The solid phase extraction column is prepared by mass polymerization with bitertanol molecularly imprinted polymer (MIP) as solid phase extraction agent. Diniconazole and tebuconazole in fruit and vegetable can be separated and enriched by the column, and their residuals are determined by high performance liquid chromatography (HPLC). The average recoveries of diniconazole and tebuconazole are 74.6%～88.6% and 66.9%～73.3%, respectively. The detection limits are 0.5 μg/mL for diniconazole and 0.25 μg/mL for tebuconazole.