The invention discloses a one-pot synthesis method of an anti-liver cancer drug selenite chitosan copper. The one-pot synthesis method comprises the following steps: 1, dissolving chitosan in a formicacid solution to obtain a chitosan acid solution; 2, adding an aqueous copper sulfate solution into the chitosan acid solution, carrying out a complexation reaction under a heating condition for 1 h,and adjusting the pH value of the obtained solution to 7 for 2-3 times until the complexation reaction is complete and the pH value does not drop in order to obtain a product I; 3, heating the product I, adding an aqueous seleninic acid solution in batches, and carrying out a reaction to generate a product II; and 4, cooling the product II, adding isopyknic ethanol until no precipitate is generated, filtering the obtained solution, and washing and drying the precipitate to obtain the selenite chitosan copper. The invention further discloses an application of the selenite chitosan copper in the preparation of antitumor drugs. The method has the advantages of no great change of the pH value of the solution, avoiding of copper ion dissociation and other side reactions, ensuring of complete esterification of 6-hydroxyl groups of chitosan, and increase of the selenization amount.