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Chitosan-based molecularly imprinted polymers (CHI-MIPs) were prepared under optimized conditions using chloramphenicol (CAP) as template molecule, acrylamide as functional monomer and CHI as functional matrix. The influences of template molecule and concentrations of functional monomer as well as imprinting conditions on the binding properties of the MIPs were analyzed. The adsorption properties were evaluated by kinetic binding experiments, static binding experiments and Scatchard analysis, which indicated that the synthesized polymers had high binding capacity. Results of competitive recognition studies showed that the imprinting factor of CAP was found to be 7.43, which was much higher than those of its analogues. Solidphase extraction (SPE) using the prepared CHI-MIPs as sorbents was then applied to selectively extract CAP from spiked honey and milk samples. The average recoveries of SPE were in the range of 94.96–100.58% with relative standard deviation less than 5%.