MEIA). Methods: Using LC-MS / MS method, blood samples were precipitated by acetonitrile and the supernatant was mixed with chlorine gas to dry the mobile phase. Column: ZorbaxExtend-C 18 column (150 mm × 4.6 mm, 5 μm); mobile phase: methanol-acetonitrile 20 mmol·L -1 acetic acid amine solution ml · m ~ (-1). Electrospray ionization quadrupole tandem mass spectrometry was used to determine the concentration of the sample. The detected ion pairs were m / z 821.7 → m / z 768.4 and m / z 821.7 → m / z 786.5, respectively. RESULTS: Tacrolimus had a good linear relationship between peak area and concentration (r = 0.999 1, n = 5) and the lowest limit of quantification was 0.5 ng · ml ~ (-1) in the range of 0.5 ~ 128 ng · ml ~ -1). The average recovery was 99.0%. The intraday precision and intraday precision RSD were 3.38% and 4.16%, respectively. MEIA analysis using standard IMX determination of two methods, the standard deviation of less than 3.0, indicating good correlation. Conclusion: The method has good sensitivity, accuracy, accuracy and specificity, which can be used for the monitoring of tacrolimus blood concentration and pharmacokinetics of human body.