The invention discloses a preparation method of superfine yak bone powder. The superfine yak bone powder of which the particle size is 500-700nm is processed by preliminary boning, enzymolysis and residual meat removal and smashing on the raw material yak bone, wherein trypsase is adopted to process the preliminarily boned yak bone in the enzymolysis and residual meat removal process; the additive amount of the trypsase according to the amount of the yak bone is 1100-1300U / g; the enzymolysis temperature is 45-55 DEG C; the enzymolysis time is 1.5-2.5 hours; and the pH value is 10-11. According to the method disclosed by the invention, through mild degradation by using biological enzyme, impurities such as yak bone meat can be effectively removed, the purpose of high biological activity of calcium ions in the yak bone powder is achieved, the method has the advantages of simplicity in operation and strong practicability, the prepared yak bone powder is free of peculiar smell, the particle size of over 97% of bone powder in the product reaches 500-700nm, the crude protein is greater than or equal to 26%, the moisture is smaller than or equal to 3.2%, the crude fat is smaller than or equal to 0.5%, calcium is greater than or equal to 30%, phosphorus is greater than or equal to 13.8%, the superfine yak bone powder is even in dispersing, good in taste, free of impurity, and the nutrition in the yak bone is completely preserved, and is easy to be directly absorbed and utilized by a human body.