Once selenium (Se) is absorbed by the body, it is excreted mostly into the urine and the major metabolite is 1beta-methylseleno-N-acetyl-d-galactosamine (selenosugar) within the required to low-toxic range. Selenosugar plateaus with a dose higher than 2.0 microg Se/ml water or g diet, and trimethylselenonium (TMSe) starts to increase, indicating that TMSe can be a biomarker of excessive and toxic doses of Se. Here, we show dose-related changes in the two urinary Se metabolites to clarify the relationship between the dose and urinary metabolites by feeding selenite to rats. It was also examined whether the metabolites are related to age, and further whether a possible exogenous source of the N-acetyl-d-galactosamine moiety, chondroitin 4-sulfate, affects the urinary metabolites. Selenite in drinking water was fed ad libitum to male Wistar rats of 36 and 5 weeks of age, and the concentrations of Se in the urine and organs were determined together with speciation of the urinary Se metabolites. In young rats, selenosugar was always the major urinary metabolite and TMSe increased with a dose higher than 2.0 microg Se/ml drinking water. On the other hand, in adult rats, TMSe increased only marginally despite that the rats suffered much more greatly from the Se toxicity, suggesting that TMSe cannot be a biomarker of Se toxicity. The results suggest that sources of the sugar moiety of selenosugar are more abundant in adult rats than in young rats. Chondroitin 4-sulfate did not affect the ratio of the two urinary metabolites, suggesting that the sugar source is of endogenous origin and that it increases with age.