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

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40 Scopus citations

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 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.

Original languageEnglish
Pages (from-to)5687-5692
Number of pages6
JournalJournal of Agricultural and Food Chemistry
Volume57
Issue number13
DOIs
StatePublished - 8 Jul 2009

Keywords

  • 3-amino-2-oxazolidinone (AOZ)
  • Eel
  • Enzyme-linked immunosorbent assay (ELISA)
  • Furazolidone
  • Nitrofurans
  • Residues

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