Adolescents' objectively assessed physical activity (PA) patterns during specific segments of the day remain unclear. In order to develop a clearer understanding this study examined country and gender differences in moderate to vigorous physical activity (MVPA) levels during specific segments of weekdays and weekend days, and explored the contribution of each segment to PA guidelines. Morpho-demographic, socio-economic and PA data were collected from a sample of 829 French and Spanish adolescents (45.0% Spanish; 55.2% females; 14.33±.73 years). Actigraph GT3X accelerometers were worn for seven days to assess adolescents' MVPA for three segments of weekdays (school-travel-time, school-time, and non-school-time), and weekend days (morning-time, afternoon-time and night-time). Data were analysed using multilevel modelling. The most active segments were non-school-time (29.2±17.5 min) and school-time (25.8±14.2 min) during weekdays, and morning-time (28.2±25.8 min) on weekend days. Except for school-time, Spanish adolescents were more significantly active than French adolescents during all segments. Significant gender differences were found in all segments. Country differences highlight the need to recognise cultural contexts that influence adolescents' PA. Common European-wide strategies may be insufficient to increase MVPA levels if cultural variability is not considered. Spanish and French PA intervention programmes should target girls and low-active boys during non-school-time and weekends.