Screening urine for exogenous testosterone by isotope ratio mass spectrometric analysis of one pregnanediol and two androstanediols
Screening urine for exogenous testosterone by isotope ratio mass spectrometric analysis of one pregnanediol and two androstanediols
复制标题
DOI：
10.1016/s0378-4347(99)00066-3
复制
发表时间：
1999-04-30
期刊：
影响因子：
--
通讯作者：
Hatton, CK
中科院分区：
文献类型：
--
作者：
Aguilera, R;Catlin, DH;Hatton, CK
提出了一种新的检测运动中睾酮（T）兴奋剂的方法。目前检测T给药的方法是基于发现尿液中T与表阿霉素的比值（T/E）超过6。T/E的困难在于T给药并不总是导致T/E>6，并且在没有T给药的情况下，罕见的个体将具有T/E>6。我们以前的研究表明，碳同位素比值有助于确定尿T的来源，因为外源性T给药后，T及其代谢产物的值降低。本研究提出了一种快速、高效的筛选样品制备方法，该方法基于三次连续的液-固萃取、E.第一次提取后的大肠杆菌β-葡萄糖醛酸酶，第二次提取后的乙酰化，和乙酸酯的最终提取。采用气相色谱-燃烧-同位素比值质谱法（GC-C-IRMS）测定了10名健康男性服用T前后10 ml尿液中两种T代谢产物（5 β-雄甾烷-3 α，17 β-二醇和5 α-雄甾烷-3 α，17 β-二醇）和一种孕甾烷（5 β-雄甾烷-3 α，20 α-二醇）的C-13/C-12。在T施用后，5 β-雄甾烷-3 α，17 β-二醇二乙酸酯和5 α-雄甾烷-3 α，17 β-二醇二乙酸酯的C-13/C-12从-26.2份/千份显著下降至-30.8份/千份，从-25.2份/千份显著下降至-29.9份/千份，而5 β-胆甾烷-3 α，20 α-二醇二乙酸酯的C-13/C-12没有变化。此外，T后尿中雄甾烷二醇与孕甾烷的比例增加。(C)1999 Elsevier Science B. V.保留所有权利。
We propose a new screening method for testosterone (T) doping in sport. The current method for detecting T administration is based on finding a T to epitestosterone ratio (T/E) in urine that exceeds six. The difficulties with T/E are that T administration does not always result in a T/E>6 and that a rare individual will have T/E>6 in the absence of T administration. Our previous studies reveal that carbon isotope ratio helps to determine the origin of the urinary T because the values for T and its metabolites decrease after the administration of exogenous T. In this study, we present a rapid and efficient screening sample preparation method based on three successive liquid-solid extractions, deconjugation with E. coli beta-glucuronidase after the first extraction, acetylation after the second extraction, and a final extraction of the acetates. The C-13/C-12 of two T metabolites (5 beta-androstane-3 alpha,17 beta-diol and 5 alpha-androstane-3 alpha,17 beta-diol) and one pregnanediol as endogenous reference (5 beta-pregnane-3 alpha,20 alpha-diol) was measured by gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) on 10 mi of urine collected from 10 healthy men before and after T administration. Following T administration, the C-13/C-12 Of 5 beta-androstane-3 alpha,17 beta-diol diacetate and 5 alpha-androstane-3 alpha,17 beta-diol diacetate declined significantly from -26.2 parts per thousand to -30.8 parts per thousand and from -25.2 parts per thousand to -29.9 parts per thousand, respectively and the C-13/C-12 of 5 beta-pregnane-3 alpha,20 alpha-diol diacetate was unchanged. In addition, the ratio of androstanediols to pregnanediol increased in the post-T urines. (C) 1999 Elsevier Science B.V. All rights reserved.