Molecularly imprinted polymeric membranes, bearing the tetrapeptide derivative H-Asp(OcHex)-Ile-Asp(OcHex)-Glu(OBzl)-CH2-, were prepared during the membrane preparation (casting) process in the presence of print molecule Boc-L-Trp. The molecularly imprinted membranes thus obtained showed adsorption selectivity toward a print molecule family, such as L-Trp, L-Phe, L-Ala, L-Arg, and L-Glu. The tetrapeptide derivative in the molecularly imprinted membranes preferentially recognized the L-amino acid from the D-isomer. Enantioselective permeation was attained with the present membrane, and the D-isomer was permeated in preference to the L-isomer by using the concentration difference as a driving force for membrane transport. Electrodialysis of racemic amino acid showed the possibility that permselectivity directly reflects its adsorption selectivity. It was made clear that the optical resolution was attained by the molecularly imprinted polymeric membranes.