This study investigated how data series length and gaps in human kinematic data impact the accuracy of Lyapunov exponents (LyE) calculations with and without cubic spline interpolation. Kinematic time series were manipulated to create various data series lengths (28% and 100% of original) and gap durations (0.05–0.20 s). Longer gaps generally resulted in significantly higher LyE% error values in each plane in noninterpolated data. During cubic spline interpolation, only the 0.20-second gap in frontal plane data resulted in a significantly higher LyE% error. Data series length did not significantly affect LyE% error in noninterpolated data. During cubic spline interpolation, sagittal plane LyE% errors were significantly higher at shorter versus longer data series lengths. These findings suggest that not interpolating gaps in data could lead to erroneously high LyE values and mischaracterization of movement variability. When applying cubic spline, a long gap length (0.20 s) in the frontal plane or a short sagittal plane data series length (1000 data points) could also lead to erroneously high LyE values and mischaracterization of movement variability. These insights emphasize the necessity of detailed reporting on gap durations, data series lengths, and interpolation techniques when characterizing human movement variability using LyE values.