A comprehensive UHPLC–MS/MS method for the analysis of endogenous and exogenous steroids in serum for anti‐doping purposes

类固醇 内生 睾酮(贴片) 二氢睾酮 化学 表雄酮 尿 雄激素 类固醇激素 合成代谢类固醇 背景(考古学) 内分泌学 内科学 色谱法 激素 药理学 合成代谢 医学 生物化学 生物 古生物学
作者
Tobias Langer,Olivier Salamin,Raul Nicoli,Silke Grabherr,Tiia Kuuranne,Alessandro Musenga
出处
期刊:Drug Testing and Analysis [Wiley]
卷期号:14 (11-12): 1904-1919 被引量:9
标识
DOI:10.1002/dta.3379
摘要

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.
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