title: Determination of 13C/12C ratios of urinary epitestosterone and its main metabolites 5alpha- and 5beta-androstane-3alpha, 17alpha-diol.
abstract: Epitestosterone (17alpha-hydroxy-androst-4-en-3-one, EpiT) belongs to the list of prohibited substances of the World Anti-Doping Agency (WADA). Although it possesses no anabolic effect, it is presumed to be misused by athletes in order to mask administration of testosterone (T) by lowering the urinary T/EpiT ratio.To improve detection, an excretion study with 40 mg of orally administered EpiT was conducted focusing on the metabolites of EpiT: 5alpha- and 5beta-androstane-3alpha,17 alpha-diol (5aEpiD and 5bEpiD). A reference population of n = 74 volunteers was investigated to elucidate the urinary concentrations of these steroids.In order to prove whether an unusual finding in urinary concentrations or ratios is due to an illicit intake of steroids or due to physiological elevation, determination of carbon isotope ratios is advisable. A method for isotope ratio determination was developed to enable (13)C/(12)C ratios of EpiT, 5bEpiD, 5aEpiD, pregnanediol and androsterone and etiocholanolone to be measured from a single urine specimen. The method's validity was tested by applying linear mixing models and specificity was ensured by means of gas chromatography/mass spectrometry analysis. delta(13)C values at natural levels were determined with a reference population and both Delta values and corresponding reference limits were calculated.Considering the implemented EpiT-metabolites, a more than twofold extension of the detection time of EpiT administration was achieved with both the urinary concentration thresholds and the (13)C/(12)C ratios.
authors.affiliation: German Sport University Cologne, Institute of Biochemistry, Am Sportpark Müngersdorf 6, Köln, Germany. t.piper@biochem.dshs-koeln.de
authors.forename: Thomas
authors.initials: T
authors.lastname: Piper
authors.forename: Phillip
authors.initials: P
authors.lastname: Riemann
authors.forename: Georg
authors.initials: G
authors.lastname: Opfermann
authors.forename: Ute
authors.initials: U
authors.lastname: Mareck
authors.forename: Hans
authors.initials: H
authors.lastname: Geyer
authors.forename: Graziela
authors.initials: G
authors.lastname: Vajiala
authors.forename: Ulrich
authors.initials: U
authors.lastname: Flenker
authors.forename: Wilhelm
authors.initials: W
authors.lastname: Schänzer
mesh_terms: D000738:Androsterone / Q000652:urine; D002247:Carbon Isotopes / Q000652:urine*; D004300:Doping in Sports; D004845:Epitestosterone / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000652:urine*; D005043:Etiocholanolone / Q000652:urine; D006801:Humans; D016014:Linear Models; D008297:Male; D011276:Pregnanediol / Q000652:urine; D012016:Reference Values; D015203:Reproducibility of Results; D015813:Substance Abuse Detection / Q000379:methods*
chemical_list: D002247:Carbon Isotopes; D004845:Epitestosterone; D005043:Etiocholanolone; D000738:Androsterone; D011276:Pregnanediol