This study investigated the validity and sensitivity of a custom-made shoelace tensile testing system. The aim was to analyze the distribution pattern of shoelace tension in different positions and under different tightness levels during running. Mechanical tests were conducted using 16 weights, and various statistical analyses, including linear regression, Bland-Altman plots, coefficient of variation, and intraclass correlation coefficient, were performed to assess the system’s validity. Fifteen male amateur runners participated in the study, and three conditions (loose, comfortable, and tight) were measured during an upright stance. The system utilized VICON motion systems, a Kistler force plate, and a Photoelectric gate speed measurement system. Results showed a linear relationship between voltage and load at the three sensors (R2 ≥ 0.9997). Bland-Altman plots demonstrated 95% prediction intervals within ± 1.96SD from zero for all sensors. The average coefficient of variation for each sensor was less than 0.38%. Intraclass correlation coefficient values were larger than 0.999 (p<0.0001) for each sensor. The peak tension of the front shoelace was greater than that of the front and middle when the shoelace was loose and tight. The rear shoelace had the highest tension force. The study also found that shoelace tension varied throughout the gait cycle during running. Overall, this research provides a novel and validated method for measuring shoelace tensile stress, which has implications for developing automatic shoelace fastening systems.