BACKGROUND: Compliance with the best surgical antibiotic prophylaxis practice is usually low despite many published guidelines. OBJECTIVE: This study investigated compliance with the Hospital Infection Control Committee guideline for antibiotic prophylaxis in a Brazilian hospital using quality indicators. METHODS: A retrospective study was carried out from November 2009 to March 2010. Medical records from adult inpatients undergoing cardiac, neurologic, and orthopedic clean surgeries were included. The full compliance index was considered 100% when the antibiotic prophylaxis showed adequacy in all evaluated attributes. Analyses were conducted with 5% significance. RESULTS: Medical records from 101 cardiac, 128 neurologic, and 519 orthopedic surgical patients were evaluated. The compliance index was 4.9%, and the compliance index according to specialty was 5.8%, 3.1%, and 3.0%, respectively, for orthopedic, neurologic, and cardiac surgeries. The attribute route of administration produced the best outcomes, whereas the attribute duration of antibiotic prophylaxis produced the worst. No association was identified between compliance to the attributes and patient characteristics. CONCLUSION: This study showed a low level of adherence to Hospital Infection Control Committee guidelines for antibiotic prophylaxis. This suggests that different strategies should be implemented to promote the best possible practice in the field of antibiotic prophylaxis with greater surgeon engagement.