Abstract：The evaluation of milk quality is very important, which has already become a research hotspot. The non-destructive, rapid and sensitive analytical technique is urgently needed for ensuring the quality of milk. In this research, we established a new analytical technique based on three-dimensional (3-D) fluorescence spectrum, which can reflect the components information of milk with their unique fluorescence fingerprint. Compared with conventional fluorescence spectrum, the 3-D fluorescence spectrum provides more detailed and comprehensive information. By utilizing this technique, the pretreatment factors of milk are optimized by analyzing the changes of fluorescent components. Furthermore, the method can detect out 0.5 mg·L-1 antibiotic residual in milk. The 3-D fluorescence technique combined with Parallel Factor Analysis (PARAFAC) was used to evaluate the heat treatment of milk. A good consistency of 97.63% was calculated, which indicated that the developed 3-D fluorescence fingerprint technique was credible and could be utilized as a powerful tool to examine the quality of milk.
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