The invention discloses a drink for supplementing human body energy and recovering people from fatigue as well as a preparation method and application thereof. The beverage is characterized by containing glucose-6-phosphate. The preparation method comprises the following steps: mixing glucose, adenosine triphosphate (ATP) and magnesium chloride; and obtaining a mixed solution containing glucose-6-phosphate in the presence of hexokinase serving as a catalyst. Besides the glucose-6-phosphate, the beverage provided by the invention also contains a defined amount of components such as glucose, sodium ions, potassium ions, magnesium ions, and the like. The components, after being combined, can fully perform a synergistic action, and effectively perform functions of quickly supplementing human body energy, and recovering people from fatigue. Meanwhile, the beverage can supplement the potassium ions, the sodium ions and the magnesium ions, and achieve a function of maintaining acid-base balance of a human body and improving the immunity. The nutrient solution containing glucose-6-phosphate is mild, suitable for long-term consumption and free of side effects.