Sesame seeds (Sesamum indicum L.) are unique because of potent and various physiological activities imparted by their bioactive lignans. This investigation studied the intestinal distribution and excretion of sesaminol in Sprague-Dawley (SD) rats. To investigate the distribution of sesaminol (per oral 220 mg/kg), the changes in concentration of sesaminol and its metabolites were determined in the intestines and plasma within the 24 h period after tube feeding of sesaminol to SD rats. Results show that the epimerization of sesaminol appeared to be catalyzed by acid in the simulated gastric fluids. The major sesaminol epimer was characterized as 2-episesaminol using 2D-NMR. These findings indicate that sesame sesaminol and its epimer are poorly absorbed prior to reaching the rectum and that substantial amounts pass from the small to the large intestine, where they are metabolized by the colonic microflora to tetrahydrofuranoid metabolites. Sesaminol in plasma was largely present as phase II conjugates, and the seven metabolites were detected as the 2-episesaminol, sesaminol-6-catechol, methylated sesaminol-catechol, R,R-hydroxymethylsesaminol-tetrahydrofuran, S,R-hydroxymethylsesaminol-tetrahydrofuran, enterolactone, and enterodiol. Excretions of sesaminol in urine and feces within the 24 h period were equivalent to 0.02 and 9.33% of the amount ingested, respectively.