The effect of different gamma-ray irradiations on the functional groups and the mechanical properties of chitosan, collagen, and chitosan/collagen blend have been studied. Commercially available chitosan and extract of collagen from bovine tendon by acid dissolution method were employed. Chitosan, collagen and chitosan/collagen blend membranes were prepared using the solvent evaporation method. The dried membranes were subjected to gamma-ray irradiation using 0, 15, and 25 kGy. The membranes then characterized by Infrared spectroscopy and measured in tensile and elongation at break. The irradiated chitosan membranes displayed reduced intensity of -OH, -CH, -NH and -C-O-C groups, whereas, the collagen membranes revealed amide I, II and III groups. The irradiated chitosan/collagen (50/50%) blend membranes demonstrated the decreased intensities in the –OH and amide groups. The application of gamma-ray irradiation had produced chitosan/collagen blend membranes with a tensile strength and elongation at break between the chitosan and collagen membranes. It was further determined that the decrease in the mechanical properties of the membranes was likely to be due to some changes of the functional groups.