Total phenolics,flavonoids, anthocyanins and antioxidant activity following simulated gastro-intestinal digestion and dialysis of apple varieties: Bioaccessibility and potential uptake

In the present study, an in vitro model simulating gastrointestinal (GI) digestion, including dialysability, was adapted to assess free soluble polyphenols from apples (four varieties). Results indicated that polyphenol release was mainly achieved during the gastric phase (ca. 65% of phenolics and flavonoids), with a slight further release (<10%) during intestinal digestion. Anthocyanins present after the gastric phase (1.04–1.14 mg/100 g) were not detectable following intestinal digestion. Dialysis experiments employing a semipermeable cellulose membrane, presenting a simplified model of the epithelial barrier, showed that free soluble dialysable polyphenols and flavonoids were 55% and 44% of native concentrations, respectively, being approximately 20% and 30% lower than that of the GI digesta. Similar results were found for the antioxidant capacity of dialysable antioxidants, being 57% and 46% lower compared to total antioxidants in fresh apples (FRAP and ABTS test, respectively). It is suggested that some polyphenols are bound to macromolecular compounds that are non-dialysable, that the presented method allowed the study of free soluble polyphenols available for further uptake, and that both chemical extraction and concentrations in final digesta would overestimate polyphenol availability.     

In vitro digestion of dairy and egg products enriched with grape extracts: Effect of the food matrix on polyphenol bioaccessibility and antioxidant activity

The aim of this study was to evaluate the effect of the food matrix on polyphenol bioaccessibility and antioxidant activity during the in-vitro digestion of dairy and egg products enriched with grape extracts (GE). Four GE-enriched matrices produced under industrial conditions (custard dessert, milkshake, pancake and omelet) and the GE dissolved in water (control solution) were submitted to in vitro digestion and the bioaccessibility of the major classes of polyphenols as well as the evolution of the antioxidant activity of the matrices were monitored at oral, gastric and intestinal level. Apart from the digestion effect itself, the release, stability and solubility of polyphenols was governed by mainly two factors: 1 — the composition and structure of the food matrices and 2 — the class of polyphenol. Results showed that the inclusion of the GE extracts into the different egg and dairy food matrices greatly impacted the release and solubility of anthocyanins and proanthocyanidins during digestion, especially in the solid food matrices and during the oral and gastric phases of digestion. Also, the presence of the food matrices protected anthocyanins from degradation during the intestinal phase. However, if the total phenolic content is considered at the end of the whole digestion process, the proportion of soluble (bioaccessible) and insoluble phenolics delivered by the enriched-matrices was quite similar to that of the control solution. On the contrary, the food matrix effect did not affect the antioxidant activity of the matrices, which remained constant during the oral and gastric phases but greatly increased during the intestinal phase of digestion. Among the GE-enriched matrices, omelet presented higher recoveries of total phenolics and antioxidant activity at the end of digestion.     

Intake and bioaccessibility of total polyphenols in a whole diet

The knowledge of dietary intake of polyphenols and their bioaccessibility in the human gut are key factors in assessing their significance in human health. The aim of this work was to estimate the amount of total polyphenols consumed in a whole diet (Spanish Mediterranean diet) and their intestinal bioaccessibility. Total polyphenols were determined, as the sum of the polyphenols present in methanol:acetone:water extracts (extractable polyphenols) of plant foods and condensed tannins and hydrolysable polyphenols (non-extractable polyphenols) in the corresponding residues. The polyphenols intestinal bioaccessibility was estimated by an in vitro gastrointestinal model where food polyphenols are released by enzyme digestion and colonic fermentation. The mean daily intake of polyphenols in the Spanish diet was estimated between 2590 and 3016 mg/person/day. The amount of non-extractable polyphenols was almost double that of extractable polyphenols. It was estimated that about 48% of dietary polyphenols are bioaccessible in the small intestine, while 42% become bioaccessible in the large intestine.     

Bioactive dietary polyphenols inhibit heme iron absorption in a dose-dependent manner in human intestinal Caco-2 cells

Abstract: Although heme iron is an important form of dietary iron, its intestinal absorption mechanism remains elusive. Our previous study revealed that (‐)‐epigallocatechin‐3‐gallate (EGCG) and grape seed extract (GSE) markedly inhibited intestinal heme iron absorption by reducing the basolateral iron export in Caco‐2 cells. The aim of this study was to examine whether small amounts of EGCG, GSE, and green tea extract (GT) could inhibit heme iron absorption, and to test whether the inhibitory action of polyphenols could be offset by ascorbic acid. A heme‐⁵⁵Fe absorption study was conducted by adding various concentrations of EGCG, GSE, and GT to Caco‐2 cells in the absence and presence of ascorbic acid. Polyphenolic compounds significantly inhibited heme‐⁵⁵Fe absorption in a dose‐dependent manner. The addition of ascorbic acid did not modulate the inhibitory effect of dietary polyphenols on heme iron absorption when the cells were treated with polyphenols at a concentration of 46 mg/L. However, ascorbic acid was able to offset or reverse the inhibitory effects of polyphenolic compounds when lower concentrations of polyphenols were added (≤ 4.6 mg/L). Ascorbic acid modulated the heme iron absorption without changing the apical heme uptake, the expression of the proteins involved in heme metabolism and basolateral iron transport, and heme oxygenase activity, indicating that ascorbic acid may enhance heme iron absorption by modulating the intracellular distribution of ⁵⁵Fe. These results imply that the regular consumption of dietary ascorbic acid can easily counteract the inhibitory effects of low concentrations of dietary polyphenols on heme iron absorption but cannot counteract the inhibitory actions of high concentrations of polyphenols. Practical Application: Bioactive dietary polyphenols inhibit heme iron absorption in a dose‐dependent manner. The small amounts of polyphenolic compounds present in foods are capable of reducing heme iron transport across the intestinal enterocyte. However, the inhibitory effects of dietary polyphenolic compounds on heme iron absorption can be offset by ascorbic acid and can possibly be avoided by decreasing the consumption of polyphenols while simultaneously taking ascorbic acid.     

多酚化合物体外模拟消化的稳定性分析

选取7 种不同种类的酚类化合物,研究体外模拟胃肠道消化对其稳定性的影响。结果表明：不同酚类化合物在胃肠道消化过程中的稳定性不同。高效液相色谱分析结果显示,胃消化过程对酚酸类化合物的影响较小,其含量无显著变化,但类黄酮化合物在胃消化过程中稳定性较差,儿茶素含量下降了16.6%、表儿茶素含量下降了约6%。经模拟肠液处理后,除阿魏酸、对羟基苯甲酸外,其他酚类化合物含量均显著降低,此外,绿原酸在肠液消化过程中生成了新的物质。体外模拟胃肠液消化对混合标准品的作用结果有所不同,模拟胃液消化对混合标准品含量均无显著影响,模拟肠液消化后,仅表儿茶素、没食子酸含量分别降低为76.96%和50.30%,其他酚类化合物含量均无显著变化。     

Polyphenolic profile and antioxidant activity of five apple cultivars grown under organic and conventional agricultural practices

The polyphenols and total antioxidant activities of five apple cultivars, grown by organic and conventional agricultural methods in neighbouring farms, were determined and compared. Total polyphenols in the whole fruit and in the peel were determined by the Folin-Ciocalteu method, and the total antioxidant activity was determined by three established methods, diphenyl picrylhydrazyl (DPPH), azinobis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and ferric reducing/antioxidant power (FRAP). Polyphenolic content for the whole fruit was in the range of 80–196 and for the peel 165–400 (mg Gallic Acid Equivalent (GAE)/100 g fresh weight) for both types of agricultural practices. Antioxidant activities of fruit extracts were also relatively similar and well correlated to their polyphenolic content. High-performance liquid chromatography (HPLC) analysis of the most important polyphenolics (chlorogenic acid, catechin, epicatechin, procyanidin B1 and B2, cyaniding 3-galactoside, phloridzin, quercetin 3-galactoside and quercetin 3-arabinoside) also showed that concentrations do not differentiate significantly between the organic and conventional apples. Statistical significance of differences in antioxidant activities among the same cultivars was relatively small (flesh + peel or peel only) for both types of apples. These results indicate that organic apples do not present higher antioxidant or nutritional value compared with conventionally grown ones, as far as polyphenolic content and total antioxidant activities are concerned.     

Bioaccessibility and antioxidant activity of phenolic compounds in cooked pulses

The aim of this study was to evaluate the bioaccessibility of polyphenols and antioxidant activity in cooked pulses and to study the effect of cooking on their total phenolic content (TPC) and antioxidant capacity. Cooked faba beans showed the highest TPC, followed by soybeans and lentils or peas. TPC ranged from 10.4 ± 0.2 to 52.9 ± 0.3 mg/100 g and was positively correlated with antioxidant activity. Cooking resulted in increased TPC and antioxidant activity of the methanolic extracts, caused by cell disruption and improved extraction of polyphenols. Although polyphenols were lost in the cooking water, boiled legumes had more polyphenols than those resulting cooking broths. In vitro gastrointestinal digestion resulted in increased TPC and antioxidant capacity of the extracts. Soybeans showed the highest amount of bioaccessible polyphenols. The release of phenolics from cooked legumes was mainly achieved during the intestinal phase. Literature data may underestimate the TPC and antioxidant capacity of pulses.     

Effect of in vitro‐simulated gastrointestinal digestion on the stability and antioxidant activity of blueberry polyphenols and their cellular antioxidant activity towards HepG2 cells

In this study we investigated the influence of in vitro‐simulated gastrointestinal digestion on the bioaccessibility and antioxidant activity of polyphenols from blueberries (Vaccinium spp.). Total phenolic, anthocyanin, and flavonoid content was determined, and extract and digesta compositions were analysed by HPLC‐ESI‐MS/MS. The phenolic compounds were relatively stable under a gastric environment, whereas polyphenols and anthocyanins were unstable under an intestinal environment. The bioaccessibility of polyphenol, anthocyanin, and flavonoid was greatly decreased after the intestinal digestion, and the recoveries were only 13.93%, 1.95%, and 15.68% (the IN sample), respectively. Polyphenolic profile alteration occurred during in vitro‐simulated gastrointestinal digestion. Changes of phenolic compound antioxidant activity during digestion correlated with polyphenol, flavonoid, and caffeic acid concentrations. Digested extract cellular antioxidant activity was lower than non‐digested extract activity (P < 0.05). Polyphenol dose–response correlations with cellular antioxidant activity were observed. These results indicated that in vitro‐simulated gastrointestinal digestion significantly impact polyphenols and their antioxidant activity.     

体外模拟胃肠消化中山楂多酚及抗氧化活性的变化

本实验研究了山楂多酚、黄酮及抗氧化活性在体外模拟胃肠道环境下的变化规律,并利用高效液相色谱法测定了胃肠消化对山楂中5 种主要单体酚类化合物的影响。结果表明：山楂多酚在模拟胃液消化中稳定性较好,总多酚、黄酮含量及抗氧化能力均无显著性变化（P＞0.05）;而在模拟肠液消化中山楂多酚易发生降解,总多酚、黄酮含量在0.5 h内分别降低16.26%和7.46%,抗氧化能力也随之降低10.25%。高效液相色谱测定结果显示,5 种单体酚类化合物含量在胃液消化中无显著性变化（P＞0.05）;而在肠液消化中变化情况不相一致,其中金丝桃苷和异槲皮苷含量无显著性变化（P＞0.05）,表儿茶素在0.5 h内完全降解,绿原酸和原花青素B2在0.5 h内分别显著下降40.78%和28.99%（P＜0.05）。     

Polyphenol and nutrient release from skin of almonds during simulated human digestion

The bioaccessibility of nutrients and phytochemicals from almond skin has not been previously evaluated. We quantified the release of lipid, protein and polyphenols during simulated human digestion from natural (NS) and blanched (BS) skins, the latter being a by-product of the almond industry. Higher percentages of polyphenols were released from NS compared to BS during in vitro digestion. Most of the limited release of lipid and protein occurred during gastric digestion, with no significant differences between NS and BS. The total dietary fibre content was 45% for NS and 46% for BS, glucose and galacturonic acid being the major sugars present. No changes in dietary fibre composition and distribution of autofluorescent phenolics were observed in the cell walls of almond skin after simulated digestion. In the GI tract, the cell walls may therefore function as a useful source of fermentable fibre with beneficial implications for gut health.     

柑橘在体外模拟胃肠消化过程中总多酚、总黄酮及总抗氧化活性的变化规律

通过体外模拟胃肠消化法模拟柑橘在胃肠道的消化过程,测定模拟胃肠消化过程中总多酚、总黄酮的释放量及总抗氧化活性的变化规律。结果显示,柑橘多酚主要是游离酚,有机化学提取法释放的多酚高于模拟胃肠消化法。模拟胃消化3 h后,与消化0 h相比,宽皮柑、脐橙和金橘的多酚最大释放量是其1.02、0.93、1.21 倍,黄酮最大释放量是其1.46、2.37、2.16 倍,最大抗氧化活性是其1.10、1.23、1.62 倍。体外模拟肠消化4 h后,与胃消化0 h和肠消化0 h相比,宽皮柑、脐橙和金橘的多酚释放量分别为其0.99、0.80、1.29 倍和91%、81%、90%,黄酮的最大释放量分别为其73%、39%、64%和51%、17%、39%,最大抗氧化活性分别为其1.67、1.09、1.76 倍和1.31、0.88、1.22 倍。这表明柑橘在模拟胃肠消化过程中,胃蛋白酶、胰酶、胆汁及胃酸均可以促进抗氧化活性物质释放。     

Kinetics of Polyphenol Losses During Soaking and Drying of Cider Apples

A regular consumption of fresh or processed fruits can contribute to preventing cardiovascular diseases and cancer. Polyphenols, which are naturally present in fresh apples (Malus x domestica), are widely acknowledged as antioxidants. Cider apples contain high concentrations of polyphenols, but their composition differs markedly as a function of cultivar. Two of which were studied in this respect. The first cultivar was Marie Menard, which is highly concentrated in polyphenols and has a standard profile showing high levels of hydroxycinnamic acids and monomeric catechins, widely involved in oxidation reactions catalyzed by the polyphenol oxidase; the second one was Guillevic, which has an atypical phenolic profile for a cider apple, with highly polymerized procyanidins and no monomeric catechins, which thus induces a low sensitivity to enzymatic browning. Both cultivars were subjected to osmotic dehydration at two temperatures (45 and 60 °C) or to convective air-drying, and a combination of the two processes was also tested. Phenolic compounds were quantified by high performance liquid chromatography for various processing times and regardless of the quantity of impregnated sugar. The results revealed different behaviors depending on the polyphenolic groups of compounds. Procyanidins were better preserved by the processes than hydroxycinnamic acids or monomeric catechins. Indeed, these latter groups of polyphenols were first of all involved in enzymatic browning but could also diffuse more easily as their molecular weight was lower. Polyphenol retention was also dependent on the process applied. Polyphenols were better retained by convective drying than by osmotic dehydration, and when soaking was applied as a pretreatment, polyphenol losses were limited during subsequent drying. Moreover, the level of sucrose impregnation could mask the astringency of procyanidins.     

Polyphenols bioaccessibility and bioavailability assessment in ipecac infusion using a combined assay of simulated in vitro digestion and Caco-2 cell model

In this report, we investigated for the first time the total polyphenols content (TPC) and antioxidant activity before and after digestion of Carapichea ipecacuanha root infusion, better known as ipecac, prepared at different concentrations. An in vitro digestion system coupled to a Caco-2 cell model was applied to study the bioavailability of antioxidant compounds. The ability of ipecac bioaccessible fractions to inhibit reactive oxygen species (ROS) generation at cellular level was also evaluated. The findings revealed that water volume of 50 mL g⁻¹ of sample provided the maximum yield of extraction of TPC and antioxidant activity. Polyphenols increased in content and activity after digestion and they were highly bioavailable (75% of intestinal absorption). Polyphenols were also present in the residual parts which indicate a possible local activity. Results also suggest that ipecac infusion could represent a promising source of effective bioavailable antioxidants to be exploited in functional foods field.     

柑橘模拟胃肠消化后的多酚生物有效性和抗HepG2细胞增殖活性

本文通过模拟胃肠消化法评价柑橘类水果(宽皮柑、脐橙和金桔)的多酚生物有效性和抗肿瘤细胞增殖的活性。柑橘水果经过模拟胃肠消化后,采用高效液相色谱法测定主要多酚单体的释放量,亚甲基蓝法测定柑橘水果提取物抗HepG2细胞增殖的活性。结果表明,释放量较大(20mg/kg FW)的柑橘多酚是新橙皮甙、根皮苷、柚皮素、香草酸和原儿茶酸等。模拟胃消化过程可以促进根皮苷、香草酸和原儿茶酸的释放,而所检测的大部分多酚在肠消化过程显著降解(p0.05)。宽皮柑、脐橙和金桔模拟胃肠消化后其产生细胞毒性的浓度显著降低(p0.05),模拟胃消化及模拟肠消化后的提取物(浓度分别为84.56、93.74、32.26mg/mL和121.23、107.14、40.03mg/mL)作用于HepG2细胞72h后,细胞的存活率分别为65.46%、37.20%、24.19%和21.32%、43.52%、12.31%,表明模拟胃肠消化过程可以促进抗HepG2细胞增殖的活性物质释放。     

Tannase improves gallic acid bioaccessibility and maintains the quality of mango juice

The effect of tannase on gallic acid (GA) bioaccessibility and auto-oxidative browning of mango juice was investigated. After 2 h of simulated gastric digestion, the concentration of bioaccessible GA increased (P < 0.05) 94.3 ± 7.0% in juice treated with 0.5 U mL⁻¹ tannase while juice not treated with tannase had only a 6.3 ± 3.4% increase in GA. During 2–10 h of simulated intestinal digestion, tannase treated juice continued to have a higher concentration (P < 0.05) of bioaccessible GA in comparison to juice that was not treated with tannase. The use of 167 U 100 mL⁻¹ tannase while processing mango juice did not result in any differences (P < 0.05) in browning measured at 420 nm, yet there was significantly higher (P < 0.05) GA in mango juice post-storage. Processing mango juice with tannase can help improve the bioaccessibility of mango polyphenols without hindering the quality of juice during storage.     

In vitro digestion of protein-enriched restructured beef steaks with pea protein isolate,rice protein and lentil flour following sous vide processing

The effect of plant protein inclusion in cooked meat upon in vitro gastro-intestinal (GI) digestion was investigated. Pea protein isolate, rice protein and lentil flour were used to increase the protein content in a meat model system restructured using two transglutaminase enzymes [Activa®EB (TG) and Transgluseen™-M (TS)]. Restructured beef steaks were subjected to simulated GI digestion using the static INFOGEST method. Samples taken at different digestion times were analysed using SDS-PAGE, size exclusion-HPLC, free amino acid analysis and microscopy. SDS-PAGE analysis revealed significant protein hydrolysis during GI digestion. Most soluble peptides had a molecular weight smaller than 500 Da, corresponding to peptides of <5 amino acids, regardless of food treatment. The amounts of released, free amino acids isoleucine, lysine, phenylalanine and valine were higher (P < 0.05) in lentil-enriched restructured beef steaks following GI digestion. Confocal laser scanning microscopy (CSLM) revealed pronounced aggregation in digested samples. In vitro digestates of protein-enriched restructured beef steaks showed lower production of small molecular weight peptides. This study demonstrated how the bioaccessibility of protein-enriched restructured beef steaks are influenced by formulation and processing.     

茶多酚在模拟胃肠消化过程中含量及活性的变化规律

通过模拟消化模型分析茶多酚在胃肠消化过程中成分和抗氧化活性的变化规律,及其对肠道细菌生长的影响。结果表明,与胃部相比,茶多酚在肠部降解能力更为显著;经过模拟胃消化,茶多酚含量降低了12.00%,GCG显著降低了32.50%,EGC增加了13.80%;模拟肠消化后,茶多酚含量大幅度地降低了35.90%～46.10%,EGCG显著降低了98.40%～98.70%,GC则显著增加了97.00%～114.00%;模拟胃肠消化后,茶多酚的总抗氧化能力、DPPH·和ABTS~+·的清除能力均大幅度降低(p0.05),除肠消化阶段清除ABTS~+·的能力外,其余消化阶段的抗氧化能力均与茶多酚含量呈正相关;此外,研究模拟消化前后茶多酚对肠道细菌生长的影响,发现消化前其具有显著抑制大肠杆菌和增殖嗜酸乳杆菌生长的作用,模拟胃肠消化后,茶多酚对大肠杆菌抑菌效果增强,但对嗜酸乳杆菌无显著作用。     

苹果模拟胃肠消化后的多酚释放和抗HepG2细胞增殖活性研究

分别采用80%丙酮提取以及模拟胃肠消化法提取苹果中的多酚,测定提取物的多酚释放量以及抗HepG2细胞增殖的活性。结果显示,在水晶富士、嘎啦果和青苹果的80%丙酮提取物中,绿原酸的释放量最高,分别为30.72±1.54、22.37±1.12、8.70±0.44 mg/kg FW(Fresh weight,FW)。模拟胃消化后释放量较高的为原儿茶酸、对羟基苯甲酸、没食子酸和绿原酸等酚酸;模拟肠消化后释放量较高的为槲皮苷、槲皮素-3-葡萄糖苷、扁蓄苷等黄酮醇。水晶富士、嘎啦果和青苹果的模拟胃消化及模拟肠消化后的提取物(浓度分别为297、99、255 mg/mL和198、154、106 mg/mL)作用于HepG2细胞72 h后,细胞的存活率分别为26%、42%、44%和23%、28%、28%。这表明,模拟胃消化主要促进酚酸的释放,模拟肠消化主要促进黄酮醇的释放;在一定浓度范围内,苹果模拟胃肠消化提取物对HepG2细胞有较强的抗增殖活性。     

体外模拟胃肠道消化下刺梨抗氧化成分的释放

通过体外模拟胃肠消化来研究贵州刺梨多酚、黄酮及抗氧化活性的变化规律,探讨其对刺梨抗氧化成分及其活性的影响。对刺梨进行体外模拟胃消化和体外模拟肠消化,分别使用Na NO2-Al(NO3)3比色法、福林-酚比色法以及DPPH自由基清除率与普鲁士蓝法测还原力来测定贵州刺梨在体外模拟胃肠消化中多酚、黄酮含量和抗氧化活性的变化。实验结果显示,相较于空白实验,贵州刺梨在模拟胃消化中多酚、黄酮含量、DPPH自由基清除率以及还原力数值都有明显上升的趋势,模拟胃消化组0.5h内有明显上升的趋,最大分别增加1.48、1.35、1.14、1.48倍,之后趋于平稳。胃酸对照组最大分别增加1.28、1.21、1.07、1.41倍,之后趋于平稳,模拟肠消化组1h内有明显上升的趋势,最大分别增加1.07、1.65、1.28、1.07倍,之后趋于平稳。研究结果表明胃蛋白酶、胃酸、胆盐以及胰酶能促进贵州刺梨中抗氧化成分的释放及其活性的增加。     

In vitro bioaccessibility of peptides and amino acids from yogurt made with starch,pectin, or β-glucan

The bioaccessibility of peptides and amino acids (AA) during in vitro gastrointestinal digestion of proteins from acid dairy gel (or yogurt) containing different polysaccharides (PS) was investigated. The nature of added PS was shown to modulate the kinetics of proteins gastric digestion. Yogurts with β-glucan and with pectin showed faster proteolysis, lower release of large peptides, and a higher proportion of free AA than those with starch or without PS. Segregation between β-glucan and dairy proteins was hypothesized to explain faster proteolysis by concentration of enzymes and proteins in a separated phase considered as “micro-reactors”. Kinetics of protein digestion during the buccal phase was not affected by the presence of PS in yogurts. At the beginning of duodenal phase yogurts with starch and with pectin showed a higher degree of hydrolysis; at the end of this phase proteins were fully digested (no native protein left and bioaccessible AA was increased).