Monomeric Flavanols Research Articles

Cocoa Flavanols Protect Human Endothelial Cells from Oxidative Stress

Oxidative stress may cause functional disorders of vascular endothelia which can lead to endothelial apoptosis and thus alter the function and structure of the vascular tissues. Plant antioxidants protect the endothelium against oxidative stress and then become an effective option to treat vascular diseases. Cocoa flavanols have been proven to protect against oxidative stress in cell culture and animal models. In addition, epidemiological and interventional studies strongly suggest that cocoa consumption has numerous beneficial effects on cardiovascular health. The objective of this study was to test the chemo-protective effect of realistic concentrations of a cocoa phenolic extract and its main monomeric flavanol epicatechin on cultured human endothelial cells submitted to an oxidative challenge. Both products efficiently restrained stress-induced reactive oxygen species and biomarkers of oxidative stress such as carbonyl groups and malondialdehyde, and recovered depleted glutathione, antioxidant defences and cell viability. Our results demonstrate for the first time that a polyphenolic extract from cocoa and its main flavonoid protect human endothelial cells against an oxidative insult by modulating oxygen radical generation and antioxidant enzyme and non-enzyme defences.

LC-MS/MS based molecular networking approach for the identification of cocoa phenolic metabolites in human urine

Dietary phenolic compounds are often transformed by gut microbiota prior to absorption. This transformation may modify their structures, producing novel gut flora metabolites associated with numerous health benefits. Traditional mass spectrometry (MS) based approaches for assessing dietary exposure of cocotea (cocoa, coffee and tea) products provided very little information about the modification and fate of dietary phenolics after ingestion, mainly due to limitation of complex sample nature and their data analyses. Mass spectrometry techniques are well-suited to a high-throughput characterization of natural products, however, analyzing MS based data of complex biological matrix is still considered a challenge. In order to overcome such limitations and simplify the analysis of complex MS data, a cocotea based human trial was conducted where MS based molecular networking approach was implemented.To demonstrate the utility of this approach in one of the specific cocotea diets, we have applied it to a diverse collection of human (n = 15) urine samples, who consumed cocoa rich in polyphenols over a 48-h period. This approach illustrated the power of the new strategy, allowing the rapid identification of new analogues of cocoa metabolites after human consumption.Analysis of human urine samples after cocoa consumption revealed (by assignment of unknown metabolites based on the network similarities) that monomeric flavanols are mainly absorbed and transformed directly into their glucuronide and sulfated moieties. Subsequently, the hydroxy and methoxy phenyl-g-velerolactone as well as their smaller metabolites (such as hydroxyphenyl valeric acids, hydroxy and methoxy phenyl propionic acids and their derivates) are indicative of bacterial metabolism of cocoa major flavanols. For the first time, our study exemplifies and highlight the implementation of MS based molecular networking approach in illustrating the tracking of various structural motifs of ingested cocoa phenolics in human based study.

Reliable, accessible and transferable method for the quantification of flavanols and procyanidins in foodstuffs and dietary supplements

Flavanols and procyanidins are plant-derived bioactives that are receiving increasing attention because of their potential health benefits. Analytical tools that can accurately identify and reproducibly quantify these bioactives are critical to researchers for test material characterization, as well as to the food industry and regulators, notably for product labeling. However, the chemical complexity of procyanidins, and the absence of analytical standards have prevented the development of methods that could serve the needs of these different sectors. This report describes the development and validation of a reliable, accessible and transferable method for the quantification of flavanol monomers and procyanidins in cocoa-derived dietary supplements and foodstuffs. To accomplish this, flavanols and procyandins from cocoa, one of the most studied dietary sources of these compounds, were used as a model system. To overcome limitations related to the absence of analytical standards, a cocoa extract was thoroughly characterized for use as a calibrant. It was then used in the development and validation of a method based on reliable and accessible instrumentation, namely HPLC coupled with fluorescence detection. The resulting method permitted the quantification of flavanols and procyanidins in amounts ranging from 2 to 500 mg g-1, with high precision (%RSD 0.2 to 1.9%) and accuracy (100.7 to 102.9%). The method was successfully applied to assess the flavanol and procyanidin content of different cocoa-based commercial products. Furthermore, the high precision of the methods showed the feasibility of using principal component analysis of flavanol and procyanidin profiles to discriminate cocoa-derived products by origin and manufacturing processes. A feature that offers advantages in monitoring product authenticity/adulteration. Overall, these findings support the application of this method for the routine analysis of cocoa flavanols and procyandins.

Flavanol Glycoside Content of Grape Seeds and Skins of Vitis vinifera Varieties Grown in Castilla-La Mancha, Spain.

Glycosylated flavanols (monoglycosides and diglycosides) in skin and seed extracts of Vitis vinifera grapes grown in Castilla-La Mancha (Spain) were investigated using ultra-high-performance liquid chromatography—tandem mass spectrometry (UHPLC-ESI-QQQ-MS/MS). Six grape varieties (Airén, Tempranillo, the recently identified Albillo Dorado, Montonera del Casar, Moribel, and Tinto Fragoso) were studied over two consecutive years (2016 and 2017). A total of twenty monomeric flavanol monoglycosides, four diglycosylated monomers, and three dimeric flavanol monoglycosides were detected in all grape samples. The diversity observed in the composition of glycosylated flavanol in the grape berries suggests a strong influence of variety and grape tissue (skin or seed). Monomeric flavanol glycosides were more abundant in grape seed extracts, in contrast with monoglycosylated dimeric forms. In addition, the glycosylated flavanol content was related to berry color in grape skins, with higher concentrations measured in black grape varieties.

Influence of barley variety and growing locality on the profile of flavonoid polyphenols in malt

Some polyphenol substances are important for beer production as well as for the health benefits of beer consumption. Hop polyphenols have been relatively well researched in the last decade, less attention has been paid to malt. Our study was aimed to determine the influence of the barley variety and growing locality on the profile of proanthocyanidins (PAC), monomeric flavanols, flavonols and their glycosides in malt. Laboratory malts from three barley cultivars grown in seven countries of the European Union were analyzed by the developed LC-HR / MS method with sample preparation by the QuEChERS technique. Dimers of (epi)catechin (37.0%) and dimers (epi)catechin – (epi) gallocatechin (36.4%) dominate the PAC profile of malt, monomer content being low. Flavanols (epi)catechin and (epi)catechin-O-glucoside have been identified and quantified in malts. The flavonols were quercetin and myricetin, their glycosides were not detected. The results suggest varietal dependence of the PAC, monomeric flavanols and flavonols profile and the lesser impact of the growing locality.

Chemical composition and sensory profile of Syrah wines from semiarid tropical Brazil – Rootstock and harvest season effects

This study aims to characterise the chemical compositions and sensorial profiles of wines made with Syrah grapes over two harvest seasons (first and second semester), over in different calendar years, from vines on two different rootstocks (1103P and IAC 313). Wine chemical composition and sensory profile were influenced by both rootstock and harvest season. Syrah wines on IAC 313 were favoured with higher concentrations of, petunidin 3-O-acetylglucoside (3.7 mg/L), malvidin 3-O-acetylglucoside (17.8 mg/L), malvidin 3-O-coumarylglucoside (4.4 mg/L), petunidin 3-O-coumarylglucoside (2.0 mg/L), peonidin 3-O-coumarylglucoside (1.4 mg/L), monomeric flavanols (23.6 mg/L), oligomeric tannins (183.0 mg/L), total condensed tannins (1037.7 mg/L), dimer B1 (9.8 mg/L), B4 (5.0 mg/L), trimer C1 (3.1 mg/L) and calcium (80 mg/L). Syrah wines on 1103P had higher concentrations of total anthocyanins (375.6 mg/L, catechin (8.6 mg/L), epicatechin (12.6 mg/L), dimer B2 (21.0 mg/L), B1 3-O-gallate (1.8 mg/L), B2 3′-O-gallate (6.0 mg/L) and total flavanols (53.4 mg/L). The sensory profiles of the Syrah wines were influenced by rootstock followed by, mainly in relation to aromatic attributes. Syrah wines on IAC 313 were scored more highly by the tasters.

The Effect of Cocoa Flavanols and Metabolites on β‐cell Proliferation and Survival

Currently, an estimated 30.3 million Americans have diabetes, and experts predict 54.9 million Americans will have diabetes by 2030. The increasing prevalence of Type 2 diabetes (T2D) accounts for much of this expected growth. T2D is characterized by insulin resistance, which eventually leads to a reduction in functional β‐cell mass. Thus, treatments that increase β‐cell proliferation, survival, and function have the potential to reverse the effects of diabetes. One treatment that has shown promise is the use of dietary cocoa flavanols. These compounds have shown the ability to prevent the onset of diabetes in mice. Furthermore, a recent study conducted by our lab demonstrated that monomeric cocoa flavanols improved insulin secretion by increasing mitochondrial respiration. In addition to stimulating insulin secretion, we hypothesize that cocoa flavanols may also confer antidiabetic effects by promoting β‐cell proliferation and protecting β‐cells from lipotoxicity‐mediated cell death. To that end, we present data that shows the effect of epicatechin (monomeric flavanol), oligomeric, and polymeric cocoa flavanols on β‐cell proliferation and survival. Our results demonstrated that epicatechin enhances β‐cell proliferation. This finding is important because only monomeric flavanols have been found at significant levels in circulation. Notably, several gut bacteria‐derived cocoa metabolites have also been observed and identified. Consequently, we tested the effects of several cocoa metabolites on β‐cell proliferation and survival as well. We found that the cocoa metabolite hippuric acid, also enhances β‐cell proliferation. Additionally, we suggest a mechanism for the observed increase in proliferation caused by epicatechin and hippuric acid. These findings strengthen the growing body of evidence that dietary cocoa flavanols can ameliorate the effects of T2D.Support or Funding InformationDiabetes Action Research and Education Foundation grant #475 to JSTThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Evolution of Flavanol Glycosides during Red Grape Fermentation.

Monomeric and dimeric flavanol glycosides were quantified by UHPLC-MRM in Syrah (SYR) and Grenache (GRE) grapes and in their corresponding wines for the first time. Quantities were extremely variable depending on grape tissue (seeds or skins) and during fermentation. Overall, 22 monomeric and dimeric mono- and diglycosides were determined with concentrations ranging from 0.7 nanograms to 0.700 micrograms per gram of grape tissue, and 0 to 60 micrograms per liter for wines. The evolution of the glycosides’ composition during winemaking suggests that almost all these compounds originate in the grapes themselves and display different extraction kinetics during winemaking. One isomer of the monomeric (epi) flavanol monoglycosides seemed to be biosynthesized by yeasts during wine fermentation. The sharp decrease in concentration of some isomers at the late stages of fermentation or after pressing suggests that some grape glycosidase activities convert these compounds into non-glycosylated flavanols.

Assessing the respective contributions of dietary flavanol monomers and procyanidins in mediating cardiovascular effects in humans: randomized, controlled, double-masked intervention trial

ABSTRACTBackgroundFlavanols are an important class of food bioactives that can improve vascular function even in healthy subjects. Cocoa flavanols (CFs) are composed principally of the monomer (−)-epicatechin (∼20%), with a degree of polymerisation (DP) of 1 (DP1), and oligomeric procyanidins (∼80%, DP2–10).ObjectiveOur objective was to investigate the relative contribution of procyanidins and (−)-epicatechin to CF intake–related improvements in vascular function in healthy volunteers.DesignIn a randomized, controlled, double-masked, parallel-group dietary intervention trial, 45 healthy men (aged 18–35 y) consumed the following once daily for 1 mo: 1) a DP1–10 cocoa extract containing 130 mg (−)-epicatechin and 560 mg procyanidins, 2) a DP2–10 cocoa extract containing 20 mg (−)-epicatechin and 540 mg procyanidins, or 3) a control capsule, which was flavanol-free but had identical micro- and macronutrient composition.ResultsConsumption of DP1–10, but not of either DP2–10 or the control capsule, significantly increased flow-mediated vasodilation (primary endpoint) and the concentration of structurally related (−)-epicatechin metabolites (SREMs) in the circulatory system while decreasing pulse wave velocity and blood pressure. Total cholesterol significantly decreased after daily intake of both DP1–10 and DP2–10 as compared with the control.ConclusionsCF-related improvements in vascular function are predominantly related to the intake of flavanol monomers and circulating SREMs in healthy humans but not to the more abundant procyanidins and gut microbiome–derived CF catabolites. Reduction in total cholesterol was linked to consumption of procyanidins but not necessarily to that of (−)-epicatechin. This trial was registered at clinicaltrials.gov as NCT02728466.

Untargeted Metabolomics Analytical Strategy Based on Liquid Chromatography/Electrospray Ionization Linear Ion Trap Quadrupole/Orbitrap Mass Spectrometry for Discovering New Polyphenol Metabolites in Human Biofluids after Acute Ingestion of Vaccinium myrtillus Berry Supplement.

In this work, liquid chromatography, coupled with an electrospray ionization hybrid linear ion trap quadrupole/Orbitrap mass spectrometry, has been used to accurately identify polyphenol metabolites in human serum and urine after acute ingestion of a V. myrtillus berry supplement. The supplement was obtained by cryo-milling of bilberries, which were freeze-dried within 1week after their harvesting, to maintain the berry native composition. Thirty-six derivatives of benzoic acids, hydroxyhippuric acids, cinnamic acids, phenylpropionic acids, phenylvaleric acids, phenylpentenoic acids and abscisic acid, together with two berry-native anthocyanins, one flavonol metabolite and two catechol derivatives were putatively identified in the investigated biofluids. The annotated compounds included 13 metabolites, among glucuronides and sulphates of phenylvaleric and phenylpentenoic acids, which have been identified for the first time in human biofluids after ingestion of V. myrtillus berries. It should be emphasized that the presence of phenylvaleric and phenylpentenoic acid derivatives is in agreement with their origin from fruit native flavanol monomers and oligomers, which are widely distributed in Vaccinium berries, but usually overlooked in metabolomics studies regarding bilberry. The identification of these compounds confirmed the key-role of untargeted metabolomics approach in the discovery of new metabolites which could be biologically active. Graphical Abstract.

Lack of acute or chronic effects of epicatechin-rich and procyanidin-rich apple extracts on blood pressure and cardiometabolic biomarkers in adults with moderately elevated blood pressure: a randomized, placebo-controlled crossover trial

The reported effects of flavanol-rich foods such as cocoa, dark chocolate, and apples on blood pressure and endothelial function may be due to the monomeric flavanols [mainly (-)-epicatechin (EC)], the oligomeric flavanols [procyanidins (PCs)], or other components. Reports of well-controlled intervention studies that test the effects of isolated oligomeric flavanols on biomarkers of cardiovascular health are lacking. We studied the acute and chronic effects of an EC-rich apple flavanol extract and isolated apple PCs on systolic blood pressure (BP) and other cardiometabolic biomarkers. Forty-two healthy men and women with moderately elevated BP completed this randomized, double-blind, placebo-controlled, 4-arm crossover trial. Participants ingested a single dose of an apple flavanol extract (70 mg monomeric flavanols, 65 mg PCs), a double dose of this extract (140 mg monomeric flavanols, 130 mg PCs), an apple PC extract (130 mg PCs, 6.5 mg monomeric flavanols), or placebo capsules once daily for 4 wk, in random order. Biomarkers of cardiovascular disease risk and vascular function were measured before and 2 h after ingestion of the first dose and after the 4-wk intervention. Compared with placebo, none of the isolated flavanol treatments significantly (P<0.05) changed systolic or diastolic BP (peripheral and aortic), plasma nitric oxide (NO) reaction products, or measures of arterial stiffness (carotid femoral pulse-wave velocity, brachial-ankle pulse-wave velocity, or Augmentation Index) after 2 h or 4 wk of the intervention. There were no changes in plasma endogenous metabolite profiles or circulating NO; endothelin 1; total, HDL, or LDL cholesterol; triglycerides; fasting glucose; fructosamine; or insulin after 4 wk of the intervention. Our data suggest that, in isolation, neither monomeric flavanols nor PCs affect BP, blood lipid profiles, endothelial function, or glucose control in individuals with moderately elevated BP. The reported benefits of consuming flavanol-rich cocoa, chocolate, and apple products appear to be dependent on other components, which may work in combination with monomeric flavanols and PCs. This trial was registered at www.clinicaltrials.gov as NCT02013856.

Identification and Quantification of Flavanol Glycosides in Vitis vinifera Grape Seeds and Skins during Ripening.

Monomeric and dimeric flavanol glycosides were analyzed in Vitis vinifera grapes and seeds during ripening. An analytical method using ultra-high performance liquid chromatography coupled with a triple quadrupole mass spectrometry (UHPLC-ESI-QQQ-MS/MS) in multiple reaction monitoring (MRM) mode was employed. Three grape varieties (Merlot, Syrah and Tannat) were chosen and grape berries were sampled at different stages of development. Ten monoglycosylated and six diglycosylated flavanol monomers were detected. Twelve monoglycosylated and three diglycosylated flavanol dimers were also detected for all three grape varieties. All diglycosides were detected for the first time in Vitis vinifera grapes, though some of these compounds were only detected in skins or seeds. Furthermore, the evolution of all these compounds was studied, and a decrease in monomeric (epi) catechin monoglycosides was observed during ripening for Tannat, Merlot and Syrah grape skins. The dimers would appear to accumulate in skin tissues up to mid-summer (after veraison) and decrease when grape berries reached maturity.

The Impact of Epicatechin on Human Cognition: The Role of Cerebral Blood Flow.

Epicatechin is a monomeric flavanol found in food sources such as tea, apples, berries and cocoa. A number of large-scale epidemiological studies have demonstrated an association between the consumption of these foods and cognitive function, as well as improved blood flow. The aim of this review is to summarise the evidence from intervention studies to clarify the effect of epicatechin on cognition and to consider the role of increased cerebral blood flow as a mechanism for any effects. The effects of epicatechin as consumed in cocoa are, therefore, reviewed here as this represents the only dietary source where it is purported to be the major active component. Our main findings are that a) the positive modulation of tasks that involve memory, executive function and processing speed in older adults; b) the cognitive benefits are more often shown in studies containing more than 50 mg epicatechin/day; and c) all studies with a duration of 28 days or longer in populations >50 years old demonstrate a cognitive improvement. However, as highlighted by this review, it is not currently possible to attribute effects solely to epicatechin without consideration of synergies. In order to overcome this issue, further studies examining the cognitive effects of epicatechin in isolation are required. The role of cerebral blood flow also requires further investigation through simultaneous measurement alongside cognitive function.

Prospective study of flavonoid intake and risk of primary open-angle glaucoma.

To evaluate the association between flavonoid intake and incident primary open-angle glaucoma (POAG). We followed 65516 women from the Nurses' Health Study (from 1984) and 42156 men from the Health Professionals Follow-up Study (from 1986) biennially to 2012, who were 40+ years old, free of POAG, and reported eye examinations. Dietary flavonoid intake was assessed with validated repeated semi-quantitative food frequency questionnaires. Incident POAG cases (n=1575) were confirmed with medical record review. Cohort-specific multivariable-adjusted relative risks (RRs) and 95% confidence intervals (CIs) were calculated and meta-analysed. Total flavonoid intake was not associated with risk of POAG development [RR for highest (Q5: median ~645mg/day) versus lowest quintile (Q1: ~130mg/day)=0.91 (95% CI=0.77, 1.08); p for trend (p-trend)=0.19]; the flavonoid subclasses of flavones, flavanones, polymeric flavanols or anthocyanidins were also not associated (Q5 versus Q1 comparison p-values ≥0.05 and p-trend ≥0.09). Higher intakes of flavonols and monomeric flavanols were nominally associated with lower POAG risk, based on the Q5 versus Q1 comparisons or p-trends. The Q5 versus Q1 comparison RRs were: for flavonols, 0.82 (95% CI=0.69, 0.97; p-trend=0.05; ~28 versus ~8mg/day), and for monomeric flavanols, 0.86 (95% CI=0.72, 1.02; p-trend=0.04; ~110 versus 10mg/day). The food/beverage that contributed most to both the variation of flavonols and monomeric flavanols was tea; consuming ~2 cups/day was associated with 18% lower POAG risk (RR=0.82; 95% CI=0.68, 0.99; p-trend=0.02). Total flavonoid intake was not associated with POAG risk. Greater intakes of flavonols and monomeric flavanols and of tea showed suggestive modest associations with lower risk; these results need confirmation.

Effect of the harvest season on phenolic composition and oenological parameters of grapes and wines cv. ‘Touriga Nacional’ (Vitis viniferaL.) produced under tropical semi-arid climate, in the state of Pernambuco, Brazil

The northeastern region of São Francisco Valley is the third largest wine producer in Brazil, differentiated by the production of at least two harvests per year, generally in the first and second semesters, respectively. The productive cycle of the vine in the first semester is higher than in the second semester, mainly due to differences in temperature, with maximum and average temperatures of approximately 30 ºC and 26 ºC, respectively. Second semester is characterized by higher temperatures (summer season) and that promote a rapid maturation of the grapes. The objective of this work was to evaluate the phenolic composition, other oenological parameters and the sensorial profile of grapes and wines of cv. 'Touriga Nacional' (Vitis viniferaL.), characterizing grapes and wines from four harvest seasons, two of them referring to the first half of the year and two referring to the second half of the year, using a broad analytical approach. The experimental design consisted of three randomized blocks, with sixty plants marked in different lines and positions (twenty plants in each block at the beginning, middle and end of the vineyard). The results showed that there was a harvesting effect for some of the grape chemical compounds (monomeric anthocyanins, organic acids, condensed tannins and monomeric flavanols, for example) as well as in wines (calcium, monomeric anthocyanins, condensed tannins and monomeric flavanols). Regarding the sensorial profile of the wines, it was possible to observe that higher scores were attributed to floral aroma in wines from the first harvest (first half of the year). Fruity, spice and empyreumatic aromas were higher in wines from second harvest season (second half of the year). For other attributes, there was a tendency for higher variations between the period of study (2014 – 2017) than between the harvest seasons.

The matrix of fruit & vegetables modulates the gastrointestinal bioaccessibility of polyphenols and their impact on dietary protein digestibility

Fruit and vegetables (F&V) polyphenols have numerous positive health effects, ascribed either to their antioxidant activity within the gastrointestinal tract (GIT) or to bioactivity of their absorbed metabolites. The effect of the F&V matrix on the gastrointestinal bioaccessibility of polyphenols was investigated along with its possible interaction with protein digestion. Minipigs were fed a complete meal with either cubed F&V (apple, plum, artichoke) added, or the corresponding phenolic extract (PE). Gastric and ileal chymes were kinetically collected over the postprandial period. The overall polyphenol bioaccessibility in the stomach was found to be 1.5% and 3.1% after F&V and PE consumption, respectively. The lower release rate from artichoke than from apple showed evidence of a plant effect. Flavanol monomers and glucoside conjugates were not recovered in the ileum in agreement with their absorption in the upper GIT. Interestingly, PE, but not F&V, significantly decreased the speed and efficiency of dietary protein digestion.

Degradation kinetics of encapsulated grape skin phenolics and micronized grape skins in various water activity environments and criteria to develop wide-ranging and tailor-made food applications

Abstract Micronized grape skin powder (GS) and maltodextrin-encapsulated grape skin phenolics (eGSP) were recovered from winemaking byproducts as potential food ingredients. Hygroscopicity was higher in eGSP than in GS. Both eGSP and GS had intense color and less fermented odor than the wet GS. Phenolic content, antioxidant activity and inhibitory effectiveness towards enzymes related to hyperglycemia damage were ~ double in eGSP than in GS. During storage, the rate of phenolic degradation diminished with decreasing aw from 0.75 to 0.11. Anthocyanins and proanthocyanidins were less stable than monomeric flavanols and flavonols. The rate of decrease in antioxidant activity was lower compared to the extent of phenolic degradation. At aw 0.11 no degradation was observed in eGSP, while anthocyanin and proanthocyanidin contents slightly decreased in GS (k ∗ 103 in the range 0.69–2.94 d− 1). Criteria for GS and eGSP storage were defined in relation to their final uses. Industrial relevance The conversion of winemaking by products into value added products is considered the unique strategy to overcome the cost of not recycling, including waste disposal and decontamination of affected areas. As winemaking is a seasonal activity, long-term stability of recovered byproducts is needed for their further utilization. GS and eGSP represent potential value-added food ingredients for wide-ranging applications (antioxidant, colorant, phenolic sources) and tailor-made functionalities (inhibitors of enzymes related to hyperglycemia). The results obtained led to the definition of criteria for GS and eGSP storage, which depend on their final use in foods, as illustrated by two discussed scenarios.

Subchronic treatment with grape-seed phenolics inhibits ghrelin production despite a short-term stimulation of ghrelin secretion produced by bitter-sensing flavanols.

Grape-seed phenolic compounds have recently been described as satiating agents in rats when administered as a whole phenolic extract (GSPE). This satiating effect may involve the release of satiating gut hormones such as GLP-1, although a short-term increase in the orexigenic hormone ghrelin was also reported. In this study, we investigated the short- and long-term effects of GSPE in rats, focusing on the role of the main grape-seed phenolics in ghrelin secretion. GSPE produced a short-term increase in plasma ghrelin in rats after an acute treatment. A mouse ghrelinoma cell line was used to test the effects of the main pure grape-seed phenolic compounds on ghrelin release. Monomeric flavanols stimulated ghrelin secretion by activating bitter taste receptors. In contrast, gallic acid (GA) and oligomeric flavanols inhibited ghrelin release. The ghrelin-inhibiting effects of GA were confirmed in rats and in rat duodenal segments. One day after the last dose of a subchronic treatment, GSPE decreased plasma ghrelin in rats, ghrelin secretion in intestinal segments, and ghrelin mRNA expression in stomach. The sustained satiating effects of GSPE are related to a long-term decrease in ghrelin expression. GA and oligomeric flavanols play a ghrelin-inhibiting role in this process.

Optimization of Ultrasound-Assisted Extraction and Rapid Resolution Analysis of Flavanols and Methylxanthines for the Quality Control of Cocoa-Derived Products

The aim of this study was to develop a rapid and reliable method to quantify cocoa methylxanthines, flavanol monomers, and its major procyanidins. To accomplish this goal, an ultrasound-assisted extraction process was optimized by response surface methodology. In addition, an improved separation of these compounds was carried out on a sub-2-μm particles column while employing a conventional high-performance liquid chromatography (HPLC) instrument. Our results show that the highest extraction yields are obtained by using a 66:44 (v/v) ethanol/water mixture and performing the extraction at least during 20 min. The qualitative and quantitative analyses of cocoa metabolites were achieved in less than 12 min. Moreover, the optimized analytical approach was fully validated and applied to the analysis of cocoa samples collected at five different processing stages. The results show that the approach developed herein significantly increased the extraction efficiency, reaching concentrations approximately 20 % higher than that of the official Association of Analytical Community (AOAC) method for the analysis of polyphenols in cocoa-based matrices.

Impact of chemical structure of flavanol monomers and condensed tannins on in vitro anthelmintic activity against bovine nematodes.

Plants containing condensed tannins (CT) may have potential to control gastrointestinal nematodes (GIN) of cattle. The aim was to investigate the anthelmintic activities of four flavan-3-ols, two galloyl derivatives and 14 purified CT fractions, and to define which structural features of CT determine the anti-parasitic effects against the main cattle nematodes. We used in vitro tests targeting L1 larvae (feeding inhibition assay) and adults (motility assay) of Ostertagia ostertagi and Cooperia oncophora. In the larval feeding inhibition assay, O. ostertagi L1 were significantly more susceptible to all CT fractions than C. oncophora L1. The mean degree of polymerization of CT (i.e. average size) was the most important structural parameter: large CT reduced larval feeding more than small CT. The flavan-3-ols of prodelphinidin (PD)-type tannins had a stronger negative influence on parasite activity than the stereochemistry, i.e. cis- vs trans-configurations, or the presence of a gallate group. In contrast, for C. oncophora high reductions in the motility of larvae and adult worms were strongly related with a higher percentage of PDs within the CT fractions while there was no effect of size. Overall, the size and the percentage of PDs within CT seemed to be the most important parameters that influence anti-parasitic activity.