RESUMEN
The detection of metabolites of the anti-estrogenic substance cyclofenil, listed on the World Anti-Doping Agency (WADA) Prohibited List since 2004 is described. Target substances are hydroxylated metabolites, bearing an aliphatic hydroxyl group either in the 2-, 3- or 4-position of the aliphatic ring, in addition to the phenolic functions on the aromatic rings. Structural identification used NMR as well as high-resolution mass spectrometry after nano-electrospray ionisation (ESI). Unambiguous detection of all three synthesised cyclofenil metabolites M1-M3 was done using gas chromatography for separation and electron ionisation mass spectrometry for detection of the per-silylated compounds in comparison with a reference urine deriving from an excretion study within the WADA 2007 Educational Programme.
Asunto(s)
Ciclofenil , Doping en los Deportes , Moduladores de los Receptores de Estrógeno , Drogas Ilícitas/síntesis química , Detección de Abuso de Sustancias/métodos , Cromatografía de Gases , Ciclofenil/análogos & derivados , Ciclofenil/química , Ciclofenil/orina , Moduladores de los Receptores de Estrógeno/química , Moduladores de los Receptores de Estrógeno/orina , Humanos , Hidroxilación , Drogas Ilícitas/orina , Nanotecnología , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Using gas chromatography/electron impact-mass spectrometry (GC/EI-MS) and high performance liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry (HPLC/APCI-MS/MS), the structures of cyclofenil metabolites in human urine have been assigned. The hydroxyl metabolites liberated from the glucuronide conjugates after acid hydrolysis were characterized as the trimethylsilyl (O-TMS) derivatives using GC/MS. The conjugate glucuronide forms were detected without hydrolysis by HPLC/MS. Cyclofenil was not observed in urine. Tentative structures for the two metabolites are proposed.