Investigation on possible transformations of cortisol,cortisone and cortisol glucuronide in bovine faecal matter using liquid chromatography–mass spectrometry |
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| Authors: | Francesco Arioli Marco Fidani Alessio Casati Maria L Fracchiolla Giuseppe Pompa |
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| Institution: | 1. Department of Veterinary Sciences and Technologies for Food Safety, University of Milan, Via Celoria 10, 20133 Milan, Italy;2. U.N.I.R.E. Lab. S.r.l., Via Gramsci 70, 20019 Settimo Milanese (MI), Italy;1. VMRD Global Discovery, Zoetis, 333 Portage Street, Kalamazoo, MI 49007, USA;2. Department of Chemistry and Chemical Biology, Rutgers University, 610 Taylor Road, Piscataway, NJ 08854, USA;1. Norwegian University of Life Sciences (NMBU), Department of Animal and Aquacultural Sciences, Faculty of Biosciences, P.O. Box 5003, NO-1432, Aas, Norway;2. University of Veterinary Medicine, Department of Biomedical Sciences, Veterinärplatz 1, A-1210, Vienna, Austria;3. Norwegian University of Life Sciences (NMBU), Faculty of Veterinary Medicine, Department of Production Animal Clinical Sciences, P.O. Box 8146, Dep N-0033, Oslo, Norway;4. Department of Animal Biosciences, University of Guelph, 50 Stone Rd. East, Guelph, ON, N1G 2W1, Canada;1. Ghent University, Faculty of Veterinary Medicine, Department of Veterinary Public Health & Food Safety, Laboratory of Chemical Analysis, Salisburylaan 133, B-9820 Merelbeke, Belgium;2. Ghent University, Faculty of Veterinary Medicine, Department of Pharmacology, Toxicology and Biochemistry, Laboratory of Pharmacology and Toxicology, Salisburylaan 133, B-9820 Merelbeke, Belgium;3. CER Groupe, Département Santé, Rue du Point du Jour 8, B-6900 Marloie, Belgium |
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| Abstract: | Given the close resemblance of the ring A structure of prednisolone and prednisone on the one hand, and of androstadienedione on the other, the transformation of cortisol and cortisone into prednisolone and prednisone in cattle faeces was evaluated. A simple method that does not involve extraction but only the 1:100 dilution of cattle faeces, spiking with 400 ng/mL cortisol, cortisone or cortisol glucuronide and incubation of the suspension, was used. The analyses were performed by HPLC–MS3 to detect the supposed Δ1 dehydrogenation of the glucocorticoids. The decision limits (CCα) and detection capabilities (CCβ) were 2.0 and 3.0 ng/mL for cortisol, cortisone and prednisolone, 3.0 and 4.0 ng/mL for cortisol glucuronide and 7.0 and 10.0 ng/mL for prednisone, respectively. Intra-day and inter-day coefficients of variation (CV%), were 5.6–6.2 and 5.2–6.6 for cortisol glucuronide, cortisol, cortisone and prednisolone, and 16.0 and 16.2 for prednisone, respectively. The recoveries were in the range 110–143% for all analytes. Regression coefficients (R2) were in the range 0.996–0.999 for all analytes. The results show the hydrolysis of the conjugated form and the dehydrogenation in ring A in diluted faeces. It is therefore predicted that urine contaminated with faeces may be positive for prednisone and prednisolone in the same way as they are positive for boldenone, i.e. as a result of microbiological dehydrogenase activity on cortisol and cortisone. |
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