Information regarding the menstrual knowledge and practices of Pakistani girls is sparse. Therefore, we aimed to evaluate the knowledge and practices of Pakistani girls regarding menstruation. This cross-sectional study was conducted among four categories of university female students of medical, pharmacy, nursing, and arts & humanities during a period of 5 months (November 2016-March 2017. The data were acquired using a selfadministered questionnaire from 3 medical institutes, 2 pharmacy institutes, 3 nursing institutes and 2 arts & humanities institutes at Lahore, Pakistan. The mean age of participants (N ¼ 1777) was 20.38 AE 2.39 years. Overall 39.5%, 76.1% and 29.3% respondents' provided correct answers to the questions concerning menstrual cycle length, duration of menstrual bleeding and source of bleeding, respectively. Regarding the understanding of menstruation, around 27% understood it as monthly bleeding happening with every woman, whereas 18% stated that it was a sign of adulthood. However, a small proportion believed that it was removal of dirt from the body. Regarding the practices-related to menstruation, there were some malpractices regarding diet and dietary intake (avoiding certain foods, eating less, and less fluid intake). However, the hygienic practices were found to be satisfactory. In conclusion, Pakistani university girls have some misconceptions, misperceptions and malpractices related to menstruation. Our findings draw attention to conduct awareness programs in order to eradicate these misconceptions, myths and malpractices.