用液相色谱-串联质谱法检测人尿中三种普坦类兴奋剂控制药物

Determination of Tolvaptan, Conivaptan and Lixivapta in Human Urine by LC-MS/MS for Doping Control

目的：建立使用液相色谱一串联质谱（LC—MS／MS）同时测定尿样中三种普坦类药物（托伐普坦、考尼伐坦和利希普坦）的方法。方法：样品用5％乙酸铅水溶液沉淀蛋白质并离心后．在电喷雾正离子模式下，以多反应监测（MRM）方式采集数据进行定性和定量分析。结果：托伐普坦、考尼伐坦、利希普坦的最低检测限（LOD）分别为15ng／mL、1ng／mL和2ng／mL，线性范围（r2〉0．99）分别为45～3000、3～2000和6，400ng／mL。3种药物在低、中、高3个浓度水平下，日内变异系数均小于8．2％，日间变异系数均小于13．6％．检测的准确度大于90％。结论：该方法具有样品前处理简单、检测快捷、灵敏度高、重现性好、定性定量准确等特点，已用于我们实验室的兴奋剂控制样品常规检测中。

Objective To introduce the development and validation of simultaneous determination of Tolvaptan, Conivaptan and Lixivapta in human urine using liquid chromatography-tandem mass spectrometry approach. Method The proteins in urine samples were precipitated with 5% of aqueous lead acetate and then centrifugated. The obtained supematant was analyzed by electrospray positive ion mode using multiple reaction monitoring （MRM）technique. Result The limits of detection for Tolvaptan,Conivaptan and Lixivaptan were 15,1 ,and 2 ng/mL,respectively. The linear ranges （with r2 〉 0.99） for the substances were 45-3000, 3-2000 and 6-400 ng/mL,respectively. The intra- and inter-day precisions were favorable （RSD〈13.6%） and the analysis bias for the substances is less than 10% at three different concentration levels （low, medium and high）. Conclusion The method is highly precise ,sensitive ,reproducible ,and accurate with minimum sample preparation, and has been employed in our laboratory for routine doping control.