Cellulose acetate-based molecularly imprinted polymeric membranes were prepared using vanillin as template molecule. The microscopic structure of the resultant polymeric membranes was characterized by SEM and FTIR spectroscopy, and the selective binding properties and separation capacity of the membranes for vanillin and o-vanillin were tested with binding experiments and separate experiments, respectively. The results showed that the vanillin-imprinted polymeric membranes displayed higher binding and separation capacity for vanillin in comparison with the non-imprinted polymeric membranes; the maximum binding amount Qmax for vanillin is 387.3 µmol/g and the selectivity factor is 3.82 for vanillin/o-vanillin. In addition, the effect of cross-linker concentration on the binding of vanillin on the imprinted polymeric membranes was also evaluated and the optimal concentration is 320 mM.