液相色谱串联质谱法检测马尿中32种蛋白同化激素

Detection of 32 Anabolic Steroids in Horse Urine by LC-MS/MS

目的：建立马尿中32种蛋白同化激素的液相色谱串联质谱（LC-MS/MS）检测方法。方法：利用Agilent 6410A LC-MS/MS仪器;色谱柱采用Agilent Zorbax XDB-C18柱（2.1×100 mm,3.5μm）;流动相采用0.1%甲酸水溶液（A）和乙腈（B）进行梯度洗脱;离子检测方式为多反应离子检测（MRM）;正离子检测。结果：建立了马尿中32种蛋白同化激素的检测方法,药物浓度线性方程的相关系数均大于0.992,日内和日间精密度基本小于20%,提取回收率在84%以上且基质效应多在80%~120%之间,LOD和LOQ满足相关马术检测机构的要求。结论：本方法操作简单,经方法学验证符合相关标准,能满足马术项目的兴奋剂检查要求,可以应用于常规的兴奋剂检测。

Objective To develop a method for detection of 32 anabolic steroids in horse urine. Methods An Agilent 64 liquid chromatography tandem mass spectrometry（LC-MS/MS） instrument and a C18 column（2.1×50mm,3.5μm） were used. The mobile phase was a mixture of acetonitrile and the solution（0.1% formic acid） at a flow rate of 0.2m L/min. The mass spectrometer for the detection of the steroids was operated in the positive electrospray ionisation mode with multiple reaction monitoring. Results Horse urine samples mixed with 32 anabolic steroids at low ng/m L levels were detected. The correlation coefficient of linear equation were all higher than 0.992. The inter-day and intra-day RSD were mostly lower than 20% and the recovery rate of the extraction were mostly higher than 84%. The matrix effect of this method mostly ranged from 80%to 120%. Conclusions The developed analytical method could be used as routine screening of 32 anabolic steroids in horse urine. 【Key Words】：