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Development of a Monoclonal Antibody‐based ELISA for Detection of Sulfamethazine and N 4 −acetyl Sulfamethazine in Chicken Breast Muscle Tissue
Author(s) -
He Jihong,
Shen Jianzhong,
Suo Xun,
Jiang Haiyang,
Hou Xiaolin
Publication year - 2005
Publication title -
journal of food science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.772
H-Index - 150
eISSN - 1750-3841
pISSN - 0022-1147
DOI - 10.1111/j.1365-2621.2005.tb09012.x
Subject(s) - chemistry , monoclonal antibody , residue (chemistry) , metabolite , chromatography , high performance liquid chromatography , detection limit , sulfadimidine , antibody , ic50 , maximum residue limit , microbiology and biotechnology , biochemistry , biology , in vitro , immunology , pesticide residue , pesticide , agronomy
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 atlevels 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 2 residue.