With ever-developing analytical capabilities, the protection of the dairy chain from contamination by residues of veterinary drugs is improving. Legislative requirements are an inherent part of this process. Of antibiotics in dairy farming, representatives of the beta-lactams, tetracyclines, sulfonamides, (fluoro)quinolones, aminoglycosides, or polypeptide antibiotics are the most widely used. Due to the typically low levels of antibiotic residues in milk, mass spectrometry is the most commonly used detection technique. However, the interference of the sample matrix is one of its main limiting factors, and therefore, it needs to be eliminated. In the first step, the lipid fraction is removed and proteins are precipitated, followed by solid-phase or liquid–liquid extraction. The current trends include the reduction in the consumption of organic solvents (to reduce occupational hazards and burden to the environment) and automation, eliminating the influence of human error and optimizing the workflow. These trends lead to the development of new microextraction and automated techniques as well as the use of new sorbents and/or (green) solvents of natural origin. To capture the latest developments in the field and the relatively recent aforementioned trends, this review focuses on papers investigating antimicrobial residues in milk that were published between 2015 and 2024, providing an overview of emerging techniques applicable to antibiotic residue detection in milk samples.
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