Starch films are developed as the biodegradable packaging materials to replace the petroleum-based materials in recent years. Thus, it is extremely beneficial to improve the hydrophobicity and mechanical strength of starch films, through a novel approach of co-cross-linking in neutral conditions, with glyoxal and AZC. In this work, systematic studies have been conducted to assess the performance of the co-cross-linked starch along with the control starch and starch cross-linked by glyoxal or AZC alone. Results showed that the co-cross-linked starch films exhibited significantly improved hydrophobicity and strength and the wet stress reached 1.53 MPa, compared to the control, glyoxal or AZC cross-linked starch films. More interestingly, the co-cross-linked film also demonstrated excellent UV shielding capacity and transmittance at visible wavelength range. The reaction mechanism was revealed based on the findings from UV, FT-IR and NMR spectra. This work established an innovative approach to improving the performance of starch film in neutral conditions for packaging applications.