Abstract: New Zealand White rabbits were immunized with three artificial immunizing antigens that modeling the common structure of sulfonamides, and the immune effect were detected by ELISA. A new Ci-ELISA method against sulfonamides and standard curves were established using two polyclonal antisera because of their good inhibitory effect. The IC10 of polyclonal antisera from group A against SMD, SMM, SMZ, SDM, SDM′ and SD was 0.010, 0.046, 0.010, 0.067, 0.019 μg/mL respectively, which was less than the MRLs(0.1 μg/mL). The IC10 of polyclonal antisera from group C against SMD, SMM, SMZ, SDM, SQ, SD, sulfachloropyridazine and sulfachaoropyrazine were 0.009 1, 0.051, 0.058, 0.088, 0.020, 0.010, 0.014, 0.011 μg/mL respectively, less than the MRLs. The recoveries of two polyclonal antibodies ranged 54.5%111.8%, 54.2%-130.3% with CV of 3.6%-10.9% and 3.7%-5.9%, respectively.