High intakes of fruits and vegetables and of carotenoids are associated with a lower risk for a variety of chronic diseases. It is therefore important to test the validity of dietary questionnaires that assess these intakes. We compared intakes of five carotenoids, as calculated from responses to the Willett 126-item food-frequency questionnaire, with corresponding biochemical measures. Subjects included 346 women and 201 men, aged 67-93 y, in the Framingham Heart Study. Unadjusted correlations were higher among women than men as follows: alpha-carotene 0.33 and 0.18, beta-carotene, 0.36 and 0.25; beta-cryptoxanthin, 0.44 and 0.32; lycopene, 0.35 and 0.21; and lutein + zeaxanthin, 0.27 and 0.10, respectively. Adjustment for age, energy intake, body mass index (BMI, kg/m2), plasma cholesterol concentrations and smoking reduced the gender differences, respectively, to the following: alpha-carotene 0.30 and 0.28; beta-carotene, 0.34 and 0.31; beta-cryptoxanthin, 0.45 and 0.36; lycopene, 0.36 and 0.31; and lutein + zeaxanthin, 0.24 and 0.14. Plots of adjusted mean plasma carotenoid concentration by quintile of respective carotenoid intake show apparent greater responsiveness among women, compared with men, to dietary intake of alpha- and beta-carotene and beta-cryptoxanthin, but similar blood-diet relationships for lycopene and lutein + zeaxanthin. Reported daily intake of fruits and vegetables correlated most strongly with plasma beta-cryptoxanthin and beta-carotene among women and with plasma alpha- and beta-carotene among men. With the exception of lutein + zeaxanthin, this dietary questionnaire does provide reasonable rankings of carotenoid status among elderly subjects, with the strongest correlations for beta-cryptoxanthin. Appropriate adjustment of confounders is necessary to clarify these associations among men.