Fe deficiency is a public-health problem worldwide, and effective measures for preventing Fe deficiency are needed. The aim of the present study was to determine the bioavailability of Fe in cocoa using the Hb regeneration efficiency (HRE) method. Thirty-five F344/N male weanling rats were fed a low-Fe diet for 4 weeks to deplete body Fe stores. Then, four groups of seven animals each were repleted for 20 d using a modified AIN-93G diet fortified with ferrous sulphate, ferric citrate or two brands of cocoa powder to provide a total dietary Fe concentration of 20 mg/kg. As a negative control, seven rats were maintained on the low-Fe diet. The HRE were 0·733, 0·350, 0·357 and 0·336 for ferrous sulphate, ferric citrate and the two brands of cocoa powder, respectively. The relative biological values (RBV), defined as the ratio of the sample HRE to that of ferrous sulphate, were 0·478, 0·488 and 0·459 for ferric citrate and the two brands of cocoa powder, respectively. The Fe bioavailability of cocoa was significantly less than that of ferrous sulphate and was similar to that of ferric citrate. The difference in Fe bioavailability between the two brands of cocoa powder was negligible. When the negative control was used to correct the data, estimates of the RBV derived from Hb gain were similar to those derived from the HRE. These results suggest that cocoa is a significant source of moderately bioavailable Fe.

The effect of different food matrices on the metabolism and excretion of polyphenols is uncertain. The objective of the study was to evaluate the possible effect of milk on the excretion of ( − )-epicatechin metabolites from cocoa powder after its ingestion with and without milk. Twenty-one volunteers received the following three test meals each in a randomised cross–over design with a 1-week interval between meals: (1) 250 ml whole milk as a control; (2) 40 g cocoa powder dissolved in 250 ml whole milk (CC–M); (3) 40 g cocoa powder dissolved in 250 ml water (CC–W). Urine was collected before consumption and during the 0–6, 6–12 and 12–24 h periods after consumption. ( − )-Epicatechin metabolite excretion was measured using liquid chromatography–MS. One ( − )-epicatechin glucuronide and three ( − )-epicatechin sulfates were detected in urine excreted after the intake of the two cocoa beverages (CC–M and CC–W). The results show that milk does not significantly affect the total amount of metabolites excreted in urine. However, differences in metabolite excretion profiles were observed; there were changes in the glucuronide and sulfate excretion rates, and the sulfation position between the period of excretion and the matrix. The matrix in which polyphenols are consumed can affect their metabolism and excretion, and this may affect their biological activity. Thus, more studies are needed to evaluate the effect of these different metabolite profiles on the body.

The aims of the present study were to determine the level of (-)-epicatechin (EC) and its metabolites in rat plasma after oral administration of cocoa powder and to evaluate the protective effect of cocoa powder in terms of suppressing the oxidation of plasma components. Rats were orally administered 1 g cocoa powder/kg body weight, containing 7·80 mg EC, and their blood was collected before administration and at designated time intervals thereafter. The EC and its metabolites in plasma were treated with β-glucuronidase and/or sulfatase, then analysed by HPLC and by liquid chromatography–MS. Several EC-related compounds were detected in plasma such as free EC, and glucuronide, sulfate, and glucuronide–sulfate conjugates of non-methylated or methylated EC. All EC metabolites showed a maximum concentration in plasma at 30–60 min post-administration. Glucuronide conjugates of both non-methylated and methylated EC were found in high concentration in plasma. Moreover, administration of cocoa powder significantly reduced the accumulation of lipid peroxides in plasma and significantly reduced the consumption of α-tocopherol in plasma oxidized by treatment with 2,2′-azobis-(2-amidinopropane) dihydrochloride (AAPH (25 mmol/l)) or CuSO4 (100 μmol/l) compared with that in the case of plasma obtained before administration. The total EC concentration in plasma was negatively correlated with the level of accumulation of lipid peroxides in plasma oxidized by treatment with AAPH (25 mmol/l) and was positively correlated with the level of residual α-tocopherol in plasma oxidized by treatment with CuSO4 (100 μmol/l). These results indicate that EC in cocoa powder was absorbed from the digestive tract, that various conjugated forms of EC were generated in the digestive tract and distributed to the plasma, and that these enhanced the antioxidative activity of plasma.