Physical fitness is an important indicator reflecting the mobility and health status of students. The assessment of the physical fitness of first-year female students is significant in identifying the limited components and providing a practical basis for organizing physical education in universities. The research aims to assess the physical fitness status of first-year female students at Saigon University. A cross-sectional descriptive study was conducted on 435 18-year-old female students enrolled in Physical Education I in the first semester of the 2024–2025 academic year. The research results show that, according to Decision No. 53/2008/QĐ-BGDĐT, 90.11% of students did not meet the overall physical fitness standards; among them, 81.84% did not pass the 5-minute run test, and 38.39% did not pass the 4 × 10 m shuttle run. Compared to the reference data for Vietnamese women aged 18, 5 out of 6 tests showed statistically significant differences; female students at Saigon University performed better in some components but worse in quick movement and agility. Conclusion: The physical fitness of first-year female students at Saigon University is not developing evenly; endurance and the ability to move and change direction are notable limitations. The results indicate the need to enhance the requirements for endurance and coordination development in the Physical Education program.