Abstract: In order to determine the optimal extraction conditions of amino acid flavor substances in Hypsizygus marmoreus, by using the extraction rate of amino acid nitrogen as the index, the ultrasonic extraction technology of amino acid flavor substances of Hypsizygus marmoreus was optimized by using the single factor experiment and orthogonal experimental design. The results of the single factor experiment showed that the extraction temperature of 70℃, the ratio of material and water of 1:14, and the extraction time of 45 min were more suitable. The results of the orthogonal experiment showed that the factors affecting the extraction rate of amino acid nitrogen were the temperature > the time > the ratio of material and water, and the temperature had the biggest effect, followed by the time, and the ratio of material and water had the least effect. The optimum technological conditions for the ultrasonic extraction of amino acid flavor substances in Hypsizygus marmoreus were as follows:the temperature was 80℃, the time was 30 min, the ratio of material and water was 1:16. Under the optimum conditions, the extraction rate of amino acid nitrogen was (0.915±0.007)mg·g^-1.