Hand-washing is one of the most important factors in infection control and in preventing crosscontamination. The objective of this study was to investigate female university students’ awareness of hand-washing, their hand washing practices, and the difference between their awareness and practices. A self-administered questionnaire survey and direct observation in restrooms were separately carried out in a university campus and over four weeks’ period. A total of 97.4% of the survey respondents claimed to wash their hands after using toilet, and 98.2% of the observed students actually did so according to the unnoticed observational study. However, only 6.3% of the students who washed their hands in the direct observation washed for more than 10 seconds, although 46.4% of respondents in the survey reported that they usually wash their hands for more than 10 seconds. Among the observed students who washed their hands, only 0.9% used soap, and 0.9% washed four parts of their hands. Paper towel was the most common hand-drying method in the direct observation and also in the survey. Significant differences were found in duration, use of soap, part of washing, and hand-drying method between the questionnaire survey and the direct observation (p<0.05). This study indicates that there is a noticeable difference between the awareness of handwashing and hand-washing practices among female university students. Further research should examine hand-washing practices of female university students in restrooms outside the university campus.