Reportedly, daytime napping affects the physical fitness of athletes. However, results of these studies are conflicting, and may not be generalizable to all populations. Early adulthood is an important period linking adolescents and adults, during which building good physical fitness is crucial for their remaining lives. Thus, we investigated whether daytime napping duration is associated with physical fitness among Chinese university students. This study was based on an annual physical health examination for all university students and included 11,199 participants (6690 males; 4509 females). The daytime napping duration was assessed using a self-report questionnaire. Physical fitness was measured with a 50 m sprint; 1000 m (for males) and 800 m (for females) runs; standing long jump, sit-and-reach, pull-up (for males), and sit-up (for females) tests; and vital capacity. The adjusted association was evaluated using analysis of covariance. Of the participants, 86% napped regularly. After covariate adjustment was performed, significant V-shaped associations were observed between the daytime napping duration and the 50 m sprint and 800 m run results in males and females. Inverted V-shaped associations were observed between the daytime napping duration and the sit-and-reach, standing long jump, and pull-up test performances and vital capacity in males and between the daytime napping duration and the standing long jump test performance in females. Daytime napping for <30 min may have beneficial effects on physical fitness among university students.