A simple and efficient extraction and purification procedure for Aflatoxin was developed by means of the immunoaffinity column (IAC-SEP®AFLA-B1) as a cleanup tool．AflatoxinB1 content in Grain, Food, Feeds, Nuts, Peanuts, Soy sauce, Vinegar, Chili, Pepper, Medicinal herbs and Wine samples are cleaned up by IAC and determined by HPLC or LC-MS/MS. It is a fast, simple, safe and highly accurate method for quantitatively measuring aflatoxin-B1.