Background: Studies do not show consistent relationships between self-reported intake of sugar and outcome of disease. To overcome the drawbacks of self-reported intake methods, we investigated whether there is an agreement in ranking of individuals between their self-reported sugar intake and urinary sucrose and fructose. Methods: We used data of 198 Dutch adults (106 women) from the DUPLO study. Sugar intake of all foods and drinks consumed over 24-hour period was estimated by collecting duplicate portions (DP) and 24-hour recalls (24hR), telephone (24hRT) and Web-based (24hRW), while sugar excretion was based on 24-hour urine samples. Sugar content of 24hR was calculated using a newly developed sugar database and sugar content of DPs and urine samples was calculated using high-performance liquid chromatography-atomic emission spectrometry and LC/MS-MS, respectively. Measurement error models assessed validity coefficients (VC) and attenuation factors (AF). Coefficients were compared with those of protein biomarker. Results: The VC for the marker, using DP as reference, showed comparability with substantially better ranking of participants (0.72 for women and 0.93 for men), than 24hRT (0.57 and 0.78) or 24hRW (0.70 and 0.78) as reference in the sucrose models. The VC of the sucrose models was within 10% of the protein models, except for the model with 24hRT as reference, among women. The AF started at higher values and increased by a greater factor compared with the VC. Conclusions: Repeated measurements of urinary sucrose and fructose as a marker of daily sucrose intake had a ranking performance comparable to urinary nitrogen as marker of protein intake in free-living Dutch adults. Impact: The validation of the sugar biomarker in a free-living population with three different dietary assessment methods and its comparable ranking ability with a good recovery biomarker (i.e., protein biomarker) have important research applications. The biomarker may be used for validating dietary assessment methods, for monitoring compliance in human feeding studies, for monitoring the effect of public health interventions, and as a surrogate for ranking subjects according to sucrose intake when information on sucrose in food composition databases is lacking.