Bioaccessibility of phenolic compounds using the standardized INFOGEST protocol: A narrative review.

The INFOGEST protocol creation was a watershed for phenolic bioaccessibility studies. Because of this important initiative to standardize bioaccessibility studies, data comparisons between different laboratories are now expedited. It has been eight years since the INFOGEST protocol creation, and three from the latest update. However, the current status in terms of phenolic bioaccessibility and how far different laboratories are from reaching a consensus are still unrevealed. In this sense, this narrative review considered an evaluation of different studies that applied the INFOGEST protocol to investigate the bioaccessibility of phenolic compounds. The central objective was to compile the main findings and consensus and to identify possible gaps and future opportunities. This approach intends to further facilitate the use of this protocol by professionals in the field of food science and technology and related areas, generating a reflection on the actual level of standardization of the method. Despite the differences in phenolic compounds from diverse food matrices, and their peculiar behavior, some trends could be elucidated, in terms of phenolic release, stability, and/or transformation upon in vivo digestion. In contrast, there was no general consensus regarding sample preparation, how to report results and the form to calculate bioaccessibility, making it difficult to compare different studies. There is still a long road to effectively standardize the results obtained for phenolic bioaccessibility using the INFOGEST protocol, which is also an opportunity in terms of food analysis that can impact the food industry, especially for the development of nutraceuticals and functional foods.

Effect of guarana seed powder on cholesterol absorption in vitro and in Caco-2 cells.

Cardiovascular disease risk is related to oxidative stress and hypercholesterolemia. Guarana seed powder contains flavanols that possess antioxidant properties and cholesterol-lowering effects. However, the molecular mechanism through which guarana seed powder may decrease cholesterol uptake from the intestinal lumen remains unclear. Thus, this study aimed to investigate the effect of guarana powder aqueous extract on cholesterol absorption mechanisms. After simulated gastrointestinal digestion, we performed assays to determine enzymatic inhibitory capacity, bile acid binding capacity, and cholesterol micellar solubilization. Caco-2 cells were used for permeation and protein identification assays. Digested guarana powder extract inhibited pancreatic lipase in a dose-dependent manner (half-maximal inhibitory capacity: 1.033 µg/mL) and, at a concentration of 1 mg/mL, bound 45.63 % of sodium taurocholate and decreased cholesterol micellar solubilization by 10.14 %. Moreover, incubation with the extract reduced cholesterol absorption by Caco-2 cells and decreased intracellular cholesterol transporter levels. These results indicate that guarana seed powder have potential applications for blood cholesterol management, presenting hypocholesterolemic effects owing to the presence of bioacessible polyphenols.

Vitamin C and Phenolic Antioxidants of Jua (Ziziphus joazeiro M.) Pulp: A Rich Underexplored Brazilian Source of Ellagic Acid Recovered by Aqueous Ultrasound-Assisted Extraction.

Jua (juá in Portuguese) is an underexplored fruit from Brazil's northeast. This fruit is rich in antioxidant substances. However, there is a dearth of information about jua's bioactive potential. The present study evaluated two extraction methods (continuous agitation and ultrasound-assisted extraction-UAE) and employed three different solvents (water, ethanol, and acetone) to efficiently recover soluble phenolic compounds. Aqueous extracts obtained by UAE showed the highest total phenolic content (TPC) and antiradical activity. Besides being an eco-friendly procedure, extraction and/or solubility in an aqueous medium is also important for food application. Ellagic acids were the predominant phenolics (80%) found in aqueous jua pulp extract obtained by UAE, as determined by HPLC, while its TPC was 405.8 gallic acid equivalent per gram of fruit. This extract also exhibited a higher scavenging activity towards peroxyl radicals when compared to that of several other fruits from the literature, including grape, strawberry, cranberry, and walnuts, which are known references in terms of antioxidants. This is the first report that demonstrates jua pulp's potential as an alternative source of ellagic acid and other phenolic acids and flavonoids. Therefore, the outcome of this study provides new information that can be useful for functional food and nutraceutical industries.

Chickpeas from a Chilean Region Affected by a Climate-Related Catastrophe: Effects of Water Stress on Grain Yield and Flavonoid Composition.

The Valparaiso region in Chile was decreed a zone affected by catastrophe in 2019 as a consequence of one of the driest seasons of the last 50 years. In this study, three varieties ('Alfa-INIA', 'California-INIA', and one landrace, 'Local Navidad') of kabuli-type chickpea seeds produced in 2018 (control) and 2019 (climate-related catastrophe, hereafter named water stress) were evaluated for their grain yield. Furthermore, the flavonoid profile of both free and esterified phenolic extracts was determined using liquid chromatography-mass spectrometry, and the concentration of the main flavonoid, biochanin A, was determined using liquid chromatography with diode array detection. The grain yield was decreased by up to 25 times in 2019. The concentration of biochanin A was up to 3.2 times higher in samples from the second season (water stress). This study demonstrates that water stress induces biosynthesis of biochanin A. However, positive changes in the biochanin A concentration are overshadowed by negative changes in the grain yield. Therefore, water stress, which may be worsened by climate change in the upcoming years, may jeopardize both the production of chickpeas and the supply of biochanin A, a bioactive compound that can be used to produce dietary supplements and/or nutraceuticals.

Effects of the consumption of guarana on human health: A narrative review.

Guarana (Paullinia cupana) is a plant from the Amazon region with cultural importance. Despite its early ancestral use by indigenous tribes, the first reports regarding the benefits of guarana consumption for human health were published in the 19th century. Since then, the use of guarana seed in powder and extract forms has been studied for its diverse effects on human health, such as stimulating, anti-inflammatory, antioxidant, anticancer, hypocholesterolemic, and anti-obesity effects. These effects are attributed to the high content of bioactive compounds found in guarana seeds, especially methylxanthines and flavonoids. In fact, the Brazilian Food Supplement Law has officially acknowledged guarana as a source of bioactive compounds. The number and diversity of studies focused on guarana and human health are increasing; thus, organizing and describing the available evidence on guarana and its applications is necessary to provide a framework for future studies. In this narrative review, we have organized the available information regarding guarana and its potential effects on human health. Guarana produces unique fruits with great potential for human health applications. However, the available evidence lacks human studies and mechanistic investigations. Future studies should be designed considering its applicability to human health, including intake levels and toxicity studies.

Protection against indomethacin-induced loss of intestinal epithelial barrier function by a quercetin oxidation metabolite present in onion peel: In vitro and in vivo studies.

Oxidative stress is directly implicated in the loss of intestinal epithelial barrier function (IEBF) induced by non-steroidal anti-inflammatory drugs (NSAIDs). Previous studies by our research team demonstrated that 2-(3,4-dihydroxybenzoyl)-2,4,6-trihydroxy-3(2H)-benzofuranone (BZF), a quercetin oxidation metabolite that naturally occurs in onion peels, exhibits an antioxidant potency notably higher than quercetin. Thus, we assessed the potential of BZF and a BZF-rich onion peel aqueous extract (OAE) to protect against the loss of IEBF in Caco-2 cell monolayers and in rats exposed to indomethacin. In vitro, pure BZF and OAE standardized in BZF (100 nM), protected against the drop in transepithelial electrical resistance by 70 - 73%. Likewise, it prevented the increase in fluorescein-isothiocyanate labelled dextran (FITC-dextran) paracellular transport by 74% and oxidative stress by 84 - 86%. In vivo, BZF, given orally at a dose 80 µg/Kg bw as OAE, totally abolished a 30-fold increase in FITC-dextran serum concentration induced by indomethacin. This effect was dose-dependent and largely conserved (85%) when OAE was given 180-min prior to indomethacin. The IEBF-protective effect of OAE was accompanied by a full prevention of the NF-ĸB activation, and the increases in interleukine-8 secretion and myeloperoxidase activity induced by indomethacin. The protection was also associated with a 21-fold increase in Nrf2, and a 7-fold and 9-fold increase in heme oxygenase-1 and NAD(P)H-quinone oxidoreductase 1, respectively. The IEBF-protecting effect of OAE involves, most likely, its dual capacity to activate Nrf2 while inhibiting NF-ĸB activation. The extremely low doses of BZF needed to promote such actions warrants extending its IEBF-protective effects to other NSAIDs.

Vitamin E as an essential micronutrient for human health: Common, novel, and unexplored dietary sources.

Vitamin E comprises a group of vitamers that includes tocopherols and tocotrienols. They occur in four homologues according to the number and position of methyl groups attached to the chromanol ring. Vitamin E, a liposoluble antioxidant, may participate as an adjuvant in the prevention and treatment of cardiovascular, neurological, and aging-related diseases. Furthermore, vitamin E has applications in the food industry as a natural additive. In this contribution, the most recent information on the dietary sources of vitamin E, including common, novel, and unexplored sources, is presented. Common edible oils, such as those of corn, olive, palm, rice bran, and peanut, represent the most prominent sources of vitamin E. However, specialty and underutilized oils such as those obtained from tree nuts, fruit seeds, and by-products, emerge as novel sources of this important micronutrient. Complementary studies should examine the tocotrienol content of vitamin E dietary sources to better understand the different biological functions of these vitamers.

Quercetin Oxidation Metabolite Present in Onion Peel Protects Caco-2 Cells against the Oxidative Stress, NF-kB Activation, and Loss of Epithelial Barrier Function Induced by NSAIDs.

The potential of 2-(3,4-dihydroxybenzoyl)-2,4,6-trihydroxy-3(2H)-benzofuranone (BZF), a quercetin oxidation metabolite, and that of a BZF-rich onion peel aqueous extract (OAE) to protect Caco-2 monolayers against the oxidative stress (OS) and an increased permeability (IP) induced by five nonsteroidal anti-inflammatory drugs (NSAIDs) (indomethacin, diclofenac, piroxicam, ibuprofen, and metamizole) were investigated. Under identical OS conditions, the NSAIDs substantially differed in their ability to induce an IP and/or NF-kB activation. The OAE (100 nM BZF) protected in identical magnitude (84-86%) against OS but in a highly dissimilar manner against the IP (18-73%). While all NSAIDs activated NF-kB, the OAE prevented only that induced by indomethacin. Results reveal that the IP has no direct relationship with the OS and that with the exception of indomethacin, the prevention of NSAIDs-induced OS and/or NF-kB activation plays no fundamental role in the IP-protecting effect of OAE. These results warrant the in vivo evaluation of OAE against indomethacin-induced loss of intestinal barrier function.

Phytoene synthase 1 (Psy-1) and lipoxygenase 1 (Lpx-1) Genes Influence on Semolina Yellowness in Wheat Mediterranean Germplasm.

Phytoene synthase 1 (Psy1) and lipoxygenase 1 (Lpx-1) are key genes involved in the synthesis and catalysis of carotenoid pigments in durum wheat, regulating the increase and decrease in these compounds, respectively, resulting in the distinct yellow color of semolina and pasta. Here, we reported new haplotype variants and/or allele combinations of these two genes significantly affecting yellow pigment content in grain and semolina through their effect on carotenoid pigments. To reach the purpose of this work, three complementary approaches were undertaken: the identification of QTLs associated to carotenoid content on a recombinant inbred line (RIL) population, the characterization of a Mediterranean panel of accessions for Psy1 and Lpx-1 genes, and monitoring the expression of Psy1 and Lpx-1 genes during grain filling on two genotypes with contrasting yellow pigments. Our data suggest that Psy1 plays a major role during grain development, contributing to semolina yellowness, and Lpx-1 appears to be more predominant at post-harvest stages and during pasta making.

Insoluble-Bound Polyphenols Released from Guarana Powder: Inhibition of Alpha-Glucosidase and Proanthocyanidin Profile.

The Brazilian Food Supplement Law recently recognized that guarana (Paullinia cupana) contains bioactive substances, hence supporting its role as a functional food ingredient. The health benefits of guarana are associated, at least in part, to its phenolic compounds. However, to the best of our knowledge, there is no literature addressing the presence of phenolic compounds in the fraction containing insoluble-bound compounds and its contribution in terms of alpha-glucosidase inhibition. The concentration of phenolic extracts released from the insoluble-bound fraction required to inhibit 50% of alpha-glucosidase (IC50) activity was 5.8-fold lower than that present in the soluble counterpart. Both fractions exhibited a mixed inhibition mode. Fourteen proanthocyanidins (dimers to tetramers) present in the insoluble-bound fraction were tentatively identified by MALDi-TOF-MS. Future studies aiming at increasing the concentration of the soluble counterpart are deemed necessary. The results presented here enhance the phenolic database of guarana and have a practical impact on the procurement of nutraceuticals and functional ingredients related to the prevention and/or management of type 2 diabetes. The Brazilian normative on food supplements has been recently revised. This study lends support to the future inclusion of guarana powder in the list of sources of proanthocyanidins for the industry of food supplements.

Should we ban total phenolics and antioxidant screening methods? The link between antioxidant potential and activation of NF-κB using phenolic compounds from grape by-products.

Free radical imbalance is associated with several chronic diseases. However, recent controversies have put in check the validity of colorimetric methods to screen the functionality of polyphenols. Therefore, in this study two antioxidant methods, based on chemical reactions, were tested for their ability in anticipating the reduction of the activation of NF-κB using LPS-activated RAW 264.7 macrophages, selected as a biological model. Grape processing by-products from winemaking showed higher total phenolic content (TPC), antioxidant capacity towards peroxyl radical (31.1%) as well as reducing power (39.5%) than those of grape juice by-products. The same trend was observed when these samples were tested against LPS-activated RAW 264.7 macrophages by reducing the activation NF-κB. Feedstocks containing higher TPC and corresponding ORAC and FRAP results translated to higher reduction in the activation of NF-κB (36.5%). Therefore, this contribution demonstrates that colorimetric methods are still important screening tools owing their simplicity and widespread application.

Phenolics from Winemaking By-Products Better Decrease VLDL-Cholesterol and Triacylglycerol Levels than Those of Red Wine in Wistar Rats.

Winemaking by-products account for more than 30% of the grape production, but this inexpensive feedstock has not yet been fully exploited. Accordingly, we evaluated the potential biological activity of winemaking by-products produced with Syrah grapes in comparison with those of the wine produced using the same grape cultivar. Winemaking by-products showed higher contents of total anthocyanins, flavonols, stilbenes, and flavanols than red wine as evaluated by HPLC-DAD-FD (on a dry weight basis). In contrast, red wine was a better source of phenolic acids. However, the contribution of phenolic acids was minor for both samples. Furthermore, equivalent concentration of winemaking by-products (100 mg/kg/d) showed greater biological activity by than that of red wine by decreasing the levels of VLDL-cholesterol and triacylglycerols in Wistar rats. Therefore, this study supports the use of winemaking by-products as an economical source of bioactive phenolics with potential use in the food and nutraceutical industries.

Phenolic acids and flavonoids of peanut by-products: Antioxidant capacity and antimicrobial effects.

Peanut skin (PS) and meal from dry-blanched peanuts (MDBP) were evaluated as sources of phenolic compounds. PS rendered the highest total phenolic content, antioxidant capacity towards ABTS radical cation, DPPH and hydroxyl radicals as well as reducing power. Phenolic acids were present in PS and MDBP whereas proanthocyanidins and monomeric flavonoids were found only in PS as identified by HPLC-DAD-ESI-MSn. Procyanidin-rich extracts prevented oxidation in non-irradiated and gamma-irradiated fish model system. Both extracts inhibited the growth of gram-positive (Bacillus cereus, Staphylococcus aureus, Listeria monocytogenes, Geobacillus stearothermophilus) and gram-negative bacteria (Pseudomonas aeruginosa, Pseudomonas fluorescens, Salmonella Enteritidis, Salmonella Typhimurium, Escherichia coli). Regardless of the strain, phenolic acid-rich extracts showed the lowest minimum inhibitory capacity (MIC); therefore presenting higher antibacterial effect. The MIC of phenolic acid-rich extracts (24-49µgphenolics/mL) was higher but comparable to Ampicillin (10µg/mL). Thus, phenolics in PS and MDBP may serve as antioxidants and antimicrobial compounds.

Identification of phenolic antioxidants and bioactives of pomegranate seeds following juice extraction using HPLC-DAD-ESI-MSn.

Phenolics from free and hydrolyzed fractions of pomegranate juice (PJ) and seeds (PS) were evaluated. In general, total phenolic contents and scavenging of ABTS+, DPPH and hydroxyl radicals, as well as metal chelation of the soluble fraction from PS, were higher than those for PJ. Insoluble-bound phenolics from PS accounted for up to 27% of total scavenging capacity (free+esterified+insoluble-bound). Phenolic acids (13), monomeric flavonoids (8), hydrolysable tannins (12), proanthocyanidin (1) and anthocyanins (12) were tentatively characterized using HPLC-DAD-ESI-MSn. Several compounds were identified for the first time in PJ or PS. The inhibition of DNA damage (induced by hydroxyl and peroxyl radicals), copper-induced LDL-cholesterol peroxidation, as well as alpha-glucosidase and lipase activities were demonstrated, therefore supporting the potential exploitation of PJ and PS as sources of bioactive compounds.

Tocopherols and Tocotrienols in Common and Emerging Dietary Sources: Occurrence, Applications, and Health Benefits.

Edible oils are the major natural dietary sources of tocopherols and tocotrienols, collectively known as tocols. Plant foods with low lipid content usually have negligible quantities of tocols. However, seeds and other plant food processing by-products may serve as alternative sources of edible oils with considerable contents of tocopherols and tocotrienols. Tocopherols are among the most important lipid-soluble antioxidants in food as well as in human and animal tissues. Tocopherols are found in lipid-rich regions of cells (e.g., mitochondrial membranes), fat depots, and lipoproteins such as low-density lipoprotein cholesterol. Their health benefits may also be explained by regulation of gene expression, signal transduction, and modulation of cell functions. Potential health benefits of tocols include prevention of certain types of cancer, heart disease, and other chronic ailments. Although deficiencies of tocopherol are uncommon, a continuous intake from common and novel dietary sources of tocopherols and tocotrienols is advantageous. Thus, this contribution will focus on the relevant literature on common and emerging edible oils as a source of tocols. Potential application and health effects as well as the impact of new cultivars as sources of edible oils and their processing discards are presented. Future trends and drawbacks are also briefly covered.

Phenolic Compounds of Pomegranate Byproducts (Outer Skin, Mesocarp, Divider Membrane) and Their Antioxidant Activities.

Pomegranate peel was separated into outer leathery skin (PS), mesocarp (PM), and divider membrane (PD), and its phenolic compounds were extracted as free (F), esterified (E), and insoluble-bound (B) forms for the first time. The total phenolic content followed the order PD > PM > PS. ABTS(•+), DPPH, and hydroxyl radical scavenging activities and metal chelation were evaluated. In addition, pomegranate peel extracts showed inhibitory effects against α-glucosidase activity, lipase activity, and cupric ion-induced LDL-cholesterol oxidation as well as peroxyl and hydroxyl radical-induced DNA scission. Seventy-nine phenolic compounds were identified using HPLC-DAD-ESI-MS(n) mainly in the form of insoluble-bound. Thirty compounds were identified for the first time. Gallic acid was the major phenolic compound in pomegranate peel, whereas kaempferol 3-O-glucoside was the major flavonoid. Moreover, ellagic acid and monogalloyl-hexoside were the major hydrolyzable tannins, whereas the dominant proanthocyanidin was procyanidin dimers. Proanthocyanidins were detected for the first time.

Enzyme-assisted extraction of phenolics from winemaking by-products: Antioxidant potential and inhibition of alpha-glucosidase and lipase activities.

Phenolics in food and agricultural processing by-products exist in the soluble and insoluble-bound forms. The ability of selected enzymes in improving the extraction of insoluble-bound phenolics from the starting material (experiment I) or the residues containing insoluble-bound phenolics (experiment II) were evaluated. Pronase and Viscozyme improved the extraction of insoluble-bound phenolics as evaluated by total phenolic content, antioxidant potential as determined by ABTS and DPPH assays, and hydroxyl radical scavenging capacity, reducing power as well as evaluation of inhibition of alpha-glucosidase and lipase activities. Viscozyme released higher amounts of gallic acid, catechin, and prodelphinidin dimer A compared to Pronase treatment. Furthermore, p-coumaric and caffeic acids, as well as procyanidin dimer B, were extracted with Viscozyme but not with Pronase treatment. Solubility plays an important role in the bioavailability of phenolic compounds, hence this study may assist in better exploitation of phenolics from winemaking by-products as functional food ingredients and/or supplements.

Antioxidants and bioactivities of free, esterified and insoluble-bound phenolics from berry seed meals.

Phenolic compounds present in the free, soluble ester and insoluble-bound forms of blackberry, black raspberry and blueberry were identified and quantified using high-performance liquid chromatography-diode array detection-electrospray ionisation multistage mass spectrometry. The total phenolics, scavenging activity against hydroxyl and peroxyl radicals, the reducing power and chelating capacity were, in general, in the decreasing order of blackberry>black raspberry>blueberry. Amongst fractions, the order was insoluble-bound>esterified>free. These trends were the same as those found against copper-induced LDL-cholesterol oxidation and supercoiled plasmid DNA strand breakage inhibition induced by both peroxyl and hydroxyl radicals. Extracts were found to contain various levels of phenolic compounds that were specific to each berry seed meal type. Berry seed meals should be considered as a good source of phenolics with potential health benefits. Their full exploitation may be helpful for the food industry and consumers.

Low molecular weight phenolics of grape juice and winemaking byproducts: antioxidant activities and inhibition of oxidation of human low-density lipoprotein cholesterol and DNA strand breakage.

Bioactive compounds belonging to phenolic acids, flavonoids, and proanthocyanidins of grape juice and winemaking byproducts were identified and quantified by HPLC-DAD-ESI-MS(n). The concentration of phenolic compounds in different grape cultivars was in the order Tempranillo > Cora > Syrah > Isabel. The insoluble-bound fraction was most prominent, contributing 63 and 79% to the total for Isabel and Tempranillo, respectively. Juice-processing byproducts had a higher content of free than esterified phenolics, but the opposite was noted for winemaking byproducts. Insoluble-bound phenolics were up to 15 and 10 times more effective as antioxidants than those of free and esterified fractions, respectively, as evaluated by the DPPH, ABTS, and H2O2 scavenging activities and reducing power determinations. In general, insoluble-bound phenolics (100 ppm) were more effective in inhibiting copper-induced human LDL-cholesterol oxidation than free and esterified phenolics, exhibiting equal or higher efficacy than catechin. Phenolic extracts from all fractions inhibited peroxyl radical-induced DNA strand breakage. These findings shed further light for future studies and industrial application of grape byproducts, which may focus not only on the soluble phenolics but also on the insoluble-bound fraction.

Gamma radiation induced oxidation and tocopherols decrease in in-shell, peeled and blanched peanuts.

In-shell, peeled and blanched peanut samples were characterized in relation to proximate composition and fatty acid profile. No difference was found in relation to its proximate composition. The three major fatty acids were palmitic acid, oleic acid, and linoleic acid. In order to investigate irradiation and storage effects, peanut samples were submitted to doses of 0.0, 5.0, 7.5 or 10.0 kGy, stored for six months at room temperature and monitored every three months. Peanuts responded differently to irradiation, particularly with regards to tocopherol contents, primary and secondary oxidation products and oil stability index. Induction periods and tocopherol contents were negatively correlated with irradiation doses and decreased moderately during storage. α-Tocopherol was the most gamma radiation sensitive and peeled samples were the most affected. A positive correlation was found among tocopherol contents and the induction period of the oils extracted from irradiated samples. Gamma radiation and storage time increased oxidation compounds production. If gamma radiation is considered an alternative for industrial scale peanut conservation, in-shell samples are the best feedstock. For the best of our knowledge this is the first article with such results; this way it may be helpful as basis for future studies on gamma radiation of in-shell crops.