The aim of this study was to evaluate regional differences in knowledge, attitudes, and practice in emergency contraception use among Brazilian university students. A sample of university students answered a semi-structured questionnaire on knowledge, attitudes, and practice related to emergency contraception and sexual behavior. Fisher's exact test and ANOVA were used for statistical analysis. Statistical significance was set at p < 0.05. Ninety-six percent (n = 588) of the students had heard of emergency contraception, and 19% (n = 111) knew all the situations in which emergency contraception is indicated, with statistical differences between regions of the country. Forty-two percent of sexually active women in the sample had already used emergency contraception; 35% (n = 207) of students equated emergency contraception with abortion; and 81% (n = 473) thought emergency contraception involves health risks. No significant difference was observed between regions of the country regarding use and attitudes towards emergency contraception. Inter-regional differences in knowledge had no impact on students' attitudes and practice in emergency contraception. National awareness-raising campaigns are needed to improve knowledge on emergency contraception.