Cyclododecyl 2,4-dihydroxybenzoate (CDHB) is very similar to zearalenone (ZEN) in terms of size, shape, steric conformation and property. It is commonly used as a template of ZEN for the preparation of molecularly imprinted polymers. N’N′-carbonyldiimidazole (CDI) and 1,8-diazabicyclo [5.4.0] Decacarbonyl-7-ene (DBU) are added in the reaction substrate as the activator and catalyst respectively to ensure the yield of CDHB. This study compares three different purification methods of column chromatography, high-speed countercurrent chromatography and preparative liquid chromatography. The method of preparative liquid chromatography obtained the highest purity product (≥98%), followed by the method of high-speed countercurrent chromatography. Based on different production requirements, all three methods can be used for the systematic preparation process. Structure of CDHB was confirmed by FTIR, UV-Vis, 1H NMR, 13C NMR and MS (ESI), which proved the method of CDHB synthesis was repeatable and stable. The yield of CDHB was over 70%. In terms of application, CDHB was introduced into graphene carriers during the preparation of molecularly imprinted polymers, which can greatly improve the selective adsorption capacity of molecularly imprinted polymer to ZEN. The kinetic adsorption behavior of adsorbent followed the pseudo-second-order kinetic model.