Rethinking the Mechanism of the Health Benefits of Proanthocyanidins: Absorption,Metabolism, and Interaction with Gut Microbiota

Proanthocyanidins, as the oligomers or polymers of flavan‐3‐ol, are widely discovered in plants such as fruits, vegetables, cereals, nuts, and leaves, presenting a major part of dietary polyphenols. Although proanthocyanidins exert several types of bioactivities, such as antioxidant, antimicrobial, cardioprotective, and neuroprotective activity, their exact mechanisms remain unclear. Due to the complexity of the structure of proanthocyanidins, such as their various monomers, different linkages and isomers, investigation of their bioavailability and metabolism is limited, which further hinders the explanation of their bioactivities. Since the large molecular weight and degree of polymerization limit the bioavailability of proanthocyanidins, the major effective site of proanthocyanidins is proposed to be in the gut. Many studies have revealed the effects of proanthocyanidins from different sources on changing the composition of gut microbiota based on in vitro and in vivo models and the bioactivities of their metabolites. However, the metabolic routes of proanthocyanidins by gut microbiota and their mutual interactions are still sparse. Thus, this review summarizes the chemistry, absorption, and metabolic pathways of proanthocyanidins ranging from monomers to polymers, as well as the mutual interactions between proanthocyanidins and gut microbiota, in order to better understand how proanthocyanidins exert their health‐promoting functions.     

Absorption,metabolism, and excretion of green tea flavan‐3‐ols in humans with an ileostomy

Green tea containing 634 μmol of flavan‐3‐ols was ingested by human subjects with an ileostomy. Ileal fluid, plasma, and urine collected 0–24 h after ingestion were analysed by HPLC‐MS. The ileal fluid contained 70% of the ingested flavan‐3‐ols in the form of parent compounds (33%) and 23 metabolites (37%). The main metabolites effluxed back into the lumen of the small intestine were O‐linked sulphates and methyl‐sulphates of (epi)catechin and (epi)gallocatechin. Thus, in subjects with a functioning colon substantial quantities of flavan‐3‐ols would pass from the small to the large intestine. Plasma contained 16 metabolites, principally methylated, sulphated, and glucuronidated conjugates of (epi)catechin and (epi)gallocatechin, exhibiting 101–256 nM peak plasma concentration and the time to reach peak plasma concentration ranging from 0.8 to 2.2 h. Plasma pharmacokinetic profiles were similar to those obtained with healthy subjects, indicating that flavan‐3‐ol absorption occurs in the small intestine. Ileostomists had earlier plasma time to reach peak plasma concentration values than subjects with an intact colon, indicating the absence of an ileal brake. Urine contained 18 metabolites of (epi)catechin and (epi)gallocatechin in amounts corresponding to 6.8±0.6% of total flavan‐3‐ol intake. However, excretion of (epi)catechin metabolites was equivalent to 27% of the ingested (?)‐epicatechin and (+)‐catechin.     

Human Colonic Bacterial Degradability of Dietary Fibres from Sea-Lettuce (Ulva sp)

Sea-lettuce (Ulva sp) is one of the commonly consumed seaweeds which contains 16·5% of water-soluble and 13·3% insoluble dietary fibres. Since physiological effects of fibres are partly related to their colonic bacteria fermentability, Ulva sp and its constitutive soluble and insoluble fibres were incubated with faecal bacteria in an in vitro batch fermenter system. After 24 h of incubation, 32·0±0·4%, 25·9±0·4% and 50·9±7·4% of Ulva, soluble and insoluble fibres constitutive sugars, respectively, were degraded. Consequently, Ulva and its soluble fibre, ulvan, are poorly fermented by colonic bacteria. The constitutive sugars, rhamnose and glucuronate and the aldobiouronate β-D -glucuronosyluronate-(1,4)-L -rhamnose of the glucuronoxylorhamnan sulphate present in the soluble fibre are highly fermented. Chemical desulphation and/or carboxyl group reduction did not modify this fermentation behaviour. Thus, the particular chemical structure of ulvan is responsible for the resistance of this polysaccharide and of Ulva to colonic bacterial fermentation. As a physiological consequence of this particular behaviour, consumption of dietary fibres from sea-lettuce could be expected to act mainly as bulking agents with little effect on nutrient metabolism due to colonic bacterial fermentation products (short-chain fatty acids). © 1997 SCI.     

Acerola and cashew apple as sources of antioxidants and dietary fibre

Several tropical fruits have been described as natural sources of dietary fibre (DF) and phenolic compounds, associated with different health effects. The aim of this work was to ascertain the DF, phenolic compounds content (including non‐extractable polyphenols, mostly associated with DF) and antioxidant capacity in acerola fruits and cashew apples from selected clones. ‘BRS 236’ acerola fruits presented a high antioxidant capacity because of the combination of both extractable polyphenols and l ‐ascorbic acid (providing together a Folin value of 170 kg^?1 g d.m.). ‘CCP 76’ cashew apples contained 28 g kg^?1 d.m. of extractable polyphenols and 13 g kg^?1 d.m. of ascorbic acid as well as a high amount of non‐extractable condensed tannins (52 g kg^?1 d.m.). DF content was of 260 g kg^?1 d.m. in acerola fruit and of 209 g kg^?1 d.m. in cashew apple. Acerola fruits and cashew apple should therefore be considered as new natural sources of DF and phenolic compounds.     

Profile of urinary and fecal proanthocyanidin metabolites from common cinnamon (Cinnamomum zeylanicum L.) in rats

Cinnamon (Cinnamomum zeylanicum L.) bark is widely used as a spice and in traditional medicine. Its oligomeric and polymeric proanthocyanidins are believed to be partly responsible for the beneficial properties of the plant. We describe here the metabolic fate of cinnamon proanthocyanidins in the urine and feces of rats fed a suspension of the whole bark. The metabolites include ten mono‐, di‐, and tri‐ conjugated (epi)catechin phase II metabolites and more than 20 small phenolic acids from intestinal microbial fermentation. Some of these are sulfated conjugates. Feces contain intact (epi)catechin and dimers. This suggests that free radical scavenging species are in contact with the intestinal walls for hours after ingestion of cinnamon. The phenolic metabolite profile of cinnamon bark in urine is consistent with a mixture of proanthocyanidins that are depolymerized into their constitutive (epi)catechin units as well as cleaved into smaller phenolic acids during their transit along the intestinal tract, with subsequent absorption and conjugation into bioavailable metabolites.     

Metabolic pathways of the colonic metabolism of flavonoids (flavonols,flavones and flavanones) and phenolic acids

Flavonoids are metabolized by the gut microflora giving rise to a wide range of phenolic acids that may exert systemic effects in the body. Nevertheless, the colonic metabolism pathways and the function of the metabolites formed are poorly studied. In the present study, the individual colonic metabolism of three subclasses of flavonoids (flavonols, flavones and flavanones) and phenolic acids was evaluated. For this, seven standards of flavonoids (quercetin, quercetin–rhamnoside, quercetin–rutinoside, myricetin, luteolin, naringenin and kaempferol–rutinoside) and two phenolic acids (protocatechuic acid and gallic acid) were submitted to an in vitro fermentation model using rat colonic microflora. Simultaneously, a nuts–cocoa cream enriched with these standards of flavonoids was incubated and the colonic metabolism of these compounds was evaluated. The results showed that the greatest number of colonic metabolites came from the fermentation of quercetin and quercetin–rhamnoside, and the maximum concentration of fermentation products was observed after 48 h of fermentation. On the other hand, a smaller number of fermentation products were observed after the colonic fermentation of kaempferol–rutinoside, naringenin, luteolin and myricetin. The phenolic acids were slightly metabolized by the colonic microflora.     

Mexican lime peel: Comparative study on contents of dietary fibre and associated antioxidant activity

High dietary fibre (DF) powders from Persian and Mexican lime peels were prepared and their dietary fibre composition and antioxidant capacities determined. The total dietary fibre (TDF) contents of both varieties were high; 70.4% and 66.7%, respectively. Both lime peel varieties had an appropriate ratio of soluble/insoluble fractions. The water-holding capacities (WHC) of DF concentrates are high (6.96–12.8 g/g). The WHC was related to the soluble dietary fibre (SDF) which was higher in the DF concentrate of Mexican lime. As part of this analysis, the antioxidant activity (AA) of total extractable polyphenols (TEP) was studied, using three methods: azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) ABTS radical-scavenging activity, α,α-diphenyl–picrylhydrazyl (DPPH) and β-carotene-linoleic acid antioxidant assay. DF concentrates of Persian lime peel had greater polyphenol contents than those of Mexican lime peel. The polyphenols associated with the DF in both lime peel varieties showed a good AA. From a nutritional standpoint, DF lime concentrates may be suitable as food additives.     

Simulated gastrointestinal digestion and in vitro colonic fermentation of date (Phoenix dactylifera L.) seed polyphenols

Date seeds are a by‐product of date fruit industry and a rich source of polyphenols. In this study, in vitro bioaccessibility and colonic fermentation of major polyphenols from date seed powder (DSP) and DSP‐fortified yoghurt (DSPY) were investigated using HPLC. Catechin, epicatechin and procyanidin A2, B1 and B2 were stable during simulated gastric and sequential intestinal digestion. Bioaccessibility was significantly (P < 0.05) higher for all compounds from DSPY compared with DSP. After in vitro colonic fermentation of insoluble digestion materials, most of the target compounds were metabolised by faecal bacteria to ferulic acid, 3‐hydroxyphenylacetic acid, 3‐phenylpropionic acid and 3‐(4‐hydroxyphenyl) propionic acid. DSPY contained significantly (P < 0.05) higher level of free polyphenols as indicated by higher bioaccessibility; however, the stability of the polyphenols and their fermentation products from DSPY were similar to that of DSP alone. These data would be useful in product developments incorporating DSP as a source of polyphenols in food products.     

Effect of a hybrid process,high hydrostatic pressure treatment combined with mixed-strain fermentation,on the quality of the dietary fibre in pickled vegetables

A new hybrid processing technology, mixed-strain (Lactobacillus plantarum and Pediococcus acidilactici; mixed-LAB) fermentation combined with high hydrostatic pressure (HHP) treatment, was used to modify the dietary fibre (DF) in Sichuan pickled vegetables. This process significantly improved the swelling and water holding capacities of the DF, while the oil holding capacities first increased and then decreased sharply with elevated pressure. Physio-functional properties of DF, including heavy metal adsorption, cholate adsorption and α-amylase inhibitory activity, were also enhanced, and antioxidant activity assays revealed an increase in the free radical scavenging ability and the ferric reducing adsorption capacity. The DF structure analysis showed that the hybrid process increased the contents of soluble dietary fibre and monosaccharides of the DF. Moreover, the microstructure is loosened and the surface pore size is expanded. These results indicate that combining mixed-LAB fermentation with HHP treatment is a potential tool that would enable the production of a better quality of DF.     

谷物膳食纤维与结肠健康

膳食纤维(DF)为微生物生长提供均衡的能量和营养,这是维持结肠生态系统平衡所必需的。另外,膳食纤维的发酵,特别是丁酸发酵,有利于结肠健康。目前,结肠癌、炎症性肠炎和其他结肠紊乱疾病已经严重影响身体健康。作者综述了谷物膳食纤维中非淀粉谷物多聚糖、抗性淀粉,通过发酵产物短链脂肪酸和菌群的调节对于结肠健康的影响。     

Proanthocyanidin content in foods is largely underestimated in the literature data: An approach to quantification of the missing proanthocyanidins

The content of proanthocyanidins (PA) in foods is usually determined by HPLC analysis of aqueous-organic extracts. However, the appreciable amounts of PA that remain in the residues of extraction usually are not considered for analysis. Attempts to determine these non-extractable proanthocyanidins (NEPA) by enzymatic-chemical methods have obtained partial results. The general aim of this work was the estimation of the total content of PA, including both extractable PA (EPA) and NEPA in some foods included in the USDA database. A specific procedure based on depolymerization by HCl/BuOH treatment was used to quantify NEPA. The content of NEPA in analyzed samples ranged from 11 mg/100 g for pear to 6.7 g/100 g dry weight for red grape pomace. The results showed that literature data underestimate the actual content of PA in foods. NEPA may have a significant contribution to the healthy effects associated to PA-rich foods and diets.     

Broad bean and pea by‐products as sources of fibre‐rich ingredients: potential antioxidant activity measured in vitro

BACKGROUND By‐products generated during the processing of plant food can be considered a promising source of dietary fibre as a functional compound. The dietary fibre composition, soluble sugars and antioxidant activity of the extractable polyphenols of pea and broad bean by‐products have been analysed in this study. RESULTS: Total dietary fibre using AOAC methods plus hydrolysis (broad bean pod: 337.3 g kg^?1; pea pod: 472.6 g kg^?1) is higher (P < 0.05) in both by‐products than with the Englyst method (broad bean pod: 309.7 g kg^?1; pea pod: 434.6 g kg^?1). The main monomers are uronic acids, glucose, arabinose and galactose in broad bean pods. However, pea pods are very rich in glucose and xylose. The soluble sugars analysed by high‐performance liquid chromatography in both by‐products have glucose as the most important component, followed by sucrose and fructose. The ferric reducing antioxidant power (broad bean pod: 406.4 µmol Trolox equivalents g^?1; pea pod: 25.9 µmol Trolox equivalents g^?1) and scavenging effect on 2,2‐diphenyl‐1‐picrylhydrazyl radical (EC₅₀ of broad bean pod: 0.4 mg mL^?1; EC₅₀ of pea pod: 16.0 mg mL^?1) were also measured. CONCLUSIONS: Broad bean and pea by‐products are very rich in dietary fibre, particularly insoluble dietary fibre and their extractable polyphenols demonstrate antioxidant activity. Therefore they might be regarded as functional ingredients. Copyright © 2011 Society of Chemical Industry     

Effect of chemical treatments of sugar beet fibre on their physico-chemical properties and on their in-vitro fermentation

Chemically treated and dried sugar beet fibres were fermented in vitro in order to study the effects of chemical and physico-chemical parameters of dietary fibre on their colonic fermentation. Sugar beet fibre was treated with dilute alkali, removing mainly acetyl and methyl ester groups, and/or with dilute acids eliminating arabinose, galactose and certain uronic acid residues. The chemical treatments led to an increase in the hydration properties and fermentability by improvement of the accessibility of the remaining polysaccharides. However, if the chemically treated fibres were dried under harsh conditions (100°C), their hydration properties and their fermentability were limited, probably because of structural collapse of the fibre matrix. Whatever the conditions for chemical treatments and drying of the sugar beet fibres, it was possible to predict their fermentability from the water-binding capacity. Because of the relationship between the physiological effects of dietary fibres and the extent to which they are fermented, this result underlines the importance of the physico-chemical characterisation of the fibre in order to acquire a better knowledge of their physiological effects.     

The effect of betaine on in vitro fermentation of carbohydrate and protein combinations under osmotic stress in pigs

BACKGROUND: There is evidence that dietary betaine might improve nutrient digestibilities in pigs due to its osmoprotective properties. This in vitro study was performed to assess the potential of supplemented betaine sources to affect fermentation characteristics of different carbohydrate and protein combinations, under conditions of osmotic stress. In vitro fermentation was performed by use of the modified Hohenheim gas test with pig faeces as microbial inoculum. RESULTS: At the end of in vitro fermentation under osmotic stress conditions, differences (P < 0.05) in gas production, ammonia concentration, and concentration of short‐chain fatty acids were observed for the different carbohydrates, while for the two proteins only differences (P < 0.05) in ammonia and short‐chain fatty acid concentration were obtained. Supplementation of a native betaine source rather than addition of pure betaine increased propionic acid (P < 0.05) and butyric acid (P < 0.05) production. CONCLUSION: These results suggest that only supplementation of a native betaine source but not of pure betaine products affects microbial fermentation under osmotic stress. Furthermore, both the carbohydrate and protein source play an important role in modifying microbial activity under osmotic stress conditions. It can also be concluded that the modified Hohenheim gas test is suitable to scrutinise fermentation activity of different assay substrates under osmotic stress. Copyright © 2012 Society of Chemical Industry     

The effect of drying treatment on the fodder quality and tannin content of two provenances of Calliandra calothyrsus Meissner

The effects of freeze drying and of air drying at 30 °C on the nutritive value of Calliandra calothyrsus leaves were evaluated, using material from two provenances grown together under controlled greenhouse conditions. Freshly harvested leaf of each provenance was compared with the dried treatments in terms of in vitro digestibility, content of extractable, protein‐bound and fibre‐bound proanthocyanidin (condensed tannin) and protein precipitation activity of the extractable fraction, as well as crude protein and fibre content. The effect of long‐term (12 months) storage of fresh‐frozen and dried leaf was also evaluated. Provenance was found to have a major effect on most of the traits, and large differences between the provenances in proanthocyanidin structure were found by high‐performance liquid chromatography, with the tannins of one provenance consisting mainly of catechin/epicatechin subunits (producing cyanidin on treatment with butanol/HCl), and those of the other mainly gallocatechin/epigallocatechin subunits (producing delphinidin with butanol/HCl). This also resulted in large differences in colour yield in the butanol/HCl assay for proanthocyanidins, necessitating separate standards for each provenance. Both drying treatments tended to increase digestibility, though the effect was only significant for one provenance. Air drying, but not freeze drying, increased proanthocyanidin binding to protein, but not to fibre, in both provenances. Long‐term freezing reduced proanthocyanidin content by about half and should be avoided in tannin research. The study suggests that careful air drying, without excessive heating, may not reduce the nutritive value of C calothyrsus, though this needs validation through feeding studies with ruminants. © 2000 Society of Chemical Industry     

Feeding value,in vitro digestibility and in vitro gas production of different by‐products for ruminant nutrition

Nine by‐products used in animal nutrition were examined for their nutritive value by determining the chemical composition (crude protein, crude fat (ether extract), neutral detergent fibre, acid detergent fibre, lignin), in vitro organic matter digestibility and rumen fermentation kinetics (from gas production curves measured in vitro). The by‐products studied were giant pumpkin, red pepper, stem broccoli, brewer's grain, fresh artichoke, scalded artichoke, lemon peel, orange peel and melon. The nutritive value was very variable, depending on the by‐product and on the process applied to the material during industrial processing. In vitro gas production was measured for 500 mg dry matter in quadruplicate at 39 °C after 6, 12, 24, 36, 48 and 72 h of incubation and fitted to single‐pool exponential equations. The fermentation kinetics indicated that brewer's grain was the only by‐product which fitted well with the p = a + b(1 ? e^?ct) equation model; all other by‐products had a very fast degradation rate and their gas production fitted the equation p = b(1 ? e^?c(t?L)), because in the other exponential model a was negative. © 2002 Society of Chemical Industry     

Antioxidant Activities of Polyphenolic Compounds Isolated from Antidesma thwaitesianum Müll. Arg. Seeds and Marcs

ABSTRACT: Antidesma thwaitesianum Müll. Arg. or mao is widely used as commercial products of juice and wine in Thailand. As a result, waste products from the mao plant, such as mao seeds (MS) and mao marcs (MM), are plentiful. We aimed to purify and analyze polyphenolic content in both MS and MM and to investigate the radical scavenging activities of these polyphenolics against 1,1‐diphenyl‐2‐picryl‐hydrazyl (DPPH) and 2,2′‐Azinobis (3‐ethylbenzothiazoline 6‐sulphonate) (ABTS) radicals and thiobarbituric acid reactive products (TBARP). The results showed MS and MM to be an abundant source of polyphenols (97.32 to 130 mg gallic acid equivalents [GAE]/g) and proanthocyanidins. The radical scavenging activities of MS/MM against DPPH and ABTS radicals (IC₅₀ of 0.85 to 1.21 μg/mL) were significantly higher (P < 0.05) than that of standard trolox (IC₅₀ of 5.05 μg/mL). Activity of MS/MM extracts were 3.74 and 3.80 μg/mL trolox eq/g f.w. for the DPPH and ABTS assays, respectively. The oxidation of erythrocyte membranes using 2‐thiobarbituric acid demonstrated that the protective effect of MS/MM on lipid peroxidation is as strong as grape seed proanthocyanidin extract. These findings suggest that polyphenolic compounds and proanthocyanidins isolated from these mao extracts had much higher antioxidant activities than those of standard trolox and exhibited similar antioxidant potential to grape seed proanthocyanidin extract. These findings may also increase value of mao waste products and allow development of commercial health products.     

Prebiotic effects of fermentable carbohydrate polymers may be modulated by faecal bulking of non‐fermentable polysaccharides in the large bowel of rats

This study investigated the impact of supplementary fermentable polysaccharides inulin (a prebiotic) and barley β‐glucan on the large bowel, when consumed against a background high or low intake of mixed non‐starch polysaccharide. Rats were fed for 28 days with diets containing high (15%) and low (5%) background dietary fibre (BDF; cellulose plus sugar beet fibre), with or without 5% supplementary fermentable fibre (SFF; inulin or barley β‐glucan). High‐BDF diets unsupplemented with inulin or barley β‐glucan caused greater improvement than supplemented low‐BDF diets in several parameters linked to gut health: they increased colonic Bifidobacterium spp., butyric acid concentration in the caecum, and colonic crypt depth more than inulin and barley β‐glucan. For these parameters, inulin and barley β‐glucan added little to the effects of the high‐BDF diet alone. Furthermore, by allowing for dilution due to the greater faecal bulk induced by the mixed fibre compared with inulin or barley β‐glucan, the total quantity of butyric acid in the colon of BDF‐fed rats was greatly amplified. Interpretation of the effects of prebiotics in diets containing dietary fibre requires concurrent analysis of the multiple effects of non‐fermentable bulk.     

Evaluation of potential mechanisms by which dietary fibre additions reduce the predicted glycaemic index of fresh pastas

A range of commercially available dietary fibres (DFs), representing both insoluble and soluble forms, were used in the production of pasta. Addition of fibre was at a range of levels (2.5–10%). The potential glycaemic index of these pastas was then evaluated using an in vitro model system to determine starch digestibility and sugar release. Scanning electron microscopy was used to relate structural properties of the pasta to starch degradation. The resulting predicted glycaemic indexes illustrated that the inclusion of DFs into pasta can reduce the glycaemic index of an already low GI food by up to 40%. As such, compared to the predicted glycaemic index value for the control fresh pasta (GI = 45), inclusion of pea fibre, guar gum and locust bean gum yielded pasta with GI values of 39.2, 37.9 and 37 respectively. Thus the type of dietary fibre used was important in the rate of starch digestion observed during the in vitro process. Equally, the level of fibre inclusion was of importance with an inclusion rate of 2.5% generally resulting in pasta with a predictive GI of 42.1, whereas an inclusion of 10% yielded pasta with a GI value of 37.2. Such reductions could be achieved through a number of possible mechanisms including antagonistic and synergistic relationships existing between DF’s and other food components, the effect of DF’s in entrapping pasta particles and thus inhibiting starch degradation, and the restriction of water movement during the cooking of pasta products, thus indicating that starch swelling is impaired.     

Comparative study of colorectal health related compounds in different types of bread: Analysis of bread samples pre and post digestion in a batch fermentation model of the human intestine

Seven different types of wheat and rye bread were analysed for colorectal health related compounds, pre and post digestion, in batch fermentation model of the human intestine. Pre digestion, higher amounts of colorectal health-related dietary fibre compounds (soluble/insoluble/total dietary fibre, arabinoxylans, β-glucans) and phytochemicals (mono-/di-phenolic acids, phytic acid, hydroxymethylfurfural) were detected in wholemeal than in refined flour types of bread, as well as in rye flour types than in wheat flour types of bread. Post digestion, faecal bacterial metabolites of colorectal health promoting (acetate/propionate/butyrate, lactate, free mono-/di-phenolic acids) and impairing (amino metabolites, bile acid metabolites) activities were found in fermentation supernatants of bread samples. All types of bread positively affected faecal bacterial metabolism; among the different types of bread, the highest stimulation of organic acid production (acetate/propionate/butyrate, lactate) and the lowest detrimental bacterial enzyme activities (β-glucuronidase, urease) were detected for wheat flour bread, whereas the strongest retardation of bacterial bile acid degradation and the strongest stimulation of phenolic acid metabolite release (phenylpropionic/phenylpropenoic acid derivatives) were induced by wholemeal rye bread. This study for the first time presents a qualitative and quantitative overview over the broad spectrum of colorectal health related compounds in high- and low-fibre types of bread, pre and post in vitro digestion, and highlights the significance of bread for the preventive nutritional intervention of colorectal cancer.