Development of a Competitive ELISA for the Detection of Furaltadone Marker Residue, 3-Amino-5-Morpholinomethyl-2-Oxazolidinone (AMOZ), in Cultured Fish Samples

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This report describes an enzyme-linked immunosorbent assay (ELISA) for tissue-bound metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) and the application to residue analysis in cultured fish samples. The residue is monitored as a marker for the drug furaltadone. The assay enables the detection of protein bound AMOZ in the form of 2-nitrophenyl derivative (2-NP-AMOZ) in sample supernatant or extract after acid hydrolysis and derivatisation with o-nitrobenzaldehyde. Polyclonal rabbit antibodies were produced with a new immunogen hapten, 2-NP-HXA-AMOZ. The new ELISA had adequate analytical sensitivity (IC50 value 0.325 μg kg−1 ; limit of detection 0.1 μg kg−1 ; to determine a trace of AMOZ residue and had a high selectivity. Recoveries of AMOZ fortified at the levels of 0.1, 0.5 and 1.0 μg kg−1 ranged from 89.8 to 112.5% with coefficients of variation of 12.4–16.2% over the range of AMOZ concentrations studied. The results obtained with the ELISA correlated well with those obtained by commercial test kits for 150 tested samples (r=0.984). The results suggest that the developed ELISA is highly specific, accurate, and sensitive method, which is suitable for high throughput screening for AMOZ residues.