Slow starch hydrolysis of non-glutinous rice flour and potato starch heated with taxifolin: contribution of proteins to the former and longer amylose to the latter.

Taxifolin (dihydroquercetin), which has various pharmacological functions, is contained in edible plants. Some taxifolin-containing foodstuffs such as adzuki bean and sorghum seeds are cooked by themselves and with other starch-containing ingredients. In this study, non-glutinous rice flour (joshin-ko) and potato starch were heated with taxifolin. The heating resulted in the slowdown of pancreatin-induced hydrolysis of suspendable starch in joshin-ko and soluble starch in potato starch. The products of taxifolin formed by the heating such as quercetin were combined with starch during the heating and/or retrogradation, which was converted into the suspendable starch in joshin-ko and the soluble starch in the potato. Taking the difference in protein content and amylose chain length between joshin-ko and potato starch into account, the slowdown is discussed to be due to the binding of the reaction products of taxifolin to proteins in suspendable starch in joshin-ko and to soluble amylose in potato starch.

Slow hydrolysis of amylose in soluble starch and amylopectin in suspendable starch liberated from non-glutinous rice flour heated with a sorghum extract.

Polyphenols in plant can interact with amylose and amylopectin in different ways affecting their hydrolysis by α-amylase. Pancreatin liberated starch from non-glutinous rice flour heated with and without an aqueous extract of sorghum seeds, and hydrolyzed the liberated starch. The hydrolysis of the liberated starch was slowed down by the sorghum extract. Then, the liberated starch was fractionated into soluble starch and suspendable starch. In the soluble starch, amylose hydrolysis was slowed down more significantly than amylopectin hydrolysis, and in the suspendable starch, the hydrolysis of amylopectin was slowed down efficiently by the sorghum extract. It is discussed that (i) the slowdown in the former might be due to the binding of sorghum components including procyanidins to amylose, and that (ii) the slowdown in the latter might be due to the complex formation between amylopectin and shorter amylose combined with the sorghum components. The contribution of amylose to the slowdown was supported by the result that the sorghum extract inhibited the starch hydrolysis only slightly in glutinous rice flour, the starch of which was almost composed of amylopectin. It was proposed a possible mechanism of the slowdown of amylopectin hydrolysis in suspendable starch by shorter amylose combined with the sorghum components.

Further slowing down of hydrolysis of amylose heated with black soybean extract by treating with nitrite under gastric conditions.

Black soybean (BSB), which contains cyanidin-3-O-glucoside (C3G) and procyanidins, is cooked with rice in Japan. The color of the cooked rice is purplish red due to the binding of C3G and reddish oxidation products of procyanidins. These components can slowdown pancreatin-induced hydrolysis of amylose more significantly than the hydrolysis of amylopectin, and can react with nitrous acid in the stomach. This manuscript deals with the effects of nitrous acid on pancreatin-induced hydrolysis of amylose heated with BSB extract. The hydrolysis of amylose heated with BSB extract was slow, and the slowdown was due to the binding of C3G/its degradation products and degradation products of procyanidins. The amylose hydrolysis was slowed down further by treating with nitrite under gastric conditions. The further slowdown was discussed to be due to the binding of the products, which were formed by the reaction of procyanidins with nitrous acid, to amylose. In the products, dinitroprocyanidins were included. In this way, the digestibility of amylose heated with BSB extract can be slowed down further by reacting with nitrous acid in the stomach.

Pancreatin-induced liberation of starch/cyanidin 3-O-glucoside complexes from rice cooked with black soybean that exhibit slow hydrolysis.

Cyanidin 3-O-glucoside (C3G), which has various health-promoting functions, is contained in black soybean (BSB). In Japan and Korea, BSB is cooked with rice and the cooked rice appears purplish in colour. In this study, BSB was cooked with glutinous rice, non-glutinous rice, and high-amylose rice. The amount of C3G detected in high-amylose rice was greater than that detected in glutinous rice, suggesting that C3G combined more efficiently with amylose than with amylopectin. Pancreatin induced the liberation of starch/C3G complexes from the purplish cooked rice, and rate of the liberation was in the following order; glutinous rice < non-glutinous rice < high-amylose rice. The amylose/C3G complexes liberated from high-amylose rice was hydrolysed slowly, while the amylopectin/C3G complexes liberated from glutinous rice were hydrolysed into smaller amylopectin/C3G complexes that were difficult to further hydrolysis. Thus, C3G may be useful for preparing foods whose starch hydrolysis is slow.

The Procyanidin C1-Dependent Inhibition of the Hydrolysis of Potato Starch and Corn Starch Induced by Pancreatin.

Procyanidins are contained in various foods, and their effects on starch hydrolysis have been reported. In Japan, black soybeans, which contain a trimeric procyanidin, procyanidin C1 (proC1), are cooked with rice and used to prepare dumplings. In this study, the effects of proC1 on the pancreatin-induced formation of reducing sugars and starch hydrolysis were studied using potato starch and corn starch. ProC1 inhibited both reactions; the inhibition was greater in potato starch than corn starch when added to heated potato starch and corn starch. When heated with proC1, its inhibitory effects decreased, especially in potato starch, suggesting the important role of proC1 itself for the inhibition of potato starch hydrolysis. ProC1 also inhibited the hydrolysis when added to heated, longer amylose (average molecular weight: 31,200), and the inhibition decreased when heated with the amylose. On the other hand, proC1 could not inhibit the hydrolysis when added to heated, shorter amylose (average molecular weight: 4500), but could when heated with the amylose, suggesting the important role of the degradation products of proC1 for the inhibition. We discuss the mechanism of the proC1-dependent inhibition of amylose hydrolysis, taking the molecular weight into account.

Contribution of amylose-procyanidin complexes to slower starch digestion of red-colored rice prepared by cooking with adzuki bean.

Combining high-carbohydrate food with polyphenol-rich food is a possible way of producing slowly digestible starch with beneficial health properties. In Japan, non-glutinous and glutinous rice are cooked with adzuki bean and the colour of the cooked rice is pale red. In this article, we show that (1) the red colour of rice could be attributed to the oxidation of adzuki bean procyanidins, (2) pancreatin-induced starch digestion of the red-coloured non-glutinous rice was slower than white rice and (3) the digestion of amylose and potato starch but not amylopectin became slower by heating with procyanidin B2. Furthermore, the rate of starch digestion of red-coloured rice was not affected by nitrite treatment under simulated gastric conditions. The above results show that procyanidins could bind to amylose independent of the starch source by heating and could suppress starch digestion by α-amylase in the intestine.

Procyanidins in rice cooked with adzuki bean and their contribution to the reduction of nitrite to nitric oxide (•NO) in artificial gastric juice.

In Japan, adzuki bean is cooked with rice. During the cooking, the colour of rice becomes pale red. It is postulated that the red pigment is produced from procyanidins and that the ingestion of red rice causes the production of nitric oxide (•NO) in the stomach by reacting with salivary nitrite. The increase in colour intensity accompanied the decrease in the amounts of procyanidins, suggesting the conversion of procyanidins into the red pigment during the cooking. In addition, the red pigment combined with rice strongly. The red-coloured rice produced •NO by reacting with nitrite in artificial gastric juice, and the amounts were dependent on the contents of procyanidins and the equivalents. It is suggested that although adzuki procyanidins were oxidised during cooking with rice, procyanidins and the equivalents bound to rice still have the ability to produce bioactive •NO in the stomach using nitrite in mixed whole saliva.

Slow starch digestion in the rice cooked with adzuki bean: Contribution of procyanidins and the oxidation products.

Adzuki bean is often cooked with non-glutinous rice in Japan, and the dish is called adzuki-meshi. By the cooking, flavonoids in adzuki bean are transferred to rice, and the color of the rice becomes pale red. However, it has not been reported on starch digestion of the rice of adzuki-meshi. The purpose of this study is to elucidate that the transferred flavonoids, especially procyanidins could slow down the digestion of rice starch. The principal results obtained are (1) that pancreatin-induced starch digestion, which was observed as the liberation of reducing sugars and starch fragments from the rice, was slower in the pale red rice of adzuki-meshi than the rice cooked without adzuki bean, (2) that the starch fragments liberated from the rice of adzuki-meshi were digested slowly, and (3) that procyanidins and the oxidation products, which were not extracted by methanol, were present in the pale red rice. From the results, it was concluded that adzuki bean procyanidins and the oxidation products, which bound to rice starch during cooking, could contribute to slow down the starch digestion.

Suppression of Pancreatin-Induced Digestion of Starch in Starch Granules by Starch/Fatty Acid and Starch/Flavonoid Complexes in Retrograding Rice Flour.

Adzuki beans are used to prepare foods with glutinous and non-glutinous rice in Japan, and adzuki bean pigments are able to color rice starch a purplish red. This study deals with the adzuki bean extract-dependent suppression of starch digestion of non-glutinous rice flour (joshinko in Japanese), which was gelatinized in boiling water and then cooled to 37 °C. Accompanying the treatment of joshinko with pancreatin, amylose and amylopectin were released from the joshinko particles, and the released amylose and amylopectin were further digested. The adzuki extract suppressed the release and digestion by binding to amylose and amylopectin, which were present in the particles and at the surfaces of the particles. Fatty acids and flavonoids in the adzuki extract contributed to the suppression. In addition, the starch digestion in the joshinko particles appeared to be suppressed if the amylose/fatty acid complexes and amylose/flavonoid and amylopectin/flavonoid complexes, which are poor substrates of α-amylase, surrounded the particles. It is discussed that the suppression was due to the prevention of α-amylase access to the particles.

Interactions of flavonoids with α-amylase and starch slowing down its digestion.

Starch is digested to glucose in the intestine and absorbed into the body. If the increased blood concentrations of sugar after meals decrease slowly or are maintained for a long time, various adverse effects are induced. Therefore, it is important to decrease the rate of the digestibility of starch in the intestine in the patients of hyperglycemia. One of the ways to effect a decrease is the inhibition of α-amylase secreted from the pancreas. Flavonoids are a group of compounds that can inhibit this enzyme's activity, and many investigators have studied the flavonoid-dependent inhibition of this enzyme and presented mechanisms for the inhibition of its activity. Starch containing foods, however, cooked or ingested with flavonoid containing foods are mixed with saliva and gastric juice in the stomach. Thus, flavonoids in the foods can interact with starch and can react with nitrous acid derived from the oral cavity in the stomach before being transported to the intestine. This review mainly deals with: (i) the inhibition of α-amylase activity by flavonoids suggesting the mechanisms of the inhibition, (ii) suppression of starch digestion by flavonoids by forming starch-flavonoid complexes by hydrophobic interactions, and (iii) formation of starch not easily digested by α-amylase by the formation of covalent bonds between flavonoids and starch during cooking and in the stomach. In addition, the cooperation of flavonoids with fatty acids are discussed taking their binding to amylose into account.

Possible Reactions of Dietary Phenolic Compounds with Salivary Nitrite and Thiocyanate in the Stomach.

Foods are mixed with saliva in the oral cavity and swallowed. While staying in the stomach, saliva is contentiously provided to mix with the ingested foods. Because a salivary component of nitrite is protonated to produce active nitrous acid at acidic pH, the redox reactions of nitrous acid with phenolic compounds in foods become possible in the stomach. In the reactions, nitrous acid is reduced to nitric oxide (•NO), producing various products from phenolic compounds. In the products, stable hydroxybezoyl benzofuranone derivatives, which are produced from quercetin and its 7-O-glucoside, are included. Caffeic acid, chlorogenic acid, and rutin are oxidized to quinones and the quinones can react with thiocyanic acid derived from saliva, producing stable oxathiolone derivatives. 6,8-Dinitrosocatechis are produced from catechins by the redox reaction, and the dinitrocatechins are oxidized further by nitrous acid producing the quinones, which can make charge transfer complexes with the dinitrosocatechin and can react with thiocyanic acid producing the stable thiocyanate conjugates. In this way, various products can be produced by the reactions of salivary nitrite with dietary phenolic compounds, and reactive and toxic quinones formed by the reactions are postulated to be removed in the stomach by thiocyanic acid derived from saliva.

Inhibition of Pancreatin-Induced Digestion of Cooked Rice Starch by Adzuki (Vigana angularis) Bean Flavonoids and the Possibility of a Decrease in the Inhibitory Effects in the Stomach.

Flavonoids of adzuki bean bind to starch when the beans are cooked with rice. The purpose of this study is to show that adzuki flavonoids can suppress pancreatin-induced digestion of cooked rice starch. The diethyl ether extract of water boiled with adzuki bean inhibited starch digestion, and quercetin and a cyanidin-catechin conjugate (vignacyanidin) but not taxifolin in the extract contributed to the inhibition. The order of their inhibitory effects (taxifolin < quercetin < vignacyanidin) suggested that the effects increased with an increase in their hydrophobicity. The diethyl ether extract also inhibited the starch digestion of cooked rice preincubated in artificial gastric juice, and the level of inhibition was decreased by nitrite. The decrease was due to nitrite-induced consumption of quercetin and vignacyanidin. Taking these results into account, we discuss mechanisms of quercetin- and vignacyanidin-dependent inhibition of starch digestion and the possibility of the decrease in their inhibitory effects by nitrite in the stomach.

Reactions of polyphenols in masticated apple fruit with nitrite under stomach simulating conditions: Formation of nitroso compounds and thiocyanate conjugates.

By the ingestion of fresh apple fruit, it is masticated squeezing apple juice into the oral cavity and the juice is mixed with saliva. The mixture of saliva and apple juice is swallowed into the stomach where the pH is around 2. This paper deals with the reactions of polyphenols in the juice obtained by mastication of apple fruit with salivary nitrite under acidic conditions. The concentrations of catechins, procyanidins, and chlorogenic acid in the apple juice were approximately 55, 55, and 170µM, respectively, and the polyphenols were oxidized by salivary nitrite under conditions of the stomach. Rates of the oxidation increased in order chlorogenic acid

Quercetin 7-O-glucoside suppresses nitrite-induced formation of dinitrosocatechins and their quinones in catechin/nitrite systems under stomach simulating conditions.

Foods of plant origin contain flavonoids. In the adzuki bean, (+)-catechin, quercetin 3-O-rutinoside (rutin), and quercetin 7-O-ß-D-glucopyranoside (Q7G) are the major flavonoids. During mastication of foods prepared from the adzuki bean, the flavonoids are mixed with saliva and swallowed into the stomach. Here we investigated the interactions between Q7G and (+)-catechin at pH 2, which may proceed in the stomach after the ingestion of foods prepared from the adzuki bean. Q7G reacted with nitrous acid producing nitric oxide (˙NO) and a glucoside of 2-(3,4-dihydroxybenzoyl)-2,4,6-trihydroxy-3(2H)-benzofuranone. (+)-Catechin reacted with nitrous acid producing ˙NO and 6,8-dinitrosocatechin. The production of the dinitrosocatechin was partly suppressed by Q7G, and the suppression resulted in the enhancement of Q7G oxidation. 6,8-Dinitrosocatechin reacted further with nitrous acid generating the o-quinone, and the quinone formation was effectively suppressed by Q7G. In the flavonoids investigated, the suppressive effect decreased in the order Q7G≈quercetin>kaempferol>quercetin 4'-O-glucoside>rutin. Essentially the same results were obtained when (-)-epicatechin was used instead of (+)-catechin. The results indicate that nitrous acid-induced formation of 6,8-dinitrosocatechins and the o-quinones can be suppressed by flavonols in the stomach, and that both a hydroxyl group at C3 and ortho-hydroxyl groups in the B-ring are required for efficient suppression.

Interactions between (+)-catechin and quercetin during their oxidation by nitrite under the conditions simulating the stomach.

When foods that contain catechins and quercetin glycosides are ingested, quercetin glycosides are hydrolyzed to quercetin during mastication by hydrolytic enzymes derived from oral bacteria and the generated quercetin aglycone is mixed with catechins in saliva. The present study deals with the interactions between (+)-catechin and quercetin during their reactions with nitrous acid under the conditions simulating the gastric lumen. Nitrous acid reacted with (+)-catechin producing 6,8-dinitrosocatechin, and quercetin partially suppressed the dinitrosocatechin formation. Nitric oxide, which was produced by not only (+)-catechin/nitrous acid but also quercetin/nitrous acid systems, was used to produce 6,8-dinitrosocatechin. Furthermore, 6,8-dinitrosocatechin was oxidized by nitrous acid to the quinone form. The quinone formation was significantly suppressed by quercetin. Quercetin-dependent suppression of the above reactions accompanied the oxidation of quercetin, which was observed with the formation of 2-(3,4-dihydroxybenzoyl)-2,4,6-trihydroxy-3(2H)-benzofuranone. Taking the above results into account, we proposed a possible mechanism of 6,8-dinitrosocatechin formation and discuss the importance of quercetin to prevent the quinone formation from 6,8-dinitrosocatechin in the gastric lumen, taking the interactions between quercetin and catechins into account.

Interactions of starch with a cyanidin-catechin pigment (vignacyanidin) isolated from Vigna angularis bean.

A cyanidin-catechin pigment isolated from adzuki bean (vignacyanidin) interacted with starch. The pigment had absorption maxima at 530 and 540 nm at pH 2.0 and 6.8, respectively, and starch (10 and 100 mg ml(-1)) increased the absorbance, shifting the absorption maxima to longer wavelengths. Nitrite oxidised vignacyanidin at pH 2.0, and the oxidation resulted in the production of nitric oxide (NO). Rates of the oxidation and the NO production were enhanced by starch. Vignacyanidin inhibited α-amylase-catalysed digestion of starch at pH 6.8, and amylose digestion was more effectively inhibited than amylopectin digestion. The above results suggest (i) that binding of the pigment to starch increased the accessibility of nitrous acid to the pigment, and (ii) that the binding reduced the digestibility of starch by α-amylase. Possible functions of the pigment in the stomach and the intestine are postulated, taking the above results into account.

Isolation and characterization of a cyanidin-catechin pigment from adzuki bean (Vigna angularis).

Adzuki bean is used to prepare many kinds of foods in east Asia, and the seed coat contains water-soluble anthocyanins, catechins, and flavonols. In the present study, ethyl acetate-soluble purplish pigments were isolated from adzuki bean. Pigments of soaked adzuki bean were extracted with 1% HCl in methanol. Ethyl acetate-soluble purple pigments were obtained from the methanol soluble components. Purple pigments 1 and 2 were purified from the ethyl acetate-soluble pigments by Sephadex LH-20 column chromatography and preparative reversed-phase HPLC. NMR and mass spectra suggested that pigment 1 was a condensation product of cyanidin and (+)-catechin, in which 5-hydroxy and C-4 positions of the cyanidin moiety were substituted by the addition of 5-hydroxy and C-6 positions of the (+)-catechin moiety, respectively. Pigment 2 was an isomer of pigment 1. It is suggested that pigments 1 and 2 contribute to the purplish-red colour of foods prepared using adzuki bean.

Effects of starch on nitrous acid-induced oxidation of kaempferol and inhibition of α-amylase-catalysed digestion of starch by kaempferol under conditions simulating the stomach and the intestine.

Kaempferol glycosides can be hydrolyzed to their aglycone kaempferol during cooking under acidic conditions and in the oral cavity and the intestine by glycosidases. Kaempferol was oxidised by nitrite under acidic conditions (pH 2.0) to produce nitric oxide (NO), and the nitrite-induced oxidation of kaempferol was enhanced and inhibited by 10 and 100mg of starch ml(-1), respectively. The opposite effects of starch were discussed by considering the binding of kaempferol to starch and starch-dependent inhibition of the accessibility of nitrous acid to kaempferol. Kaempferol inhibited α-amylase-catalysed starch digestion by forming starch/kaempferol complexes, and the inhibitory effects increased in the order of amylopectin

Effects of the food additive sulfite on nitrite-dependent nitric oxide production under conditions simulating the mixture of saliva and gastric juice.

The food additive sulfite is mixed with saliva, which contains nitrite, in the oral cavity, and the mixture is mixed with gastric juice in the stomach. In the stomach, salivary nitrite can be transformed to nitric oxide (NO). In this study, the effects of sulfite on nitrite-dependent NO production were investigated using acidified saliva (pH 2.6) and acidic buffer solutions (pH 2.0). Sulfite enhanced NO production in acidified saliva and acidic buffer solutions, and the enhancement increased with the increase in sulfite concentration from 0 to 0.1 mM, whereas suppressed NO production and the suppression increased as the concentration was increased over 0.2 mM. The enhancement was due to the increase in reaction rate between nitrous acid and nitrososulfonate (ONSO(3)(-)) that was formed by the reaction of nitrous acid with hydrogen sulfite, and the suppression was due to the increase in hydrogen sulfite-dependent consumption rate of ONSO(3)(-). A salivary component SCN(-) (1 mM) enhanced and suppressed NO production induced by 1 mM nitrite when sulfite concentrations were lower and higher than 1 mM, respectively. ONSO(3)(-) formed from hydrogen sulfite and nitrosyl thiocyanate (ONSCN), which was produced by the reaction of nitrous acid with SCN(-), seemed to contribute to the enhancement and suppression. NO production induced by nitrite/ascorbic acid systems was suppressed by sulfite, and the suppressive effects were decreased by SCN(-), whereas sulfite-induced suppression of NO production in nitrite/rutin systems was increased by SCN(-). During reactions of nitrite with sulfite in the presence and absence of SCN(-), oxygen was taken up. The oxygen uptake is discussed to be due to autoxidation of NO and radical chain reactions initiated by hydrogen sulfite radicals. The results of the present study suggest that sulfite can enhance and suppress nitrite-dependent NO production. It is discussed that radicals including hydrogen sulfite radicals can be formed through the reactions of nitrite and sulfite in the stomach.

Enhancement of iron(II)-dependent reduction of nitrite to nitric oxide by thiocyanate and accumulation of iron(II)/thiocyanate/nitric oxide complex under conditions simulating the mixture of saliva and gastric juice.

Iron(III) ingested as a food component or supplement for iron deficiencies can react with salivary SCN(-) to produce Fe(SCN)(2+) and can be reduced to iron(II) by ascorbic acid in the stomach. Iron(II) generated in the stomach can react with salivary nitrite and SCN(-) to produce nitric oxide (NO) and FeSCN(+), respectively. The purpose of this investigation is to make clear the reactions among nitrite, SCN(-), iron ions, and ascorbic acid under conditions simulating the mixture of saliva and gastric juice. Iron(II)-dependent reduction of nitrite to NO was enhanced by SCN(-) in acidic buffer solutions, and the oxidation product of iron(II) reacted with SCN(-) to produce Fe(SCN)(2+). Almost all of the NO produced was autoxidized to N(2)O(3) under aerobic conditions. Iron(II)-dependent production of NO was also observed in acidified saliva. Under anaerobic conditions, NO transformed Fe(SCN)(2+) and FeSCN(+) to Fe(SCN)NO(+) in acidic buffer solutions. Fe(SCN)NO(+) was also formed under aerobic conditions when excess ascorbic acid was added to iron(II)/nitrite/SCN(-) systems in acidic buffer solutions and acidified saliva. The Fe(SCN)NO(+) formed was transformed to Fe(SCN)(2+) and iron(III) at pH 2.0 and pH 7.4, respectively, by O(2). Salivary glycoproteins could complex with iron(III) in the stomach preventing the formation of Fe(SCN)(2+). Ascorbic acid reduced iron(III) to iron(II) to react with nitrite and SCN(-) as described above. The above results suggest (i) that iron(II) can have toxic effects on the stomach through the formation of reactive nitrogen oxide species from NO when supplemented without ascorbic acid and through the formation of both reactive nitrogen oxide species and Fe(SCN)NO(+) when supplemented with ascorbic acid, and (ii) that the toxic effects of iron(III) seemed to be smaller than and similar to those of iron(II) when supplemented without and with ascorbic acid, respectively. Possible mechanisms that cause oxidative stress on the stomach through Fe(SCN)NO(+) are discussed.