title: Metabolism of androsta-1,4,6-triene-3,17-dione and detection by gas chromatography/mass spectrometry in doping control.
abstract: The urinary metabolism of the irreversible aromatase inhibitor androsta-1,4,6-triene-3,17-dione was investigated. It is mainly excreted unchanged and as its 17beta-hydroxy analogue. For confirmation, 17beta-hydroxyandrosta-1,4,6-trien-3-one was synthesized and characterized by nuclear magnetic resonance (NMR) in addition to the parent compound. In addition, several reduced metabolites were detected in the post-administration urines, namely 17beta-hydroxyandrosta-1,4-dien-3-one (boldenone), 17beta-hydroxy-5beta-androst-1-en-3-one (boldenone metabolite), 17beta-hydroxyandrosta-4,6-dien-3-one, and androsta-4,6-diene-3,17-dione. The identification was performed by comparison of the metabolites with reference material utilizing gas chromatography/mass spectrometry (GC/MS) of the underivatized compounds and GC/MS and GC/tandem mass spectrometry (MS/MS) of their trimethylsilyl (TMS) derivatives. Alterations in the steroid profile were also observed, most obviously in the androsterone/testosterone ratio. Even if not explicitly listed, androsta-1,4,6-triene-3,17-dione is classified as a prohibited substance in sports by the World Anti-Doping Agency (WADA) due to its aromatase-inhibiting properties. In 2006 three samples from human routine sports doping control tested positive for metabolites of androsta-1,4,6-triene-3,17-dione. The samples were initially found suspicious for the boldenone metabolite 17beta-hydroxy-5beta-androst-1-en-3-one. Since metabolites of androst-4-ene-3,6,17-trione were also present in the urine samples, it is presumed that these findings were due to the administration of a product like 'Novedex Xtreme', which could be easily obtained from the sport supplement market.
authors.affiliation: Center for Preventive Doping Research, Institute of Biochemistry, German Sport University Cologne, Cologne, Germany. m.parr@biochem.dshs-koeln.de
authors.forename: Maria K
authors.initials: MK
authors.lastname: Parr
authors.forename: Gregor
authors.initials: G
authors.lastname: Fusshöller
authors.forename: Nils
authors.initials: N
authors.lastname: Schlörer
authors.forename: Georg
authors.initials: G
authors.lastname: Opfermann
authors.forename: Thomas
authors.initials: T
authors.lastname: Piper
authors.forename: Grigory
authors.initials: G
authors.lastname: Rodchenkov
authors.forename: Wilhelm
authors.initials: W
authors.lastname: Schänzer
mesh_terms: D000733:Androstatrienes / Q000652:urine*; D019587:Dietary Supplements / Q000032:analysis*; D004300:Doping in Sports / Q000517:prevention & control*; D004791:Enzyme Inhibitors / Q000032:analysis; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D013287:Illicit Drugs / Q000652:urine*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D015813:Substance Abuse Detection / Q000379:methods*; D016482:Urinalysis / Q000379:methods*
chemical_list: D000733:Androstatrienes; D004791:Enzyme Inhibitors; D013287:Illicit Drugs; C011658:androsta-1,4,6-triene-3,17-dione