Dissertation
Dissertation > Medicine, health > Preventive Medicine,Health > Nutrition, hygiene,food hygiene > Food hygiene and food inspection > Food hygiene and inspection

Enzyme-Linked Immunosorbent Assay for the determination of Furazolidone and Its Metabolite

Author LiuZuoZuo
Tutor WangShuo
School Tianjin University of Science and Technology
Course Nutrition and Food Hygiene
Keywords furazolidone AOZ ELISA antibody feed edible tissue
CLC R155.5
Type Master's thesis
Year 2010
Downloads 85
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A sensitive polyclonal ELISA was developed for the determination of furazolidone and its tissue metabolite3-amino-2-oxazolidinone (AOZ). The antibodies were compared from immunogens of CPAOZ-KLH-1and CPAOZ-KLH-2preparatived via the active ester method and the mixed acid anhydride method. In the indirect competition ELISA (ciELISA), the heterologous coating antigen of FZ-NH2-OVA was produced by the glutaraldehyde method. The IC50and limit of detection (IC15) of the icELISA using antibodies against CPAOZ-KLH-1were2.5μg/L and0.25μg/L of3-{[(2-nitrophenyl) methylene] amino}-2-oxazolidinone (NPAOZ),2.0μg/L and0.09μg/L of furazolidone. The direct competition ELISA (cdELISA) showed lower LOD and non-significant cross-reactivity with the other nitrofuran drugs and their metabolites. The IC50and limit of detection (IC15) of the dcELISA were2.1μg/L and0.02μg/L of NPAOZ,2.8(μg/L and0.05μg/L of furazolidone. Considering illegal use of furazolidone in animal feed and the carcinogenicity and mutagenicity of its metabolite AOZ, we measured the residues of furazolidone in animal feed and AOZ (AOZ in the form of NPAOZ) in animal tissues of swine muscle, swine liver, swine kidney, chicken muscle, chicken liver and shrimp muscle. The recoveries at the three levels of1,5,20μg/kg were in the range of76%-109%, the CV were in the range of4.7%-11%. The correlations of the AOZ (R2=0.9976) and furazolidone (R2=0.9999) between the datas obtained using the ELISA and HPLC were very well. This study is the first publication for the simultaneous detection of furazolidone drug in the animal feed and metabolite in the animal tissues.

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