The invention discloses a fructus cannabis xanthine oxidase inhibitory peptide and a preparation method thereof. The sequence of the inhibitory peptide is shown as SEQ ID NO: 1. The method comprises the following steps: performing virtual enzymolysis on fructus cannabis protein by using a computer technology, and evaluating the biological activity of polypeptide to obtain a fructus cannabis polypeptide library; evaluating the combination mode of the polypeptide and xanthine oxidase through molecular docking to obtain a candidate hemp seed polypeptide sequence; chemically synthesizing the xanthine oxidase inhibitory peptide and measuring the inhibitory activity of the xanthine oxidase inhibitory peptide; the xanthine oxidase inhibitory peptide is obtained by performing enzymolysis on the fructus cannabis protein by using corresponding protease and identifying the fructus cannabis protein. The inhibitory peptide has the characteristics of safety, no toxic or side effect and better water solubility, and can continuously and stably inhibit the activity of xanthine oxidase. The compound has a wide application prospect in the fields of medicines for preventing and treating gout and hyperuricemia, functional food additives and long-term treatment and health care of patients.