Langer Tobias, Salamin Olivier, Nicoli Raul, Grabherr Silke, Kuuranne Tiia, Musenga Alessandro
Swiss Laboratory for Doping Analyses, University Center of Legal Medicine Lausanne-Geneva, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland.
University Center of Legal Medicine Lausanne-Geneva, Lausanne University Hospital, University of Geneva, Geneva, Switzerland.
Drug Test Anal. 2022 Nov;14(11-12):1904-1919. doi: 10.1002/dta.3379. Epub 2022 Oct 21.
In the context of steroid analyses, the use of blood could represent a valuable complement to urine. While the blood steroid profile is currently being established to aid unveiling testosterone (T) doping, this matrix is also well suited for detection of exogenous anabolic steroids and steroid esters. In this study, a method to determine a simplified blood steroid profile in combination with the direct detection of exogenous anabolic steroids and steroid esters using just one serum aliquot was developed to obtain a comprehensive analytical workflow. Following the first chromatographic analysis of endogenous and exogenous steroids, samples were derivatised with Girard's reagent T (GT) to improve the ionisation of steroid esters and re-injected. The quantitative performance for T, androstenedione (A4) and 5α-dihydrotestosterone (DHT) was evaluated and the method was validated for qualitative analysis of exogenous analogues with estimated limits of detection (LOD) between 50 and 500 pg/ml. To demonstrate the applicability of the method, samples collected from a clinical study with an oral administration of testosterone undecanoate (TU) to 19 male volunteers were then analysed. The individual serum steroid profiles with the endogenous markers T, A4 and DHT were established as well as the concentrations of TU. TU was detected in all 19 volunteers up to 24 h, while DHT represented the most promising biomarker in endogenous steroid profile for the detection of oral TU administration. These results showed that the selected approach to combine exogenous and endogenous steroid analysis has the potential to strengthen T doping detection in the future.
在类固醇分析的背景下，血液检测可作为尿液检测的重要补充。目前，血液类固醇谱已用于辅助揭露睾酮（T）兴奋剂使用情况，该样本基质也非常适合检测外源性合成代谢类固醇及其酯类。在本研究中，开发了一种方法，仅用一份血清样本即可同时测定简化的血液类固醇谱，并直接检测外源性合成代谢类固醇及其酯类，从而获得全面的分析流程。在内源性和外源性类固醇的首次色谱分析之后，样本用吉拉德试剂T（GT）进行衍生化处理，以改善类固醇酯的离子化，然后重新进样。评估了T、雄烯二酮（A4）和5α-双氢睾酮（DHT）的定量性能，并对该方法进行了验证，以对外源性类似物进行定性分析，估计检测限（LOD）在50至500 pg/ml之间。为了证明该方法的适用性，随后分析了从一项对19名男性志愿者口服十一酸睾酮（TU）的临床研究中采集的样本。确定了内源性标志物T、A4和DHT的个体血清类固醇谱以及TU的浓度。在所有19名志愿者中，直到24小时都检测到了TU，而DHT是内源性类固醇谱中检测口服TU给药最有前景的生物标志物。这些结果表明，所选的结合外源性和内源性类固醇分析的方法未来有可能加强对T兴奋剂使用的检测。
