Determination of 3-Amino-2-oxazolidinone in Animal Tissue by an Enzyme-Linked Immunosorbent Assay and a Time-Resolved Fluoroimmunoassay

Abstract

An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and a time-resolved fluoroimmunoassay (TR-FIA) for the determination of the furazolidone metabolite 3-amino-2-oxazolidinone (AOZ) were developed. 3-Amino-2-oxazolidinone was detected using its derivative, 3-[[(2-nitrophenyl)-methylene]-amino]-2-oxazolidinone (NPAOZ). 3-([(3-Carboxyphenyl)methylene]-amino)-2-oxazolidinone (CPAOZ) was used as the immunizing hapten for the production of monoclonal antibodies against NPAOZ. The ic-ELISA had a 50 percent inhibition (IC50) value of 0.82 microgram per liter and a detection limit of 0.11 microgram per liter. The recoveries from animal tissues spiked with AOZ from 0.2 to 5 micrograms per kilogram were 82.3–112.8 percent with coefficients of variation values of 3.5–12.5 percent. The time-resolved fluoroimmunoassay (TR-FIA) showed a 50 percent inhibition of 0.56 microgram per liter and a detection limit of 0.07 microgram per liter. The recoveries and coefficients of variation were 87.2–117.4 percent and 3.6–8.5 percent, respectively. The analysis of fortified samples by both methods provided similar results to those obtained by a standard high-performance liquid chromatography-tandem mass spectrometry method. These results indicate the ic-ELISA and TR-FIA methods are suitable for the determination of 3-amino-2-oxazolidinone in animal tissue.