Antigens synthesis and antibodies preparation for furazolidone and its metabolite 3-amino-2-oxazolidinone

Two haptens of 3-[(5-amino-furan-2-ylmethylene) amino]oxazolidin-5-one (FZ-NH₂) and 3-{[(4-carboxypbenyl) methylene]-amino}-2-oxazolidinone(CPAOZ)were synthesized. For FZ-NH₂, immunogens were prepared by glutaraldehyde and diazo salt methods. For CPAOZ, immunogens were connected by the methods of the active ester and mixed acid anhydride. Compared with the combination, indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed with coating antigen of FZ-NH₂-OVA via the glutaraldehyde method and immunogen of CPAOZ-KLH via active ester method. For furazolidone and its metabolite AOZ (NPAOZ as derivative), the sensitivities (IC₅0) were 2.0 μg/L and 2.5 μg/L, limits of detection (IC₁₅) were0.09 μg/L and 0.25 μg/L, respectively. A sensitive method was developed for the simultaneous determination of furazolidone in feed and its metabolite AOZ in tissue.