The permeabilities of three kinds of solutes with similar sizes such as anionic benzenesulfonic acid, neutral styrene glycol, and cationic theophylline in the chitosan membrane were investigated. The chitosan membrane becomes cationic below its pKa value. Benzenesulfonic acid showed the highest permeability, whereas theophylline showed the lowest, although these solutes have almost the same size. This can be explained by the electrostatic attraction or repulsion between the solute and the membrane instead of the size exclusion effect. The permeabilities of benzenesulfonic acid and theophylline increased and decreased, respectively, with the decrease of pH from 7.4 to 4.0 because of the increase of the charge density of the membrane. Thus, the selectivity of benzenesulfonic acid to theophylline increased and reached about 30 at pH 4.0, in contrast to the selectivity of nearly unity in poly(vinyl alcohol) (PVA) membrane. The partition coefficients for three solutes showed the similar tendencies to those in the permeabilities. Contrary to the results of the permeabilities, the partition coefficients increased with the increase of the degree of the crosslinking in the membrane. This is probably due to the increase of the electrostatic interaction between the solute and the membrane brought about by the smaller mesh size and also due to the increase of the bound water fraction in the membrane.