The invention relates to a preparation method of an amphoteric cellulose adsorbent. The preparation method comprises the following steps: adding cellulose into a sodium hydroxide-urea aqueous solution, and stirring uniformly to obtain a cellulose suspension; freezing and unfreezing the cellulose suspension, and stirring and mixing uniformly to obtain a cellulose solution; centrifuging the cellulose solution; taking the supernatant; sequentially dropwise adding bis(2-acetoxy)amine derivative aqueous solution, bis(pyridine ethyl)amine derivative aqueous solution into the supernatant; stirring and reacting for 2-12h at 50-100 DEG C to obtain a reaction product; washing the reaction product with deionized water, acetone and ethyl ether successively; performing suction filtration and drying toobtain the amphoteric cellulose adsorbent. According to the amphoteric cellulose adsorption material prepared by the method provided by the invention, the surface is connected with ionic type groups with a clamp structure, and the ionic type groups are more than the active groups of single-chain ion group modified cellulose while the adsorption action is stronger; moreover, the method has the advantages of few reaction steps, mild reaction conditions and easiness in operation and is beneficial to large-scale production.