To improve the hydrophobicity of bagasse hemicellulose-based films, glutaraldehyde was applied when preparing films of original and cationic bagasse hemicellulose with the addition of polyvinyl alcohol and sorbitol. The results showed that the cationic modification could increase the hydrophobicity of the hemicellulose-based film, and the hydrophobicity of hemicellulose-based films crosslinked with glutaraldehyde also increased. However, cationic modification of hemicellulose decreased the stress of the hemicellulose-based film. While crosslinking with glutaraldehyde increased the stress of both the original and cationic hemicellulose-based films. Macrophotography indicated that the film formability of the original hemicellulose was better than that of cationic hemicellulose. Through SEM observation, the degree of bonding of different components of the films was found to be increased due to crosslinking with glutaraldehyde. The crosslinking reaction between glutaraldehyde and hemicellulose was further confirmed by FT-IR spectroscopy.