Background: Acute altitude has a relevant impact on exercise physiology and performance. Therefore, the positive impact on the performance level is utilized as a training strategy in professional as well as recreational athletes. However, ventilatory thresholds (VTs) and lactate thresholds (LTs), as established performance measures, cannot be easily assessed at high altitudes. Therefore, a noninvasive, reliable, and cost-effective method is needed to facilitate and monitor training management at high altitudes. High Alt Med Biol . 25:94-99, 2024. Methods: In a cross-sectional setting, a total of 14 healthy recreational athletes performed a graded cycling exercise test at sea level (Munich, Germany: 512 m/949 mbar) and high altitude (Zugspitze: 2,650 m/715 mbar). Anaerobic thresholds (ATs) were assessed using a novel method based on beat-to-beat repolarization instability (dT) detected by Frank-lead electrocardiogram (ECG) monitoring. The ECG-based ATs (ATdT°) were compared to routine LTs assessed according to Dickhuth and Mader. Results: After acute altitude exposure, a decrease in AT was detected using a novel ECG-based method (ATdT°: 159.80 ± 52.21 W vs. 134.66 ± 34.91 W). AtdT° levels correlated significantly with LT Dickhuth and LT Mader , at baseline (r Dickhuth/AtdT° = 0.979; p < 0.001) (r Mader/AtdT° = 0.943; p < 0.001), and at high altitude (r Dickhuth/AtdT° = 0.969; p < 0.001) (r Mader/AtdT° = 0.942; p < 0.001). Conclusion: Assessment of ATdT is a reliable method to detect performance alterations at altitude. This novel method may facilitate the training management of athletes at high altitudes.