Abstract: A chemiluminescent enzyme immunoassay based on magnetic beads was developed for quantitative determination of chloramphenicol (CAP) residues in pork samples. Sample, anti-chloramphenicol antibody, and magnetic beads coated with goat anti-mouse antibody were mixed on the microwell plate. After reacting for a certain period of time, CAP-horseradish peroxidase (HRP) was added and incubated. Then chemiluminescence substrate solution was added after washing the magnetic beads three times. Ten minutes later, the results were recorded by a microplate reader. Besides, the amount of magnetic beads coated with goat anti-mouse antibody, CAP-HRP, antibody concentration and reaction pattern were optimized. A sensitive and accurate magnetic bead-based chemiluminescent enzyme immunoassay for CAP was successfully developed. The limit of detection was 0.013 μg/kg, and good linear relationship was observed in the concentration range between 0.060 and 2.421 μg/kg. The recoveries of spiked pork samples varied from 85.24% to 93.57%, with relative standard deviation of less than 11.5%.

Key words: chemiluminescent enzyme immunoassay, chloramphenicol, magnetic beads, swine muscle