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Background: The validation of dietary assessment instruments is critical in the evaluation of diet as a chronic disease risk factor., Objective: The objective was to assess the validity of a self-administered food-frequency questionnaire by comparison with dietary recall, urinary nitrogen excretion, and total energy expenditure data., Design: Over a 1-y period, data from twelve 24-h dietary recalls, a food-frequency questionnaire, and four 24-h urine samples were obtained from 134 study participants of the European Prospective Investigation into Cancer and Nutrition (EPIC) Study in Potsdam, Germany. In a substudy of 28 participants, total energy expenditure from doubly labeled water measurements was assessed., Results: Energy-adjusted, deattenuated correlation coefficients between the questionnaire and the recalls ranged from 0.54 for dietary fiber to 0.86 for alcohol. Cross-classification of quintiles of nutrient intakes from the questionnaire and recalls indicated severe misclassification to be <4%. Reported protein intake correlated with estimated protein excretion (r = 0.46). Energy intake and total energy expenditure were also significantly correlated (r = 0.48); however, all but one subject underreported their energy intake. The magnitude of underreporting varied considerably, by 22% on average, and increased slightly with increasing energy intake. A similar pattern of underreporting was observed when energy intakes from the 24-h dietary recalls were compared with total energy expenditure., Conclusions: These data indicate an acceptable relative validity of the food-frequency questionnaire in this study population. Compared with measurements of total energy expenditure and protein excretion, however, only moderate agreement with both the food-frequency questionnaire and the 24-h dietary recalls was observed.