Purpose: Caffeine is the most widely consumed psychostimulant of the methylxanthine class. Among adolescents, high-dose of caffeine consumption has increased rapidly over the last few decades due to the introduction of energy drinks. However, little is known about the time-dependent effect of high doses of caffeine consumption in adolescents. The present study aims to examine the short- and long-term influence of high-dose caffeine on behavior of adolescence. Methods: The animals were divided into three groups: a “vehicle” group, which was injected with 1 ml of phosphate-buffered saline for 14 days; a “Day 1” group, which was injected with caffeine (30 mg/kg), 2 h before the behavioral tests; and a “Day 14” group, which was infused with caffeine for 14 days. An open-field test, a Y-maze test, and a passive avoidance test were conducted to assess the rats’activity levels, anxiety, and cognitive function. Results: High-dose caffeine had similar effects in short-and long-term treatment groups. It increased the level of locomotor activity and anxiety-like behavior, as evidenced by the increase in the number of movements and incidences of rearing and grooming in the caffeine-treated groups. No significant differences were observed between the groups in the Y-maze test. However, in the passive avoidance test, the escape latency in the caffeine-treated group was decreased significantly, indicating impaired memory acquisition. Conclusion: These results indicate that high-dose caffeine in adolescents may increase locomotor activity and anxiety- like behavior and impair learning and memory, irrespective of the duration of administration. The findings will be valuable for both evidence-based education and clinical practice.