In recent years， the problem of antibiotic residues in food is one of the food safety problems that people are concerned about. To determine the residue of semicarbazide (SEM) in crayfish， this study provides a method for the determination of SEM residues in crayfish based on Excitation-emission matrix fluorescence combined with chemometrics. After simple pretreatment， the sample was derivatized with 2-hydroxy-1-naphthylaldehyde and SEM to produce 2-hydroxy-naphthylaldehyde nitrofuran metabolites derivative (SEM-HN) with fluorescence properties. The fluorescence spectrometer collects the fluorescence spectra of the sample to obtain the three-dimensional fluorescence data matrix. After the blank sample was deducted， the excitation-emission matrix fluorescence data matrix was analyzed using PARAFAC and ATLD in chemometrics. Both algorithms can get better quantitative analysis results. In order to evaluate the accuracy of the two algorithms， the quality factors were calculated: sensitivity (SEN)， selectivity (SEL)， detection limit (LOD) and predicted root mean square error (RMSEP). The results show that the accuracy of the two algorithms is good. In order to further verify the performance of PARAFAC and ATLD， EJCR test analysis is used. The results show that ATLD algorithm is better than PARAFAC algorithm. Excitation-emission matrix fluorescence combined with ATLD algorithm can quickly determine SEM residues in crayfish.