BACKGROUND For alcohol-based hand rubs, the currently recommended application time of 30 seconds is longer than the actual time spent in clinical practice. We investigated whether a shorter application time of 15 seconds is microbiologically safe in neonatal intensive care and may positively influence compliance with the frequency of hand antisepsis actions. METHODS We conducted in vitro experiments to determine the antimicrobial efficacy of hand rubs within 15 seconds, followed by clinical observations to assess the effect of a shortened hand antisepsis procedure under clinical conditions in a neonatal intensive care unit (NICU). An independent observer monitored the frequency of hand antisepsis actions during shifts. RESULTS All tested hand rubs fulfilled the requirement of equal or even significantly higher efficacy within 15 seconds when compared to a reference alcohol propan-2-ol 60% (v/v) within 30 seconds. Microbiologically, reducing the application time to 15 seconds had a similar effect when compared to 30-second hand rubbing, but it resulted in significantly increased frequency of hand antisepsis actions (7.9±4.3 per hour vs 5.8±2.9 per hour; P=.05). CONCLUSION Time pressure and workload are recognized barriers to compliance. Therefore, reducing the recommended time for hand antisepsis actions, using tested and well-evaluated hand rub formulations, may improve hand hygiene compliance in clinical practice. Infect Control Hosp Epidemiol 2017;38:1430-1434.