Abushareeda Wadha, Lyris Emmanouil, Kraiem Suhail, Wahaibi Aisha Al, Alyazidi Sameera, Dbes Najib, Lommen Arjen, Nielen Michel, Horvatovich Peter L, Alsayrafi Mohammed, Georgakopoulos Costas
Anti-Doping Lab Qatar, Sports City, P.O. Box. 27775, Doha, Qatar.
Sandoz GmbH, Biochemiestrasse 10, A-6250 Kundl/Tirol, Austria.
J Chromatogr B Analyt Technol Biomed Life Sci. 2017 Sep 15;1063:74-83. doi: 10.1016/j.jchromb.2017.08.019. Epub 2017 Aug 19.
This paper presents the development and validation of a high-resolution full scan (FS) electron impact ionization (EI) gas chromatography coupled to quadrupole Time-of-Flight mass spectrometry (GC/QTOF) platform for screening anabolic androgenic steroids (AAS) in human urine samples. The World Antidoping Agency (WADA) enlists AAS as prohibited doping agents in sports, and our method has been developed to comply with the qualitative specifications of WADA to be applied for the detection of sports antidoping prohibited substances, mainly for AAS. The method also comprises of the quantitative analysis of the WADA's Athlete Biological Passport (ABP) endogenous steroidal parameters. The applied preparation of urine samples includes enzymatic hydrolysis for the cleavage of the Phase II glucuronide conjugates, generic liquid-liquid extraction and trimethylsilyl (TMS) derivatization steps. Tandem mass spectrometry (MS/MS) acquisition was applied on few selected ions to enhance the specificity and sensitivity of GC/TOF signal of few compounds. The full scan high resolution acquisition of analytical signal, for known and unknown TMS derivatives of AAS provides the antidoping system with a new analytical tool for the detection designer drugs and novel metabolites, which prolongs the AAS detection, after electronic data files' reprocessing. The current method is complementary to the respective liquid chromatography coupled to mass spectrometry (LC/MS) methodology widely used to detect prohibited molecules in sport, which cannot be efficiently ionized with atmospheric pressure ionization interface.
本文介绍了一种用于筛查人尿样中合成代谢雄激素类固醇（AAS）的高分辨率全扫描（FS）电子轰击电离（EI）气相色谱-四极杆飞行时间质谱联用（GC/QTOF）平台的开发与验证。世界反兴奋剂机构（WADA）将AAS列为体育运动中的禁用兴奋剂，我们开发的方法符合WADA的定性规范，可用于检测体育反兴奋剂禁用物质，主要针对AAS。该方法还包括对WADA运动员生物护照（ABP）内源性甾体参数的定量分析。尿样的预处理步骤包括酶解II相葡萄糖醛酸结合物、通用液-液萃取和三甲基硅烷基（TMS）衍生化。对少数选定离子采用串联质谱（MS/MS）采集，以提高几种化合物的GC/TOF信号的特异性和灵敏度。对已知和未知的AAS的TMS衍生物进行全扫描高分辨率分析信号采集，为反兴奋剂系统提供了一种检测设计药物和新型代谢物的新分析工具，在对电子数据文件进行重新处理后，可延长AAS的检测时间。目前的方法是对广泛用于检测体育赛事中违禁分子的液相色谱-质谱联用（LC/MS）方法的补充，后者无法通过大气压电离接口有效电离。
