The consumption of anthocyanins has been shown to prevent certain chronic diseases. However, anthocyanin metabolism has not yet been fully elucidated. The aim of this study was to evaluate anthocyanin urinary excretion in humans receiving a meal containing blackberries and to identify possible metabolites in urine. Five healthy volunteers were fed 200 g of blackberries (960 mumol of anthocyanins). Urine samples were collected and rapidly treated by solid-phase extraction. Anthocyanin metabolites were identified and quantified by HPLC-ESI-MS-MS and HPLC with UV-vis detection, respectively. In addition to native cyanidin 3-glucoside, several other anthocyanin metabolites were identified in the urine: methylated glycosides, glucuronides of anthocyanidins and anthocyanins, a sulfoconjugate of cyanidin, and anthocyanidins. Total urinary excretion of blackberry anthocyanin metabolites was 0.160 +/- 0.020% (n = 5) of the amount of anthocyanins ingested. Monoglucuronides of anthocyanidins represented >60% of this excretion. Urinary excretion of anthocyanins was maximal between 2 and 4 h after the meal, but continued during the 24 h of the experiment. This study highlighted the influence of aglycon structure on anthocyanin urinary excretion. It demonstrated that anthocyanins are not only methylated but also glucuroconjugated and sulfoconjugated in humans and that the main metabolites of blackberry anthocyanins in human urine were anthocyanidin monoglucuronides.