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Hand-washing is a process of removing the dirt and debris from skin of both hands mechanically by using soap and running water. The purpose of hand-washing is removing the dirt and organisms from the skin and reducing the microbes. The study was conducted to describe the knowledge level of health students at Universitas Muhammadiyah Magelang on hand-washing tool according WHO. The method used was descriptive analytic research by distributing a questionnaire to respondents revealing the knowledge level of health students about hand-washing. The samples were 124 health students taken by total sampling technique. The data were presented as percentage of frequency distribution. The Analysis using chi-square test, the result between the knowledge level and age (p=0,000), the knowledge level and gender (p=0,359) , the knowledge level and study program (p=0,965) , the knowledge level and year (p= 0,148). This study showed that there was a positive relationship between the knowledge level and age, but there was no relationship between the knowledge level and gender, year or study program. The students in aged 17-20 have a good level of hand-washing knowledge with a percentage of 51.6%.