Global urine fingerprinting by LC-ESI(+)-HRMS for better characterization of metabolic pathway disruption upon anabolic practices in bovine

Abstract

The use of anabolic agents in meat producing animals is forbidden in Europe since 1988. Nevertheless, the possibility of widespread abuse of hormonal substances still exists, mainly due to economical benefits. The use of “omics” approach is a promising strategy to highlight anabolic hormone abuse by indirectly detecting their physiological action. In this context, the aim of this work was to set up a liquid chromatography – high resolution mass spectrometry based metabolomics workflow for the screening of a combined trenbolone acetate/estradiol implant abuse in cattle urine. Therefore, an untargeted metabolomics approach combining the information provided by reverse-phase liquid chromatography and hydrophilic interaction chromatography both coupled to high resolution mass spectrometry was developed and applied to characterize and compare cattle urinary metabolic profiles from non-implanted and implanted animals. The combination of both separation modes improved the metabolite information richness. Discrimination between treated and untreated animals was observed by application of multivariate statistical analysis. Moreover, OPLS models permitted to highlight the candidate biomarkers appearing as the ions which contribute the most in the observed discrimination. From the results obtained, metabolomics approaches can be considered as a powerful strategy for the detection of fraudulent anabolic treatments in cattle since global urinary metabolic response provides helpful discrimination.