The invention discloses a preparation method of water-soluble chitosan/polyvinyl alcohol composite films capable of being used as biological package materials. Water-soluble chitosan and polyvinyl alcohol are respectively dissolved in deionized water, the solution is subjected to mixing casting for forming films, then, the films are soaked into one kind of solution or several kinds of mixed solution from glutaraldehyde solution, borax solution, epichlorohydrin, glyoxal and sodium tripolyphosphate to be subjected to crosslinking for 40 to 60 minutes, peeling and natural drying or baking are carried out, and the water-soluble chitosan/polyvinyl alcohol composite films are obtained. The composite films have good antibacterial activity, biocompatibility and biodegradability, in addition, special gas blocking performance and excellent mechanical performance are realized, and the composite films have the advantages of chitosan and the polyvinyl alcohol. In addition, the production cost is low, the preparation process is simple, the production efficiency is high, and the environment is friendly.