Thevis Mario, Guddat Sven, Schänzer Wilhelm
Center for Preventive Doping Research, Institute of Biochemistry, German Sport University Cologne, Am Sportpark Müngersdorf 6, 50933 Cologne, Germany.
Steroids. 2009 Mar;74(3):315-21. doi: 10.1016/j.steroids.2008.10.004. Epub 2008 Oct 21.
Trenbolone (17beta-hydroxy-estra-4,9,11-trien-3-one) and its derivatives such as 17alpha-methyltrenbolone represent a class of highly potent anabolic-androgenic steroids, which are prohibited in sports according to the regulation of the World Anti-Doping Agency (WADA). Due to marginal gas chromatographic properties of these compounds but excellent proton affinities resulting from a large and conjugated pi-electron system, liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been the method of choice for the detection of these analytes in sports drug testing. Recent findings of trenbolone and methyltrenbolone in doping control urine samples of elite athletes demonstrated the importance of a sensitive and robust analytical method, which was based on an enzymatic hydrolysis of target compounds, liquid-liquid extraction and subsequent LC-MS/MS measurement. Diagnostic product ions obtained after collision-induced dissociation of protonated molecules were found at m/z 227, 211, 199 and 198, which enabled targeted screening using multiple reaction monitoring. Using 7 model compounds (trenbolone, epitrenbolone, methyltrenbolone, ethyltrenbolone, propyltrenbolone, 17-ketotrenbolone and altrenogest), the established method was validated for specificity, lower limits of detection (0.3-3ng/mL), recovery (72-105%), intraday and interday precision (< or =20%).
群勃龙（17β-羟基-4,9,11-三烯-3-酮）及其衍生物，如17α-甲基群勃龙，属于一类高效的合成代谢雄激素类固醇，根据世界反兴奋剂机构（WADA）的规定，这些物质在体育运动中是被禁止的。由于这些化合物的气相色谱性质不佳，但因其大的共轭π电子体系而具有出色的质子亲和力，液相色谱-串联质谱法（LC-MS/MS）已成为体育药物检测中检测这些分析物的首选方法。最近在精英运动员的兴奋剂检测尿样中发现群勃龙和甲基群勃龙，这表明了一种灵敏且稳健的分析方法的重要性，该方法基于目标化合物的酶水解、液液萃取以及随后的LC-MS/MS测量。质子化分子经碰撞诱导解离后获得的诊断产物离子在m/z 227、211、199和198处被发现，这使得能够使用多反应监测进行靶向筛查。使用7种模型化合物（群勃龙、表群勃龙、甲基群勃龙、乙基群勃龙、丙基群勃龙、17-酮群勃龙和烯丙孕素），对所建立的方法进行了特异性、检测下限（0.3 - 3 ng/mL）、回收率（72 - 105%）、日内和日间精密度（≤20%）的验证。
