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Antibiotic residues in milk are a serious health and technological problem in dairy processes. This study aims to verify the absence of administered antimicrobials after therapeutic treatments, taking into consideration the withdrawal period, and to evaluate the reliability of screening tests under field conditions after confirmatory HPLC-HRMS (High Performance Liquid Chromatography-High-Resolution Mass Spectrometry) Orbitrap analysis. Moreover, the presence of expected or non-targeted metabolites was investigated using the new Compound Discoverer approach. The presence of antimicrobial drugs was shown in 29% of the samples, and also sometimes their metabolites (for enrofloxacin and lincomycin), despite the fact that samples were collected at the seventh milking. Moreover, in 9% of the samples, undeclared treatments were revealed due to the presence of both parent drugs and metabolites. Lastly, the putative identification of two new enrofloxacin metabolites, ENRO-N-methylacetamide and ENRO-ornithine, was proposed. In the light of this evidence, it must be borne in mind that metabolites, some of which are pharmacologically active, may also pose a risk to consumers and for the entire processing of milk in the cheese industries.

Antibiotics and Non-Targeted Metabolite Residues Detection as a Comprehensive Approach toward Food Safety in Raw Milk