Development of a Monoclonal Antibody-based ELISA for Detection of Sulfamethazine and N~4-acetyl Sulfamethazine in Chicken Breast Muscle Tissue

摘要

An anti-sulfamethazine monoclonal antibody was developed in a BALB/c mouse immunized with Sulfamethazine (SM_2)-human serum albumin (HSA). Using this monoclonal antibody, an indirect competitive enzyme-linked immunosorbent assay (cELISA) was developed to detect SM_2 and its metabolites in chicken breast muscle tissue. The 50% inhibition value (IC_(50)) was 9.3 ng/mL. When SM_2 was spiked at levels of 20 to 200 ng/g, recoveries ranged from 81.3% to 104.2% with coefficients of variation (CVs) of 4.3% to 19.3%. The metabolite N~4-acetyl SM_2 was also evaluated by the same assay. When it was fortified at levels of 20 to 200 ng/g, recoveries ranged from 80.4% to 100.8% with CVs of 3.0% to 14.2%. The results were confirmed with analysis by high-performance liquid chromatography (HPLC). In an actual residue study, the results obtained by cELISA did not correlate well with those obtained by HPLC (P < 0.05). This might be due to the coextraction of cross-reactive SM_2-related residues that were not quantified by the HPLC method. The study indicated that the presence of residues should be anticipated when considering the maximum residue limit of SM., residue.