The study evaluates findings on the impact of γ-irradiation on the single-use, multilayer plastic film that is manufactured for the biotechnological and biopharmaceutical environments. It is comprised of a multilayer film that offers durability and gas (O 2 , CO 2 and H 2 O) shielding properties. Polyethylene (PE) and ethylene vinyl alcohol (EVOH) are the parts within this film, which as a whole, is PE/EVOH/PE. The impact of γ-irradiation on the nonirradiated and γ-irradiated (from 30 kGy up to 270 kGy) PE multilayer film is investigated by colorimetry measurement, Fourier transform infrared and Raman spectroscopies, pH measurement, ion chromatography and conductivity measurement, tensile tests, X-ray photoelectron spectroscopy, differential scanning, and oxygen and water vapor transmission rate. AComDim and Principal Components Analysis are two chemometric tools that are utilized to understand both physical and chemical changes, along with the main factors contributing to polymer degradation in the PE multilayer film while under γ-irradiation.