Objective: To validate the fruit and vegetable intake estimated from the Food Frequency Questionnaire (FFQ) used in the Danish National Birth Cohort (DNBC). Subjects and setting: The DNBC is a cohort of 101,042 pregnant women in Denmark, who received a FFQ by mail in gestation week 25. A validation study with 88 participants was made. A seven-day weighed food diary (FD) and three different biomarkers were employed as comparison methods. Results: Significant correlations between FFQ and FD-based estimates were found for fruit (r=0.66); vegetables (r=0.32); juice (r=0.52); fruit and vegetables (F&V) (r=0.57); and fruit, vegetables, and juice (F&V&J) (r=0.62). Sensitivities of correct classification by FFQ into the two lowest and the two highest quintiles of F&V&J intake were 58-67% and 50-74%, respectively, and specificities were 71-79% and 65-83%, respectively. F&V&J intake estimated from the FFQ correlated significantly with urinary flavonoids ( r=0.39, p