LC-MS/MS法测定全血中他克莫司浓度及对肝移植患者药动学研究  

Determination of Tacrolimus in Whole Blood by Liquid Chromatographic-Tandem Mass Spectrometry and Its Application in the Pharmacokinetic Study in Liver Transplant Patients

作　　者：刘晓雪[1] 胡珊珊[1] 陈中坚[1] 陈冰[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院药剂科,上海200025

出　　处：《药学与临床研究》2013年第4期329-333,共5页Pharmaceutical and Clinical Research

摘　　要：目的：建立LC-MS／MS法测定人全血中他克莫司浓度。以此法研究中国肝移植患者体内的药动学特征。方法：100μL全血标本采用硫酸锌破裂血细胞，加入乙醚液液萃取，分离有机相，以氮气吹干后流动相复溶进样。色谱柱为AgilentEclipseXDB—C。柱（3．5μm，2．1mmX100mm），流动相为2mmol·L-1乙酸铵水溶液和甲醇（5：95，v／v），流速0．3mL·min-1，采用多反应监测进行定量，ESI正离子方式进行检测，他克莫司与内标子囊霉素用于定量分析的检测离子对分别为m／z821．8—768．6和m／z809．8—756．7。，采用常规监测移植患者的他克莫司标本．比较LC—MS／MS法与MEIA法检测结果。采集19例肝移植患者服用他克莫司后第一周和第三周全血，采用本法测定浓度并计算主要药动学参数。结果：本法线性范围为0．46±92ng·mL-1（r=0．9997），最低检测浓度为0．46ng·mL-1。低、中、高三个浓度的日内和日间相对标准差（RSD）均〈15％，平均提取回收率为（55．46±4．13）％。LC-MS／MS法与MEIA法检测结果具有较好的相关性（r2=0．7701）。肝移植患者第一周及第三周AUC。12分别为（71．3±39．6）ng·h·mL-1和（116．1±62．2）ng·h·mL-1，AUG”。分别为（137．3±90．5）ng·h·mL-1和（183．3±95．5）ng·h·mL-1，G。分别为（9．0±5．2）ng·mLl和（13．5±9．6）ng·mL-1，分别为（2．2±1．2）h和（4．2±2．7）h，t1／2分别为（9．7±3．5）h和（7．0±2．2）h，G1分别为（3．9±2．6）ng·mL-1和（6．4±3．6）ng·mL-1。结论：本研究所建立的方法快速准确、灵敏、专属性强，适用于他克莫司血药浓度监测和人体药动学研究。Objective： To establish an LC-MS/MS method for the determination of tacrolimus in human whole blood and investigate the pharmacokinetic characteristics of tacrolimus in Chinese liver-transplanted patients. Methods： The zinc sulfate was added to 100 μL whole blood to lyse the blood cells, then the sample was extracted by diethyl ether. The organic phase was evaporated to dryness by nitrogen, the residues were redissolved by methanol. The resultant was eluted by water （2 mmol-L-1 ammonium acetate）： methanol （5：95, V/V） through an Agilent Eclipse XDB-C18 （3.5 μm, 2.1 mmxl00 mm） column with a flow rate of 0.3 mL min-1. Electrospray ionization （ESI） source was applied and operated in the positive ion mode. Multiple reaction monitoring （MRM） mode with the transition of m/z 821.8---768.6 and m/z 809.8-- 756.7 was used to quantify tacrolimus and internal standard ascomycin. The consistency of LC-MS/MS and MEIA assay was compared by using conventional TDM samples from transplant patients. The total blood samples of 19 liver transplanted patients receiving tacrolinms were collected and tacrolimus concentrations were determined by the established method at the 1st and 3sd week of therapy, the pharmacokinetic pa- rameters were determined. Results： The assay was linear in the range of 0.46-92 ngmL-1 （r=0.9997）. The lower quantitative limit of tacrolimus in whole blood was 0.46 ng-mL-1 The intra and inter-day RSD were within 15%. The mean extract recoveries were （55.46±4.13）%. The tacrolimus concentrations determined by LC-MS/MS were in good consistency with those by MEIA assay （r2=0.7701）. The pharmacokinetic parame ters of liver-transplanted patients in the 1st and 3rd week were respectively as follows： AUC02, （71.3±39.6） and （116.1±62.2）ng-h.mL-1; AUCo, （137.3±90.5） and （183.3±95.5） ng-h＇mL-1; C, （9.0±5.2） and （13.5± 9.6） ng＇mL-1; To, （2.2±1.2） and （4.2±2.7）htjn, （9.7±3.5） and （7.0±2.2）h; Co, （3.9±2.6） and （6.4±3.6�

关 键 词：LC—MS MS法 他克莫司 药动学 MEIA 

分 类 号：R969.1[医药卫生—药理学]