Studies on the presence of natural and synthetic corticosteroids in bovine urine

Ferranti, Carolina; Quadri, F. delli; Palleschi, Luca; Marchiafava, C.; Pezzolato, Marzia; Bozzetta, Elena; Caramelli, Maria; Draisci, Rosa

doi:10.1016/j.steroids.2011.02.044

Cited by 32 publications

(40 citation statements)

References 36 publications

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“…administration of a synthetic analogue of the adrenocorticotropic hormone caused the urinary cortisol level to abruptly rise up to 163-238 µg l -1 coincident with the appearance of remarkable prednisolone concentrations (1.01-3.51 µg l -1 ) as soon as two hours after the treatment. Similar results were observed also in urines from experimental untreated veal calves, showing that the stress induced by slaughtering procedures may entail a fourfold increase in cortisol concentration, accompanied by the detection of measurable prednisolone levels (up to 1.4 µg l -1 ) in 5 out of 9 individuals (Ferranti et al 2011). From a quantitative point of view, however, a significant difference exists between our study and the previous ones, as the concentrations of prednisolone that we occasionally detected in urines never exceeded the CC level, i.e.…”

Section: Relationship Between Alleged Stress and The Recovery Of Tracsupporting

confidence: 79%

“…The biochemical formation of prednisolone was demonstrated in artificially stressed cows by Pompa et al (2011), and in cattle urine samples contaminated by fecal bacteria by Arioli et al (2010). The finding of some non compliances for prednisolone in urine samples collected from officially untreated veal calves (Ferranti et al 2011) apparently corroborated the conclusions drawn in the studies mentioned above. Taking into account that the misuse and abuse of prednisolone in cattle is well established, it becomes important also from a forensic viewpoint to make a clear distinction between the conditions that may favour the alleged endogenous production of prednisolone and cases when the presence of prednisolone in urine samples is expected to reflect an exogenous administration.…”

Section: Introductionsupporting

confidence: 76%

“…Finally, prednisolone at 0.5 µg l -1 concentration was also detected in one out of seven urine samples from experimental Friesian veal calves before starting the administration of the same glucocorticoid (Ferranti et al 2011) .…”

Section: Occurrence Of Prednisone or Prednisolone In Cow Urinesmentioning

confidence: 87%

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A field survey on the presence of prednisolone and prednisone in urine samples from untreated cows

Vincenti

Leporati

Capra

et al. 2012

Food Additives & Contaminants: Part A

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A field survey on the presence of prednisolone and prednisone in urine samples from untreated cows Prednisolone is a synthetic glucocorticoid widely employed in bovine clinical practice, which may be also illegally used as a growth promoter. Recent in vitro and in vivo studies lend support to the hypothesis that prednisolone could be synthesized from cortisol in untreated cattle subjected to stressful events. To verify such a hypothesis, a field survey was conducted on urine samples collected from 131 guaranteed untreated cows and analyzed for the presence of prednisolone and prednisone -in some instances also for cortisol and cortisone-with a validated HPLC/MS-MS method. None of the examined samples exhibited either prednisolone levels higher than the CC limit (around 0.70 g l -1 ) or prednisone, being therefore officially compliant for both analytes. Trace amounts of prednisolone, approximately estimated in the range 0.1-0.3 g l -1 , were found only in 7 samples from cows also showing urinary cortisol and cortisone levels higher than those detected in negative specimens, as the result of a probable stress condition. KEYWORDS:Prednisolone; cows; urine; cortisol, illicit growth promoters; liquid chromatography-tandem mass spectrometry Such modifications result in both an extended duration of GC therapeutical effects and a severalfold increase in their pharmacological potency, particularly in the anti-inflammatory action. These features, combined with the absence of a parallel increase of sodium-retaining effects, enhance the suitability of synthetic GCs for therapeutic purposes (Ferguson and Hoenig 1995). A number of commercial preparations containing different prednisolone esters are currently available for administration to cattle, covering a wide range of therapeutical applications, including primary ketosis, disorders of tendons and the musculoskeletal system, allergic reactions, skin diseases, and shock. In addition, prednisolone is included in a number of antibiotic preparations for intramammary administration, indicated for the treatment of cow mastitis (McDonald et al. 2007).Besides legal treatments, prednisolone and other GCs are frequently employed just before the animals are sold, to mask more or less severe pathologies, especially in the case of old cows at the end of their productive cycle. Another common law infringement is the administration of intramammary infusions without declaring nor applying an appropriate withdrawal time. In the last decade, synthetic GCs, including prednisolone, have been increasingly employed as illicit growth promoters in cattle (Stephany 2001;Cannizzo et al. 2011; European Commission 2012). When administered at low dosages for an extended period of time, GCs are known to significantly improve the feed conversion ratio, to increase the area of longissimus dorsi, one of the most valuable muscles in bovine meat breeds, and to enhance the overall carcass quality traits, yielding pale and tender meat ( apparently corroborated the conclusions drawn in the studies mentioned...

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Section: Relationship Between Alleged Stress and The Recovery Of Tracsupporting

confidence: 79%

Section: Introductionsupporting

confidence: 76%

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A field survey on the presence of prednisolone and prednisone in urine samples from untreated cows

Vincenti

Leporati

Capra

et al. 2012

Food Additives & Contaminants: Part A

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“…prednisone, prednisolone)'. A study on the presence of corticosteroids in bovine urine reports the presence of prednisolone at an average concentration of 0.6 ng/ml in live control animals or live animals before treatment (5 of 72 samples; Ferranti et al, 2011). Moreover, Andersen et al (2008) showed the detection of prednisolone in 3 of 19 'blank' bovine urine samples at concentrations of 13.0, 5.9 and 4.2 ng/ml, not specifying, however, whether the sampling was made on live or slaughtered bovines.…”

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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“…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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Studies on the presence of natural and synthetic corticosteroids in bovine urine

Cited by 32 publications

References 36 publications

A field survey on the presence of prednisolone and prednisone in urine samples from untreated cows

A field survey on the presence of prednisolone and prednisone in urine samples from untreated cows

Prednisolone and prednisone neo-formation in bovine urine after sampling

Determination of prednisolone metabolites in beef cattle

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