The purpose of this study was to examine agreement between the Walk4Life DUO pedometer (W4L; Walk4Life, Plainfield, Illinois, USA) and two criterion instruments in the measurement of physical activity. Participants (N = 189, M = 16.74 years, SD = .99) in high school physical education concurrently wore the DUO (i.e., comparison instrument) and Yamax SW651 pedometers (Yamax Corp., Japan; i.e., criterion instrument), and were video recorded for off-site systematic behavioral observation (i.e., criterion instrument). Physical activity measures were steps/minute (i.e., DUO and SW651) and physical activity time (minutes; i.e., DUO and behavioral observation). Relative agreement between physical activity measures was strong (r >= .97, p <= .05). However, overall absolute agreement suggests that the DUO pedometer underestimates steps/minute (mean difference score [M-diff]= 3.13, SD = 4.73, t = 9.07, p =. 0001), overestimates physical activity time (M-diff = -10.76, SD = 6.46, t = - 17.22, p = .0001), and does not produce clinically acceptable steps/minute (mean absolute percent error [M-APE] = 10.82%, SD = 9.82) and physical activity time (M-APE = 76.13%, SD = 37.04) outcomes. In conclusion, it is recommended that researchers and practitioners cautiously interpret DUO steps/minutes and not employ the DUO physical activity time function during physical activity programs, such as high school physical education, to estimate physical activity time.