Investigation of the origin of prednisolone in cow urine

Pompa, G.; Arioli, Francesco; Casati, Anna; Fidani, Marco; Bertocchi, L.; Dusi, Guglielmo

doi:10.1016/j.steroids.2010.09.005

Cited by 52 publications

(45 citation statements)

References 6 publications

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“…The first was that prednisolone appears or increases to detectable concentrations when the animal is stressed. In a previous study (Pompa et al, 2011), the role of stress, both pharmacologically induced or caused by slaughter operations, on inducing the presence of prednisolone in bovine urine together with an increase in cortisol and cortisone concentrations was shown.…”

Section: Discussionmentioning

confidence: 98%

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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Section: Discussionmentioning

confidence: 98%

Prednisolone and prednisone neo-formation in bovine urine after sampling

Arioli

Casati

Fidani

et al. 2012

Animal

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“…[14 , 18 , 19] The application of HPLC single quadrupole MS technique to the determination of corticosteroids in biological samples has received only limited attention. [1 ] According to literature, ESI is suitable for glucocorticoids, which can undergo ionization both in positive and negative modes. [20] Although the positive ionization mode gave a stronger absolute response, the negative ionization was selected due to more specific and more characteristic frag-mentation with less background noise.…”

Section: Results and Discussion Conditions For Hplc And Esi-msmentioning

confidence: 99%

“…Prednisolone (PL), a glucocorticosteroid used as a therapeutic agent in livestock, has also been recently found in cow urine as an endogenous product of corticosteroid metabolism. [1]Prednisone (PN) can be considered an inactive precursor of PL; therefore, it can be linked to the same metabolism. Effects of the administration of synthetic corticosteroids on urinary F profile were studied in more detailed in humans.…”

Section: Introductionmentioning

confidence: 99%

Determination of Cortisol, Cortisone, Prednisolone and Prednisone in Bovine Urine by Liquid Chromatography–electrospray Ionisation Single Quadrupole Mass Spectrometry

Pavlović

Chiesa

Soncin

et al. 2012

Journal of Liquid Chromatography & Related Technologies

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A quantitative LC-ESI single quadrupole MS method for the determination of cortisol (F), cortisone (E), prednisolone (PL), and prednisone (PN) in bovine urine has been developed and validated. After addingflumethasone as internal standard, the samples were subjected tofiltration, deconjugation, and solid-phase extraction, while the chromatographic separation was achieved using a Restek Ultra II Allure Biphenyl column with isocratic mobile phase. The analytes were detected after negative electrospray ionization using SIM mode. In order to obtain spectra with maximum intensities of at least one of the three characteristic ions, (Mþformate) , (M-H), and [(M-H) -CH 2 0]~, an individual optimization of MS parameters for each corticosteroid was set up. MS data was acquired in the three-ion selected monitoring mode and the ion ratios between chosen diagnostic ions were used in order to increase the specificity. Calibration graphs were linear and the intra-day and intermediate precision was estimated as RSD values which were less than 17%. For F and E, obtained values indicated negligible absolute matrix effects (10 3 % and 98%, respectively). The method was applied to real samples, and basal levels ofFandE were preliminarily evaluated, while PL and PN were not detected.

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“…Unlike other analogues, such as dexamethasone, prednisolone does not contain any halogen atom and its structure closely resembles that of other endogenous steroids, including cortisol. From this structural similarity arises the hypothesis that in meat cattle prednisolone could be generated by physiological metabolic processes, possibly under extremely stressful conditions, such as transport and slaughtering, or by faecal contamination, as is supported by a few recent studies (Arioli et al 2010;Ferranti et al 2011;Pompa et al 2011). On the basis of these results and the opinion of the European Union Reference Laboratory, the Italian Ministry of Health has recently enacted a new disposition: a bovine urine sample is considered non-compliant for prednisolone only when its concentration exceeds 5.0 ng mL l (Circular of Ministry of Health 2012).…”

Section: Introductionmentioning

confidence: 91%

Determination of prednisolone metabolites in beef cattle

Leporati

Capra

Cannizzo

et al. 2013

Food Additives & Contaminants: Part A

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Prednisolone is a synthetic corticosteroid acting on both hydrosaline balance and metabolism that is liable to fraudulent administration to meat-producing animals for growth-promoting purposes. Its use outside strict therapeutic control and prescription is banned by the European legislation, but official controls are hampered by its negligible direct excretion into the urinary matrix. Recent studies reported on a potential endogenous origin of prednisolone in animals subjected to stressful conditions, accounting for its occasional detection in control urines. The objective of the present study was the identification and quantification of prednisolone urinary metabolites to be used as illicit treatment biomarkers in place of the parent drug. An LC-MS/MS screening was conducted on urine samples collected from a bullock intramuscularly administered with prednisolone acetate by using a therapeutic protocol (2 x 0.52 mg kg l at 48-hour interval). Four prednisolone metabolites were identified: 20β-dihydroprednisolone, 20α-dihydroprednisolone, 6β-hydroxyprednisolone and 20β-dihy-droprednisone; the first was detected at relatively high concentrations. An existing quantitative LC-MS/MS method was expanded and revalidated to include these metabolites. The new analytical method proved sensitive (LODs: 0.35-0.42 ng mL l ) and specific and was applied to urine samples collected from eight beef cattle subjected to low-dosage oral administration of prednisolone acetate for a 35-day period, as in standard growth-promoting treatments. 20β-Dihydroprednisolone was detected in all urine samples collected during the treatment, at relatively high concentration (1.2-27 ng mL -1 ), whereas the prednisolone concentration was virtually negligible (<0.7 ng mL -1 ). 20β-Dihydroprednisolone was no longer present in almost all samples collected 6 days after the end of the treatment, but trace amounts of this metabolite were found in two urine samples from control animals. 20β-Dihydroprednisolone is proposed as an effective biomarker to test illegal growth-promoting treatments with prednisolone in meat cattle, alternatively to the parent drug.

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Investigation of the origin of prednisolone in cow urine

Cited by 52 publications

References 6 publications

Prednisolone and prednisone neo-formation in bovine urine after sampling

Prednisolone and prednisone neo-formation in bovine urine after sampling

Determination of Cortisol, Cortisone, Prednisolone and Prednisone in Bovine Urine by Liquid Chromatography–electrospray Ionisation Single Quadrupole Mass Spectrometry

Determination of prednisolone metabolites in beef cattle

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