Background Adolescent girls are one of the most sedentary demographic groups. A better understanding of their accumulation of sedentary time is needed to inform future interventions. The purpose of this study was to examine the longitudinal levels and bouts of objectively measured sedentary time accumulated during different days of the week and periods of the weekday among a large sample of adolescent girls. Methods The results are based on 655 adolescent girls from the Girls in Sport Intervention and Research Project. Levels and bouts of sedentary time were derived from accelerometer data collected at baseline and 18-month follow-up. Total, weekday, weekend, school (i.e., morning bell to afternoon bell), after school (i.e., afternoon bell to 19:00), and evening (i.e. 19:01 to 23:59) sedentary time levels and bouts were calculated. Repeated-measures ANCOVAs were conducted to examine differences in sedentary time levels and bouts between days and time periods after adjusting for wear time, accelerometer model, and intervention group. Results Cross-sectional analyses revealed that levels and bouts of sedentary time were higher on weekdays compared to weekend days at baseline. Similar trends were observed at follow-up. In addition, percentage of wear time spent sedentary and bouts/hr of sedentary time were highest in the evening compared to the school and after school periods at both baseline and follow-up. Longitudinal analyses revealed that levels and bouts of sedentary time were higher at follow-up compared to baseline across the different days of the week and periods of the weekday examined, with the biggest increase (15%) occurring in the school period. Conclusions Future interventions targeting sedentary time among adolescent girls should consider developing strategies to reduce and break up prolonged sedentary time during the school day and in the evening.