The invention discloses a preparation method of edible bird's nest polypeptide powder. The method comprises: selecting edible bird's nest, and performing air drying, pulverization wall breaking, sieving, enzymatic hydrolysis, ultrafiltration and drying to obtain edible bird's nest pure polypeptide powder; through cooperation of Chinese dates, Chinese wolfberry fruits, poria, glossy privet fruits, rose-boots, palmleaf raspberry fruits, Chinese angelica, Korean ginseng, American ginseng, tangshen, milkvetch roots, glossy ganoderma, pilose antler, musk, Chinese caterpillar fungus, saussurea involucrate, pearl powder and honey, and through cleaning, drying, grinding, microwave decoction, microfiltration, drying and grinding, making auxiliary material powder; and performing mixing according to a ratio to obtain the edible bird's nest polypeptide powder. The edible bird's nest polypeptide powder is fast in solution rate, good in solution effect, and easy to be absorbed by the human body. Through cooperation with multiple rare traditional Chinese medicines, the edible bird's nest polypeptide powder has the health-care function and the beautifying effect.