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.     

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.     

Antiacetylcholinesterase and antioxidant activities of Plectranthus barbatus tea,after in vitro gastrointestinal metabolism

This study describes the in vitro metabolism, by the gastrointestinal tract, together with the biological activity of the herbal tea obtained from Plectranthus barbatus leaves. The results showed that there was no appreciable degradation and that the activity was kept constant after gastric juice digestion. In the presence of pancreatic juice, degradations varied from negligible, in the case of flavonoid glucuronides, to almost complete, in the case of the abietane diterpenoid. The digested decoction contained only 50% of its initial biological activity, after pancreatic digestion. The action of Caco-2 cells on the extract revealed that neither rosmarinic acid, the main compound of the extract, nor the other components present in minute quantities were metabolised by the intact cells. Rosmarinic acid could be found inside Caco-2 cells although in trace amount. Glucoronidase from Escherichia coli, a gut bacterium, was able to hydrolyse the flavonoid derivatives, thus the aglycones were formed and permeate the Caco-2 cells.     

The in vitro and in vivo antioxidant properties of seabuckthorn (Hippophae rhamnoides L.) seed oil

The antioxidant capacity of seabuckthorn (Hippophae rhamnoides L.) seed oil was investigated with a number of established in vitro assays and in an in vivo study of carbon tetrachloride (CCl₄)-induced oxidative stress in mice. The results showed that DPPH radical scavenging activity, ferrous ion chelating activity, reducing power and inhibition of lipid peroxidation activity all increased with increasing concentrations of seabuckthorn seed oil. Moreover, the EC₅₀ values of seabuckthorn seed oil from the hydrogen peroxide, superoxide radical, hydroxyl radical scavenging assays were 2.63, 2.16 and 0.77 mg/ml, respectively. In the in vivo study, seabuckthorn seed oil inhibited the toxicity of CCl₄, as seen from the significantly increased activities of the antioxidant enzymes superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase. The GSH content in the liver was also increased, whereas hepatic malondialdehyde was reduced. Taken together, these results clearly indicate that seabuckthorn seed oil has significant potential as a natural antioxidant agent.     

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.     

Neuroprotective effect of blackberry (Rubus sp.) polyphenols is potentiated after simulated gastrointestinal digestion

Blackberry ingestion has been demonstrated to attenuate brain degenerative processes in rodents with the benefits ascribed to the (poly)phenolic components. The aim of this work was to assess the efficacy of blackberry polyphenolics in a neurodegeneration cell model before and after simulated gastrointestinal digestion.Digested blackberry metabolites protected neuroblastoma cells from H₂O₂-induced death at low, non-toxic levels that approach physiologically-relevant serum concentrations. However, the original extracts were not protective even at fivefold higher concentrations. This potentiation may reflect alterations in the polyphenolic composition caused by the digestion procedure, as detected by liquid-chromatography-mass spectrometric analysis. This protection was not caused by modulation of the intracellular antioxidant capacity or through alteration of glutathione levels, although the original extract influenced both of these parameters. This work reinforces the importance of evaluating digested metabolites in disease cell models and highlights the possible involvement of other mechanisms beyond antioxidant systems.     

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.     

Changes in antioxidant capacity,isoflavone profile,phenolic and vitamin contents in soymilk during extended fermentation

In this study, the influence of lactic acid bacteria fermentation on the antioxidant capacity, phenolic compounds and vitamins profile of soymilk with different final fermentation pH values (pH 4.55, 4.15 and 3.85) was examined. Fermented soymilk (FSM) with a final pH of 3.85 exhibited relatively higher antioxidant capacity than samples terminated at pH 4.55 or 4.15, as shown from results of Folin–Ciocalteu assay (57.3–63.8 mg GAE/100 mL FSM), DPPH-radical scavenging efficiency (30.0–36.5%) and ferricyanide reducing power (1.13–1.58 mg AAE/100 mL FSM). Among all products, Lactobacillus rhamnosus WQ2-fermented soymilk showed the highest antioxidant level among the four Lactobacillus species, while Lactobacillus acidophilus CSCC 2400 possessed the best ability to deglycosylate isoflavone glucoside (IG), reducing IG concentration by 60.8%. Four vitamins, three phenolic acids, one flavanol, and six isoflavones were detected in fermented and non-fermented soymilk by HPLC-DAD. The contents of antioxidant compounds in FSM were closely related to reducing power and anti-radical ability. In addition, antioxidant capacity was highly correlated with proteolytic activity. This study also demonstrated the potential of extended fermentation to enhance the overall health-promoting property of the products.     

Profiling of in vitro neurobiological effects and phenolic acids of selected endemic Salvia species

The ethyl acetate and methanol extracts from 16 Salvia L. species were screened for their inhibitory activity against acetylcholinesterase, butyrylcholinesterase, lipoxygenase, and tyrosinase; the enzymes linked to neurodegeneration. Their antioxidant activity was also tested using DPPH radical scavenging, metal-chelation, and ferric-reducing antioxidant power (FRAP) assays. Total flavonoid content of the extracts was determined by AlCl₃ reagent, while HPLC technique was applied for analysis of various phenolic acids in the extracts. The extracts exerted weak cholinesterase and tyrosinase inhibition, and remarkable inhibition against lipoxygenase (13.07 ± 2.73-74.21 ± 5.61%) at 100 μg ml⁻¹. The methanol extracts showed higher antioxidant activity in DPPH radical scavenging and FRAP assays. The extracts were analyzed for their gallic, protocateuchic, p-hydroxy-benzoic, vanillic, caffeic, chlorogenic, syringic, o- and p-coumaric, ferulic, rosmarinic, and tr-cinnamic acid contents and the methanol extract of Salvia ekimiana (153.50 mg 100 g⁻¹) was revealed to be the richest in terms of rosmarinic acid.     

Phenolic profiles of in vivo and in vitro grown Coriandrum sativum L.

Coriandrum sativum L. is a source of a variety of polyphenols and other phytochemicals, related to its high antioxidant activity and to its use for indigestion, rheumatism, and prevention of lipid peroxidation damage. Plant cell cultures are a means to study or to produce some active metabolites, such as polyphenols. This technique was applied to the investigation of coriander, and a detailed analysis of individual polyphenols in vivo and in vitro grown samples was performed. The in vivo vegetative parts showed quercetin derivatives as the main flavonoids and quercetin-3-O-rutinoside (3296 mg/kg dw) was the main polyphenol found in this part of coriander. The fruits revealed only phenolic acids and derivatives, caffeoyl N-tryptophan hexoside (45.33 mg/kg dw) being the most abundant phenolic derivative. In vitro samples also gave a high diversity of polyphenols, being C-glycosylated apigenin (2983 mg/kg dw) the main compound. Anthocyanins were only found in clone A, which was certainly related to its purple pigmentation, and peonidin-3-O-feruloylglucoside-5-O-glucoside was the major anthocyanin found (1.70 μg/kg dw). In vitro culture can be used to explore new industrial, pharmaceutical, and medicinal potentialities, such as the production of secondary metabolites like flavonoids.     

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.     

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 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.     

In vitro bioaccessibility of carotenoids and tocopherols from fruits and vegetables

Aim of the study

To assess the in vitro bioaccessibility of carotenoids, including xanthophyll esters, and tocopherols from fruits and vegetables.

Results

Stability for carotenoids and tocopherols was over 70%. Xanthophyll esters were cleaved by cholesterol esterase but not by human pancreatic lipase. Less than 40% of the β-cryptoxanthin initially present was hydrolyzed and the amount of free xanthophylls recovered was higher when liquid was used than when fresh homogenized matrix was employed. cis-Isomers of β-carotene and lutein did not significantly increase during the process. Xanthophylls were more efficiently transferred into supernatants than tocopherols and β-carotene. cis-Carotenoids, epoxy-xanthophylls and ester forms were also transferred.

Conclusion

The results are consistent with observations in other in vitro digestion models and human studies and support the usefulness of in vitro assessment to study food-related determinants of the bioavailability of carotenoids and tocopherols from fruits and vegetables.     

Antioxidant activity and phenolic compounds of fractions from Portulaca oleracea L.

Portulaca oleracea fractions obtained from the crude extract of the plant by reversed-phase separation were investigated for their antioxidant activities and phenolic compounds content. Five fractions were classified based on optical absorption between the wavelength range of 200–400 nm. Fraction 3 displayed higher absorption than the other fractions. The total amount of quantified phenolics in this fraction was found to be quite high compared to that of crude extract. Chlorogenic, caffeic, p-coumaric, ferulic and rosmarinic acids were the free phenolic acids, and quercetin and kaempferol were the free flavonoids detected in Fraction 3. IC₅₀ value of crude extract was found to be 511.8 μg/ml whereas Fraction 3 exhibited an IC₅₀ value of 154.1 μg/ml. TEAC of Fraction 3 was found to be almost four times higher than that of the crude extract. Fraction 3 was also found to show the highest lipid peroxidation inhibiting capacity, determined by TBARS assay.     

Effect of alkaline pH-shift processing on in vitro gastrointestinal digestion of herring (Clupea harengus) fillets

The effect of alkaline pH-shift processing on herring (Clupea harengus) protein oxidation, salt solubility and digestibility, has been evaluated. For the latter, herring mince and pH-shift produced herring protein isolate, both raw and heat-treated, were digested using a static gastrointestinal in vitro model. The pH-shift process resulted in drastically lowered protein salt solubility and increased lipid oxidation while protein carbonyl formation was unaffected. Yet, no significant differences in the degree of hydrolysis (DH) were observed between mince and isolates after completed gastrointestinal digestion, something which was confirmed by a similar release of proteinaceous material <3 kDa and similar free amino acid profiles. The polypeptide profiles of digested samples however revealed that two peptides (33 and 36 kDa) were present in larger amounts in the digested protein isolate compared to the digested herring mince. The results indicate that alkaline pH-shift processing had limited quantitative influence on the gastrointestinal digestibility of herring proteins despite its negative effects on protein salt solubility and lipid oxidation.