Anthocyanin metabolites in human urine and serum

Abstract

In the present study we investigated the metabolic conversion of cyanidin glycosides in human subjects using solid-phase extraction, HPLC–diode array detector, MS, GC, and enzymic techniques. Volunteers consumed approximately 20 g chokeberry extract containing 1·3 g cyanidin 3-glycosides (899 mg cyanidin 3-galactoside, 321 mg cyanidin 3-arabinoside, 51 mg cyanidin 3-xyloside and 50 mg cyanidin 3-glucoside). Blood samples were drawn at 0, 0·5, 1, and 2 h post-consumption of the extract. Urine samples were also collected at 0, 4–5, and 22–24 h. We have confirmed that human subjects have the capacity to metabolise cyanidin 3-glycosides, as we observed at least ten individual anthocyanin metabolites in the urine and serum. Average concentrations of anthocyanins and anthocyanin metabolites in the urine reached levels of 17·9 (range 14·9–20·9) μmol/l within 5 h post-consumption and persisted in 24 h urine samples at levels of 12·1 (range 11·1–13·0) nmol/l. In addition, average total levels of anthocyanins and anthocyanin metabolites detected in the serum were observed at 591·7 (range 197·3–986·1) nmol/l within 2 h post-consumption. Cyanidin 3-galactoside accounted for 55·4 % (9·9 (range 7·2–12·6) μmol/l) and 66·0 % (390·6 (range 119·4–661·9) nmol/l) of the detected anthocyanins in the urine and serum samples, respectively. The metabolites were identified as glucuronide conjugates, as well as methylated and oxidised derivatives of cyanidin 3-galactoside and cyanidin glucuronide. Conjugation probably affects the biological activity of anthocyanins and these metabolic products are likely in part responsible for the reported health benefits associated with the consumption of anthocyanins.