Considered as a xenobiotic for many years, 19-nortestosterone has been extensively studied. Analyses developed to control the illegal use of this steroid in meat-producing animals led researchers to demonstrate the endogenous presence of 19-nortestosterone in several species. In this paper, the natural occurrence of 19-nortestosterone in its alpha form (epinandrolone) in the urine of pregnant sheep was demonstrated using gas chromatography-mass spectrometry. This reference method allowed 17 alpha-nandrolone to be detected and identified, and the absence of the beta epimer in the urine of pregnant sheep to be demonstrated. 17 alpha-Nandrolone was accurately determined at different stages of pregnancy. The analyses showed that epinandrolone, which was not detectable in the urine of non-pregnant sheep, was excreted in small amounts (leading to a < 0.5 ppb concentration) during the first 4 months of pregnancy. 17 alpha-Nandrolone concentrations then increased during the last month until parturition. The origin of this molecule was not determined.
The fate of salbutamol sulphate given orally has been investigated in calves. The urinary excretion rate and the tissue distribution of this beta-agonistic drug were studied by capillary gas chromatography coupled to low resolution mass spectrometry (GC-LRMS) under electron impact (EI) ionization mode, using an hexadeuterated salbutamol analogue as the internal standard. The parent drug and metabolites were extracted via solid phase extraction (SPE) mixed-phase-containing disposable columns and analysed as their trimethylsilyl derivatives. A more efficient clean-up had to be carried out for tissue samples. An acidic precipitation followed by a liquid-liquid extraction were therefore performed before the SPE. Moreover, the problem of tissue digestion was elucidated by means of an ultrasonic probe. Samples were also analysed before and after enzymic hydrolysis using purified beta-glucuronidase and a mixture of beta-glucuronidase and arylsulphatase, to obtain evidence of phase II conjugation mechanisms. Both free salbutamol and conjugated metabolites were detected in urine and tissue samples. Except for liver or kidney, salbutamol was rapidly cleared from most tissues after a withdrawal period. The possible excretion of some phase I metabolites was also investigated, using further analyses under positive chemical ionization LRMS and high resolution mass spectrometry (HRMS).
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