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This study focuses on detection of antimicrobial residues in milk through Fourier transform near-infrared spectroscopy. Simulated and real samples were considered. The simulated ones take into account veterinary drugs added in milk samples in the following concentrations: enrofloxacin 100 μg/L, terramycin 100 μg/L, and penicillin 4 μg/L. The statistical tool used to discriminate the samples was the principal component analysis (PCA). Our results show that, with this experimental procedure, it is possible to discriminate different types of antimicrobials dissolved in milk. Moreover, the methodology was able to detect real sample milked on different days after the injection of ceftiofur hydrochloride which is in principle a zero grace period antimicrobial. The methodology proved to be fast and accurate within the maximum residue limits allowed by European Agency for Medicinal Products and Ministry of Agriculture Livestock and Food Supply from Brazil.