Purpose:To investigate the relationships of objectively measured physical activity (PA) and sedentary time (ST) to arterial stiffness in prepubertal children.Method:Altogether 136 children (57 boys, 79 girls) aged 6–8-years participated in the study. Stiffness index (SI) was assessed by pulse contour analysis based on photoplethysmography. ST, light PA, moderate PA, and vigorous PA were assessed using combined acceleration and heart rate monitoring. We investigated the associations of ST (<1.5METs) and time spent in intensity level of PA above 2–7METs in min/d with SI using linear regression analysis. We studied the optimal duration and intensity of PA to identify children being in the highest quarter of SI using Receiver Operating Characteristics curves.Results:Moderate PA, vigorous PA, and cumulative time spent in PA above 3 (β=–0.279, p = .002), 4 (β =–0.341, P<0.001), 5 (β =–0.349, P<0.001), 6 (β =–0.312, P<0.001), and 7 (β =–0.254, p = .005) METs were inversely associated with SI after adjustment for age, sex, and monitor wear time. The cutoffs for identifying children being in the highest quarter of SI <68 min/d for PA exceeding 5 METs and <26 min/d for PA exceeding 6 METs.Conclusion:Lower levels of PA exceeding 3–6 METs were related to higher arterial stiffness in children.