Menstruation and menstrual hygiene have been a sensitive concept to be discussed in public. It is necessary to address the neglect of menstrual hygiene as an issue because puberty and menstruation have significant effect on women's quality of life like causing reproductive tract infections, dropout from school, absenteeism from work, and others. A descriptive correlational design was used to assess and compare the knowledge regarding menstrual hygiene practices among women of reproductive age in rural, urban, and tribal areas of Vellore, Tamil Nadu. Ninety women of the reproductive age who visited the rural, urban and tribal PHCs were selected using purposive sampling technique. Data was collected using demographic and menstrual variable proforma. Knowledge regarding menstrual hygiene practices was assessed using a structured questionnaire. Major findings of the study reflected 67% of the urban and 60% of the rural women had moderately adequate knowledge and less than half of the tribal women had inadequate knowledge (47%) regarding menstrual hygiene practices. There was a significant difference (F ratio - 3.44, p<0.05) in knowledge scores obtained by rural, urban, tribal women of the reproductive age. Reproductive tract infections were reported by 20% of tribal women. There was significant association between selected demographic variables like age (?² - 7.53, p<0.05), educational status (?² - 6.21, p<0.05) and level of knowledge regarding menstrual hygiene of mothers residing in urban community. This indicated the need for creating awareness regarding menstrual hygiene to ensure protection against reproductive tract infections caused by poor menstrual hygiene.