A method of ELISA for the determination of furazolidone metabolites residues in aquatic products is established in this paper. There were no literatures about determination of AOZ residues in aquatic products by Enzyme linked Immunosorbent assay (ELISA) at home. Samples were derivatived with 2nitrobenzaldehyde and extracted with ethyl acetate. Then they were purified with nhexane and determinated by ELISA. In the paper, effects of different disposal conditions including dilution factors, extraction times, illumination conditions and shelf time to recovery were researched. By determination on Eriocheir sinensis, Micropterus salmoides and Procambarus clarkii spiked with AOZ, recovery, precision and detection limit of the method were researched by single laboratory validation approach as well as between laboratory comparisons. Results show that there is a remarkable effect of dilution factors on the recovery while no such effect is displayed for extraction times by ethyl acetate, illumination conditions and shelf time. So the best disposal condition is that dilution factor and extraction time are both one. The average recoveries are 99.2%，96.0%and 100.0% respectively when the samples are spiked with 0.60 μg·kg^-1，2.00 μg·kg^-1 and 6.00 μg·kg^-1 of AOZ. The detection limit is 0.3 μg·kg^-1. The relative standard deviations(n=3) for intraassay are 1.76%-12.57% and coefficients of variation for interassay are 5.43%-8.58%. Three positive samples analyzed by ELISA are confirmed by LCMS and no false positive samples are found. The method is sensitive with a fairly good rep roducibility and suitable for the determination of furazolidone metabolites(AOZ) in aquatic products.