The invention provides a preparation method of a cellulose acetate membrane surface crosslinked chitosan hydrophilic membrane and belongs to the technical field of membrane surface modification. A cellulose acetate membrane is activated by an acid solution, and has coupling crosslinking with the acetic acid water solution of the chitosan, so that a hydrophilic membrane material of the cellulose acetate membrane surface crosslinked chitosan can be prepared. The preparation method of the invention is simple in technology, low in cost, natural, and environmentally-friendly. The cellulose acetate membrane modified by the method is good in hydrophillic nature and binding capability, is good in mechanical strength and mechanical toughness, can be widely applicable to the field of biochemistry, molecular biology and environment, such as the immunoassay, the protein and DNA (deoxyribonucleic acid) analysis, the colloidal gold detection card, the absorption and removal of the toxic and harmful substances in the environment such as the environmental hormone and the like, and can be taken as biomedical material due to the improvement of the mold resistance of a cellulose acetate membrane.