Study Of The Visual Andrapid Immunosorbent Methods For The Detection Of Furaltadone Metabolites

In this paper, the colloidal gold immunochromatographic rapid test strip (GlCA) and immuno-affinity test column (IATC) were developed for the determination of furaltadone metabolites in animal derived food.The furaltadone metabolites antibodies were purified by protein A-sepharose4B. AMOZ was conjugate to ovalbumin (OVA) to prepare coating-antigen. Gold chloride was reduced with sodium citrate and sodium borohydride to make17,20,25nm colloidal golds, which were coupled with the polyclonal antibodiesas immuno-colloidal gold. Select the17nm colloidal golds as the best colloidal golds. Optimize and study the factors related the experience such as nitrocellulose membrane, pH, antigen and colloidal-gold labeled antibody dilution times, temperature and time incubating coating antigen, et al. The limit of detection of NPAMOZ colloidal gold immunochromatographic assay was3μg/L. The cross-reactivity against other nitrofuran and metabolite is very low, except for furaltadone. Eight kind of animal products determined to be free of the AMOZ, were spiked a certain concentration standards. After derivatized and concentrated pre-treatment, the samples were directly used for analysis by colloidal gold immunochromatographic assay. The visual detection limit of the AMOZ in animal derived food were0.5μg/kg.This paper proposes an immuno-affinity test column (IATC). The enzyme labeled antigen was prepared by the active ester method. In order to prepare antibodies gel,’ respectivel, antibody with Ago-Gel activated by hydrogen bromide, combine with the antibody of HRP to make their HRP gel, seal the Ago-Gel activated by hydrogen bromide to obtain the sealing gel. Antibody gel and HRP gel are mixed with sealing gel to achieve Detection layer and Control layer respectively and put into chromatography column, optimize and study the factors related the experience such as pH, substrate addition mode, establish a immune chromatography method with time-saving convenience. The limit of detection of NPAMOZ immuno-affinity test column was20μg/L. The cross-reactivity against other nitrofuran and metabolite is very low, except for furaltadone. The visual detection limite of the AMOZ in animal derived food were3μg/kg.The results obtained by G1CA assay and IATC assay were coherence with the results obtained by ELISA. And the detection step is simple, visual results can be obtained, without the aid of instruments, the entire detection time of less than10min, suitable for rapid screening of large numbers of samples, with good prospects for development.