Novel polypyrrole (PPy)/polydimethylsiloxane (PDMS) composite membrane was prepared by interfacial polymerization to make a very effective gas separation membrane. We found that Polymerized PPy films as thin as ~200 nm could be chemically synthesized as freestanding membranes by using the interfacial polymerization technique. Additionally, we show that difference morphology of PPys films was obtained by controlling polymerization rate and more dense films were formed at low polymerization rate. Wide X-ray diffraction study showed the d spacing value of the PPy film decreased from 4.89 Å to 3.67 Å by the rate of polymerization decreases. According to d spacing value decrease, selectivity of a PPy composite membrane was increased dramatically and permeability was reduced gradually. This high selectivity was derived from d spacing closed to the kinetic diameter of nitrogen. These results indicated that the permeability is controlled by the diffusion coefficient, reflecting the packed structure of the PPy film. The highest selectivity value of composite membrane that was prepared by interfacial polymerization was O2/N2=17.2 and permeability for O2 was 40.2 barrer.