Abstract: Antibiotic residues in animal tissues and products could bring risks to the health of consumers.The commonly employed methods for detecting antibiotic residues often involve time consuming protocols, whereas the fluorescence method was characterized as rapid and sensitive.In this study, a method had been developed for rapid screening of sulfamethoxazole residues in milk with fluorescence spectroscopy and the chemometrics tools.The established model was applied to 6 brands of commercial milk including pasteurized and UHT milk (whole, skimmed, semi-skimmed, high calcium) .The results showed a significant discrimination by partial least squares discriminant analysis (PLS-DA) between the negative and positive milk samples with a 3.12% of false negative probability, which was below 5% required by the European Union.And quantification could be acquired by further analysis with partial least squares (PLS) with a recovery between 87.9%～131.6%.The correlation coefficient was 0.9992 in the linear range of 0～210μg/kg, and the detection capability (CCβ) and detection limit (CCα) were 2.54μg/L and 5.07μg/L, respectively.The average relative standard error of the prediction model was 3.24%.This method showed potential in rapid screening and analysis of sulfamethoxazole residues in milk.