Anthocyanins, kind of flavonoids (FL) found in plants and vegetables, are known to have varieties of physiological functions. In the present study, we examined absorption and metabolism of delphinidin 3-O-beta-D-glucoside (Dp3G) in rats. Dp3G appeared in the plasma at 15 min after oral administration as an intact glucosidic form. The plasma level also showed another peak at 60 min. One metabolite peak was detected in the plasma and the structure was assigned as 4'-O-methyl Dp3G (MDp3G) by NMR and MS. The metabolite was also identified in several tissues as a major metabolite especially in the liver. No 3'-O-methyl Dp3G was detected in any tissues, therefore, 4'-OH methylation is the main path of Dp3G metabolism in rats. This finding generalized the metabolic formation of FL having pyrogallol B ring because it has been reported that FL having catechol structure produced 3'-O-methyl-derivatives, but FL having pyrogallol structure produced 4'-O-methyl-derivatives.