Lu Jianghai, Fernández-Álvarez María, Yang Sheng, He Genye, Xu Youxuan, Aguilera R
National Anti-Doping Laboratory, China Anti-Doping Agency, 1st Anding Road, ChaoYang District, Beijing, 100029, China.
J Mass Spectrom. 2015 Jan;50(1):191-7. doi: 10.1002/jms.3517.
In this study, clostebol metabolic profiles were investigated carefully. Clostebol was administered to one healthy male volunteer. Urinary extracts were analyzed by liquid chromatography quadrupole time-of-flight mass spectrometry (MS) using full scan and targeted MS/MS techniques with accurate mass measurement for the first time. Liquid-liquid extraction and direct injection were applied to processing urine samples. Chromatographic peaks for potential metabolites were found by using the theoretical M-H as target ion in full scan experiment, and their actual deprotonated ions were analyzed in targeted MS/MS mode. Fourteen metabolites were found for clostebol, and nine unreported metabolites (two free ones and seven sulfate conjugates) were identified by MS, and their potential structures were proposed based on fragmentation and metabolism pathways. Four glucuronide conjugates were also first reported. All the metabolites were evaluated in terms of how long they could be detected and S1 (4ξ-chloro-5ξ-androst-3ξ-ol-17-one-3ξ-sulfate) was considered to be the long-term metabolite for clostebol misuse detected up to 25 days by liquid-liquid extraction and 14 days by direct injection analysis after oral administration. Five conjugated metabolites (M2, M5, S2, S6 and S7) could also be the alternative biomarkers for clostebol misuse.
在本研究中，对氯睾酮的代谢谱进行了仔细研究。给一名健康男性志愿者施用了氯睾酮。首次采用全扫描和靶向质谱/质谱技术并进行精确质量测量，通过液相色谱四极杆飞行时间质谱（MS）对尿液提取物进行分析。液-液萃取和直接进样被应用于处理尿液样本。在全扫描实验中，以理论M-H作为目标离子来发现潜在代谢物的色谱峰，并在靶向质谱/质谱模式下分析其实际去质子化离子。发现了氯睾酮的14种代谢物，通过质谱鉴定出9种未报道的代谢物（2种游离型和7种硫酸酯共轭物），并根据裂解和代谢途径提出了它们的潜在结构。还首次报道了4种葡萄糖醛酸共轭物。对所有代谢物的可检测时长进行了评估，S1（4ξ-氯-5ξ-雄甾-3ξ-醇-17-酮-3ξ-硫酸酯）被认为是氯睾酮滥用的长期代谢物，口服给药后通过液-液萃取可检测到25天，通过直接进样分析可检测到14天。5种共轭代谢物（M2、M5、S2、S6和S7）也可能是氯睾酮滥用的替代生物标志物。
