Development of an indirect competitive ELISA for ciprofloxacin residues in food animal edible tissues.
An indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed to detect ciprofloxacin (CPFX) in food animal edible tissues. CPFX was converted by an active ester method into conjugates CPFX-bovine serum albumin (CPFX-BSA) and CPFX-human serum albumin (CPFX-HSA), which both allowed production of CPFX-specific rabbit antisera. In the ELISA, CPFX-HSA was coated onto the microtiter plate, followed by incubation with standard CPFX and anti-CPFX antibody. The indirect competitive ELISA revealed that the antisera have no cross-reactivity with penicillin, gentamicin, neomycin, sulfadiazine, and chlortetracycline. The antisera cross-reacted with enrofloxacin and norfloxacin about 69.8 and 44.6% as much as they did with CPFX. This ELISA was highly sensitive (0.32 ng/mL) to CPFX determination. Recovery of CPFX at 40 microg/kg was 75.58% in pork, 81.29% in chicken, and 84.30% in milk. The coefficients of variation varied from 3.7 to 9.2% over the range of CPFX concentrations studied. The linear detection range was between 1.6 and 1000 ng/mL. The results suggest that this ELISA is a specific, accurate, and convenient method for the detection of CPFX residues in food animal edible tissues.
Duan
J
J
Institute of Veterinary Pharmaceuticals, Huazhong (Central China) Agricultural University, Wuhan, Hubei 430070, People's Republic of China.
Yuan
Z
Z
D000818:Animals; D000829:Animals, Domestic; D002417:Cattle; D002939:Ciprofloxacin / Q000032:analysis*; D003429:Cross Reactions; D004350:Drug Residues / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D008460:Meat / Q000032:analysis*; D009643:Norfloxacin / Q000032:analysis; D012709:Serum Albumin; D012710:Serum Albumin, Bovine
D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D002939:Ciprofloxacin; D009643:Norfloxacin