Abstract
The use of anabolic steroid hormones as growth promoters in feed for farm animals has been banned in the European Union since 1988 on the basis of Council Directive 96/22/EC. However, there is still ongoing monitoring and reporting of positive findings of these banned substances in EU countries. The aim of this work was to investigate the efficacy and discriminatory ability of metabolic fingerprinting after the administration of 17β-testosterone esters to pigs. Plasma and urine samples were chromatographically separated on a Hypersil Gold C18 column. High resolution mass spectrometry metabolomic fingerprints were analysed on a hybrid mass spectrometer Q-Exactive. Three independent multivariate statistical methods, namely principal component analysis, clustre analysis, and orthogonal partial least squares discriminant analysis showed significant differences between the treated and control groups of pigs even 14 days after the administration of the hormonal drug. Plasma samples were also analysed by a conventional quantitative analysis using liquid chromatography with tandem mass spectrometry and a pharmacokinetic curve was constructed based on the results. In this case, no testosterone residue was detected 14 days after the administration. The results clearly showed that a metabolomics approach can be a useful and effective tool for the detection and monitoring of banned anabolic steroids used illegally in pig fattening.
Highlights
The use of hormones as growth promoters for fattening purposes in livestock has been banned in the European Union since 1988 by Council Directive 96/22/EC
Anabolic Effect of 17β-Testosterone (Esters). All animals from both groups were weighed at regular weekly intervals during the treatment experiment, and the body weight gain (BW) in kg is shown in Tables 1 and 2
The present study confirmed the ability of metabolomics approaches and techniques to significantly differentiate pigs administered an androgenic anabolic steroid which is on the list of banned substances
Summary
The use of hormones as growth promoters for fattening purposes in livestock has been banned in the European Union since 1988 by Council Directive 96/22/EC. The banned substances are still reported as positive in the European residue monitoring plans [1]. One of these banned substances is the anabolic and androgenic steroid testosterone, which naturally occurs in animal organisms. Testosterone is secreted into the bloodstream where it primarily (98%) binds to a specific protein beta-globulin termed sex hormone binding globulin (SHBG) and to a lesser extent to albumin. By this binding, testosterone is biologically protected from inactivation in the liver, and is subsequently transported to the target tissues via the bloodstream. Testosterone is reduced to 5α-dihydrotestosterone (DHT) by the cytochrome P450 enzyme 5α-reductase, an enzyme
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