The enzyme linked immunosorbent assay of aflatoxin has been adopted in Chinese Pharmacopoeia(2020 edition). Based on high-throughput screening of monoclonal antibodies technology, monoclonal antibodies that can specifically recognize the aflatoxin B_1 and the total amount of aflatoxin B_1, B_2, G_1, and G_2 in Chinese herbal medicines were prepared. By optimizing the concentration of coating antibody, enzyme-labeled antigen, and the reaction system of enzyme-linked immunosorbent assay, the enzyme linked immunosorbent assay(ELISA) were developed for detection of aflatoxins in Chinese herbal medicines, decoction pieces, and preparation of Chinese medicine. In this method, the recovery test of actual samples is 60%-120%, and the relative standard deviation is less than 15%. In addition, in view of the complicated and expensive pretreatment methods for the determination of aflatoxin in Chinese herbal medicine, we developed a highly efficient pretreatment method of liquid-liquid extraction of aflatoxin in Chinese herbal medicine without immunoaffinity column. As an effective method for the detection of aflatoxin, the ELISA can effectively reduce the aflatoxins testing cost of traditional Chinese medicine, and promote the detection ability at earlier stages of production, and strengthen the quality supervision of traditional Chinese medicine.
Keywords: Chinese Pharmacopoeia(2020 edition); aflatoxin; enzyme linked immunosorbent assay.