Development of sensitive and fast immunoassays for amantadine detection

A sensitive indirect competitive enzyme-linked immunosorbent assay(ic-ELISA) method and a gold nanoparticle immunochromatographic strip were developed to detect amantadine (AM) in foods. A novel AM hapten was prepared, and a sensitive monoclonal antibody against AM was developed. The optimum ic-ELISA conditions included a pH of 7.4 and an ionic strength of 1.6% with no organic solvents. The half-inhibition concentration (IC50) value of ic-ELISA was 1.92ng/ml, with a limit of detection of 0.62ng/ml. The immunochromatographic test strip method had a visual cut-off value at 5g/kg. Chicken samples were spiked with three concentrations of AM (1, 2, and 5 mu g/kg) and analysed by ic-ELISA. Good recoveries were obtained (92.3% at 1 mu g/kg AM, 116.1% at 2 mu g/kg AM, and 91.8% at 5 mu g/kg AM).