A phenylalanine imprinted membrane without a sponge layer containing macrovoids was prepared by the wet phase inversion method. The structure of the phenylalanine imprinted membrane prepared by an in-situ implanting procedure was denser than that of the membrane prepared by post implanting. Consecutive washings with distilled water and acetic acid solution removed nearly all of the template molecules from the membrane prepared by the post implanting procedure, although much of the template molecules remained in the membrane prepared by in-situ implanting. The removal of the template molecules from the membrane increased the population of the free COOH groups but reduced that of the dimerized COOH groups in the membrane. A D-Phenylalanine imprinted membrane prepared by post implanting selectively adsorbed D-phenylalanine from a racemate solution, where the adsorption selectivity reached 4.79. A D-phenylalanine imprinted membrane prepared by in-situ implanting exhibited an inverse adsorption affinity.