The invention belongs to the technical field of Chinese herbal medicines, and discloses an anti-fatigue preparation and a preparation method thereof. The anti-fatigue preparation is prepared from monarch drugs of rhodiola rosea, Chinese olive and sea-buckthorn, and is sweet in taste and warm in nature; the effects of invigorating spleen, tonifying qi, clearing away lung-heat, relieving cough, promoting blood circulation and removing blood stasis are achieved; polygonatum sibiricum, wrinkled gianthyssop herb, orange peel and seville orange flower are used as ministerial drugs and have the effects of nourishing yin, moistening lung, tonifying spleen, replenishing qi, resolving turbidity with aromatics, nourishing blood, strengthening body and supplementing essential amino acids for human bodies, so that the oxygen content of in-vivo cells is increased; and mint, liquorice and spina date seed are used as adjuvant drugs for clearing head and eyes, soothing the liver, promoting the circulation of qi, replenishing vital essence, improving eyesight and regulating immunity. All the Chinese herbal medicines are mutually fused, and the nature and flavor of the Chinese herbal medicines are applied according to the symptoms, so that the effects of refreshing and restoring consciousness and increasing the oxygen content of body cells are achieved, the cerebral cortical layer cell viability is further improved, and the anti-fatigue purpose is achieved. The retention rate of effective components in the preparation is guaranteed in the preparation process, granules are convenient to take and have no toxic or side effect, and the content of the effective components in all the raw materials is lower than the mouse intragastric administration LD50 standard amount.