Abstract: A method for determination of deoxynivalenol （DON） in wheat by high performance liquid chromatography （HPLC） is reported. The sample was extracted with PEG8000-water. Filtrate was cleaned up by DON test HPLC affinity cartridges. The DON was separated on Symmetry C18 column, detected by UV-detector and quantified by external standard. Wheat seeds samples were fortified with DON at different levels. This procedure could contribute to a DON recoveries of 86.11%-95.24 %, variation coefficients of 0.88 %-2.59 % and detection limit of 0.1mg/kg.