The invention discloses a shiitake mushroom chili sauce. The shiitake mushroom chili sauce comprises the following components in percentage by weight: 15%-17% of chilies, 40%-50% of a soybean sauce, 15%-17% of garlic, 15%-17% of shiitake mushrooms, 1.5%-1.7% of fresh ginger, 1.5%-1.7% of pod peppers, 1%-2% of sesame seeds, 0.5%-1% of white sugar, 1%-1.5% of onions, 5%-10% of peanuts, 0.5%-1% of sesame oil, 1%-1.5% of clean water, an appropriate amount of edible salt and an appropriate amount of edible oil. The invention further discloses a preparation method of the shiitake mushroom chili sauce. According to the shiitake mushroom chili sauce disclosed by the invention, the garlic, the sesame seeds, the peanuts, the onions and the like are added, so that the shiitake mushroom chili sauce has good effect of restraining bacterium breeding; in the canning process, the cans are disinfected with X rays, so that the bacterium breeding can be well reduced; an effect component, namely capsaicine, of the chilies in the shiitake mushroom chili sauce is an anti-oxidizing substance, can prevent relevant cells from generating metabolism, and can reduce the incidence rate of cancer cells; and theshiitake mushroom chili sauce has strong spicy taste, so that the shiitake mushroom chili sauce can stimulate the secretion of saliva and gastric juice, and can increase the appetite, promote intestinal peristalsis, promote digestion, reduce blood lipid and reduce weight.