Background/Objectives: This study addresses the growing interest in nutritional supplements that improve athletic performance in endurance sports. Previous research suggests that nitrates in beetroot juice enhance blood vessel dilation and oxygen delivery to muscles. However, the effects of these nitrates on cardiopulmonary performance in female athletes remain underexplored. The aim of this study was to evaluate the effect of beetroot juice supplementation on aerobic work capacity in female endurance athletes. Methods: A cardiopulmonary exercise test (CPET) was conducted to assess aerobic work capacity. Eighteen healthy female endurance athletes (22.9 ± 5.6 years) participated in the study. The participants were randomly assigned to two groups: the control group (placebo group n = 9), which received a nitrate-free placebo beverage, and the experimental group (beetroot juice group n = 9), which consumed 50 mL of beetroot juice concentrate (~6.2 mmol nitrate) two and a half hours before the second test. Results: The results showed that the beetroot juice group demonstrated significant improvements in minute ventilation (VE), respiratory equivalents (VE/VO2 and VE/VCO2), and heart rate (HR) (p < 0.05). Maximal oxygen consumption (VO2 max) increased by 4.82% in the beetroot juice group (from 35.24 ± 5.07 to 36.94 ± 4.91 mL·min−1·kg−1), whereas a small decrease was observed in the placebo group. Conclusions: These findings indicate that beetroot juice may be an effective ergogenic aid, enhancing oxygen utilization and energy production during exercise in female athletes. In terms of practical applications, beetroot juice could contribute to improved athletic performance and serve as a valuable addition to athletes’ nutritional plans. Future studies should explore the long-term effects, optimal dosages, and duration of supplementation in larger and more diverse populations.