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

Abstract

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 Nhydroxysuccinimide-activated ester method as immunogen, while it was conjugated to ovalbumin by mixed anhydride reaction as coating antigen. The ELISA for FF showed an IC 50 value of 2.5 ng/mL, and negligible crossreactivities with other amphenicol family of antibiotics. The inter-assay recoveries of FF from fortified fish feed at levels of 2Á20 mg/kg ranged from 98.8 to 117.6% with coefficients of variation (CV) of 9.3Á14.2%, intra-assay recoveries ranged from 102.5 to 121.2% with CV of 10.9Á13.8%. The developed ELISA were then validated by liquid chromatography method and liquid chromatographyÁtandem mass spectrometry method, the results indicated this ELISA could be used as convenient method for detection of FF in fish feed.

Development and Validation of a Sensitive Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Screening of Florfenicol and Thiamphenicol in Edible Animal Tissue and Feed

“…Wu et al (Wu et al 2008) produced a pAb against FFA and developed an ic-ELISA to detect FFA with an IC 50 of 3.34 μg L −1 in muscle of swine and chicken. The group of Luo et al developed a panel of ELISA methods to detect FFA in swine muscle (Luo et al 2009b), FF in fish feed (Luo et al 2009a), and FF and TAP in swine feed (Luo et al 2011). Fodey et al (2013 produced a panel of pAbs, which the most sensitive antibody could bind TAP (IC 50 = 0.75 μg L −1 ), FF (IC 50 = 0.61 μg L −1 ), and FFA (IC 50 = 0.93 μg L −1 ).…”

“…Recently, FF and TAP are even administered to fish by incorporation into feed for control of bacterial diseases (Hao et al 2006;Luo et al 2009a). Therefore, the need for an effective residue monitoring programme in place for FF and TAP is clear.…”

Current trends in sample preparation for growth promoter and veterinary drug residue analysis

“…developed an ELISA method for screening FF residues in fish feed [231]. Sample were extracted with EtOAc, concentrated and purified by simple LLP.…”

Amphenicols stability in medicated feed – development and validation of liquid chromatography method

“…However, a limited number of papers dealing with animal feed formulations were found in available sources. The published methods for determination of amphenicols in feeds used the techniques of enzyme-linked immunosorbent assay (ELISA) (16,17), planar chromatography (30), liquid chromatography (13,14), and liquid chromatography tandem mass spectrometry (LC-MS-MS) (3,25). The microbiological technique (9), ELISA (16,17), and a procedure utilising biosensors (15) had to be chemically confirmed.…”

“…The published methods for determination of amphenicols in feeds used the techniques of enzyme-linked immunosorbent assay (ELISA) (16,17), planar chromatography (30), liquid chromatography (13,14), and liquid chromatography tandem mass spectrometry (LC-MS-MS) (3,25). The microbiological technique (9), ELISA (16,17), and a procedure utilising biosensors (15) had to be chemically confirmed. Single quadrupole gas chromatography-mass spectrometry (GC-MS) was also adopted for the determination, confirmation, and concentration measurement of CAP, TAP, FF, and florfenicol amine (FFA) (11,24).…”