Background: Iron and zinc are important trace elements in humans. Deficiencies in these elements have been reported in several studies. However, few studies have investigated blood examination results in conjunction with human behavior. Thus, this study aimed to analyze students’ exercise, daily habits, and behavior in combination with blood laboratory examinations over a two-year study period.

Methods: Healthy female junior high and high school students were recruited between 2020 and 2021. Body weight and height were measured during annual physical and blood examinations. A survey was conducted on the type and frequency of exercise, and daily exercise habits. The data obtained were analyzed using a t-test, and logistic analysis was performed to compare quantitative characteristics.

Results: A total of 102 female students participated in this study. Among them, 51 students were examined twice over two consecutive years. The prevalence of iron, asymptomatic zinc (60–79 µg/dL), and symptomatic zinc (<60 µg/dL) deficiencies was 7.8, 47.6, and 1.2%, respectively. Only one student had symptomatic zinc deficiency based on the serum zinc concentration. This participant reported having rough skin and prolonged injury healing. Serum iron concentrations were significantly higher in the extreme exercise group than in the non-exercise group. No significant correlation was observed between serum zinc concentration and exercise habits.

Conclusions: Adolescent girls who exercise regularly may be motivated to maintain their iron intake. Serum zinc concentrations may affect skin-disordered behaviors in adolescent girls.