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Multiplexed Fluorometric ImmunoAssay Testing Methodology and Troubleshooting
Published on: December 12, 2011
Multidetermination of four nitrofurans in animal feeds by a sensitive and simple enzyme-linked immunosorbent assay
Jun Li1, Ju Xiang Liu, Jian Ping Wang
1College of Animal Science & Technology, Agricultural University of Hebei, Baoding Hebei, China.
Abstract:
In this study, the polyclonal antibody against furazolidone was produced with furazolidone coupling to protein carriers by a diazotization method and glutaraldehyde reaction, respectively. The antibody obtained showed good specificity toward furazolidone and various cross-reactivity toward nitrofurantoin, nitrofurazone, and furaltadone. Then, an indirect competitive enzyme-linked immunosorbent assay (ELISA) based on the antibody was first developed for multidetermination of four nitrofurans in animal feeds. The limit of detection (LOD) of the method was 0.2-2.1 ng/g depending on the component. After simple extraction, the fortified swine and broiler chicken feed samples were detected with recovery ranges of 75.9-86.4%. Results obtained from ELISA were confirmed by high-performance liquid chromatography (HPLC) with ultraviolet detection. Analysis of the unknown feed samples indicates that ELISA can be a practical tool for screening of nitrofurans in animal feeds before confirmation by HPLC.

