Two major flavonol glycosides [quercetin 3,4‘-diglucoside (F1) and quercetin 4‘-monoglucoside (F2)] and a flavonol aglycon quercetin were mainly localized in the abaxial epidermis of scales. Their contents increased on aging. Peroxidase in scales oxidized flavonols in the order quercetin ≫ F2 > isoquercetin ≫ F1, and the activity was higher in the outer than in the inner scales. These results suggest that the enzyme can participate in the formation of defense substances against infection and brown compounds in the dry skin from quercetin. Contents of F1 and F2 in scales were decreased by cooking by boiling. This decrease was due to the release of F1 and F2 into cooking water and their oxidation. F2 was oxidized more rapidly than F1 during cooking. The difference in the stability between F1 and F2 was due to the presence or absence of a hydroxyl group at the C-3 position of the glucosides. Keywords: Flavonols; peroxidase; distribution in tissues; thermostability; onion (Allium cepa)

Outer scales of yellow onion bulbs turn brown during maturing. The brown outer scales contain an antifungal component, 3,4-dihydroxybenzoic acid. An aim of the present study is to elucidate the mechanism of formation of the benzoic acid. In a browning scale, the scale was divided into three areas; fleshy, drying and dried brown areas. Levels of quercetin glucosides in dried brown areas were less than 10% of the glucosides in fleshy and drying areas, whereas levels of quercetin were high in dried brown areas. This result suggests that quercetin was formed by deglucosidation of quercetin glucosides on the border between drying and dried brown areas. Peroxidase (POX) activity of dried brown areas was about 10% of those of fleshy and drying areas. Quercetin was oxidized by autooxidation, and cell-free extracts of drying areas and POX isolated from onion scales enhanced the oxidation even in the absence of externally added hydrogen peroxide. The enhancement of quercetin oxidation was suppressed by catalase. No tyrosinase-like activity was detected in the cell-free extracts and the POX preparation. These results suggest that, during the enhanced oxidation of quercetin, hydrogen peroxide is formed. 3,4-Dihydroxybenzoic acid and 2,4,6-trihydroxyphenylglyoxylic acid, which were the oxidation products of quercetin, were found in dried brown area. These results suggest that an antifungal agent 3,4-dihydroxybenzoic acid is formed by POX-dependent oxidation of quercetin on browning of onion scales.

Under acidic conditions, nitrite is protonated to nitrous acid (pK(a) = 3.2-3.4) that can be transformed into nitric oxide by self-decomposition and reduction. When sodium nitrite was mixed with quercetin at pH 1-2, quercetin was oxidized producing nitric oxide. In addition to quercetin, kaempferol and quercetin 4'-glucoside were also oxidized by nitrous acid, but oxidation of apigenin, luteolin, and rutin was slow compared to oxidation of the above flavonols. These results suggested that flavonols, which have a free hydroxyl group at carbon position 3, can readily reduce nitrous acid to nitric oxide. When the pH of saliva was decreased to 1-2, formation of nitric oxide was observed. The nitric oxide formation was enhanced by quercetin, and during this process quercetin was oxidized. These results indicate that there is a possibility of reactions between phenolics and nitrous acid derived from salivary nitrite in the stomach.

Glycemic indexes of bread made from mixtures of wheat flour and buckwheat flour are lower than those made from wheat flour. To discuss the mechanism of the buckwheat flour-dependent decrease in glycemic indexes, the formation of a starch-iodine complex and amylase-catalyzed digestion of starch were studied using buckwheat flour itself and buckwheat flour from which fatty acids, rutin, and proanthocyanidins including flavan-3-ols had been extracted. Absorbance due to the formation of a starch-iodine complex was larger in extracted than control flour, and starch in extracted flour was more susceptible to pancreatin-induced digestion than starch in control flour. Fatty acids, which were found in the buckwheat flour extract, bound to amylose in the extracted flour, inhibiting its digestion by pancreatin. Rutin and epicatechin-dimethylgallate, which were also found in the extract, bound to both amylose and amylopectin in the extracted flour, inhibiting their digestion induced by pancreatin. We discussed from these results that the lower glycemic indexes of bread made from mixtures of wheat flour and buckwheat flour were due to binding of fatty acids, rutin, and epicatechin-dimethylgallate, which were contained in buckwheat flour, to wheat flour starch.