Objectives: This study was performed to investigate hand washing awareness of females and load of indicator bacteria on their hands. This study focused on the variation according to their age. Methods: A self-administered questionnaire survey and bacterial analysis of indicator bacteria were carried out for 100 Korean women in their age from 20 s to 60 s. Hand samples were collected through a modified glovejuice method. Results: In the survey, significant difference (p < 0.05) was found among the age groups in the use of hand washing agents. The levels of aerobic colony count (ACC) were the highest in both hands among the 20s (p < 0.05). The levels of Escherichia coli were higher in both hands in their 20s and 30s. No significant difference was found in the levels of Staphylococcus aureus and Salmonella spp. However, the positive rates of S. aureus (left hand, 37.5~47.1%; right hand, 58.5~62.5%) and Salmonella spp. (left hand, 25.0~52.9%; right hand, 37.5~64.7%) were higher in the hands of the 20s and 30s, and then showed decreasing trend according to increase of age. The effect of hand washing frequency on the ACC level of hands was significant (p < 0.001). Conclusions: These results indicate that there was no remarkable difference of hand hygiene awareness among female age groups. The detection of S. aureus and Salmonella spp. on the hands of some females in each age group revealed poor hand hygiene practices. The significant effect of hand washing frequency on the ACC level suggests that frequent hand washing is helpful to reduce hand contamination.