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Development of an enzyme-linked immunosorbent assay for the detection of florfenicol in fish feed

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A sensitive enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibody was developed for routine screening of florfenicol (FF) in fish feed. FF succinate was synthesised and conjugated to human serum albumin by N-hydroxysuccinimide-activated ester method as immunogen, while it was conjugated to ovalbumin by mixed anhydride reaction as coating antigen. The ELISA for FF showed an IC50 value of 2.5 ng/mL, and negligible cross-reactivities with other amphenicol family of antibiotics. The inter-assay recoveries of FF from fortified fish feed at levels of 2-20 mg/kg ranged from 98.8 to 117.6% with coefficients of variation (CV) of 9.3-14.2%, intra-assay recoveries ranged from 102.5 to 121.2% with CV of 10.9-13.8%. The developed ELISA were then validated by liquid chromatography method and liquid chromatography-tandem mass spectrometry method, the results indicated this ELISA could be used as convenient method for detection of FF in fish feed.
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Keywords: ELISA; fish feed; florfenicol; monoclonal antibody

Document Type: Research Article

Affiliations: 1: College of Animal Science and Technology, Agricultural University of Hebei, Baoding, People's Republic of China 2: Beijing Wanger Biotechnology Co. Ltd, China Agricultural University, Beijing, People's Republic of China

Publication date: March 1, 2009

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