The invention relates to a wheat germ protein source antioxidative peptide, and a preparation method and an application thereof. The wheat germ protein source antioxidative peptide comprises sequences shown as SEQ ID NO.1 and SEQ ID NO.2. The preparation method comprises the steps of making fresh wheat germs into defatted wheat germ protein powder; hydrolysizing with an alkaline protease, inactivating enzyme, centrifuging at a high speed, subjecting supernatant to ultrafiltration so as to remove large molecular substances, freeze-drying filtrate to obtain a wheat germ protein hydrolysate, passing a wheat germ protein hydrolysate solution through a macroporous absorbent resin column and a gel filtration chromatography column successively, and collecting components with the strongest antioxidant activity being a target product. The preparation method is simple and efficient in process. The obtained wheat germ protein source antioxidative peptide has relatively high antioxidant activity, has relatively small molecular weight and definite amino acid sequence, can clear free radicals and alleviate oxidative stress injuries, can also be used for antioxidant dietary supplements, and has the advantages of wide raw material sources, high security, low cost and the like, and has wide application prospects in the fields of foods, medicines, etc.