In vitro gastrointestinal digestion of liquid and semi-liquid dairy matrixes

Protein digestion in two liquid dairy matrixes with different heat treatments (pasteurized and sterilized milks) and in one semi-liquid dairy matrix (stirred-yogurt) was investigated using an in vitro gastrointestinal digestion model. After buccal digestion, significantly lower amount of soluble proteins were measured in yogurt than in both milks. This difference between dairy matrixes decreased during gastric digestion and disappeared at the end of the duodenal digestion upon the proteolytic action of pepsin and pancreatin. Electrophoresis pattern of digested mixtures showed that casein digestion began at the gastric phase and was slower for pasteurized milk than sterilized milk and yogurt. At the end of duodenal digestion, no more intact caseins were present in all the dairy matrixes while faint bands of whey proteins were still visible for pasteurized milk and yogurt. The release of free amino acids during the duodenal phase varied according to their nature (acid, basic, neutral or hydrophobic) and seems to be governed by the specificity of the enzymes. These results suggest that the severity of milk's heat treatment influences the kinetics of protein digestion, mainly during the gastric phase, and that the impact of processing has to be considered to study protein digestion in dairy products.     

Impact of dietary fibers on the properties and proteolytic digestibility of lactoferrin nano-particles

The application and development of biopolymer-based particulate delivery systems is receiving considerable attention from scientists and manufacturers seeking to engineer healthier and safer foods. Present work focused on the fabrication and proteolysis of bovine lactoferrin nano-particles onto which pectins, alginate and carrageenan were electrostatically deposited. DLS, CD and AFM were used to ascertain the formation of spherical structures with varying zeta potentials and modified protein folding. SDS-PAGE analyses of samples collected during simulated human gastro-duodenal digestion revealed that in the case of high and low methoxy pectin particle zeta potential values (−35.4 mV and −50.3 mV respectively) were found to be inversely linked to lactoferrin digestion rate. Moreover, the electrostatic binding of food grade iota-carrageenan onto lactoferrin nano-particles was found to yield highly charged particles (zeta = −69.2 mV) with low surface roughness and to enable a proportion of the lactoferrin to resist 1 h of simulated gastric digestion. Overall, these findings suggest that electrostatic biopolymer interactions may modulate protein digestion and possibly facilitate controlled protein delivery to the upper gastrointestinal tract or have implications to protein nutritional value.     

IgE-binding and in vitro gastrointestinal digestibility of egg allergens in the presence of polysaccharides

This work studies the IgE-binding and in vitro gastrointestinal digestibility of the main egg allergens, ovalbumin (OVA) and ovomucoid (OM), in the presence of pectin (P), gum arabic (G) and xylan (X), functional biopolymers commonly used in the food industry. To this aim, solutions of OVA or OM and P, G or X were digested by using a model that mimics physiological conditions. Gastric and duodenal digests were analysed by SDS-PAGE, RP-HPLC and SEC and the specific human-IgE binding capacity was assessed by immunoblotting and ELISA using sera from egg-sensitized patients. The reactivity towards human IgE of OVA and OM was considerably increased in the presence of the polysaccharides and their susceptibility to digestion was diminished when compared with the isolated proteins. As a result, the duodenal digests obtained in the presence of polysaccharides retained more IgE-binding than the isolated protein digests. Overall, the present results underline the importance of the food matrix in the digestibility of food allergens and in their potential to trigger an immune response.     

Tropical fruit pulps decreased probiotic survival to in vitro gastrointestinal stress in synbiotic soy yoghurt with okara during storage

The effect of mango and guava pulps on Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 viability in a soy yoghurt (SY) and on probiotic survival under simulated gastrointestinal conditions were investigated throughout 28 days of storage at 4 °C. The impact of fruit pulps on SY sensory acceptability was also assessed. Three formulations were produced from soymilk fermented with ABT-4 culture (La-5, Bb-12, and Streptococcus thermophilus) and supplemented with inulin and okara: SYC (control), SYM (with mango pulp), and SYG (with guava pulp). All formulations showed probiotic viabilities ranging from 8 to 9 log cfu/g, and fruit pulps did not affect the probiotic viabilities. However, the fruit pulps decreased probiotic survival significantly to simulated gastrointestinal stress. Acceptability was higher for SYM and this difference was significant at 21 days. Therefore, the improved acceptability of SY through the addition of fruit pulps might lead to a reduction in probiotic functionality.     

In vitro human digestion models for food applications

In vitro digestion models are widely used to study the structural changes, digestibility, and release of food components under simulated gastrointestinal conditions. However, the results of in vitro digestion models are often different to those found using in vivo models because of the difficulties in accurately simulating the highly complex physicochemical and physiological events occurring in animal and human digestive tracts. This paper provides an overview of current trends in the development and utilisation of in vitro digestion models for foods, as well as information that can be used to develop improved digestion models. Our survey of in vitro digestion models found that the most predominant food samples tested were plants, meats, fish, dairy, and emulsion-based foods. The most frequently used biological molecules included in the digestion models were digestive enzymes (pancreatin, pepsin, trypsin, chymotrypsin, peptidase, α-amylase, and lipase), bile salts, and mucin. In all the in vitro digestion models surveyed, the digestion temperature was 37 °C although varying types and concentrations of enzymes were utilised. With regard to digestion times, 2 h (the stomach, small intestine, and large intestine each) was predominantly employed. This survey enhances the understanding of in vitro digestion models and provides indications for the development of improved in vitro digestion models for foods or pharmaceuticals.     

Kiwifruit remnants from digestion in vitro have functional attributes of potential importance to health

The physicochemical properties of plant cell walls in the gut are important in modulating processes that influence health. We investigated the physicochemical properties of kiwifruit cell walls digested under gastric and gastroileal conditions in vitro. Soluble and insoluble undigested polymer fractions were measured, the hydration properties of the digested pulp, and the capacity to retard diffusion and mixing in a simulated small intestinal segment. Undigested polymer (dietary fibre) fractions differed little between “Hayward’ (‘Hayward’) and ‘Hort16A’ (gold) kiwifruit cultivars in their relative proportions, although total dietary fibre was greater in ‘Hayward’ than in the ‘Hort16A’. The polysaccharide composition of seed-free digestion-resistant polymer was similar in both cultivars and not affected by in vitro digestion. Indigestible remnants from kiwifruit had strong water retention and swelling capacities, also little affected by digesting, and retarded both glucose diffusion and mixing significantly, especially in the presence of low background viscosity. We conclude that the particulate cell wall remnants of digested kiwifruit retain substantial potential to influence the properties of gut contents.     

In vitro fermentation of polysaccharide from the seeds of Plantago asiatica L. by human fecal microbiota

In vitro fermentation of the polysaccharide (PLCP, Mw = 1.90 × 10⁶ Da) from seeds of Plantago asiatica L. and the contribution of its carbohydrates to the fermentation was investigated in this study. The polysaccharide was characterized by high contents of xylose, arabinose and glucuronic acid, and it was subjected to human fecal cultures to be fermented in vitro for 24 h. During fermentation, pH in fecal cultures decreased from 6.1 to 5.1 and the levels of total short-chain fatty acid (SCFA), acetic, propionic and n-butyric acids all significantly increased. Xylanase, arabinofuranosidase, xylosidase and glucuronidase activities were also improved. After 24 h incubation, 47.2 ± 1.6% of total carbohydrate in polysaccharide, including 42.9 ± 1.5% of arabinose, 53.2 ± 1.6% of xylose and 76.4 ± 1.2% of glucuronic acid, were consumed. In addition, relationship between carbohydrate consumption of the polysaccharide and SCFA production was also evaluated. It was found that the increase of acetic and n-butyric acid productions mainly resulted from the fermentation of glucuronic acid and xylose in polysaccharide, while the increase of propionic acid production was primarily due to the fermentation of arabinose and xylose. These results showed that the polysaccharide was physiologically active for human large bowel, and its carbohydrate composition determined its SCFA production.     

Predicting relative concentrations of bioavailable iron in foods using in vitro digestion: New developments

In vitro methods have been developed for the prediction of iron bioavailability from foods and supplements. The dialyzability method measures dialyzable iron, released during a simulated gastrointestinal digestion, as an index of iron bioavailability. A new setup, that involves six-well plates and a ring insert that holds the dialysis membrane, is proposed for the application of the dialyzability method with the objective to increase efficiency and to allow testing small-volume samples. A series of solutions (water, ascorbic acid, and phytate), liquid foods (fresh milk and condensed milk), and solid foods (bread + meat meal, corn flakes), were tested in the presence or absence of added iron and digested with the new setup and the setup previously described for the dialyzability method. In both cases, percent dialyzable iron in each treatment remained similar (P > 0.05). These results suggest that the new setup can be employed in future applications with similar food matrices of the dialyzability method.     

Effects of simulated digestion in vitro on cell wall polysaccharides from kiwifruit (Actinidia spp.)

Cell wall polysaccharides are resistant to digestion and absorption in the human small intestine and are considered to be delivered to the colon in a chemically unaltered state. In this paper, pulp from green and gold kiwifruit was subjected to in vitro upper-intestinal tract digestion and the chemical and physical changes to cell wall polysaccharides (dietary fibre) were investigated. Yields of insoluble fibre decreased slightly with simulated digestion while soluble fibre yields increased. Constituent sugar and glycosyl linkage analysis of the soluble and insoluble fibre fractions revealed that the chemical composition and structure of the non-starch polysaccharides remained largely unchanged. However, the degree of methylesterification of galacturonic acid residues present in the pectin-rich soluble fibre fractions of both fruit decreased with treatment; size-exclusion chromatography detected changes in the molecular weight profiles of these fractions. These changes may affect the physicochemical properties and fermentability of kiwifruit dietary fibre in the large intestine.     

Impact of salt and lipid type on in vitro digestion of emulsified lipids

This study examined the effects of oil type and NaCl addition on the micro-structural changes that occur to emulsified lipids as they pass through a model gastrointestinal tract (GIT). Oil-in-water emulsions stabilized by a non-ionic surfactant (Tween 20) were prepared using different kinds of lipids (3% soybean oil, corn oil, olive oil or lard). The emulsified lipids were passed through an in vitro digestion model that simulated the composition (pH, minerals, surface active components, and enzymes) of the human GIT. Prior to digestion, emulsified lipid droplets appear to be bridging flocculation in 1% NaCl added emulsified lipids, moreover lipid droplets of 1% NaCl added emulsified lipids seems to be more disrupted than no NaCl added emulsified lipids. Mean particle size prepared with lard was smaller than those of other emulsified lipids. Free fatty acid contents increased after in vitro digestion in all emulsified lipids. Especially, free fatty acid content of emulsified lipid made from lard and olive oil were significantly higher than those of other emulsions after in vitro digestion.     

Granola bars prepared with Agave tequilana ingredients: Chemical composition and in vitro starch hydrolysis

The stem of Agave tequilana is used to obtain: agave syrup (AS) and native agave fructans (NAF). Ground-agave-fiber is the by-product from fructans production. These ingredients were used to design a food ingredient: agave dietary fiber (ADF), containing NAF (30 g/100 g) as soluble dietary fiber (DF) and ground-agave-fiber (70 g/100 g) as an insoluble DF. The objective of this work was to evaluate the effect of the incorporation of A. tequilana ingredients (AS, NAF, ADF) on the proximate composition, in vitro starch hydrolysis (HI) and predicted glycemic index (pGI) of oat-based granola bars. Total DF (82.03 g/100 g) was the main component in ADF, with 22.8 g/100 g soluble DF. Granola bars were prepared by substituting honey and wheat flour by AS and ADF. A sensory test was used to select the level of sugar substitution by NAF, where 62 g NAF/100 g was the preferred one. The effect of each ingredient on the chemical composition was evaluated using a 2³⁻¹ fractional design. Soluble DF in a granola bar containing a combination of three agave ingredients (AS, NAF and ADF) was 23.35 g/100 g, with HI and pGI values of 74 and 72%, respectively, pointing this product as a moderate GI food.     

Influence of initial emulsifier type on microstructural changes occurring in emulsified lipids during in vitro digestion

The purpose of this study was to examine the impact of emulsifier type on the micro-structural changes that occur to emulsified lipids as they pass through a model gastrointestinal system. Lipid droplets initially coated by different kinds of emulsifiers (lecithin, Tween 20, whey protein isolate and sodium caseinate) were prepared using a high speed blender. The emulsified lipids were then passed through an in vitro digestion model that simulated the composition (pH, minerals, surface active components, and enzymes) of mouth, stomach and small intestine juices. The change in structure and properties of the lipid droplets were monitored by laser scanning confocal fluorescence microscopy, conventional optical microscopy, light scattering, and micro-electrophoresis. In general, there was a decrease in mean droplet diameter (d₃₂) as the droplets moved from mouth to stomach to small intestine. The electrical charge on the droplets stabilized by lecithin, Tween 20 and sodium caseinate were negative throughout the model GI system, while those stabilized by whey protein were positive in the stomach. This suggests that at least some of this globular protein remained attached to the droplet surfaces. The data was interpreted in terms of the competitive adsorption of phospholipids/bile salts with the adsorbed emulsifiers, as well as the enzymatic digestion of proteins and lipids. These results enhance our understanding of the physicochemical and structural changes that may occur to emulsified lipids within the gastrointestinal tract, which may have important consequences for the design of functional foods that alter lipid bioavailability. Nevertheless, there were appreciable differences between the behavior of emulsions within the in vitro model used in this study and literature reports of their behavior within in vivo studies, which highlights the need for more realistic in vitro digestion models.     

Soymilk phenolic compounds,isoflavones and antioxidant activity as affected by in vitro gastrointestinal digestion

The aim of this research was to evaluate changes in the phenolic compounds, isoflavones and antioxidant activity of soymilk following in vitro gastrointestinal digestion (including dialysis). Gastric digestion significantly influenced the release of bioactive substances from the soymilk matrix, increasing the concentration of total phenolic components (35% as the sum of individuals and 14% by Folin–Ciocalteu [F–C] method), total isoflavone content (22%) and total antioxidant activity (76%). The concentration of all those compounds was reduced significantly in the duodenal fraction in comparison to gastric digestion and their lowest concentration was observed in the dialysed fraction, where phenolic acids were not detected. The bioaccessibility of soymilk phenolic compounds was 15% as the sum of individuals and 20% by F–C assay; isoflavones 36% and constituents with antioxidant activity 27%. Results suggest that most of these compounds were sufficiently available to be absorbed and could contribute health benefits.     

Impact of inulin and okara on Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 viability in a fermented soy product and probiotic survival under in vitro simulated gastrointestinal conditions

The effect of inulin and/or okara flour on Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 viability in a fermented soy product (FSP) and on probiotic survival under in vitro simulated gastrointestinal conditions were investigated throughout 28 days of storage at 4 °C. Employing a 2² design, four FSP trials were produced from soymilk fermented with ABT-4 culture (La-5, Bb-12, and Streptococcus thermophilus): FSP (control); FSP-I (with inulin, 3 g/100 mL of soymilk); FSP-O (with okara, 5 g/100 mL); FSP-IO (with inulin + okara, ratio 3:5 g/100 mL). Probiotic viabilities ranged from 8 to 9 log cfu/g during the 28 days of storage, and inulin and/or okara flour did not affect the viability of La-5 and Bb-12. Bb-12 resistance to the artificial gastrointestinal juices was higher than for La-5, since the Bb-12 and La-5 populations decreased approximately 0.6 log cfu/g and 3.8 log cfu/g, respectively, throughout storage period. Even though the protective effect of inulin and/or okara flour on probiotic microorganisms was not significant, when compared to a fresh culture, the FSP matrix improved Bb-12 survival on day 1 of storage and may be considered a good vehicle for Bb-12 and could play an important role in probiotic protection against gastrointestinal juices.     

Phenolic composition and inhibitory effect against oxidative DNA damage of cooked cowpeas as affected by simulated in vitro gastrointestinal digestion

Cowpeas contain phenolic compounds with potential health benefits. The effect of simulated gastrointestinal digestion on phenolic composition of cooked cowpeas and the ability of the digests to inhibit radical-induced DNA damage was determined. A red and a cream-coloured cowpea type were used. The phenolic composition of acetone extracts and enzyme digests of cooked cowpeas was determined using UPLC-MS. Compounds such as p-hydroxybenzoic acid, p-coumaric acid, coumaroylaldaric acid and feruloylaldaric acid were present in the acetone extracts of the cooked cowpeas but were not detected in the enzyme digests. Glycosides of quercetin and myricetin decreased upon in vitro gastrointestinal digestion of cooked cowpeas whereas flavan-3-ols were hardly present except catechin glucoside. The enzyme digest of the red cowpea type was about thrice as effective as that of the cream cowpea type in protecting DNA from oxidative damage. The observation that enzyme digests of cooked cowpeas inhibited radical-induced DNA damage suggests that cowpea phenolics retain some radical scavenging activity after gastrointestinal digestion.     

Stability of polyphenols in chokeberry (Aronia melanocarpa) subjected to in vitro gastric and pancreatic digestion

Berries and red fruits are dietary sources of polyphenols with reported health benefits. As part of the diet, polyphenols are ingested as complex mixtures immersed in a food matrix which is digested in the gut. Epithelial cells lining the gut are regularly exposed to these digested mixtures. To understand the effects of dietary polyphenols on human gut health it is essential to determine their stability and fate in the lumen. In this work, we investigated the effects of an in vitro gastric and pancreatic digestion on the stability and composition of the major polyphenols in chokeberry juice. Digestion was carried out with a mixture of pepsin-HCl for 2 h, followed by a 2 h incubation with pancreatin and bile salts at 37 °C with shaking, in the absence of light and under N₂. After digestion, the chokeberry samples were acidified, filtered and HPLC-DAD/HPLC–MS–MS analysed to determine the content of total soluble recovered phenolics. Gastric digestion had no substantial effect on any of the major phenolic compounds in chokeberry, namely anthocyanins, flavan-3-ols, flavonols and caffeic acid derivatives. However, these compounds were significantly altered during the pancreatic digestion and this effect was more marked for anthocyanins (approximately 43% was lost during the 2 h treatment with pancreatin). Flavonols and flavan-3-ols decreased by 26% and 19%, respectively. Neochlorogenic acid decreased by 28% whereas chlorogenic acid was increased by 24%. In vitro digestion of standard phenolic compounds, representing each of the groups of phenolics in chokeberry, confirmed some of the observed changes. Interactions with the digestive enzymes were not responsible for the observed losses which were mostly due to the chemical conditions during pancreatic digestion. Our results, in accordance with previously published results, show that dietary polyphenols are highly sensitive to the mild alkaline conditions in the small intestine and that a good proportion of these compounds can be transformed into other unknown and/or undetected structural forms with different chemical properties and, consequently, different bioaccessibility, bioavailability and biological activity.     

Comparison of in vitro digestion characteristics and antioxidant activity of hot- and cold-pressed peanut meals

Due to the poor protein solubility, hot-pressed peanut meal (HPM) has less value than cold-pressed peanut meal (CPM) in the food industry. The objective of this study was to determine whether the denatured proteins in HPM were suitable for hydrolysis by digestive enzymes. The hydrolysis characteristics and antioxidant activity of HPM and CPM during in vitro digestion were compared. The results showed that HPM was hydrolysed more extensively than CPM. There were more free amino acids and small peptides with MW < 5 kDa in HPM hydrolysates. In addition, HPM hydrolysates displayed stronger 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,20-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical-scavenging activities, ORAC value and reducing power than CPM hydrolysates, which could be due to the higher content of small peptides, antioxidative amino acids and melanoidins in HPM hydrolysates. The above results indicated that HPM was a potential source of protein supplement for human consumption.     

Effect of food matrix and processing on release of almond protein during simulated digestion

The aims of the present work were to assess digestibility of almond protein in the upper gastrointestinal tract, evaluate the effects of food matrix on protein release and assess the persistence of immunoreactive polypeptides generated during simulated digestion. Prunin, the most abundant protein in almond flour, was sensitive to pepsin, with complete digestion after 20 min in the gastric phase. Addition of the surfactant phosphatidylcholine did not affect the rate and kinetic of digestion, as observed by SDS-PAGE analysis and HPLC, in the stomach and the small intestine of either natural or blanched almond flour. However, incorporation of almond flour into a food matrix, such as chocolate mousse and Victorian sponge cake, decreased the rate of almond protein degradation by pepsin and immunoreactivity of almond polypeptides detected by dot blots and sandwich ELISA retained better. Most of the almond protein identified by in-gel tryptic digestion and MALDI-TOF analysis corresponded to prunin, with pI values of 5–7.     

负载姜黄素/花色苷的木薯淀粉基W1/O/W2型Pickering双重乳液的制备及性质

以木薯纳米淀粉、木薯醋酸酯变性淀粉为颗粒乳化剂制备食品级W₁/O/W₂型Pickering双重乳液,分别于内层水相、油相对姜黄素与花色苷进行负载。考察不同木薯醋酸酯变性淀粉质量分数对双重乳液显微形态、粒径、Zeta电位、负载率及贮藏稳定性的影响,并分析其在体外消化过程的控释性能。结果表明,木薯醋酸酯变性淀粉质量分数为6%时,乳液综合性能最好。此时,乳滴分布均匀,呈“三相两膜”结构,粒径为3.65μm,Zeta电位为-14.50mV,姜黄素、花色苷的负载率达到95.23%、93.00%,4、25、40℃时贮藏20d均未出现明显分层;经模拟消化后乳液中姜黄素、花色苷保留率为41.59%、76.29%,与游离姜黄素、花色苷相比,二者生物利用率分别提高了约4倍、3倍。证实了木薯淀粉基双重乳液能实现在模拟消化系统中对活性物质的保护,并能有效提升其生物利用率。研究结果旨在为食品工业中淀粉基功能性乳液运载体系的构建及新型食品级Pickering双重乳液的开发提供理论参考。