ID-LC-MS/MS法同时测定全血中环孢素A、他克莫司、西罗莫司和依维莫司候选参考方法的建立  

Establishment of a candidate reference measurement procedure for the quantification of cyclosporine A,tacrolimus,sirolimus and everolimus in whole blood by ID-LC-MS/MS

作　　者：曾庆璋 张天娇[1] 龙琪琛 金利子 张莉[1] 张传宝[1] Zeng Qingzhang;Zhang Tianjiao;Long Qichen;Jin Lizi;Zhang Li;Zhang Chuanbao(National Center for Clinical Laboratories,Institute of Geriatric Medicine,Chinese Academy of Medical Sciences,Beijing Hospital/National Center of Gerontology,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100730,China)

机构地区：[1]北京医院,国家老年医学中心,国家卫生健康委临床检验中心,中国医学科学院老年医学研究院,中国医学科学院北京协和医学院,北京100730

出　　处：《中华检验医学杂志》2022年第5期472-477,共6页Chinese Journal of Laboratory Medicine

摘　　要：目的建立基于同位素稀释液相色谱串联质谱法(ID-LC-MS/MS)同时测定人全血中4种免疫抑制剂环孢素A、他克莫司、西罗莫司和依维莫司的候选参考方法。方法分别采用环孢素A、他克莫司、西罗莫司和依维莫司同位素标记物为内标,重量法取样,标准曲线法进行定量;采用蛋白沉淀和固相萃取对制备的全血样本进行前处理,超高液相色谱-三重四级杆质谱检测。对方法的特异性、基质效应、检测限、定量限、批内变异系数(CV)、实验室内CV、加标回收率和不确定度等指标进行评价。结果方法选择性和特异性良好,结构类似物和基质效应均不影响检测结果。环孢素A、他克莫司、西罗莫司和依维莫司的检测限及定量限满足临床需求,批内CV为1.4%~1.8%、1.7%~2.8%、1.3%~3.7%和2.3%~3.2%,实验室内CV为1.8%~2.9%、1.7%~3.8%、2.6%~4.7%和3.5%~4.6%,加标回收率分别为97.9%~100.3%、98.4%~103.1%、99.4%~102.0%和98.3%~99.4%。4个浓度质控品相对扩展不确定度分别为4.2%~4.4%、1.5%~2.4%、4.4%~4.9%和2.2%~2.7%。结论本研究建立了ID-LC-MS/MS测定全血中环孢素A、他克莫司、西罗莫司和依维莫司4种免疫抑制剂的方法,方法准确、精密、快速,有望成为候选参考方法。Objectives To establish a candidate reference measurement procedure based on isotope dilution liquid chromatography-tandem mass spectrometry(ID-LC-MS/MS)for cyclosporin A,tacrolimus,sirolimus,and everolimus measurements in human whole blood.Methods The isotope labeled cyclosporine A,tacrolimus,sirolimus,and everolimus were selected as the internal standards.Samples were accurately weighed while protein precipitation and solid phase extraction were selected for the sample preparation.The standard curve method was applied for quantification.The ultra-high liquid chromatography coupled with triple quadrupole mass spectrometer was used for analysis.The specificity,matrix effect,detection limit,quantification limit,precision,accuracy,and uncertainty of the method were evaluated.Results The method showed good selectivity and specificity.No apparent interferences or matrix effects were found in the target analyte measurements.The detection limits and quantification limits of cyclosporin A,tacrolimus,sirolimus and everolimus met clinical requirements.Intra-batch coefficients of variation(CV)were from 1.4%to 1.8%for CSA,from 1.7%to 2.8%for TAC,from 1.3%to 3.7%for SRL and from 2.3%to 3.2%for EVR,and total CVs were from 1.8%to 2.9%for CSA,from 1.7%to 3.8%for TAC,from 2.6%to 4.7%for SRL and from 3.5%to 4.6%for EVR.The relative recoveries were from 97.9%to 100.3%for CSA,from 98.4%to 103.1%for TAC,from 99.4%to 102.0%for SRL and from 98.3%to 99.4%for EVR,and the relative expanded uncertainties at four concentrations were from 4.2%to 4.4%for CSA,from 1.5%to 2.4%for TAC,from 4.4%to 4.9%for SRL and from 2.2%to 2.7%for EVR.Conclusion A candidate reference measurement procedure for the cyclosporine A,tacrolimus,sirolimus,and everolimus in human whole blood was established by ID-LC-MS/MS.

关 键 词：串联质谱法 环孢素A 他克莫司 西罗莫司 依维莫司 

分 类 号：R969.1[医药卫生—药理学]