SCOPE: Dietary choices modulate the risk of chronic diseases and improving diet is a central component of public health strategies. Food-derived metabolites present in urine could provide objective biomarkers of dietary exposure. To assist biomarker validation we aimed to develop a food intervention strategy mimicking a typical annual diet over a short period of time and assessed urine sampling protocols potentially suitable for future deployment of biomarker technology in free-living populations. METHODS AND RESULTS: Six different menu plans representing comprehensively a typical UK annual diet that were split into two dietary experimental periods. Free-living adult participants (n = 15 and n = 36, respectively) were provided with all their food, as a series of menu plans, over a period of 3 consecutive days. Multiple spot urine samples were collected and stored at home. CONCLUSION: We established a successful food exposure strategy following a conventional UK eating pattern, which was suitable for biomarker validation in free-living individuals. The urine sampling procedure was acceptable for volunteers and delivered samples suitable for biomarker quantification. Our study design provides scope for validation of existing biomarker candidates and potentially for discovery of new biomarker-leads and should help inform the future deployment of biomarker technology for habitual dietary exposure measurement.