Investigations of the microbial transformation of cortisol to prednisolone in urine samples

Bredehöft, Michael; Baginski, R; Parr, Maria Kristina; Thevis, Mario; Schänzer, Wilhelm

doi:10.1016/j.jsbmb.2010.04.021

Cited by 37 publications

(23 citation statements)

References 45 publications

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“…The fecal contamination of urine can also generate false positives for boldenone presence [10,11]. An analog explanation considers the in vitro formation of prednisolone from cortisol in bovine [12] and human urine [13]. Moreover, cattle that are under stress conditions [14][15][16] could produce prednisolone.…”

Section: Introductionmentioning

confidence: 99%

Detection of boldenone, its conjugates and androstadienedione, as well as five corticosteroids in bovine bile through a unique immunoaffinity column clean-up and two validated liquid chromatography–tandem mass spectrometry analyses

Chiesa

Nobile

Panseri

et al. 2014

Analytica Chimica Acta

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The presence of β-boldenone II phase metabolites and prednisolone in urine samples, owing to endogenous or natural origin or illicit treatment, is under debate within the European Union. The detection of β-boldenone conjugates, α-boldenone conjugates at concentrations higher than 2 ng mL(-1) and prednisolone above the cut-off level of 5 ng mL(-1) in urine have been, until now, critical in deciding if illegal drug use has occurred. The use of urine sometimes is not entirely satisfactory, especially when the drug is administrated at low doses or when its metabolic conversion is very fast. This subsequently would hamper its detection in urine. The introduction of a new, advantageous matrix where the illicit treatment can be investigated would be highly appreciated. In this study, we have developed and validated a simple and unique immunoaffinity clean-up procedure, which was applied to bovine bile samples, followed by two different analytical liquid chromatography-electrospray-tandem mass spectrometry methods. The first method tests androstadienedione, α- and β-boldenone sulphate, glucuronate and related free forms, while the other method assays prednisolone, prednisone, dexamethasone, cortisone, and cortisol. The methods were validated according to European Commission Decision 2002/657/EC. The evaluated parameters were linearity, specificity, precision (repeatability and intra-laboratory reproducibility), recovery, decision limit and detection capability. The decision limits (CCα) were between 0.38 and 0.45 ng mL(-1) for anabolic steroids, and 0.13 and 0.15 ng mL(-1) as far as corticosteroids were concerned. Intra- and inter-day repeatability was below 15.8 and 19.9% for all analytes, respectively. The methods were applied to the analysis of some bile samples collected from untreated young bulls in order to investigate the presence of the studied steroids in this matrix.

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Section: Introductionmentioning

confidence: 99%

Detection of boldenone, its conjugates and androstadienedione, as well as five corticosteroids in bovine bile through a unique immunoaffinity column clean-up and two validated liquid chromatography–tandem mass spectrometry analyses

Chiesa

Nobile

Panseri

et al. 2014

Analytica Chimica Acta

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“…The possibility of contamination of urine is in fact much higher when the collection is made from live animals rather than directly from the bladder. The possibility of false positives for prednisolone has also been encountered in human doping, under the aspect of microbial contaminations (Bredehö ft et al, 2010), once again demonstrating that the problem exists.…”

Section: Introductionmentioning

confidence: 99%

Prednisolone and prednisone neo-formation in bovine urine after sampling

Arioli

Casati

Fidani

et al. 2012

Animal

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The rise in the frequency of detecting prednisolone in bovine urine from northern Italy has come into focus of attention in recent years. The possibility that neo-formation of prednisolone or that prednisone may occur in urine after collection of samples was therefore investigated. Cow urine collected for official routine controls in Lombardy containing more than 80 ng/ml cortisol, and prednisolone and prednisone below the decision limit (CCa) of the method (0.4 and 0.5 ng/ml, respectively) was used. The C 1-2 dehydrogenation of naturally present cortisol and cortisone was checked by incubating urine, both contaminated and uncontaminated with faeces, at 378C and by collecting samples at 0, 1, 2, 4, 6 and 24 h. The influence of Helix pomatia juice was also investigated in order to determine whether deconjugation could influence the reliability of the results. All samples were analysed by HPLC-MS 3 for the presence of cortisol, cortisone, prednisolone and prednisone in negative electrospray ionisation mode, utilising the consecutive reaction monitoring of product ions derived from the formate molecular adduct ([M1HCOO] -). The observed neo-formation of prednisolone shows that inappropriate temperatures in sample storage and processing can result in an incorrect accusation of non-compliance. The faecal contamination of urine, performed with the aim to mimic a collection conducted without the necessary care, moreover, evoked a high increase in prednisolone concentration in two out of seven animals. Moreover, H. pomatia juice had no significant effect on the prednisolone concentration, indicating that this corticosteroid is present in its free form in cow urine.

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“…Analogous explanations are given regarding the in vitro formation of prednisolone from cortisol in bovine (Arioli, Fidani, Casati, Fracchiolla, & Pompa, 2010) and human urine (Bredehöft, Baginski, Parr, Thevis, & Schänzer, 2012). Moreover, cattle that are under stress conditions could produce prednisolone (Pompa et al, 2011), as well as a-and b-nortestosterone, as shown by Glenn Kennedy et al (2009) in a study on injured male cattle.…”

Section: Introductionmentioning

confidence: 95%

Suitability of bovine bile compared to urine for detection of free, sulfate and glucuronate boldenone, androstadienedione, cortisol, cortisone, prednisolone, prednisone and dexamethasone by LC–MS/MS

et al. 2015

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Investigations of the microbial transformation of cortisol to prednisolone in urine samples

Cited by 37 publications

References 45 publications

Detection of boldenone, its conjugates and androstadienedione, as well as five corticosteroids in bovine bile through a unique immunoaffinity column clean-up and two validated liquid chromatography–tandem mass spectrometry analyses

Detection of boldenone, its conjugates and androstadienedione, as well as five corticosteroids in bovine bile through a unique immunoaffinity column clean-up and two validated liquid chromatography–tandem mass spectrometry analyses

Prednisolone and prednisone neo-formation in bovine urine after sampling

Suitability of bovine bile compared to urine for detection of free, sulfate and glucuronate boldenone, androstadienedione, cortisol, cortisone, prednisolone, prednisone and dexamethasone by LC–MS/MS

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