人尿中6种利尿剂的液相色谱-串联质谱联用检测方法研究  

Development and Validation of a LC-MS/MS Method for the Detection of Six Diuretics in Human Urine

作　　者：赵君[1] 董颖[2] 张丽娟[1] 闫宽 马艳华[2] 

机构地区：[1]北京化工大学理学院,北京100029 [2]国家体育总局反兴奋剂中心,北京100029

出　　处：《中国运动医学杂志》2016年第10期947-951,共5页Chinese Journal of Sports Medicine

摘　　要：目的：使用液相色谱-串联质谱联用（LC-MS/MS）技术,为氯拉扎尼（chlorazanil,I）、氯索隆（cicletanine,II）、西氯他宁（chlorxolone,III）、苄氢氯噻嗪（benzylhydrochlorothiazide,IV）、芬喹唑（fenquizone,V）和美替克仑（meticrane,VI）等6种利尿剂建立在人尿中的检测方法并进行方法验证。方法：将尿样用同等体积的5%乙酸铅水溶液沉淀蛋白质并离心后,使用电喷雾电离源的正、负离子切换模式,以多反应监测（MRM）模式进行定性和定量分析。结果：上述利尿剂I-VI的最低检测限（LOD）分别为1、0.1、0.01、1、2和10 ng/m L;线性范围（R2〉0.99）分别为3~300 ng/m L、0.3~500 ng/m L、0.03~4000 ng/m L、3~1500 ng/m L、6~600 ng/m L和30~1600 ng/m L;6种药物在低、中和高三个浓度水平的日内变异系数均小于8%,日间变异系数均小于12%,检测的准确度在80%至120%之间;基质效应均在83%~115%范围内。结论：新建方法具有前处理简单、分析速度快、灵敏度高和重现性好等特点,已应用于我们实验室的兴奋剂常规检测中。Objective To develop and validate a LC-MS/MS method for the detection of 6 diuretics including chlorazanil（I）,cicletanine（II）,chlorxolone（III）,benzylhydrochlorothiazide（IV）,fenquizone（V）and meticrane（VI）. Method An equivalent volume of 5% Pb Ac2 aqueous solution was applied to precipitate the proteins in urine samples. The supernatant was directly introduced into the LC-MS/MS system using electrospray ionization in negative-to-positive switch mode. Results The limit of detections（LODs）for the above described diuretics I-VI were 1,0.1,0.01,1,2 and 10 ng/m L respectively. The assays were linear over the concentration ranges of 3-300 ng/m L（I）,0.03-4000 ng/m L（II）,0.03-500 ng/m L（III）,3-1500 ng/m L（V）,6-600 ng/m L（V）and 30-1600 ng/m L（VI）in human urine respectively. The intra- and inter-day variation coefficients of the method at three concentration levels（low,medium and high）were less than 12%.The method also afforded satisfactory results in terms of specificity,accuracy（80-120%）and matrix effect（83-115%）. Conclusions The established method is precise,sensitive and accurate with minimum sample preparation. It is a fit-for-purpose approach and has been applied in our laboratory for doping-control sample analysis.

关 键 词：液相色谱-串联质谱 利尿剂 方法验证 兴奋剂控制 

分 类 号：R446.12[医药卫生—诊断学] O657.63[医药卫生—临床医学]