Determination of Sulfameter by Chemiluminescence Enzyme Immunoassay Based on Alkaline Phosphatase-Adamantane

OBJECTIVE To establish a chemiluminescence enzyme immunoassay detection method for the quantitative assay of sulfameter, using alkaline phosphatase (ALP)-adamantane (AMPPD) system. METHODS The diazotization-coupling method was used to combine horseradish peroxidase with sulfameter. The optimal conditions were as follows. The concentration of antibody-coated solid-phase was 5.0 μg·mL^-1. The ALP-sulfameter was diluted to l∶10. Every well was added by standards or samples (50 μL) and ALP-sulfameter (50 μL), then plates were placed at 37 ℃ for 1 h. After washed, each well was added by 100 μL AMPPD. The result was obtained by luminometer after adding substrate for 25 min. RESULTS The limit of detection was 28.0 pg·mL^-1, the relative standard deviations of intra-assay and inter-assay of ten samples were 13.5%-14.6% and 16.1%-17.5%, respectively. The regression equation was Y=-0.021 5X+5.172 6 (r=0.996 1), the linear range for the method was 50-10 000 pg·mL^-1. CONCLUSION ALP-AMPPD chemiluminescence enzyme immunoassay can be used to determine the residues of veterinary drugs of sulfameter.