This feasibility study explored the contextual factors influencing office workers’ adherence to an e-health intervention targeting total and prolonged sedentary time over 12 weeks. A three-arm quasi-randomized intervention included prompts at 30 or 60 min intervals delivered via a smartphone application, and a no-prompt comparison arm. Fifty-six office workers completed baseline (64% female) and 44 completed the 12 week follow-up (80% retention). Ecological momentary assessments (EMA) captured contextual data, with 82.8 ± 24.9 EMA prompt questionnaires completed weekly. Two focus groups with n = 8 Prompt 30 and 60 participants were conducted one-month post-intervention to address intervention acceptability and feasibility. Contextual findings indicate that when working on a sedentary task (i.e., reading or screen-based work) and located at an individual workstation, hourly prompts may be more acceptable and feasible for promoting a reduction in total and prolonged sedentary time compared to 30 min prompts. Interpersonal support also appears important for promoting subtle shifts in sedentary working practices. This novel study gives a real-time insight into the factors influencing adherence to e-health prompts. Findings identified unique, pragmatic considerations for delivering a workplace e-health intervention, indicating that further research is warranted to optimize the method of intervention delivery prior to evaluation of a large-scale intervention.