In this paper, a novel biomimetic enzyme-linked immunoassay method (BELISA) was successfully established for the detection of histamine and tryptamine, based on catalytically active cupric oxide@gold nanoparticles (CuO@Au NPs) as a marker and a molecularly imprinted polymer (MIP) as the biomimetic antibody. Under optimized conditions, the detection limitations of the BELISA method for histamine and tryptamine were 0.04 mg L-1 and 0.14 mg L-1, respectively. For liquor spiked with histamine and tryptamine, the BELISA method delivered satisfactory recoveries ranging from 89.90% to 115.00%. Furthermore, the levels of histamine and tryptamine in fish, soy sauce, and rice vinegar samples were detected by the BELISA method and a high performance liquid chromatography method, with no significant difference between the two methods being found. Although the catalytic activity of nanozymes is still lower than that of natural enzymes, the BELISA method could still sensitively determine the histamine and tryptamine levels in food samples.
Keywords: biomimetic immunoassay; histamine; nanozyme; tryptamine.