The invention belongs to the technical field of biological sensors, and discloses a non-enzyme sensor based on nitrogen-doped graphene and a preparation method and application thereof. The sensor consists of a reference electrode, a counter electrode and a modified working electrode, wherein the modified working electrode consists of a working electrode and a material recognizing membrane cured on the surface of the working electrode, and the material recognizing membrane consists of a nitrogen-doped graphene composite material and perfluorinated sulfonic acid resin. The non-enzyme sensor provided by the invention is applied to the detection of hydrogen peroxide, is wide in detection range, is low in detection limit, and has stable performance and a good application prospect; a reaction is implemented in a neutral environment at the room temperature.