Background: Women make up half of the world's population, and comprise 20% of the world's one billion smokers. The aim of this study was to assess the prevalence of smoking among female medical students in comparison to female non-medical students, and to assess the importance of medical education and knowledge in decreasing the prevalence of smoking among female university students in Saudi Arabia. Method: We used a self-administered questionnaire to collect cross-sectional data from a randomly selected sample of 320 female students attending King Abdul-Aziz University, Jeddah. Medical students comprised 50% of the sample. Results: A total of 310 students (96.9%) completed and returned the questionnaire. The prevalence of smoking was higher in non-medical female students (4.2%) compared to medical female students (0.32%) (P < 0.001). Conclusion: The prevalence of smoking is low among female medical students compared to female non-medical students, presumably because of their awareness, level of education, and knowledge of the risks to health associated with smoking. Our study highlights the need for increased knowledge, health education, and awareness of the risks of smoking to reduce smoking among female university students.