Increased fruit and vegetable consumption has become a health priority in many countries. Therefore, data investigating the influence of different types of fruits and vegetables on serum antioxidant levels would be useful. The objective of the study was to assess the relationship between fruit and vegetable consumption and vitamin serum antioxidant concentrations. Specific fruit and vegetable groups are evaluated. A total of 3521 subjects (1487 men and 2034 women), aged 35–60 years, participating in the SU.VI.MAX cohort were included in this study. Blood samples of participants were analysed for β-carotene, vitamin C and α-tocopherol. Each subject had completed at least six dietary records during the first 2 years of the study. It was found that women had higher mean β-carotene and vitamin C serum concentrations than men, but lower α-tocopherol serum concentrations. Serum β-carotene and vitamin C concentrations were positively correlated with consumption of both fruit and vegetables, as well as with most of the fruit and vegetable groups tested. These relationships persisted after adjustment for confounding factors. Regression analysis showed a linear dose–response relationship. Root vegetables and citrus fruits were particularly associated with β-carotene serum status as were citrus fruits for vitamin C. Fruit and vegetable consumption was either not or weakly associated with α-tocopherol serum concentrations. These results describe antioxidant serum concentrations according to fruit and vegetable consumption in a large sample and support the findings of previous studies involving a more limited number of subjects.