The invention discloses a novel wheat antioxidant peptide and a preparation method thereof, belonging to the technical field of fine and further processing of agricultural products. According to the preparation method disclosed by the invention, a novel wheat antioxidant peptide product with cytoprotection activity is obtained through acid protease hydrolysis, different cut-off molecular weight membrane filtration, gel chromatography column separation, reverse high-efficiency liquid chromatography purification, and the like by taking wheat gluten protein (wheat gluten) as a raw material. The novel wheat antioxidant peptide disclosed by the invention has relative molecular weight in the range of 1746 Da and the structure of serine-glycine-alanine-aspartate-lysine-lysine-proline-isoleucine-lysine-alanine-asparagine-histidine; in addition, the product prepared by adopting the preparation method disclosed by the invention has high purity and higher free radical scavenging capacity, can be used for effectively repairing cell destruction and tissue damage and has great application prospect in cosmetics and functional foods or medicaments.