液相色谱-串联质谱直接进样法测定人尿中的沙丁胺醇

Dilute and shoot Method for the Quantification of Salbutamol in Human Urine through Liquid Chromatography-Tandem Mass Spectrometry

沙丁胺醇（salbutamol）为世界反兴奋剂机构（WADA）颁布的《禁用清单》中的阈值物质,兴奋剂检测工作需要对其进行定量测定。尿样加入内标后用5%醋酸铅溶液沉淀蛋白并稀释尿样,无需其它前处理步骤,直接进样于高效液相-串联质谱仪（HPLC-MS/MS）进行分析,使用ZORBAX SB-aq柱进行分离,使用多反应监测（MRM）模式进行测定,建立了阈值物质沙丁胺醇的定量检测方法并进行了方法验证。尿中定量限为0.02μg/ml,线性范围（r^2〉0.99）为0.02-10μg/ml,实验的日内精密度和日间精密度（CV）分别为2.67%和3.02%,测量不确定度为5.36%。阈值浓度时的定量偏差均小于10%。尿样无需水解和提取,操作简单,结果准确。该方法可应用于常规检测和WADA外部质量评估计划。

Objective To introduce a dilute and shoot method for quantification and validation of salbutamol. Methods The urine samples were diluted with 5% lead acetate solution and then injected their supernatant into HPLC/MS/MS directly. Results After determination of the series of spiked urine samples,the LOQ,linear ranges（r^2〉0.99）,intra- and inter-day precisions（CV）,and measurement uncertainty of salbutamol in urine were respectively 0.02 μg/m L,0.2 -10.0 μg/m L,2.67% and 3.02%,and 5.36%. The analytical biases at threshold concentration were less than 10%. Conclusion This dilute and shoot quantitative method has been successfully applied to the routine analysis and the External Quality Assessment Scheme of WADA in our laboratory.