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.     

The enhancement of gastrointestinal digestibility of β-LG by dynamic high-pressure microfluidization to reduce its antigenicity

Dynamic high-pressure microfluidization (DHPM) can increase the antigenicity of β-lactoglobulin (β-LG) in previous researches. We observed in this study that the antigenicity of DHPM-modified β-LG at pressures of 0.1, 80 and 160 MPa after in vitro digestion declined from 13.41 to 12.27 and 7.19 μg mL⁻¹, gradually. The enhancement of gastrointestinal digestibility of β-LG was related to the aggregation state and conformational changes induced by DHPM and was reflected by the electrophoretic bands of low molecular weight (5 and 10 kDa) shown in electrophoresis, the reduction of particle size, the generation of smaller peptide fragments (m/z 837.444, 955.492 and 1215.616) detected by mass spectrum and the increase in surface hydrophobicity. These changes contributed to the decrease in antigenicity. Simultaneously, a schematic diagram was proposed to demonstrate the conceivable mechanism of antigenicity changes of modified β-LG after in vitro digestion.     

Effect of in vitro gastrointestinal digestion on the antioxidant activity of protein hydrolysates prepared from Cape hake by‐products

The antioxidant activity of fish protein hydrolysates may change in the digestive tract during the human gastrointestinal digestion depending on their preparation conditions. Thus, the objective of this study was to assess the antioxidant properties of Cape hake hydrolysates prepared by three different methods (HPH‐A, HPH‐B and HPH‐C) and the effect of their in vitro gastrointestinal digestion on their antioxidant properties. HPH‐A showed the highest reducing power, while HPH‐B and HPH‐C exhibited the highest OH antiradical activity. Fe²⁺‐chelating activity was similar for all hydrolysates, but HPH‐C had a higher Cu²⁺‐chelating activity comparable to that of EDTA. The in vitro gastrointestinal digestion of hydrolysates caused a decrease in the DPPH inhibition, but an increase in scavenging ABTS in all hydrolysates. Copper‐ and iron‐chelating activities increased after digestion of hydrolysates. The results showed that the gastrointestinal digestion of HPHs generally increased their antioxidant properties.     

Physicochemical characterisation and α‐amylase inhibitory activity of tea polysaccharides under simulated salivary,gastric and intestinal conditions

Tea polysaccharides (TPS) are one of the main components of tea with various bioactivities. Digestion could result in the changes of the physicochemical properties and bioactivities of polysaccharides. The objective of this study was to determine the changes in physicochemical properties and α‐amylase inhibitory activities of TPS after simulated digestion. The structure of TPS determined by UV, IR and SEM showed that no obvious changes after salivary digestion, but significant changes after gastrointestinal digestion were observed. After intestinal digestion, the contents of reducing sugar were increased from 0.650 ± 0.011 to 1.594 ± 0.078 mm , and the centre molecular weight was decreased from 540.57 to 375.35 KDa. The molar ratio of monosaccharide was changed after digestion. And α‐amylase inhibition rate was significantly increased (P < 0.05). The results could provide information on the digestibility of TPSin vitro and contribute to the development of related functional foods or medicines.     

Spray-drying microencapsulation of pomegranate juice increases its antioxidant activity after in vitro digestion

Most of the work on pomegranate antioxidant and antibacterial activity has been carried out with solvent extracts of different plant or fruit parts. Biosensitive compounds in juice may be subject to oxidation, reducing their biological activities. Microencapsulation can be used to protect compounds, allowing its incorporation into functional foods. This study aimed at investigating antioxidant activity after in vitro digestion of microencapsulated juice. Pomegranate juice was encapsulated by spray drying its maltodextrin and gum arabic. The average diameter of the microcapsules was 10–50 µm. We evaluated the bioaccessibility of microencapsulated phenolic compounds by using an in vitro enzymatic digestion. The total phenolic content in digested microencapsulated juice was three times greater than in undigested, indicating that the compounds were made bioaccessible. Digestion also increased antioxidant activity, as measured by ABTS●+ or by DPPH●. Additionally, microencapsulated pomegranate juice showed antibacterial activity against the nine bacteria species tested.     

Stability of the phenolic compounds and antioxidant capacity of five fruit (apple,orange, grape,pomelo and kiwi) juices during in vitro‐simulated gastrointestinal digestion

The in vitro digestive stability of phenolic compounds and the antioxidant capacity of five kinds of commonly consumed fruit juices in the daily diet, including apple juice (AJ), orange juice (OJ), grape juice (GJ), pomelo juice (PJ) and kiwifruit juice (KJ), were studied. Following in vitro digestion, the total phenolic (TP) content of fruit juices decreased to different extents by 35%, 25.3%, 23.5%, 22.2% and 7.8% for KJ, OJ, PJ, GJ and AJ, respectively. The individual phenolic content showed similar changes to the TP content, showing reductions of naringenin‐trisaccharide in OJ and PJ, epicatechin in GJ, and chlorogenic acid in AJ by 43.74%, 27.59%, 47.11% and 33.28%, respectively. Conversely, the antioxidant capacity of fruit juices during digestion measured by ABTS assay increased from 4.79% to 35.53%, except in KJ, which decreased by 19.34%. These results show the health benefits of fruit juices after processing and contribute towards establishing suitable dietary recommendations.     

Optimisation of antioxidant hydrolysate production from sweet potato protein and effect of in vitro gastrointestinal digestion

In this study, response surface methodology (RSM) was applied to optimise antioxidant hydrolysate production from sweet potato protein (SPP). The effects of enzyme‐to‐substrate ratio, pH value and reaction temperature on ·OH radical scavenging activity and Fe²⁺‐chelating activity of antioxidant hydrolysates from SPP were investigated. The maximum ·OH radical scavenging activity (40.10%) and Fe²⁺‐chelating activity (74.37%) of SPP hydrolysates (SPPH) were obtained at an enzyme‐to‐substrate ratio of 4%, pH value of 7.8 and reaction temperature of 57 °C, which was in agreement with the predicted value (40.11% and 74.83%, respectively) estimated by RSM. The simulated in vitro gastrointestinal digestion (GID) dramatically modified molecular weights distribution, increased peptide (<3 kDa) concentration and highly retained antioxidant activity of SPPH, indicating potentially utilisation of SPPH as a functional supplement in food system.     

In vitro bioaccessibility of avenanthramides in cookies made with malted oat flours

This study evaluated the effects of malting and in vitro digestion on the bioaccessibility of six avenanthramides (AVNs), soluble phenols (SPs) and antioxidant capacity (ORAC) in cookies (3B, 4B, 5B) prepared with oat flour malted for three (M3), four (M4), five (M5) days, respectively. Control cookies (CTRL) were made with non-malted oat flour (M0). Five-days malting increased the AVN content of native kernels up to 10-folds, whereas SPs and ORAC tripled their values. After the in vitro digestion, AVNs showed their bioaccessibility in the following order: 5B > 4B > 3B, with a recovery of 1703 μg AVNs per 50 g portion of 5B cookies, compared to 135 μg of CTRL. Results showed that the inclusion of 27% malted oat flour was effective to formulate functional cookies with satisfactory technological properties, high AVN content and bioaccessibility.     

Effects of particle size on physiochemical and in vitro digestion properties of durum wheat bran

In order to promote the potential health benefits of high fibre products, wheat bran is the main focus of the food industry. The physiochemical and in vitro digestion properties of wheat bran containing different particle size were investigated and compared against raw bran samples. Firstly, the bran sample containing superfine particles (11.63 μm) was hydrolyzed by the α-amylase, pepsin, and pancreatin enzymes separately using a single factor and orthogonal test. Secondly, optimized hydrolysis parameters of superfine particles were employed to measure in vitro digestibility of macronutrients in raw, coarse, medium, and superfine bran particles. The maximum degree of hydrolysis obtained via α-Amylase (concentration 10 mg mL⁻¹, pH 6.6, and time 12.5 min) was 55.71%; pepsin (concentration 50 mg mL⁻¹, pH 1.2, and time 9 h) was 82.10%; and pancreatin (concentration 100 mg mL⁻¹, pH 7.0 and time 12 h) was 84.71%, respectively. The highest in vitro digestibility rate of reducing sugar, protein, fat, and soluble fibre content was observed in superfine bran samples to 33.4%, 82.55%, 91.53%, and 21.66%, respectively.     

The effect of alginates on in vitro gastric digestion of particulated whey protein

The aim of the study was to investigate how microparticulated and nanoparticulated whey proteins mixed with alginate respond to simulated in vitro gastric digestion conditions at pH 3.0. Initially, particle size distributions and zeta potential were measured in all mixtures at pH 3.0. Particle size distributions as well as SDS‐PAGE were used to investigate the rate of protein degradation by pepsin during simulated in vitro gastric digestion. The complexation of nanoparticulated and microparticulated whey protein with alginates causes formation of insoluble and soluble complexes, which can resist pepsin degradation to a different degree. These results highlight the potential of developing new food products, which can enhance satiety.     

Digestive protection of probiotic Lacticaseibacillus rhamnosus in Ricotta cheese by monoglyceride structured emulsions

This research aimed at studying the potential use of monoglyceride (MG) structured emulsions (MSEs) as delivery and protective systems for probiotic bacteria in Ricotta cheese. To this purpose, a low-fat commercial Ricotta cheese was added with MSEs formulated with milk, as water phase, and sunflower oil (MSE-SO) or anhydrous milk fat (MSE-AMF), as lipid phase. A commercial whole milk Ricotta cheese (W-RC) was considered as reference. A probiotic Lacticaseibacillus rhamnosus strain was inoculated as free cells in W-RC or embedded into the MSEs and added to the low-fat Ricotta at the same reference fat content. After physico-chemical characterisation, L. rhamnosus viability and sample destructuring behaviour upon in vitro digestion were evaluated. At the end of in vitro digestion, both W-RC and sample containing MSE-SO were unable to protect cells. By contrast, sample with AMF ensured a sufficient probiotic viability, even after 14 days of storage at 4 °C. This result was attributed to system composition and structure. During the gastric phase, the presence of caseins and MG-AMF mixed structures induced the formation of clots, entrapping and protecting cells against the acidic pH of the stomach, as confirmed by confocal micrographs and particle size. During the intestinal phase, cell viability was guaranteed by the formation of mixed micelles promoted by MG. It was demonstrated that microbial cells located near MG structures where they found protection.     

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.     

Improving antioxidant properties and probiotic effect of clementine juice inoculated with Lactobacillus salivarius spp. salivarius (CECT 4063) by trehalose addition and/or sublethal homogenisation

This study evaluates the effect of trehalose addition (10% or 20%, w/w) and/or sublethal homogenisation (25–150 MPa) on antioxidants content (vitamin C, total phenols and flavonoids) and activity (measured both by ABTS-TEAC and DPPH assays), as well as on microbial counts and survival to in vitro digestion of clementine juice inoculated with Lactobacillus salivarius spp. salivarius. Particle size, vacuum impregnation parameters and anti-Helicobacter pylori effect were also measured. Incubation with the probiotic improved the antioxidant properties of the juice. Homogenisation pressures below 100 MPa following incubation increased both the probiotic counts in the juice and its antioxidants bioaccesibility. Adding 10% (w/w) of trehalose to the juice was effective in preventing these bioactive compounds deterioration under adverse conditions. Once homogenised, liquids containing 10% (w/w) of trehalose became as able as those without trehalose to enter a food solid matrix. Inhibition of Helicobacter pylori growth was evident in all probiotic beverages.     

Solvent extraction and in vitro simulated gastrointestinal digestion of phenolic compounds from purple sweet potato

The total content of phenolic compounds in purple sweet potato (PSP) was determined and the release of such compounds from PSP in gastrointestinal digestion was studied in vitro. The extraction conditions for the maximum recovery of free phenol (FP) and bound phenol (BP) from PSP were determined by response surface methodology (RSM). The maximum recovery of FPPSP was 14.16 ± 0.87 mg GAE per g short for dry weight (DW), which was obtained using 60% (v/v) ethanol maceration with a liquid–solid ratio of 57.21:1 (mL g⁻¹) at 51.93 °C for 2.12 h. The maximum recovery for BPPSP was 7.54 mg GAE per g DW, which was obtained upon hydrolysis with 1.87 mol L⁻¹ NaOH at a liquid–solid ratio of 35.93:1 (mL g⁻¹) for 4.74 h. The maximum phenolic content was released after 1 and 2 h for the in vitro gastric and intestinal digestion respectively. The release of the phenolics was promoted by pepsin and gastric acid during gastric digestion, while it was further promoted by trypsin during intestinal digestion.     

Effect of thermal processing and digestive protease on the antioxidant capacity of fruit juice–milk beverage model systems under simulated gastrointestinal digestion

The effects of thermal processing and digestive protease on the antioxidant capacity of fruit juice–milk beverage (FJMB) models containing whey protein (WP) and chlorogenic acid or catechin (CAT) under in vitro digestion were investigated. After gastric digestion, the FJMB showed a significant (P < 0.05) increase in ABTS, but little change in ferric reducing antioxidant power (FRAP). Intestinal digestion decreased the FRAP and increased ABTS except in the model containing 0.1% CAT. There were different effects (none, masking, synergistic effect) in the antioxidant activity of the WP and phenolics under digestion. Pasteurisation (63 °C/30 min) had no significant effect on the FJMB's antioxidant capacity, while sterilisation (121 °C/10 min) significantly (P < 0.05) increased, decreased or had no significant effect on ABTS and FRAP under digestion, depending on the model. Pepsin and pancreatin significantly (P < 0.05) decreased the fruit phenolics' ABTS. Ferric reducing antioxidant power was not significantly influenced by pepsin during gastric digestion, but was significantly (P < 0.05) decreased by pancreatin during intestinal digestion. These results would be helpful to improve the efficacy of fruit phenolics in FJMB subjected to thermal processing and to maintain the health benefits of this kind of functional products in the food industry.     

Effect of processing and oil type on carotene bioaccessibility in traditional foods prepared with flour and puree from orange-fleshed sweetpotatoes

Consumption of Orange-Fleshed sweetpotato (OFSP) and products as source of provitamin A is being promoted to tackle vitamin A deficiency (VAD) in sub-Saharan Africa. However, limited information is available on β-carotene retention in foods and delivery after digestion. The study assessed carotene retention and bioaccessibility following in vitro digestion on traditional foods having OFSP among the ingredients. Sunflower oil, margarine and beef fat were evaluated on their effect on β-carotene bioaccessibility. Porridge and chapatis were prepared with either OFSP puree or flours in the formulations. Carotene retention was highest in chapatis (83%) as compared to porridge (65%). Micerallisation efficiency of all-trans β-carotene was comparable between similar products but greater in chapatis (62%) than porridge (11%). Sunflower oil had the highest all-trans β-carotene bioaccessibility compared to margarine and beef fat. The results support the promotion of consumption of OFSP-based products as good source of provitamin A to fight VAD.     

Investigation into the potential of commercially available lesser mealworm (A. diaperinus) protein to serve as sources of peptides with DPP-IV inhibitory activity

Food-derived peptides are known to possess inhibitory activity against the dipeptidyl-peptidase IV (DPP-IV) enzyme, a target in the management of type 2 diabetes. While proteins from commonly consumed food commodities have been investigated as precursors of DPP-IV-inhibiting peptides, studies on novel protein sources, such as those from insects, are sparse. This research aimed to determine if DPP-IV inhibitors can be generated upon in vitro digestion or enzymatic hydrolysis of lesser mealworm protein isolate and concentrate. Treatment of the proteins with digestive enzymes and proteases generated hydrolysates with varying potency, thermolysin being the most effective at releasing active peptides (IC₅₀ = 0.63 and 0.60 mg mL⁻¹ for the isolate and concentrate). Ultrafiltration of the thermolysin-treated hydrolysates did not significantly improve the potency. This study shows that DPP-IV inhibitors can be generated from lesser mealworm protein and provides insight on the potential of insects to serve as functional food ingredients.     

Zein/tannic acid complex nanoparticles‐stabilised emulsion as a novel delivery system for controlled release of curcumin

The present work was aimed to evaluate the potential of the gel‐like Pickering emulsions (50%, v/v, oil) stabilised by zein/tannic acid (TA) complex colloidal particles (ZTPE) as a new encapsulation system of lipophilic ingredients. Compared with sodium caseinate‐stabilised emulsions (SCE) and bulk oil, the better protective effects of ZTPE on the chemical stability of curcumin were observed when they suffered from UV light, and the lipid oxidation rate remarkably reduced in ZTPE. Also, the zein particle layers loaded with TA around the oil droplets can provide protection against harsh gastric environment, facilitating to slow down the release of free fatty acids (FFA) and curcumin during in vitro simulated digestion. These findings show that ZTPE have a good potential to act as an efficient encapsulating agent to protect functional ingredients from degradation and control their release during digestion, which can further improve the bioavailability of bioactive ingredients.     

In vitro digestion of whole chia seeds (Salvia hispanica L.): Nutrient bioaccessibility,structural and functional changes

Dietary chia seed (Salvia hispanica L.) has significant health-related benefits due to its high nutrient contents. However, few studies reported the fate of whole seeds in the gastrointestinal tract. Herein, we explored the digestive characteristics in terms of hydrolysis of nutrients, structural and functional properties with a static in vitro digestion method. After gastrointestinal digestion, the digestibility of lipid and protein was 0.46% and 11.38%, respectively. The release rates of tryptophan, tyrosine and lysine were greater than 20%, whereas the glutamic acid and aspartic acid were less than 5%. The microscopic results (optical microscopy (OM), laser scanning confocal microscopy (LSCM) and scanning electron microscopy (SEM)) demonstrated that the seeds remained intact, and the mucilage adhered tightly to the seed coat during digestion. The water holding capacity and oil holding capacity of seeds accounted for 6.37 and 3.28 g/g after intestinal digestion, which were significantly lower than gastric digestion endpoints (P < 0.05). And there were no significant differences in Fourier transform infrared spectroscopy of G-Mucilage and I-Mucilage. In general, preprocessing before being consumed is necessary for chia seeds to take full advantage of rich polyunsaturated fatty acids and proteins.     

Microencapsulation of roasted coffee oil Pickering emulsions using spray- and freeze-drying: physical,structural and in vitro bioaccessibility studies

Microcapsules produced from well-defined emulsion templates are an interesting alternative for lipid encapsulation. This work aimed to produce microcapsules by the freeze-drying (FD) and spray-drying (SD) of Pickering emulsions of roasted coffee oil (RCO) stabilised with chitosan nanoparticles produced by self-aggregation or by crosslinking with tripolyphosphate. The dried microcapsules were characterised in terms of particle size, oil retention and structure; furthermore, the in vitro bioaccessibility of polyphenols from microencapsulated RCO was investigated. The use of chitosan nanoparticles to stabilise the emulsions increased oil retention in the microcapsules giving values between 83.04% and 95.36%. SD produced spherical microcapsules with small particle sizes (˜11 μm), whereas FD microcapsules showed an irregular shape and porous structure. Although FD had the lowest impact on the bioactive compounds, SD promoted better protection for phenolic compounds and antioxidant activity during in vitro digestion.