Duodenal Phase Research Articles

Influences of freeze‐ and spray‐drying vs. encapsulation with soy and whey proteins on gastrointestinal stability and antioxidant activity of Mediterranean aromatic herbs

SummaryThe aim of the present research was to study the chemical profile of volatile and non‐volatile compounds in freeze‐ and spray‐dried oregano, thyme, sage, and rosemary as well as to study the gastrointestinal stability and the biological activity of the samples after encapsulation using two proteins (soy and whey). Chemical analysis was performed by HS‐SPME/GC‐MS and HPLC‐PDA techniques. The antioxidant activity (FRAP, DPPH and ORAC assays) and HPLC‐PDA analyses were performed before and after simulated two‐phase digestion process (gastric and duodenal) using human gastrointestinal enzymes. The effect of protein carriers was depended on the plant matrix and the class of compound. Soy protein is useful for the protection of thymol and carvacrol content in thyme and the increase of the antioxidant activity of sage. Whey protein is useful for flavanols stability in all studied herbs before and after the duodenal digestive phase.

Microplastics as Vectors of Chromium and Lead during Dynamic Simulation of the Human Gastrointestinal Tract

The human body is exposed to the ingestion of microplastics that are often contaminated with other substances, which can be released into our body. In this work, a dynamic in-vitro simulator of the gastrointestinal tract based on a membrane reactor has been used for the first time to study the release, bioaccessibility, and bioavailability of chromium (Cr) and lead (Pb) from polyethylene and polypropylene microplastics previously contaminated in the laboratory. The results showed that 23.11% of the initial Cr and 23.17% of the initial Pb present in microplastics were able to cross the tubular membrane, simulating the intestinal absorption phase. The pH evolution during the gastric phase and the duodenal phase, the interaction mechanisms with physiological fluids, and the properties of the polymers, such as specific surface, porosity, and/or surface degradation, affected the kinetics of release from the microplastics and the behavior of both heavy metals. Cr was released very early in the gastric phase, but also began simultaneously to precipitate quite fast, while Pb was released slower and in less quantity than Cr, and did not precipitate until the beginning of the duodenal phase. This study shows, for the first time, how useful the dynamic gastrointestinal simulator is to study the behavior of microplastics and some problematic heavy metals along the human gastrointestinal tract, and can serve as a reference for future studies focused on the effects of these substances in the human body.

In vitro assessment of cyanotoxins bioaccessibility in raw and cooked mussels

The oral route by ingestion of water and food contaminated with cyanotoxins is the main route of exposure to these toxins. This study addresses for the first time the bioaccessibility of some of the most common Microcystins (MC-LR, MC-RR and MC-YR) and Cylindrospermopsin (CYN) simultaneously in raw and steamed mussels spiked at 250 ng/g fresh weight of each cyanotoxin, after an in vitro digestion, including the salivary (incubation with artificial saliva, 30s), gastric (with pepsin, 2h, pH 2), duodenal (with pancreatin and bile salts, 2h, pH 6.5) and colonic phases (with lactic-acid bacteria, 48h, pH 7.2). The results obtained suggest that the potential absorption of these cyanotoxins by consumption of contaminated mussels is lower than expected. After the total effect of cooking and digestion, the mean bioaccessibility levels recorded were 24.65% (CYN), 31.51% (MC-RR), 17.51% (MC-YR) and 13.20% (MC-LR). Moreover, toxins were transferred to the steaming waters at 3.77 ± 0.24 μg L−1 CYN, 2.29 ± 0.13 μg L−1 MC-LR, 6.60 ± 0.25 μg L−1 MC-RR and 3.83 ± 0.22 μg L−1 MC-YR. These bioaccessibility results should be considered for a more accurate risk assessment related to these cyanotoxins in mussels, including the fact that the steaming waters could also represent a risk after human consumption.

Protein digestion properties of Xinong Saanen goat colostrum and mature milk using in vitro digestion model.

Xinong Saanen goat milk is a raw material for goat milk-based infant formula production. This study aims to analyze digestion properties of Xinong Saanen goat colostrum and mature milk by simulating infant gastrointestinal digestion. Zeta potential, particles size, protein profile and peptides composition of these two kinds of milk during the digestion process were studied. Zeta-potential values of the digested colostrum were lower than those of mature milk through the whole digestion. Absolute zeta potential of colostrum duodenal digestion samples showed a decrease from 16.63 ± 2.08 to 11.80 ± 2.03 mV while that of mature milk decreased sharply and then increased (P < 0.05). Colostrum had a larger particle size than mature milk and both milks showed decreased particle size with increasing digestion time but an increase for the last 30 min. Colostrum showed more high molecular weight (MW) proteins which cannot be hydrolyzed completely compared with mature milk. Digested peptides (< 10 kDa) were characterized using liquid chromatography combined with tandem mass spectrometry (LC-MS/MS). The casein-derived peptides identified in digested colostrum and mature milk accounted for 76.67% and 59.53%, respectively. β-Casein was the most abundant in colostrum while that in mature milk was αs1 -casein. Enterotoxin-binding glycoprotein PP20K, butyrophilin subfamily 1 member A1 (BTN1A1) and perilipin (PLIN) were only detected in digested mature milk. Differences in digestion properties between goat colostrum and mature milk were mainly shown in duodenal digestion phase. Data may provide useful information about utilization of goat milk for infant formula formulation. © 2019 Society of Chemical Industry.

Impact of high hydrostatic pressure and thermal treatment on the stability and bioaccessibility of carotenoid and carotenoid esters in astringent persimmon (Diospyros kaki Thunb, var. Rojo Brillante).

The carotenoid and carotenoid ester profile in astringent persimmon (Diospyros kaki Thunb., var. Rojo Brillante) was composed by 13 free xanthophylls, 8 hydrocarbon carotenes and 17 carotenoid esters. The stability and biaoccessibility of these carotenoids was determined by an adaptation of the INFOGEST protocol. Results showed that the stability of persimmon carotenoids ranged from 61 to 74%, depending on the digestion phase, being (all-E)-β-cryptoxanthin and (all-E)-antheraxanthin 3-O-palmitate the most stable carotenoids. At the final step of the digestion (oral + gastric + duodenal phase), only traces of (all-E)-antheraxanthin, (all-E)-lutein and (all-E)-β-cryptoxanthin were found in control samples due to the low efficiency of carotenoid micellization, which was affected by the high pectin content naturally present in persimmon tissues. Processing increased the overall carotenoid bioaccessibility to 54% in pressurized samples and to 25% in thermal treated ones. This effect depended on the processing technology as well as on the chemical structure of the carotenoid, being (all-E)-β-cryptoxanthin and (all-E)-β-cryptoxanthin laurate the most bioaccessible carotenoids in pressurized samples and (all-E)-β-cryptoxanthin laurate and (all-E)-antheraxanthin the most bioaccessible ones in pasteurized ones.

HPLC-UV/Vis-APCI-MS/MS Determination of Major Carotenoids and Their Bioaccessibility from "Delica" (Cucurbita maxima) and "Violina" (Cucurbita moschata) Pumpkins as Food Traceability Markers.

Carotenoids are a widespread group of fat-soluble pigments, and their major nutritional importance comes from their pro-vitamin A activity and their antioxidant capacity. In this study, two different pumpkin cultivars (Cucurbita maxima, also named ‘Delica’ and Cucurbita moschata, also known as ‘Violina’) from the southern Po Delta area were investigated in terms of carotenoid content and the influence of food processing on compositional changes and carotenoid bioaccessibility. Quali- and quantitative determination of carotenoids in sample extracts were performed on a C30 column by means of an online coupled HPLC-UV/Vis-APCI-MS/MS technique. The identification of separated compounds was tentatively achieved by merging (i) chromatographic data, (ii) UV-Vis spectra, and (iii) MS/MS fragmentation spectra. The chromatographic profiles for the two cultivars showed qualitative differences. Two major carotenoids were considered for quantification purposes and further investigations: lutein and -carotene. Quantification of target carotenoids was performed with external calibration through analytical standards. The concentration of lutein and -carotene was higher in C. maxima than in the other variety, C. moschata. Carotenoids are susceptible to degradation (isomerization and oxidation) during food processing (i.e., cooking), and the concentration of lutein and -carotene were monitored in oven-cooked and steam-cooked pumpkins. The steam-cooking process was superior in terms of limiting carotenoid loss. A complete functional profile of pumpkins as a source of carotenoids was gained with the evaluation of their in vitro bioaccessibility and their bioavailability after intake during human digestion. Bioaccessibility of lutein and -carotene were estimated by an in vitro static digestion model that involved salivary, gastric, and duodenal phases. Bioaccessibility values progressively increased from the salivary to the duodenal phase for both pumpkin varieties and cooking methods. Bioaccessibility of lutein was always lower than -carotene for both cultivars and for both cooking methods. Bioaccessibility values for lutein and -carotene changed from 1.93% to 2.34% vs. 4.94% and 8.83% in the salivary phase, from 2.7% to 4.63% vs. 7.83% and 15.60% in the gastric phase, and from 10.04% to 13.42% vs. 25.81% and 35.32% in the duodenal phase. For both target compounds, bioaccessibility in the duodenal phase was more than twice the gastric values, and it underlined that the type of cooking did not influence release from the initial matrix.

Bioaccessibility and decomposition of cylindrospermopsin in vegetables matrices after the application of an in vitro digestion model

Research on the human exposure to Cylindrospermopsin (CYN) via consumption of contaminated food is of great interest for risk assessment purposes. The aim of this work is to evaluate for the first time the CYN bioaccessibility in contaminated vegetables (uncooked lettuce and spinach, and boiled spinach) after an in vitro digestion model, including the salivar, gastric and duodenal phases and, colonic fermentation under lactic acid bacteria. The results obtained showed that the digestion processes are able to diminish CYN levels, mainly in the colonic phase, especially in combination with the boiling treatment, decreasing CYN levels in a significant way. Moreover, the potential decomposition products in a pure CYN solution and in CYN-contaminated vegetables were evaluated using UHPLC-MS/MS Orbitrap. Under the conditions assayed, only two diastereoisomers of the same fragment with m/z 292.09617 have been detected in all the analysed samples, with the exception of digested vegetables. Therefore, in terms of risk assessment, the digestion seems to play an important role in reducing the final bioaccesibility of CYN, and the consumption of cooked vegetables (spinach) would be safer in comparison to raw vegetables.

Bioaccesibility of Cylindrospermopsin from cooked fish muscle after the application of an in vitro digestion model and its bioavailability

Humans can be exposed to cyanotoxins through the ingestion of contaminated water, food or beverages. In the present work, the bioaccesibility of Cylindrospermopsin (CYN), one of the most relevant cyanotoxins, was evaluated in a pure CYN solution and cooked CYN-contaminated fish muscles (20 μg/mL). An in vitro digestion model including the salivar, gastric, duodenal and colonic phases was performed, being each fraction analyzed by HPLC-MS-MS to evaluate CYN degradation. Moreover, Caco-2/TC7 cells were exposed to the digested duodenal and colonic phases to elucidate the final bioavailability of CYN in an approximation to the real human exposure scenario. The results revealed that CYN bioaccesibility decreased after the digestive process in all the cooked fish samples. The most drastic reductions were observed after lactic acid bacteria exposure. Thus, the highest bioaccesibility values were obtained in fish cooked by steaming (12.5%) and broiling (10.9%) meanwhile CYN was not detected in the colonic phase after boiling and microwaving. Regarding the duodenal and colonic availability, only in CYN pure digested solution the cyanotoxin was identified. The results obtained showed that digestion processes plays a very important role in the degradation of CYN, which should be considered when preparing a risk assessment of CYN.

Tibiotalocalcaneal arthrodesis with a retrograde triple bending nail – Results of the first 200 cases

Humans can be exposed to cyanotoxins through the ingestion of contaminated water, food or beverages. In the present work, the bioaccesibility of Cylindrospermopsin (CYN), one of the most relevant cyanotoxins, was evaluated in a pure CYN solution and cooked CYN-contaminated fish muscles (20 μg/mL). An in vitro digestion model including the salivar, gastric, duodenal and colonic phases was performed, being each fraction analyzed by HPLC-MS-MS to evaluate CYN degradation. Moreover, Caco-2/TC7 cells were exposed to the digested duodenal and colonic phases to elucidate the final bioavailability of CYN in an approximation to the real human exposure scenario. The results revealed that CYN bioaccesibility decreased after the digestive process in all the cooked fish samples. The most drastic reductions were observed after lactic acid bacteria exposure. Thus, the highest bioaccesibility values were obtained in fish cooked by steaming (12.5%) and broiling (10.9%) meanwhile CYN was not detected in the colonic phase after boiling and microwaving. Regarding the duodenal and colonic availability, only in CYN pure digested solution the cyanotoxin was identified. The results obtained showed that digestion processes plays a very important role in the degradation of CYN, which should be considered when preparing a risk assessment of CYN.

Digestibility of Quinoa (Chenopodium quinoa Willd.) Protein Concentrate and Its Potential to Inhibit Lipid Peroxidation in the Zebrafish Larvae Model.

Quinoa protein concentrate (QPC) was extracted and digested under in vitro gastrointestinal conditions. The protein content of QPC was in the range between 52.40 and 65.01% depending on the assay used. Quinoa proteins were almost completely hydrolyzed by pepsin at pH of 1.2, 2.0, and 3.2. At high pH, only partial hydrolysis was observed. During the duodenal phase, no intact proteins were visible, indicating their susceptibility to the in vitro simulated digestive conditions. Zebrafish larvae model was used to evaluate the in vivo ability of gastrointestinal digests to inhibit lipid peroxidation. Gastric digestion at pH1.2 showed the highest lipid peroxidation inhibition percentage (75.15%). The lipid peroxidation activity increased after the duodenal phase. The digest obtained at the end of the digestive process showed an inhibition percentage of 82.10%, comparable to that showed when using BHT as positive control (87.13%).

Release of dipeptidyl peptidase IV, α-amylase and α-glucosidase inhibitory peptides from quinoa (Chenopodium quinoa Willd.) during in vitro simulated gastrointestinal digestion

As diabetes is a major cause of mortality and morbidity in epidemic rates, continuous research is being done on development of foods with anti-diabetic activity. In this study, the influence of gastrointestinal digestion of quinoa protein to release peptides with anti-diabetic potential was investigated. Quinoa protein was subjected to an in vitro simulated gastrointestinal digestion and fractionated by ultrafiltration. Gastric and gastroduodenal digests and peptide fractions were evaluated for dipeptidyl peptidase IV (DPP-IV), α-amylase and α-glucosidase inhibitory activities. Peptides released during the duodenal phase showed the highest inhibitory effects. Three novel peptides derived from 11S seed storage globulin B were identified in the most active fraction by HPLC-MS/MS. These peptides showed ability to inhibit enzymes involved in incretin degradation and digestion of dietary carbohydrates. Therefore, quinoa proteins are promising ingredients of functional foods or nutraceutical applications for the control of diabetes.

Peanut digestome: Identification of digestion resistant IgE binding peptides

Stability to proteolytic degradation in the digestive tract is considered a general feature shared by most food allergens. Current digestibility models exclusively utilize purified allergen proteins, neglecting the relevant effects of matrix that occur for foodstuff systems. In the present study, we investigated digestion stability of the major peanut allergens directly in the natural matrix using an in vitro static model that simulates the gastrointestinal digestion including the oral, gastric, duodenal and intestinal (brush border membrane enzymes) phases. Immunogenicity was evaluated by Western Blot using N=8 pooled sera of peanut allergic pediatric subjects. Immunoreactive, large-sized and fragments of Ara h 2, Ara h 6 and Ara h 3 survived hydrolysis as assessed by SDS-PAGE. Smaller resistant peptides mainly arising from Ara h 3 and also Ara h 1 were detected and further identified by LC-high resolution-MS/MS. RP-HPLC purification followed by dot-blot analysis and MS/MS-based identification demonstrated that stable IgE-binding peptides derived from Ara h 3. These results provide a more realistic picture of the potentially allergenic determinants of peanuts that survived the human digestion, taking into account the role of the food matrix, which may significantly affect gastrointestinal breakdown of peanut allergens.

Bioaccessibility and HPLC-MS/MS chemical characterization of phenolic antioxidants in Red Chicory (Cichorium intybus)

Two varieties of red chicory from southern Po Delta area have been characterized in terms of polyphenols content by HPLC-MS/MS. Seven target compounds were selected for determining their bioaccessibility from vegetable matrix. Simulated digestion process was employed for this purpose by means of an in-vitro static model. Salivary, gastric and duodenal phases were realized by using solutions of different physiological enzymes miming the stages of digestion process. Results show well defined behaviors associated with specific structures of aglycone or phenolic acid and position of O-glycosyl moiety. Dicaffeoyltartaric acid together with apigenin-7-O-glucoside and kaempferol-7-O-glucoside have similar bioaccessibility trend that is characterized by a higher value for duodenal phase than for gastric one. Quercetin-3-O-(6″-O-malonyl)-glucoside behaves in reverse way and gastric bioaccessibility turns higher than duodenal. Lastly, the difference between gastric and duodenal bioaccessibility is enhanced in the case of cyanidin-3-O-glucoside and the isomers of caffeoylquinic acid.

In vitro digestion of sodium caseinate emulsions loaded with epigallocatechin gallate

The digestion behaviour of a structured emulsion interface loaded with epigallocatechin-gallate (EGCG) was investigated. Oil in water emulsions were prepared with 0.35% sodium caseinate, 0, 2, and 6 mg/ml epigallocatechin-gallate (EGCG), and 0.45% of high methoxyl pectin (HMP). Emulsions were subjected to a static in vitro digestion, simulating the three stages of human upper gastrointestinal digestion, to investigate EGCG-salivary protein associations, and the effect of incorporated EGCG on gastric proteolysis and duodenal lipolysis. Confocal microscopy observations demonstrated that the addition of HMP reduced saliva's mucin-induced aggregation of the emulsions. Gel electrophoresis results indicated that sodium caseinate was fully hydrolysed, regardless of the presence of EGCG or HMP. The presence of EGCG reduced the extent of free fatty acid release during the duodenal phase. The findings from this study suggest that sodium caseinate-stabilized emulsions can be employed as a platform for delivery of EGCG.

Impact of in vitro digestion phases on the stability and bioaccessibility of carotenoids and their esters in mandarin pulps.

The composition of carotenoids (carotenes and free and acylated xanthophylls) and their bioaccessibilities were determined for the first time in pulps of mandarins cultivated in Brazil. Two cultivars of mandarin, Citrus reticulata Blanco cv. 'Ponkan' and Citrus reticulata × C. sinensis cv. 'Murcott', showed higher contents of most carotenoids compared to those found in C. deliciosa Tenore cv. 'Rio'. The major carotenoids in mandarin cv. 'Ponkan' and 'Murcott' were (all-E)-β-cryptoxanthin laurate (19-21%), (all-E)-β-cryptoxanthin myristate (15-17%) and (Z)-ζ-carotene (7-12%), followed by (all-E)-β-cryptoxanthin palmitate (4-7%), free (all-E)-β-cryptoxanthin (5-6%) and (all-E)-β-carotene (4-5%), while in mandarin cv. 'Rio' (all-E)-β-cryptoxanthin myristate (22%) was the major compound, followed by (all-E)-β-cryptoxanthin laurate (16%), (all-E)-β-cryptoxanthin palmitate (11%), (all-E)-β-cryptoxanthin (9%) and (all-E)-β-carotene (6%). After in vitro digestion, the qualitative carotenoid profile of the supernatant containing the micellarized carotenoids was similar to that of fresh fruits, but the contents were significantly lower. Carotenoid and mandarin physico-chemical properties influenced the bioaccessibility of carotenoids. Free (all-E)-β-cryptoxanthin showed the highest bioaccessibility in all mandarin cultivars (33-42%), while the bioaccessibilities of β-carotene (16-36%) and the major carotenoid esters (18-33%) were lower. The overall recovery of carotenoids during in vitro digestion was around 98% after the oral phase, 79% after oral + gastric phases and 77% after oral + gastric + duodenal phases, with free (all-E)-β-cryptoxanthin and (all-E)-β-carotene being the most stable ones. Besides possible E-Z isomerization and ester hydrolysis, evident losses occurred in total carotenoid contents and also in the most individual carotenoids and they were not compensated for by the former reactions.

Insight into the gastro-duodenal digestion resistance of soybean proteins and potential implications for residual immunogenicity.

Soy is an important component of the human diet thanks to its nutritional value and high protein content; however, it also represents a risk for allergenic consumers due to its potential to trigger adverse reactions in sensitized individuals. The putative correlation between immunoreactivity and resistance to the human gastrointestinal (GI) digestion has drawn attention to investigating soybean proteins digestibility. In this work, we provided further insights into this field by performing in vitro simulated GI digestion experiments directly on ground soybean seeds, to provide more realistic results obtained from the digestion of the whole food matrix. Soybean digestion products were analyzed by SDS-PAGE followed by untargeted HPLC-MS/MS analysis and the final data were software treated to enable protein/peptide identification. The latter allowed monitoring the proteolytic degradation of the main soybean proteins during the gastric and duodenal phases. In particular, β-conglycinin and trypsin inhibitors showed the highest resistance to the combined activity of GI enzymes, showing a partial degradation at the end of the duodenal phase as ascertained by the strong electrophoretic bands displayed at 50 kDa and 20 kDa, respectively. Glycinin subunits also presented, even if to a lower extent, resistance to the complete proteolytic degradation, as demonstrated by polypeptide fragments with molecular weight lower than 20 kDa displayed in the gel at the end of duodenal digestion. In addition, by bioinformatics analysis it was demonstrated that the GI resistant fragments of the allergenic proteins, β-conglycinin and glycinin, retained in their primary structure linear epitopes potentially able to trigger an immunoreaction when exposed to the intestinal mucosa. Moreover, such resistant peptides also presented a structural homology with epitope sequences recognized in other legume species, presenting a potential risk of adverse cross-reaction for a larger category of allergic consumers.

Peripheral apelin-13 administration inhibits gastrointestinal motor functions in rats: The role of cholecystokinin through CCK1 receptor-mediated pathway

Apelin is the endogenous ligand of the G protein-coupled receptor APJ. The APJ receptor is widely expressed in gastrointestinal (GI) tissues including stomach and small intestine. Apelin administration was shown to induce the release of cholecystokinin (CCK) which is a well-known alimentary hormone with its inhibitory actions on GI motor functions through CCK1 receptors on vagal afferent fibers. We investigated whether; (i) peripherally injected apelin-13 alters GI motor functions, (ii) apelin-induced changes are mediated by APJ receptor or CCK1 receptor and (iii) vagal afferents are involved in inhibitory effects of apelin. Solid gastric emptying (GE) and colon transit (CT) were measured, whereas duodenal phase III-like contractions were recorded in rats administered with apelin-13 (300μg/kg, ip). CCK1 receptor antagonist lorglumide (10mg/kg, ip) or APJ receptor antagonist F13A (300μg/kg, ip) was administered 30min prior to the apelin-13 injections. Vagal afferent denervation was achieved by systemic administration of vanilloid receptor agonist capsaicin (125mg/kg, sc). Apelin-13 administration significantly (p<0.01) increased the CCK level in portal venous plasma samples. Compared with vehicle-treated rats, apelin-13 significantly delayed both GE (p<0.001) and CT (p<0.01). Pretreatment of lorglumide or F13A completely abolished the apelin-13-induced inhibitory effects on GE and CT, moreover, apelin-13 was found ineffective in rats underwent afferent denervation. F13A administration alone significantly accelerated the basal CT. Apelin-13 noticeably disturbed the duodenal fasting motor pattern by impairing phase III-like contractions while increasing the amplitudes of phase II contractions which were prevented by pretreatment of lorglumide and capsaicin. Compared with vehicle-treated rats, lorglumide and capsaicin significantly (p<0.05) reduced the apelin-13-induced increases in phase II motility index. Peripherally administered apelin-13 inhibits GI motor functions through CCK-dependent pathway which appears to be mediated by CCK1 receptors on vagal afferents. Peripheral apelin might contribute to the motility changes occurred in postprandial period.

Stability of total phenolic concentration and antioxidant capacity of extracts from pomegranate co-products subjected to in vitro digestion.

BackgroundCo-products obtained from pomegranate juice processing contain high levels of polyphenols with potential high added values. From value-addition viewpoint, the aim of this study was to evaluate the stability of polyphenolic concentrations in pomegranate fruit co-products in different solvent extracts and assess the effect on the total antioxidant capacity using the FRAP, DPPH˙ and ABTS+ assays during simulated in vitro digestion.MethodsPomegranate juice, marc and peel were extracted in water, 50 % ethanol (50%EtOH) and absolute ethanol (100%EtOH) and analysed for total phenolic concentration (TPC), total flavonoids concentration (TFC) and total antioxidant capacity in DPPH˙, ABTS+ and FRAP assays before and after in vitro digestion.ResultsTotal phenolic concentration (TPC) and total flavonoid concentration (TFC) were in the order of peel > marc > juice throughout the in vitro digestion irrespective of the extraction solvents used. However, 50 % ethanol extracted 1.1 to 12-fold more polyphenols than water and ethanol solvents depending on co-products. TPC and TFC increased significantly in gastric digests. In contrast, after the duodenal phase of in vitro digestion, polyphenolic concentrations decreased significantly (p < 0.05) compared to those obtained in gastric digests. Undigested samples and gastric digests showed strong and positive relationships between polyphenols and the antioxidant activities measured in DPPH, ABTS+ and FRAP assays, with correlation coefficients (r2) ranging between 0.930–0.990. In addition, the relationships between polyphenols (TPC and TFC) and radical cation scavenging activity in ABTS+ were moderately positive in duodenal digests.ConclusionFindings from this study showed that concentration of pomegranate polyphenols and the antioxidant capacity during in vitro gastro-intestinal digestion may not reflect the pre-digested phenolic concentration. Thus, this study highlights the need to provide biologically relevant information on antioxidants by providing data reflecting their stability and activity after in vitro digestion.

Novel probiotic evidence of lactobacilli on immunomodulation and regulation of satiety hormones release in intestinal cells

Lactobacilli strains with probiotic traits were tested for their ability to survive to the digestion process modelled during this study. These strains managed to sustain the harsh conditions of the gastric and duodenal phases and showed good adhesion capacities to human Caco-2 cell line. These probiotic microorganisms have survived during these steps, exposed to low pH, high concentration of bile salts and enzymes occurring in the digestion and virtually reached the duodenal compartment in sufficient amount with limited population loss. These lactobacilli strains appeared to be non-cytotoxic after contact with Caco-2 cells for 24 h. Importantly, some of these strains showed immunomodulatory effect, lowering the pro-inflammatory cytokine IL-8 and promoting secretion of the anti-inflammatory IL-10. Besides, Lactobacillus gasseri CMUL34 and Lactobacillus acidophilus CMUL67 strains were able to modulate secretion and expression of two intestinal hormones: the Glucagon-Like Peptide 1 (GLP-1) and the cholecystokinin (CCK) in STC-1 cells.

Investigations on the reactions of α-dicarbonyl compounds with amino acids and proteins during in vitro digestion of biscuits.

This study investigated the interactions of reactive α-dicarbonyl compounds, particularly methylglyoxal (MGO) and 3-deoxyglucosone (3-DG), in commercial biscuits during gastrointestinal digestion. An in vitro multi-step enzymatic digestion system simulating gastric, duodenal and colon phases was used. MGO and 3-DG concentrations decreased with the progress of digestion. Model systems composed of MGO and lysine, cysteine or ovalbumin and model biscuits containing lysine, cysteine or ovalbumin were subjected to in vitro digestion. The results revealed that disappearance in dicarbonyl contents was due to interactions of reactive dicarbonyl compounds with the accumulating amino acids during digestive process. By a high resolution mass spectrometry analysis in model systems and biscuits, the formation of adducts of dicarbonyl compounds with amino or sulfhydryl groups of amino acids was confirmed.