The timing and frequency of periconceptional multivitamin use may be related to the risk of preeclampsia. Women in the Danish National Birth Cohort (1997-2003) reported multivitamin or folate-only supplement use during a 12-week periconceptional period (from 4 weeks prior to 8 weeks after the last menstrual period). Preeclampsia cases were identified by using International Classification of Diseases, Tenth Revision, codes. Cox regression was used to estimate the association of frequency (weeks of use) and timing (preconception and postconception) of use with preeclampsia risk. Overall, there were 668 cases of preeclampsia (2.3%), and 18,551 women (65%) reported periconceptional multivitamin use. After adjustment, regular use (12 of 12 weeks) was related to a reduced risk of preeclampsia among normal-weight women. Compared with nonusers with a body mass index of 22 kg/m(2), regular multivitamin users with the same body mass index had a 20% reduced risk of preeclampisa (hazard ratio = 0.78, 95% confidence interval: 0.60, 0.99). In addition, regular use in the postconception period only was associated with reduced risk, a relation that also appeared to be limited to women with a body mass index of <25 kg/m(2) (hazard ratio = 0.63, 95% confidence interval: 0.42, 0.93). Folate-only supplement use was unrelated to preeclampsia risk. Regular periconceptional multivitamin use was associated with a reduced risk of preeclampsia among normal-weight women, and the immediate postconception period appeared to be the relevant exposure window.