HPLC-MS/MS测定肝移植早期患者全血中他克莫司的浓度

HPLC-MS/MS determination of blood tacrolimus concentrations in early liver transplant recipients

目的:建立液质联用法测定人全血中他克莫司的浓度。方法:采用HPLC-MS/MS法进行分析。血样处理:采用乙腈沉淀蛋白。色谱柱为Waters TDMC18(5μm,10mm×2.1mm)柱,柱温55℃,流动相A为10mmol·L-1醋酸铵-0.5%甲酸水溶液,流动相B为0.5%甲酸甲醇溶液,流速0.25mL.min-1,采用多反应监测进行定量,采用ESI正离子方式进行检测,用于定量分析的监测离子为m/z821.30→768.20(他克莫司)和m/z809.35→756.35(内标子囊霉素)。结果:本方法线性范围为1～30ng·mL-1(r=0.9996),相对回收率在80%～120%之间,日内、日间RSD均小于10%。结论:本方法经济、简便、灵敏,用于他克莫司血药浓度监测,并进行在肝移植早期患者的药代动力学研究。

Objective：To establish an HPLC-MS/MS method for determination of tacrolimus in human blood.Methods：Proteins were precipitated by acetonitrile.The separation was carried out on a Waters TDMC18（5 μm,10 mm×2.1 mm） column using water（containing 10 mmol·L-1 ammonium acetate and 0.5% formic acid） and methanol （containing 0.5% formic acid） as mobile phase at a flow rate of 0.25 mL·min-1,and the column temperature was set at 55 ℃.ESI＋ was performed in the MRM mode using target ions at m/z 821.30→768.20（tacrolimus） and m/z 809.35→756.35（ascomycin,internal standard）.Results：The peak area of tracrolimus in blood showed good linearity in the range of 1-30 ng·mL-1（r=0.9996）.The relative recovery was 80%-120%,the intra-day and inter-day RSDs were 10%.Conclusion：This method is sensitive,simple and accurate.And it can be used for determination of blood tacrolimus concentrations and it＇s pharmacokinetic study in early liver tansplant recipients.