Identification of hydroxyl metabolites by liquid chromatography tandem mass spectrometry in urine of veal calves administered in vivo with boldenone and boldione.

Preliminary data obtained in vitro by incubating subcellular fraction from calf liver with 17b-boldenone (androsta-1, 4-dien- 17b-ol-3-one (17b-BOL)) or androsta-1,4-diene-3,17-dione (ADD or boldione) showed that some polar metabolites were produced from both compounds at detectable quantities. Indeed when ADD was incubated, HPLC analysis of the extracts showed 17b-BOL as metabolite, but another polar compound occurred in detectable amounts ; the putative identity as a hydroxyl derivative was tentatively assessed by means of LC/MS/MS fragmentation. When 17b-BOL was added to the incubation mixture two polar compounds were recovered in addition to ADD, and their nature as hydroxyl derivatives was confirmed with pure standard comparison . Because urinary hydroxyl derivatives can help confirm illegal treatment and anabolic steroids are usually completely metabolized, the main aim was the in vivo investigation of the urinary excretion profile of 17b- BOL and its metabolites, in an attempt to identify possible new in vivo markers of illicit administration.