Few food frequency questionnaires (FFQs) have been developed to assess diet in diabetes patients. This cross-sectional study examined the validity of a 45-item FFQ assessing the intake of macronutrients against three 24-h dietary recalls (24-HDRs) in Taiwan, and compared vegetable and fruit intakes with carotenoid biomarkers. We recruited 126 adults with type 2 diabetes who completed the FFQ and three 24-HDRs administered by a registered dietitian. We measured plasma carotenoids (α-carotene, β-carotene and lutein) in 71 subjects. Partial Pearson correlation coefficients derived from the FFQs and three 24-HDRs and adjusted for energy were of 0.651, 0.587, 0.639 and 0.664 for protein, fat, carbohydrate and fiber, respectively. Cross-classification analysis revealed that 71.5–81% of the macronutrients and fiber were categorized into the same or adjacent quartiles by the FFQ and 24-HDRs. Bland–Altman plots revealed good agreement for energy/macronutrients/fiber across the range of intakes. Multiple linear regression of backward elimination revealed that tertile levels of dark- or light-colored vegetables obtained by the FFQ were significantly associated with plasma α-carotene and β-carotene, but not lutein. Fruit consumption did not correlate with carotenoid biomarkers. In conclusion, this short FFQ provided a valid assessment of macronutrients and fiber intake in type 2 diabetes patients. Vegetable consumption estimated by the FFQ corresponded to plasma α-carotene and β-carotene concentrations.