The duration of sleep data collection from actigraphy is often influenced by practical factors (e.g. workdays versus non-workdays), but the impact of the variation of duration on outcome measures of interest has not been well explored. This study investigates the effect of the duration of actigraphy measurement on non-parametric measures of 24-hr sleep-wake rhythms. We examined regularity inter-daily stability and fragmentation intra-daily variation over 14 days or the first 7 days in participants (n = 41) undergoing evaluation for sleep disorders. Bland-Altman plots assessed the impact of fewer than 14 or 7 days, respectively, on inter-daily stability and intra-daily variation scores. Intra-daily variation values were also calculated for each day and compared with the 14-day intra-daily variation. Compared with the entire 14-day period, using shorter durations (< 7 days) led to a higher estimated bias and increased variance in the limits of agreement for inter-daily stability. Intra-daily variation values showed increased variation in the limits of agreement with fewer days. Similar trends were observed when comparing shorter actigraphy periods 3 or 5 days-7 days. Daily intra-daily variation calculations indicate that individuals with higher daily fragmentation experienced more pronounced day-to-day fragmentation and greater variability in the degree of fragmentation, in a linear association between daily intra-daily variation standard deviation and 14-day intra-daily variation values. Our data indicate that a minimum of 7 full days of actigraphy is recommended to reduce measurement errors, and intra-daily variation and inter-daily stability derived from less than 7 days cannot be compared with those from more than 7 days without significant error.