Purpose: To quantify the effect of acute hypoxia on muscle oxygenation and power during simulated team-sport running.Methods: Seven individuals performed repeated and single sprint efforts, embedded in a simulated team-sport running protocol, on a non-motorized treadmill in normoxia (sea-level), and acute normobaric hypoxia (simulated altitudes of 2,000 and 3,000 m). Mean and peak power was quantified during all sprints and repeated sprints. Mean total work, heart rate, blood oxygen saturation, and quadriceps muscle deoxyhaemoglobin concentration (assessed via near-infrared spectroscopy) were measured over the entire protocol. A linear mixed model was used to estimate performance and physiological effects across each half of the protocol. Changes were expressed in standardized units for assessment of magnitude. Uncertainty in the changes was expressed as a 90% confidence interval and interpreted via non-clinical magnitude-based inference.Results: Mean total work was reduced at 2,000 m (−10%, 90% confidence limits ±6%) and 3,000 m (−15%, ±5%) compared with sea-level. Mean heart rate was reduced at 3,000 m compared with 2,000 m (−3, ±3 min−1) and sea-level (−3, ±3 min−1). Blood oxygen saturation was lower at 2,000 m (−8, ±3%) and 3,000 m (−15, ±2%) compared with sea-level. Sprint mean power across the entire protocol was reduced at 3,000 m compared with 2,000 m (−12%, ±3%) and sea-level (−14%, ±4%). In the second half of the protocol, sprint mean power was reduced at 3,000 m compared to 2,000 m (−6%, ±4%). Sprint mean peak power across the entire protocol was lowered at 2,000 m (−10%, ±6%) and 3,000 m (−16%, ±6%) compared with sea-level. During repeated sprints, mean peak power was lower at 2,000 m (−8%, ±7%) and 3,000 m (−8%, ±7%) compared with sea-level. In the second half of the protocol, repeated sprint mean power was reduced at 3,000 m compared to 2,000 m (−7%, ±5%) and sea-level (−9%, ±5%). Quadriceps muscle deoxyhaemoglobin concentration was lowered at 3,000 m compared to 2,000 m (−10, ±12%) and sea-level (−11, ±12%).Conclusions: Simulated team-sport running is impaired at 3,000 m compared to 2,000 m and sea-level, likely due to a higher muscle deoxygenation.