Chitosan/natural rubber-latex film (CS/NR) vulcanized by electron beam irradiation (EB) vulcanization are an alternative material with high mechanical properties and solvent resistance in a non-polar solvent. The EB vulcanization technique can crosslink between carbon molecules of NR without the use of sulfur in the crosslinking process and generates a product that is environmentally friendly with no residual radiation and no waste from vulcanization when compared with sulfur vulcanization. The modification of NR by adding maleic anhydride and modification of CS by acetic acid to treat chitosan to become acidified chitosan in colloid form produces significant improvements in material properties. In this paper, Chitosan/natural rubber-latex film consisted of non-vulcanized NR, vulcanized NR by sulfur, and vulcanized NR by EB at doses of 150, 200, and 250 kGy with a chitosan loading of 0–15 phr. The existence of CS in the NR matrix was investigated using thermogravimetric analysis and the morphology was investigated using FE-SEM. The maximum tensile strength was found in vulcanized NR by EB at 200 kGy dose (12.4 MPa), which was 3.65 times more than that of neat NR and slightly higher than that of NR vulcanized by sulfur because the bond strength of carbon-carbon bonding was higher than in carbon-sulfur bonding. Moreover, from ozone testing, it was found that chitosan helped the degradation of this material.