Objective: To assess the acceptability, appropriateness, and feasibility of an active break designed to disrupt prolonged sitting in university students. Participants: Students attending lectures in Trinity College Dublin, Ireland. Methods: Participants took part in an active break, which consisted of following a short exercise video lasting [approximately]4 min. They then completed a validated questionnaire consisting of 12 statements with two open-ended questions capturing likes/dislikes. Results: Overall 106 (response rate 96%) predominately female (83%, n = 87), health sciences students (91%, n = 96) participated. Percentage agreement ranged from 93.4% (n = 99) to 96.2% (n = 102) for acceptability, 84.9% (n = 90) to 93.4% (n = 99) for appropriateness, and 80.2% (n = 85) to 96.2% (n = 102) for feasibility. Space constraints and warm temperatures impacted negatively. Conclusion: An active break delivered during lectures is an acceptable and feasible intervention to disrupt sitting in students. Further investigation using a broader representation of the university population is needed prior to implementation.