Labella / Arioli

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LabellaGiuseppe Federico Labella graduated in Clinical, Forensic and Sport Chemistry Master’s Degree at the University of Turin on December 2013 with the evaluation of 110/110 with honors. His thesis is “Detection of Gunshot Residues (GSR) in blowfly larvae using Scanning Electron Microscopy equipped with Energy Dispersive X-ray Microanalysis (SEM-EDX) and inductively Coupled Plasma Mass Spectrometry (ICP-MS)”. Under the same title he has published an international poster presented at the Lille Meeting of the European Association for Forensic Entomology on April 2014. His work is now focused on the presence in bovine of pseudo-endogenous compounds like prednisolone, boldenone, thiouracil.

ArioliFrancesco Arioli is Assistant Professor in Veterinary Pharmacology and Toxicology at the Department of Health, Animal Science and Food Safety of the Faculty of Veterinary Medicine of the University of Milan, Italy.

His main research field deal with the study of the presence and biotransformation of hormones and illegally drugs used (Antibiotics used for therapeutic purposes or illicitly as growth promoters, anabolic substances, halogenated and non-halogenated corticosteroid) in biological matrices collected from food producing animals. He also deals with halogenated contaminants in feed and in food of animal origin and in water.

Research Project

Origin of the presence of pseudoendogenous substances in different biological matrices from food producing animals and identification of biomarkers for their illicit use

The research of drug residues in farm animal tissues or fluids is one of the means by which the National Authorities watch over the healthy, hygienic and organoleptic properties of food of animal origin. However, some substances with doubtful effects, or with a urinary threshold level due to their natural origin, like some anabolic steroid hormones, or pharmacological principles permitted only if a proper medical cause justifies their use, e.g. corticosteroids, complicate the tasks of the Health Authorities. These compounds have been appropriately defined “grey-zone substances” and described “synthetically produced hormones that are also known to be endogenous under certain conditions, dubbed ‘pseudo-endogenous’ or ‘grey zone substances’ due to their dual synthetic/endogenous nature”. At the moment, a debate is ongoing in the European Union about the origin, the cut-off levels in urine (a matrix typically used by Control Agencies) and the use of biomarkers to discriminate between natural or endogenous origin of prednisolone, a glucocorticosteroid which differs from cortisol only for the presence of a double bond in position C1-C2, of boldenone , an anabolic steroid which differs from testosterone in the same way above described, and the thyrostatic compound thiouracil, whose endogenous formation is caused by the intake of Brassicaceae. The therapeutic use of Prednisolone is regulated by Commission Regulation (EEC) 37/2010 wich sets the MRLs in cattle tissues and in milk. Boldenone and thiouracil are banned within the European Union since 1981, due to the risk to human health of drug residues in food of animal origin.

Labella Arioli 01

The objectives of the project for each compound are here described:
- Prednisolone:
  • identification of a possible precursor of its formation in urine after collection or in faeces in the last tract of the large intestine. To this aim a saponin and a sapogenin with steroidal nucleus will be used as precursor in vitro experiments. Biological matrices (faeces, feed, etc), considered contaminants of urine during collection will be spiked with the hypothesized precursors, incubated and analysed by HPLC-MSn. The expected result is to understand if saponins and sapogenins could be responsible for the formation of prednisolone.
  • identification of a possible biomarker for its illicit use. Urine from treated and untreated bovines will be analysed for the presence of the main prednisolone and cortisol metabolites.
- Thiouracil:
  • Study on the frequency of detection of thiouracil in urine and thyroid in bovines from the food chain. Analytical methods will be developed and validated for these two matrices. As the cut-off levels are not homogeneous in the European Union, the expected result is to give a contribution to Control Organisms for the setting of an ultimate cut off level in urine or (possibly) in the thyroid.
- Boldenone:
  • Recent findings by our group show that the only presence of β-boldenone sulfate cannot be per se considered an evidence of illicit treatment with boldenone esters or its precursor androstadienedione. A cut-off level should be more appropriate and this is the aim of the research on boldenone. Other metabolites, not fully identified, have been proposed. As the topic is constantly changing, studies on bovine urine could also be carried out only if the setting of the cut-off level will not be satisfactory for the identification of illicit treatments.

The analytical methods used for this project will include different approaches for samples purification (liquid-liquid extraction, immunoaffinity column extraction, etc) and HPLC-MSn analysis.