Tetrahydro-metabolites of cortisol and cortisone in bovine urine evaluated by HPLC–ESI-mass spectrometry

Pavlović, Radmila; Cannizzo, Francesca Tiziana; Panseri, Sara; Biolatti, B.; Trutić, Nataša; Biondi, Pier Antonio; Chiesa, Luca Maria

doi:10.1016/j.jsbmb.2012.12.015

Cited by 14 publications

(9 citation statements)

References 20 publications

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“…Nebbia et al (2014) also observed a decrease in F and E levels in finishing bulls and cows after therapeutic treatment with PL acetate. This is to some extent in contrast with our preliminary study (Pavlovic et al 2013) where no differences were found in the F and E excretion time course during PL treatment in male cattle. The plausible explanation for this discrepancy is that those experimental subjects differed in age and consequently in metabolism (veal calves are not yet ruminant).…”

Section: Methods Validationcontrasting

confidence: 99%

“…Therefore, a method to identify natural corticosteroid levels in bovines could be an important tool for the control of illicit corticosteroid treatment in livestock production. Recently, we have published (Pavlovic et al 2013) that PL treatment influenced 11β-HSD, which brought up questions about the impact of PL treatment on the THF and THE metabolic rates. Accordingly, the role and importance of reduced forms of F and E and their metabolic rates during treatment with exogenous glucocorticoids are lacking and remain to be established.…”

Section: Introductionmentioning

confidence: 99%

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HPLC-ESI-MS/MS assessment of the tetrahydro-metabolites of cortisol and cortisone in bovine urine: promising markers of dexamethasone and prednisolone treatment

Chiesa

Panseri

Pavlović

et al. 2016

Food Additives & Contaminants: Part A

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The effects of long-term administration of low doses of dexamethasone (DX) and prednisolone (PL) on the metabolism of endogenous corticosteroids were investigated in veal calves. In addition to cortisol (F) and cortisone (E), whose interconversion is regulated by 11β-hydroxysteroid dehydrogenases (11βHSDs), special attention was paid to tetrahydrocortisol (THF), allo-tetrahydrocortisol (aTHF), tetrahydrocortisone (THE) and allo-tetrahydrocortisone (aTHE), which are produced from F and E by catalytic activity of 5α and 5β-reductases. A specifically developed HPLC-ESI-MS/MS method achieved the complete chromatographic separation of two pairs of diastereoisomers (THF/aTHF and THE/aTHE), which, with appropriate mass fragmentation patterns, provided an unambiguous conformation. The method was linear (r 2 > 0.9905; 0.5-25 ng ml . Recoveries were in range 75-114%, while matrix effects were minimal. The experimental study was carried out on three groups of male Friesian veal calves: group PL (n = 6, PL acetate 15 mg day -1 p.o. for 31 days); group DX (n = 5, 5 mg of estradiol (E2) i.m., weekly, and 0.4 mg day -1 of DX p.o. for 31 days) and a control group (n = 8). Urine was collected before, during (twice) and at the end of treatment. During PL administration, the tetrahydro-metabolite levels decreased gradually and remained low after the suspension of treatment. DX reduced urinary THF that persisted after the treatment, while THE levels decreased during the experiment, but rebounded substantially after the DX was withdrawn. Both DX and PL significantly interfered with the production of F and E, leading to their complete depletion. Taken together, the results demonstrate the influence of DX and PL administration on 11βHSD activity and their impact on dysfunction of the 5-reductase pathway. In conclusion, profiling tetrahydrometabolites of F and E might serve as an alternative, indirect but reliable, non-invasive procedure for assessing the impact of synthetic glucocorticosteroids administration. ARTICLE HISTORY

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Section: Methods Validationcontrasting

confidence: 99%

Section: Introductionmentioning

confidence: 99%

HPLC-ESI-MS/MS assessment of the tetrahydro-metabolites of cortisol and cortisone in bovine urine: promising markers of dexamethasone and prednisolone treatment

Chiesa

Panseri

Pavlović

et al. 2016

Food Additives & Contaminants: Part A

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“…In this paper, the negative mode was selected for further fragmentation and quantification of the compounds. This approach was used by Turpeinen et al [12], although this group measured only tetrahydrometabolites, and also by Pavlovic et al [13] who were able to measure all five steroids but the method was validated in bovine urine. In our study, all the results were generated in positive ion mode and achieved a good LLOQ of 0.1 ng/mL for F and E, and 1 ng/mL for the tetrahydrometabolites, although it has been reported that the negative ion mode produces better signal-to-noise ratio and reduced fragmentation than the positive ion mode [11].…”

Section: Discussionmentioning

confidence: 99%

LC–MS/MS Method for the Simultaneous Determination of Free Urinary Steroids

et al. 2014

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Cortisol homeostasis is implicated in hypertension and metabolic syndrome. Two enzymes modulate cortisol availability; 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) preferentially converts inactive cortisone to cortisol, whereas 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) converts cortisol to cortisone. In contrast, 5α and 5β reductases inactivate cortisol by conversion to its tetrahydrometabolites: tetrahydrocortisol, allo-tetrahydrocortisol and tetrahydrocortisone. A subtle local increase in cortisol can be detected by measuring 24-h urine metabolites, LC–MS/MS being the reference method. The 11β-HSD2 activity is assessed based on the cortisol/cortisone ratio, and the 11β-HSD1 activity on the (tetrahydrocortisol + allo-tetrahydrocortisol)/tetrahydrocortisone ratio. To better understand hypertension and/or metabolic syndrome pathogenesis a method for simultaneous determination of cortisol, cortisone, tetrahydrocortisol, allo-tetrahydrocortisol and tetrahydrocortisone was developed and validated in an LC coupled with the new detector AB Sciex QTrap® 4500 tandem mass spectrometer. The steroids were extracted from 1 mL urine, using cortisol-D4 as internal standard. The quantification range was 0.1–120 ng/mL for cortisol and cortisone, and 1–120 ng/mL for tetrahydrometabolites, with >89 % recovery for all analytes. The coefficient of variation and accuracy was <10 %, and 85–105 %, respectively. Our LC–MS/MS method is accurate and reproducible in accordance with Food and Drug Administration guidelines, showing good sensitivity and recovery. This method allows the assessment of 11β-HSD2 and 11β-HSD1 activities in a single analytical run providing an innovative tool to explain etiology of misclassified essential hypertension and/or metabolic syndrome.

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“…The therapeutic use of prednisolone in bovine is regulated by Commission Regulation (EU) No. 37/2010 (European Community 2010) which sets maximum residue limits (MRLs), even if its illicit use as a growth promoter agent cannot be discarded (Pavlovic et al 2013). No MRLs have been set for urine, but a 5 ng ml −1 cut-off level has been recommended, e.g.…”

Section: Introductionmentioning

confidence: 99%

Pseudoendogenous origin of prednisolone in pigs from the food chain

Arioli

Pasquale

Panseri

et al. 2015

Food Additives & Contaminants: Part A

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The debate about the origin of prednisolone in animal organisms has lasted for 5 years. Bovine species have been the most studied, but studies on humans and horses are also present in the literature. Even if prednisolone in pigs does not yet represent a problem for control agencies, interest has recently increased with regard to this species. To date, there has been just a single study in the literature about this topic, performed on 10 sows treated with prednisolone or a synthetic analogue of adrenocorticotropic hormone. We therefore initiated a study on 80 pigs, a number considered representative in relation to the expected frequency (prevalence) of prednisolone detection in urine collected at slaughter. Prednisolone was detected in urine both at the farm and at the slaughterhouse, with a concentration and frequency higher at slaughter. The presence of prednisolone was also studied in the adrenal glands, where the corticosteroids are produced in response to stress, and it was detected in 89% of the samples. These results, together with the similar behaviours of prednisolone and cortisol, i.e. a mutual rise in the two corticosteroids in urine collected at the slaughterhouse and the correlation between the concentrations of the two corticosteroids in the adrenal glands, seem to indicate an endogenous origin of prednisolone in pigs.

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Tetrahydro-metabolites of cortisol and cortisone in bovine urine evaluated by HPLC–ESI-mass spectrometry

Cited by 14 publications

References 20 publications

HPLC-ESI-MS/MS assessment of the tetrahydro-metabolites of cortisol and cortisone in bovine urine: promising markers of dexamethasone and prednisolone treatment

HPLC-ESI-MS/MS assessment of the tetrahydro-metabolites of cortisol and cortisone in bovine urine: promising markers of dexamethasone and prednisolone treatment

LC–MS/MS Method for the Simultaneous Determination of Free Urinary Steroids

Pseudoendogenous origin of prednisolone in pigs from the food chain

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