ABSTRACT This study aimed to characterise bilateral asymmetry in running mechanics during perceptually regulated, high-intensity intermittent running in hypoxia and normoxia and examines whether inter-limb differences in running mechanics are modified between and within intervals. Nineteen trained runners completed 4 × 4-min treadmill running bouts (3-min passive recoveries) at a perceived rating exertion of 16 on the 6–20 Borg scale in either hypoxic (FiO2 = 0.15) or normoxic (FiO2 = 0.21) conditions. Ground reaction force recordings at constant velocity (group average: 14.8 ± 1.9 km/h) allowed measurement of running kinetics/kinematics and calculation of spring-mass model characteristics at the beginning and the end of each 4-min interval. Lower limb asymmetry was assessed from the ‘symmetry angle’ (SA) score. There were no between intervals (P > 0.087), within intervals (P > 0.076) or FiO2 (P > 0.128) differences in SA scores for any of the 16 biomechanical variables. Mean SA scores were lower than 1.5% for spatio-temporal variables, ~1.5–3% for braking and push-off phase durations, peak forces and impulses and ~4–6% for mean loading rate and vertical stiffness. With preserved lower limb asymmetries both between and within intervals and with additional hypoxia, trained runners completing perceptually regulated interval treadmill runs may anticipate a maintained performance without heightened injury risk.