The invention relates to a high-quality pleurotus eryngii culture medium and a preparation method. The culture medium comprises the components in percentage by weight as follows: 58% of grain stillage, 22% of bean cakes, 12.5% of corncobs, 0.8% of glucose, 2.5% of urea, 0.15% of quick lime, 0.22% of hydrated lime, 0.15% of potassium dihydrogen phosphate and 0.15% of magnesium sulfate. In a preparation process, the bean cakes are smashed, and granular bean cake granules are obtained; the bean cake granules are soaked in hot water with a temperature higher than 60 DEG C, the water temperature is kept to be constant, soaking is performed for more than half an hour, the mixture is cooled naturally, and a bean cake soaking solution is obtained; the corncobs are placed into a sealed transparent glass or plastic container, water is added to the container, and the height of the water level is in accordance with the stacking height of the corncobs; the container is sealed and placed in a natural environment where sunlight can directly eradiate, and retting is performed for three months; and the mixture is mixed with the grain stillage, the culture medium is paved, the glucose, the urea, the quick lime, the hydrated lime, the potassium dihydrogen phosphate and the magnesium sulfate are prepared into a nutrient solution, and the surface of the culture medium is sprayed with the nutrient solution. The absorption and growth of pleurotus eryngii are facilitated, and the protein quality of the pleurotus eryngii is improved by the aid of the grain stillage.