The study aimed to investigate the usefulness of the Running-based Anaerobic Sprint Test (RAST) in anaerobic performance estimation in trained and untrained girls U12, and the effect of an 8-week training period in female U12 soccer players on anaerobic performance. A comparative study of two structurally different anaerobic tests was performed to reach the goal. The study was designed as a non-randomized, controlled before-and-after trial. Fourteen female soccer players (FSP) and twelve untrained girls (UNT) participated in the study. During that time, all participants were subjected to school’s physical education classes and the FSP additionally participated in regular soccer training. The anaerobic performance was evaluated twice, within 8-weeks period, using the traditional Wingate test (WAnT) and the RAST. A significant increase in the anaerobic performance of the FSP was noted (p < 0.05). In both tests peak, average, and relative power were significantly improved (p < 0.005). Nevertheless, strong, statistically significant (p < 0.05) correlation coefficients (0.50 < r < 0.70) were found for pre-training measurements between the WAnT and the RAST power parameters in absolute values. In UNT group significant improvement was found in peak, average and min power (p < 0.05) in the RAST. The improvement in anaerobic performance after training along with the strong correlation noted between the WAnT and the RAST power parameters prove the usefulness of the RAST in assessing anaerobic capacity in female youth athletes. Its simplicity encourages its use in monitoring anaerobic capacity in both trained and untrained girls.