title: Detection of formestane abuse by mass spectrometric techniques.
abstract: Formestane (4-hydroxy-androstenedione) is an aromatase inhibitor prohibited in sports and included, since 2004, in the list of prohibited substances updated yearly by the World Anti-Doping Agency (WADA). Since the endogenous production of formestane has been described, it is mandatory for the anti-doping laboratories to use isotope ratio mass spectrometry (IRMS) to establish the exogenous origin before issuing an adverse analytical finding. The described IRMS methods for formestane detection are time-consuming, requiring usually two consecutive liquid chromatographic sample purifications in order to have final extracts of adequate purity before the mass spectrometric analysis. After establishing a procedure for the determination of the origin of formestane by IRMS without the need of derivatization, and integrated in the overall analytical strategy of the laboratory for pseudo-endogenous steroids, a mass spectrometric analysis by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-tandem mass spectrometry (GC-MS/MS) of formestane metabolites was carried out in order to investigate whether other biomarkers of formestane abuse could be integrated in order to avoid time-consuming and expensive IRMS confirmations for formestane. From the metabolic studies performed, the inclusion of 3β,4α-dihydroxy-5α-androstan-17-one (4α-hydroxy-epiandosterone) in the routine GC-MS procedures has demonstrated to be diagnostic in order to reduce the number of unnecessary confirmations of the endogenous origin of formestane.
authors.affiliation: Laboratorio Antidoping, Federazione Medico Sportiva Italiana, Largo Giulio Onesti 1, 00197, Rome, Italy.
authors.forename: Xavier
authors.initials: X
authors.lastname: de la Torre
authors.forename: Cristiana
authors.initials: C
authors.lastname: Colamonici
authors.forename: Davide
authors.initials: D
authors.lastname: Curcio
authors.forename: Daniel
authors.initials: D
authors.lastname: Jardines
authors.forename: Francesco
authors.initials: F
authors.lastname: Molaioni
authors.forename: Maria Kristina
authors.initials: MK
authors.lastname: Parr
authors.forename: Francesco
authors.initials: F
authors.lastname: Botrè
mesh_terms: D000328:Adult; D000728:Androgens / Q000032:analysis; D000735:Androstenedione / Q000031:analogs & derivatives* / Q000032:analysis / Q000652:urine; D047072:Aromatase Inhibitors / Q000032:analysis* / Q000652:urine; D002214:Capsules / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D004300:Doping in Sports / Q000379:methods*; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007202:Indicators and Reagents; D008297:Male; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D015813:Substance Abuse Detection / Q000379:methods
chemical_list: D000728:Androgens; D047072:Aromatase Inhibitors; D002214:Capsules; D007202:Indicators and Reagents; D000735:Androstenedione; C014594:formestane
keywords: doping in sports; formestane; isotope ratio mass spectrometry (IRMS)