This study examined if accelerometer-based assessments of physical activity were responsive to changes in physical activity level commensurate with performing structured versus unstructured activity in youth. Youth (6-16 years; N = 206) participated in a simulated after-school program that included structured and unstructured games on four occasions over a 3-year period. Recruitment occurred in 2007/2008 and data collection ended in 2011. Participants wore an Actigraph GT1M accelerometer on the hip. The Evenson cut-points were used to determine the time spent in each physical activity intensity, and standardized response means (SRM) were calculated and converted to standard effect sizes to be interpreted according to Cohen's guidelines. SRMs ranged from trivial (0.16) to high (2.07), with the majority (75%) being classified as moderate or high. Our findings suggest that accelerometry was sensitive to differences in physical activity associated with structured compared to unstructured play, supporting the utility of accelerometry in evaluating activity-promoting interventions.