Approaches for the simultaneous detection of thiamphenicol, florfenicol and florfenicol amine using immunochemical techniques

• Three different haptens were employed to produce antisera to phenicol antibiotics.
• Various conjugation techniques were investigated for favouring generic antibody production.
• Antisera were compared in homologous and heterologous assays by sensitivity and specificity.
• Preferred conjugation techniques for producing generic antibody are discussed.

Thiamphenicol and florfenicol are antibacterial agents permitted for use as veterinary drugs in animals used for food production. However, as the EU has established maximum residue limits for both and the metabolite florfenicol amine, there is a requirement to monitor animal food products for their residues. In this study antisera were generated which can simultaneously detect thiamphenicol, florfenicol and florfenicol amine in an immunoassay. Details of the various coupling techniques employed to prepare immunogens and enzyme labels are provided and the antibodies produced have been assessed, in homologous and heterologous ELISA formats, with respect to sensitivity and specificity. It was found that while the antisera raised to thiamphenicol and florfenicol generally performed better in a heterologous set up, those raised to florfenicol amine were not only less affected by the assay format but also produced the most sensitive antibodies to all three target analytes. Antisera matched previous sensitivity (IC50 < 1 ng mL− 1) but had improved cross-reactivity (> 100%) to thiamphenicol and florfenicol.

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