Abstract: An indirect competitive chemiluminescent enzyme-linked immunosorbent assay (ic-CLEIA) was developed for the rapid detection of amantadine and chloramphenicol residues in poultry meat based on efficient sample pretreatment. The dilution factors of coating antigen, antibody and horseradish peroxidase (HRP)-labeled secondary antibody, and competition time were optimized. The results showed that the half-maximum inhibition concentration (IC50) and the linear range were 0.33 μg/L and 0.06–1.77 μg/L for amantadine, and 0.039 μg/L and 0.010–0.179 μg/L for CAP, respectively. Samples were sequentially extracted with ethyl acetate and potassium carbonate solution, and then the extract was blown to dryness with nitrogen gas and purified by n-hexane extraction before being analyzed. The recoveries of amantadine and chloramphenicol from spiked samples were in the ranges of 92.55%–105.14% and 92.00%–112.50%, respectively, and the coefficients of variation were both less than 10.48%. Furthermore, the results determined by ic-CLEIA were highly correlated with those of instrumental analysis method (R2 > 0.98), which demonstrates that the established ic-CLEIA method has high accuracy and good reliability. This method represents a rapid immunoassay for the detection of veterinary drug residues in poultry meat.

Key words: poultry meat; amantadine; chloramphenicol; indirect competitive chemiluminescence enzyme-linked immunosorbent assay; pretreatment