Background: In recent years, patients' safety has become high priority for health-care organizations. It has been documented that poor knowledge and compliance of health-care providers toward hand hygiene have contributed to poor patient safety outcomes. College students of health sciences may not receive adequate education and training on hand hygiene best practices. Objectives: To assess the health profession students' knowledge, attitude, and practice toward hand hygiene and to investigate the factors associated with poor hand hygiene knowledge and practice. Methods: This cross-sectional, correlational, descriptive study was conducted at King Saud Bin Abdulaziz University for Health Sciences performing clinical rotations at King Abdulaziz Medical City in Riyadh, Saudi Arabia. A proportional sampling was used to calculate the required proportions that reflect the size of student population form each of the three colleges. A total of 270 students including 140 medical, 83 nursing, and 47 respiratory therapy students who were selected using a simple random sampling method were included in the study. An adapted and validated knowledge, attitude, and practice (KAP) questionnaire was used to assess four domains: general information, knowledge, self-reported compliance to the WHO 5 Moments of Hand Hygiene, and general satisfaction on received education. Results: The overall average of knowledge score was 81.13 points out of 100. The results revealed that the knowledge score of hand hygiene was higher for nursing school (84.22 ± 12.98), followed by medical school (81.71 ± 11.31) and then respiratory therapy program (75.53 ± 11.76). The results also showed that students who took courses covering hand hygiene scored higher in knowledge score (83.28 ± 11.3) as compared to students who did not take such courses (75.16 ± 12.89). In the compliance domain, the results showed that there were no significant differences in compliance of hand hygiene between students in all categories. In terms of overall student satisfaction with hand hygiene education and training, the results showed that nursing students have higher satisfaction score (72.7%) than their counterparts in medical school (48.3%) and respiratory therapy program (49.7%). Conclusion: Hand hygiene knowledge among students was generally good, and the highest was among nursing students and the lowest among respiratory therapy students. The compliance of students toward the WHO 5 Moments of Hand Hygiene did not vary across different colleges. Improved knowledge was found to be associated with improved compliance with hand hygiene best practices. Recommendations: Increasing the academic focus on hand hygiene in both the curriculum and clinical rotations with periodic standardized educational courses and focusing on hands-on workshops could have a positive impact on the knowledge and practice of hand hygiene for current health sciences students and future health-care providers. Future studies are needed to assess the hand hygiene KAP among students in hospital settings using observation of actual practice.