The invention provides a preparation method of 2,5-furandicarboxylic acid, which comprises the following steps: under air and/or oxygen conditions, putting 5-hydroxymethylfurfural and a catalyst intoa mixed solution of water and an organic solvent, and carrying out catalytic oxidation reaction to obtain the 2,5-furandicarboxylic acid, wherein the catalyst comprises a carrier and an active component loaded on the carrier; the active component is one or more selected from ruthenium, palladium, platinum and rhodium; the carrier is one or more selected from activated carbon, graphite, fullerene and graphene oxide. The mixed solution of water and the organic solvent is used as the solvent so that solubility of the product 2,5-furandicarboxylic acid is increased, efficient conversion of 5-hydroxymethylfurfural is achieved without adding an alkaline additive to increase the solubility, high-selectivity 2,5-furandicarboxylic acid is obtained, and the method is simple in operation method, mildin reaction condition, environmentally friendly, free of pollution and good in industrial application prospect.