The invention relates to braised beef with Rhodiola rosea capable of resisting exercise-induced fatigue and a preparation method thereof, and belongs to the technical field of food processing. The braised beef with Rhodiola rosea comprises the following raw materials in parts by weight: Rhodiola rosea medicine juice 70-90, beef 150-200, scallion 0.5-1, ginger 0.5-1, monosodium glutamate 0.1-0.5, salt 0.1-0.5, white sugar 0.5-1, cooking wine 0.1-0.5, seafood sauce 0.5-1, soybean sauce 1-3, star aniseed 0.05-0.15, starch 0.5-1, and cooking oil 1-5. The preparation method comprises the following steps: pre-treatment of beef, preparation of Rhodiola rosea medicine juice, mixing of seasonings, braising, packaging, and sterilizing. The food is rich in nutrition, reasonable in match and suitable for daily diet for fatigue people, and especially suitable for people after exercise. Meanwhile, the preparation method is simple and easy to perform and suitable for large-scale production, with considerable economic benefit.