Development and Residue Screening of the Furazolidone Metabolite, 3-Amino-2-oxazolidinone (AOZ), in Cultured Fish by an Enzyme-Linked Immunosorbent Assay
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- 作者：Chu-Chen Cheng ; Kuan-Huei Hsieh ; Yi-Chih Lei ; Yung-Te Tai ; Tong-Hsuan Chang ; Shi-Yuan Sheu ; Wen-Ren Li ; Tzong-Fu Kuo
- 刊名：Journal of Agricultural and Food Chemistry
- 出版年：2009
- 出版时间：July 8, 2009
- 年：2009
- 卷：57
- 期：13
- 页码：5687-5692
- 全文大小：700K
- 年卷期：v.57,no.13(July 8, 2009)
- ISSN：1520-5118
文摘
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 μg 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 μg kg−1. According to the test preparation record, the detection limit is 0.1 μg kg−1, which is well below the minimum required performance limits (MRPLs) for tissue-bound residues of AOZ at 1 μg 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 μg 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 μg 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.