Background: The monitoring of accelerometry derived load has received increased attention in recent years. However, the ability of such measures to quantify training load during sport-related activities is not well established. Thus, the current study aimed to assess the validity and reliability of tri-axial accelerometers to identify step count and quantify external load during several locomotor conditions including walking, jogging, and running.Method: Thirty physically active college students (height = 176.8 ± 6.1 cm, weight = 82.3 ± 12.8 kg) participated. Acceleration data was collected via two tri-axial accelerometers (Device A and B) sampling at 100 Hz, mounted closely together at the xiphoid process. Each participant completed two trials of straight-line walking, jogging, and running on a 20 m course. Device A was used to assess accelerometer validity to identify step count and the test-retest reliability of the instrument to quantify the external load. Device A and Device B were used to assess inter-device reliability. The reliability of accelerometry-derived metrics Impulse Load (IL) and Magnitude g (MAG) were assessed.Results: The instrument demonstrated a positive predictive value (PPV) ranging between 96.98%–99.41% and an agreement ranging between 93.08%–96.29% for step detection during all conditions. Good test-retest reliability was found with a coefficient of variation (CV) <5% for IL and MAG during all locomotor conditions. Good inter-device reliability was also found for all locomotor conditions (IL and MAG CV < 5%).Conclusion: This research indicates that tri-axial accelerometers can be used to identify steps and quantify external load when movement is completed at a range of speeds.