R. Aguilera et al., Screening urine for exogenous testosterone by isotope ratio mass spectrometric analysis of one pregnanediol and two androstanediols, J CHROMAT B, 727(1-2), 1999, pp. 95-105
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 e
pitestosterone 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 pr
evious studies reveal that carbon isotope ratio helps to determine the orig
in of the urinary T because the values for T and its metabolites decrease a
fter the administration of exogenous T. In this study, we present a rapid a
nd efficient screening sample preparation method based on three successive
liquid-solid extractions, deconjugation with E. coli beta-glucuronidase aft
er the first extraction, acetylation after the second extraction, and a fin
al extraction of the acetates. The C-13/C-12 of two T metabolites (5 beta-a
ndrostane-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 a
lpha-diol) was measured by gas chromatography-combustion-isotope ratio mass
spectrometry (GC-C-IRMS) on 10 mi of urine collected from 10 healthy men b
efore 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 a
ndrostanediols to pregnanediol increased in the post-T urines. (C) 1999 Els
evier Science B.V. All rights reserved.