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    Please use this identifier to cite or link to this item: http://ir.lib.ncu.edu.tw/handle/987654321/28282

    Title: Development and Residue Screening of the Furazolidone Metabolite, 3-Amino-2-oxazolidinone (AOZ), in Cultured Fish by an Enzyme-Linked Immunosorbent Assay
    Authors: Cheng,CC;Hsieh,KH;Lei,YC;Tai,YT;Chang,TH;Sheu,SY;Li,WR;Kuo,TF
    Contributors: 中央大學
    Date: 2009
    Issue Date: 2010-06-29 19:45:26 (UTC+8)
    Publisher: 化學研究所
    Abstract: A sensitive and specific polyclonal enzyme-linked immunosorbent assay (ELISA) for the determination of tissue-bound metabolite 3-amino-2-oxazolidinone (AOZ) is described. The procedures allow for the detection of protein-bound AOZ in the form of a 2-nitrophenyl derivative (2-NP-AOZ) in the sample supernatant or extract after acid hydrolysis and derivatization with o-nitrobenzaldehyde. The polyclonal rabbit antibodies were produced with the immunogen hapten, 2-NP-HXA-AOZ, and the 50% inhibition values (IC50) of 0.14 mu g kg(-1) of AOZ was achieved with the most sensitive antibody A0505. The mean lower detection limit of the ELISA method is about 0.025 mu g kg(-1). According to the test preparation record, the detection limit is 0.1 mu g kg(-1), which is well below the minimum required performance limits (MRPLs) for tissue-bound residues of AOZ at 1 mu g kg(-1) in the European Communities. In the present study, we investigated the use of homemade ELISA, a new immunoassay, to monitor the presence of the furazolidone marker residue in 370 samples of cultured fish. Adopting 0.3 mu g kg(-1) AOZ as a cutoff value, the ELISA has a sensitivity of 100% and a specificity of 98.5% versus high-performance liquid chromatography-mass spectrometry (HPLC-MS) at a cutoff of 0.3 mu g kg(-1) and gives no false-negative rate results. From the practical point of view, the homemade kit could be advantageously used for the screening of large groups of animal-edible tissue samples and the kit employed has good reliability even in routine application for the control of the illegal use of the drug.
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