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The aim of the study was to assess change in physical activity (PA) and sedentary behavior (SB) in office-based employees after the implementation of a flexible work policy that allowed working at home.A total of 24 employees (62% female; 40 ± 10 years) completed an online questionnaire 4 weeks pre- and 6 weeks post-implementation of the policy. Changes in PA and SB were assessed using Wilcoxon signed rank test.There were no changes in PA after the introduction of the flexible work policy (Z = −0.29, P > 0.05). Sitting time increased on days the employees worked at home (Z = −2.02, P > 0.05) and on days they worked at the office (Z = −4.16, P > 0.001).A flexible work policy may have had a negative impact on sedentary behavior in this workplace. Future work is needed to explore the potential impact on workplace sitting time.