Background: Use of the WHO 5 Moments of Hand Hygiene (HH) by operating room (OR) anesthesia personnel has been called by some logistically unfeasible, despite evidence that HH can reduce patients’ risk of pathogen acquisition. We developed and implemented a set of 7 moments based on WHO guidance (Fig 1) with high adherence. We conducted this study 6 months later to determine whether the improvement was sustained. In addition, we sought to understand practices, beliefs, barriers, and perceptions among anesthesia providers regarding HH. Methods: We measured HH adherence by direct observation using locally developed 7 moments tailored to the anesthesia workflow during June–August 2019. Adherence was defined as the percentage of observed HH performed when a moment occurred. We used the theory of planned behavior (TPB) as a framework to conduct 11 individual interviews (8 attending anesthesiologists and 3 certified nurse anesthetists) with a semistructured instrument that included Likert scale and open-ended questions. Interview transcripts were reviewed and a codebook of themes was created through inductive thematic analysis. Resultant themes and Likert scale averages were grouped by the 3 key TPB variables. Results: In total, 294 HH moments were observed for 50 anesthesia providers during 36 cases. The average HH adherence was 21.1% with the highest adherence moment being after patient contact (61.7%). Interview participants stated universally that HH was important for patient care, but acknowledged barriers to performance. Barriers cited included interruption in workflow, a lack of evidence, lack of clarity of HH standard, and limited availability of product. Conclusions: Adherence to the 7 moments of HH for anesthesia providers was not sustained after 6 months. Providers identified numerous barriers to HH, including a lack of knowledge of a standard, as reasons for suboptimal adherence. These data suggest future interventions could be designed to address gaps in knowledge and remove barriers to improve HH adherence among OR anesthesia providers.