title: Development of an indirect competitive ELISA for the detection of furazolidone marker residue in animal edible tissues.
abstract: Due to its carcinogenicity and mutagenicity, furazolidone has been prohibited completely from being used in food animal production in the world since 1995. To monitor the illegal abuse of furazolidone, a polyclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed for the determination of tissue-bound furazolidone metabolite 3-amino-2-oxazolidone (AOZ). The highly specific antibody was targeted for PAOZ, the benzaldehyde derivative of AOZ. The 50% inhibition values (IC 50) of 0.91 microg/L for AOZ was achieved with the most sensitive antibody Ab-B1 by altering ELISA conditions. In the ELISA, sample extraction and cleanup were performed by an is MAX cartridge following combined hydrolysis of the tissue-bound AOZ and derivatization of the homogenized tissues with benzaldehyde. The limits of detection (LOD) calculated from the analysis of 20 known negative tissue samples (swine liver, swine muscle, chicken liver, chicken muscle,and fish muscle) were 0.3-0.4 microg/kg (mean+3 SD). Recoveries of AOZ fortified at the levels of 0.4, 1, and 5 microg/kg ranged from 55.8 to 96.6% in the tissues. The coefficients of variation were less than 20% over the range of AOZ concentrations studied. The linear detection range was between 0.1 and 25.6 microg/L. Validation of the ELISA method with swine muscle and liver from furazolidone-treated pigs was carried out using HPLC, resulting in a similar correlation in swine muscle (r=0.99) and in swine liver (r=0.98). The results suggest that this ELISA is a specific, accurate, and sensitive method of detecting AOZ residues in animal edible tissues.
authors.affiliation: National Reference Laboratory of Veterinary Drug Residues/MOA Key Laboratory of Food Safety Evaluation, Huazhong Agricultural University, Wuhan 430070, People's Republic of China.
authors.forename: Chao
authors.initials: C
authors.lastname: Chang
authors.forename: Da-peng
authors.initials: DP
authors.lastname: Peng
authors.forename: Jin-e
authors.initials: JE
authors.lastname: Wu
authors.forename: Yu-lian
authors.initials: YL
authors.lastname: Wang
authors.forename: Zong-hui
authors.initials: ZH
authors.lastname: Yuan
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000032:analysis; D000906:Antibodies; D000918:Antibody Specificity; D002851:Chromatography, High Pressure Liquid; D003257:Consumer Product Safety; D004350:Drug Residues / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D005664:Furazolidone / Q000032:analysis*; D006801:Humans; D008460:Meat / Q000032:analysis*; D023303:Oxazolidinones; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000890:Anti-Infective Agents; D000906:Antibodies; D023303:Oxazolidinones; D005664:Furazolidone; C000209:3-amino-2-oxazolidinone