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Development and Residue Screening of the Furazolidone Metabolite, 3-amino-2-oxazolidinone (AOZ), in Cultured Fish by an Enzyme-Linked Immunosorbent Assay

Development and Residue Screening of the Furazolidone Metabolite, 3-amino-2-oxazolidinone (AOZ), in Cultured Fish by an Enzyme-Linked Immunosorbent Assay

Chu-Chen Cheng, Kuan-Huei Hsieh, Yi-Chih Lei, Yung-Te Tai, Tong-Hsuan Chang, Shi-Yuan Sheu, Wen-Ren Li, Tzong-Fu Kuo

J Agric Food Chem. 2009 Jul 8;57(13):5687-92.

PMID: 19526989

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 (IC(50)) of 0.14 microg 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 microg kg(-1). According to the test preparation record, the detection limit is 0.1 microg kg(-1), which is well below the minimum required performance limits (MRPLs) for tissue-bound residues of AOZ at 1 microg 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 microg 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 microg 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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP80659	AOZ		80-65-9	Price

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