期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 405, 期 16, 页码 5455-5466出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-013-6949-3
关键词
Reference population; Carbon isotope ratio; Hydrogen isotope ratio; Doping control; Steroid metabolism
资金
- Federal Ministry of the Interior of the Federal Republic of Germany
- Manfred Donike Institute for Doping Analysis, Cologne
Carbon isotope ratio (CIR) analysis has been routinely and successfully applied to doping control analysis for many years to uncover the misuse of endogenous steroids such as testosterone. Over the years, several challenges and limitations of this approach became apparent, e.g., the influence of inadequate chromatographic separation on CIR values or the emergence of steroid preparations comprising identical CIRs as endogenous steroids. While the latter has been addressed recently by the implementation of hydrogen isotope ratios (HIR), an improved sample preparation for CIR avoiding co-eluting compounds is presented herein together with newly established reference values of those endogenous steroids being relevant for doping controls. From the fraction of glucuronidated steroids 5 beta-pregnane-3 alpha,20 alpha-diol, 5 alpha-androst-16-en-3 alpha-ol, 3 alpha-Hydroxy-5 beta-androstane-11,17-dione, 3 alpha-hydroxy-5 alpha-androstan-17-one (ANDRO), 3 alpha-hydroxy-5 beta-androstan-17-one (ETIO), 3 beta-hydroxy-androst-5-en-17-one (DHEA), 5 alpha- and 5 beta-androstane-3 alpha,17 beta-diol (5aDIOL and 5bDIOL), 17 beta-hydroxy-androst-4-en-3-one and 17 alpha-hydroxy-androst-4-en-3-one were included. In addition, sulfate conjugates of ANDRO, ETIO, DHEA, 3 beta-hydroxy-5 alpha-androstan-17-one plus 17 alpha- and androst-5-ene-3 beta,17 beta-diol were considered and analyzed after acidic solvolysis. The results obtained for the reference population encompassing n = 67 males and females confirmed earlier findings regarding factors influencing endogenous CIR. Variations in sample preparation influenced CIR measurements especially for 5aDIOL and 5bDIOL, the most valuable steroidal analytes for the detection of testosterone misuse. Earlier investigations on the HIR of the same reference population enabled the evaluation of combined measurements of CIR and HIR and its usefulness regarding both steroid metabolism studies and doping control analysis. The combination of both stable isotopes would allow for lower reference limits providing the same statistical power and certainty to distinguish between the endo- or exogenous origin of a urinary steroid.
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