The present study aimed to examine the differences in sleep hygiene, balance, strength, agility, and maximum aerobic speed (MAS) between two groups of female handball players aged under 14 (U14) and under 17 (U17) years. Seventy-two female handball players participated and were divided into two groups according to age: U14 (n = 36, age: 13.44 ± 0.5 years) and U17 (n = 36, age: 15.95 ± 0.76 years). Sleep hygiene was evaluated using three questionnaires: Sleep quality and sleepiness via the Pittsburgh (PSQI) and Epworth (ESS) questionnaires, and the insomnia questionnaire via the measurement of the insomnia severity index (ISI). Physical fitness was evaluated with the stork balance tests with eyes open (OEB) and closed (CEB), the vertical jump (SJ), horizontal jump (SBJ), and five jump (FJT) tests, the agility (t-test) and the maximum aerobic speed (MAS) tests. No significant differences were shown between U14 and U17 players in all PSQI, ISI, and ESS scores, and balance and strength performances. Meanwhile, the U17 players’ performances were significant better in agility quality (p = 0.003 < 0.01) and MAS (p = 0.05) compared to the U14 players. Biological gender specificity during the maturation phase may inhibit the improvement of balance, and strength performances between the age of 13 and 17 years, while agility and MAS performances are more affected by age alteration