超高效液相色谱-三重四极杆质谱法同时检测人尿中5种利尿剂

Simultaneous determination of five diuretics in human urine by UPLC-MS/MS

使用超高效液相色谱-串联质谱联用技术（UPLC-MS/MS）建立了人尿中5种利尿剂的检出方法。将尿样用同体积的5%乙酸铅水溶液沉淀蛋白质并离心后,将上清液用多反应监测（MRM）模式进行定性和定量分析。上述利尿剂1~5的检测限（LOD）在0.5~2 ng/m L之间;线性范围分别为3.0~700 ng/m L,1.5~1000 ng/m L,3.0~4000 ng/m L,6.0~5000 ng/m L和6.0~5000 ng/m L,线性良好（R2〉0.99）;在低、中和高3个浓度水平的日内和日间RSD均小于15%。

Azosemide （ 1 ）, Buthiazide （2）, Brinzolamide （ 3 ）, Dorzolamine （4） and Quinethazone （ 5 ） are diuretics which are prohibited by the World Anti-Doping Agency. The objective of this study was to develop a UPLC-MS/MS approach for the detection of 5 diuretics in human urine. An equivalent volume of 5% PbAc2 aqueous solution was applied to precipitate the proteins in urine samples and the mixture was centrifuged. The supernatant was directly introduced into the UPLC-MS/MS system for qualitative and quantitative analysis using MRM mode. The limit of detections （LODs） for the above described diuretics 1-5 were between 0. 5 -2 ng/mL. The assays were linear （rE 〉 0. 99） over the concentration ranges of 3.0 -700 ng/mL, 1.5 - 1000 ng/mL, 3.0 -4000 ng/mL, 6.0 - 5000 ng/mL and 6.0 - 5000 ng/mL, separately. The intra- and inter-day precisions of the method at three concentration levels （low, medium and high） were better than 15%. The established method is precise and sensitive, and possesses broad linear range with minimum sample preparation. It is a fit- for-purpose approach for routine sample analysis in doping-control laboratory.