Domoic acid (DA) was used as the target compound, ?structural analog 1,3,5-pentanetricarboxylic acid was used as template molecules, 4-vinyl pyridine?as the functional monomer and ethylene glycol dime-thacrylate?as the cross-linking agent for preparation of the molecularly imprinted polymers (MIPs). After Soxhlet extraction of the template 1,3,5-pentanetricarboxylic acid, binding sites were formed in the polymer with complementary size, shape, and positioning of chemical functionalities to 1,3,5-pentanetricarboxylic acid and also structural analog domoic acid. The molecularly imprinted?polymer was investigated with the aid of equilibrium binding experiments to evaluate the molecular recognition and binding characteristics of the imprinted polymer. The results show that compared with non-imprinted polymer, the imprinted polymer exhibits a much higher affinity for domoic acid.?The molecularly imprinted solid-phase extraction(MISPE) columns were prepared from a 1.0 mL glass syringe packed with 150 mg of the molecularly polymer. The MISPE procedure performed in an off-line mode coupled with reversed-phase high performance liquid chromatography was developed to extract domoic acid from Mytilus edulis and seawater samples. The mean recoveries of domoic acid from Mytilus edulis and artificial seawater spiked with 2 mg/L domoic acid were (93.4± 4.9)% and (89.7±3.2)% (n=3), respectively.