Objective To clarify the improvement effects of total flavonoids from perilla leaves on the anti-fatigue capacity of adolescent athletes and to compare the effects of different purification methods and liquid-to-solid ratios on their efficacy.Methods Total flavonoids from perilla leaves were extracted using microwave-assisted extraction, and two purification methods were employed for total flavonoid purification. Male adolescent track and field athletes aged 16 years with comparable physical fitness were selected to participate in a trial involving total flavonoid intake. Exhaustion time, maximum oxygen uptake, and serum lactate, serum urea nitrogen, and creatine kinase levels were measured in the male adolescent track and field athletes before and after total flavonoid intake.Results Total flavonoids from perilla leaves significantly prolonged the exhaustion time and increased maximum oxygen uptake in adolescent athletes, while effectively reducing post-exercise serum lactate, serum urea nitrogen, and creatine kinase levels. The chromatographic separation method for total flavonoid purification effectively retained the anti-fatigue active components in total flavonoids from perilla leaves.Conclusion Total flavonoids from perilla leaves help improve the anti-fatigue capacity and exercise recovery efficiency of adolescent athletes.