Abstract: 【Objective】 The response surface methodology was used to optimize the production process of longan， red jujube and banana vinegar herbal beverage， analyze its anti-fatigue activity， and provide reference to promote the product development of“Gui Shiwei”series.【Method】 Longan， red jujubes and banana vinegar stock solution were used the materials， the production process of longan， red jujube and banana vinegar herbal beverage was optimized by determining the amount of longan extract addition， red jujube extract addition， brown sugar addition and banana vinegar stock solution addition on the basis of single-factor Box-Behnken response surface analysis and sensory evaluation study at four factors and three levels. And the anti-fatigue activity of the beverage was studied by measuring the time of exhaustion swimming experiment， the decrease of blood lactate concentration and urea nitrogen concentration in mice， with Red Bull Vitamin energy beverage as the positive control group.【Result】 The effects of four factors on the sensory score of longan， red jujube and banana vinegar herbal beverage process were as follows： addition of red jujube extract>addition of longan>addition of banana vinegar stock solution>addition of brown sugar solution， where the interaction between the amount of longan extract and the amount of banana vinegar had a significant impact on the sensory score of herbal beverage（P<0.05）. After optimization，the optimal formula was as follows：longan extract addition 55%，red jujube extract addition 25%，brown sugar solution addition 14%，banana vinegar stock solution addition 0.4%. Under this condition， the sensory score of longan， red jujube， banana vinegar and herbal beverage was 91.87， which was consistent with the predicted value of 92.12 of the design model， and the color of the finished product was golden， transparent and without precipitation. After detection， the total number of colonies was <10 CFU/mL and the coliform group was <10 CFU/mL， and pathogenic bacteria were not detected. The results of anti-fatigue activity showed that longan， red jujubes and banana vinegar herbal beverage could effectively improve the swimming time of mice with exhaustion， reduce lactic acid and urea nitrogen content in serum， and had anti-fatigue activity.【Conclusion】 The production process of longan， red jujube and banana vinegar herbal beverage optimized by response surface is stable and reliable， the produced beverage is golden and transparent， with uniform color， longan and red jujube flavor， moderate sweet and sour. The longan， red jujube and banana vinegar herbal beverage has certain anti-fatigue activity and can be developed into functional products.