A new glucose-responsive polymeric composite membrane that provided pulsatile insulin release was developed in our laboratory previously. To develop a clinically useful insulin delivery system, this study was designed to investigate factors influencing insulin stability during delivery by this membrane. The effects of stirring, release duration, insulin concentration and surfactant on insulin stability were studied under both incubation and delivery conditions in a buffer solution at 37 degrees C. The structural change of insulin was characterized by reverse-phase HPLC and circular dichroism. Hydrophobicity of various contact surfaces was determined by contact angle measurement. The results indicated that insulin concentration played an important role in the insulin stability, followed by stirring. Treating the membrane with a non-ionic surfactant prevented insulin denaturation during delivery through the membrane.