Purpose: To obtain relationship between retention behaviors of solutes on immobilized artificial membrane (IAM) and their absorption and distribution character in vivo.Methods: Retention factors of structure-diversely drugs under two pH conditions (6.0 and 7.0) on IAM were determined (log k1AM).Sigmoid and multiple stepwise regression were pertbrmed to obtain relationship between log k1AM and intestinal absorption and blood-brain barrier permeation.Results: Sigmoid relationship was observed between log (k1AM/MW3) and fraction of intestinal absorption (FA) for drugs with transcellular diffusion.Satisfying multiple linear relationship was obtained between log k1AM modified by molecular weight and blood-brain barrier permeation (log BB).Conclusion: Retention behavior of drugs on immobilized artificial membrane can be used to predict their intestinal absorption and blood-brain barrier permeation.