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Objective: We examined the association between lifestyle factors and menstrual cycle characteristics among nulliparous Danish women aged 18–40 years who were participating in an Internet-based prospective cohort study of pregnancy planners. Methods: We used cross-sectional data collected at baseline to assess the association of age, body mass index (BMI), physical activity, alcohol and caffeine consumption, and smoking with the prevalence of irregular cycles, short (?25 days) and long (?33 days) cycles, and duration and amount of menstrual flow. We used log-binomial and multinomial logistic regression to estimate prevalence ratios and 95% confidence intervals. Results: Low physical activity and heavy alcohol consumption were associated with an increased prevalence of irregular periods. High BMI, smoking, and caffeine and alcohol consumption were related to an increased prevalence of short menstrual cycles and heavy menstrual bleeding. Women in their mid-to-late thirties had shorter and lighter menstrual flow, but a lower prevalence of irregular cycles, compared with women 18–25 years of age. Discussion: In this study, increased age, high BMI, and sedentary behavior were associated with menstrual-pattern irregularities. These factors may influence the balance and level of endogenous hormones conducive to optimal menstrual function.