The invention provides a preparation method of pinctada fucata martensii meat anti-oxidizing oligopeptides, which comprises the following steps: (1) selecting pinctada fucata martensii meat to be put into cold water with cold homogenate to obtain a pinctada fucata martensii meat homogenate solution; (2) adding alkali protease in the pinctada fucata martensii meat homogenate solution, mixing uniformly, and carrying out ultrasonic complex enzymolysis; (3) further conducting enzymolysis for the homogenate solution obtained by the ultrasonic complex enzymolysis to continue carrying out constant temperature enzymolysis in water bath, eliminating enzyme after finishing the enzymolysis, and obtaining supernate containing the pinctada fucata martensii meat anti-oxidizing oligopeptides by centrifugal separation after cooling; and (4) gradually separating the supernate containing the pinctada fucata martensii meat anti-oxidizing oligopeptides by adopting 5-10kDa and 1kDa ultrafiltration membranes through membrane separation and purification treatment and collecting the filter liquor to obtain the inctada fucata martensii meat anti-oxidizing oligopeptides solution, and carrying out purification treatment to obtain powder thereof. The method has simple operation and new technique, can greatly improve the hydrolysis rate of the pinctada fucata martensii meat by ultrasonic auxiliary enzymolysis, shortens hydrolysis time and reduces production cost.