Chlorpromazine is extensively used as a sedative hypnotic in veterinary application, however, improper and over usage of chlorpromazine pose a serious threat to human health and animal production. Due to the difficulty in monitoring trace level of chlorpromazine residues in complicated biological matrices, specific adsorption materials for preconcentration and clean-up of chlorpromazine are indispensable for sample preparation before further determination. In this study, a new molecularly imprinted polymer (MIP) for recognition of chlorpromazine was obtained by bulk polymerization. Then the imprinting effect of MIP was confirmed via chromatographic evaluation by compared with the corresponding non-imprinted polymer (NIP). Furthermore, the adsorption selectivity and capacity analysis revealed that the MIP had high specification for chlorpromazine. Finally, with the optimized protocol of molecularly imprinted solid-phase extraction (MISPE), the target analyte in pig urine at different spiked levels were selectively separated and purified for routine HPLC-UV analysis, with the improved recoveries above 73.3% and the intraday RSD of less than 8.7%, proving that the obtained MIP as specific SPE sorbents for extraction of chlorpromazine is applicable. Crown Copyright (C) 2008 Published by Elsevier Ltd. All rights reserved.