Search

Tyrosinases are essential metal-containing enzymes in the biosynthesis of melanin, therefore responsible for pigmentation of the skin. The upregulation of tyrosinase enzyme activities leads to hyperpigmentation that will become a health problems and interfere psychosocially. Inhibition of tyrosinase enzyme activity, both competitive and non-competitive become widely developed for most anti hyperpigmentation agent. Natural antioxidants are one of the potential compounds for this purpose. Red grape seeds contain high levels of antioxidant compounds, such as procyanidin, prodelphinidin, and propelargonidin. In this research in silico studies, including molecular docking, molecular dynamics simulations, and toxicity predictions, were used to assess the activity of the three molecules of polyphenolic compounds on macromolecules of the tyrosinase enzyme. Molecular docking studies show that the compound propelargonidin has the highest affinity against the macromolecule of the tyrosinase enzyme, with a binding free energy value of −32.87 kJ/mol. These results were confirmed in molecular dynamics simulations that show strong interactions at the macromolecular active site of the tyrosinase enzyme. Toxicity prediction results show that the three polyphenolic compound molecules were classified in the High-Class Category, which shows that safety is not guaranteed, but is likely, not carcinogenic and nongenotoxic. Therefore, the compound propelargonidin is predicted to be able to interact strongly with the tyrosinase enzyme. The results in this research are useful for further study in the development of tyrosinase enzyme inhibitors.

The purpose of this research was to identify metabolite change during barley (Hordeum vulgare) germination and reveal active principles for the anti-wrinkle activity. Barley was germinated with deionized water (DW) and mineral-rich water (MRW) for the comparison of the effect of mineral contents on the metabolites changes during germination. The effects of germinated barley extracts (GBEs) on collagen production and collagenase inhibition were evaluated in vitro using human dermal fibroblasts (HDFs). A pronounced anti-wrinkle activity was observed in the test group treated with the MRW-GBEs. In order to find out the active components related to the anti-wrinkle activity, an orthogonal projection to latent structure-discriminant analysis (OPLS-DA) was performed, using the data from secondary metabolites profiling conducted by UPLC–PDA–ESI–MS. The anti-wrinkle activity of MRW-GBEs was revealed to be associated with the increase of oligomeric compounds of procyanidin and prodelphinidin, indicating that it can be used as an active ingredient for anti-wrinkle agents.

In this study, the combined effect of partial postharvest dehydration and long-term ozone treatment was evaluated at 10 and 20% weight loss as a strategy to induce compositional changes in grape skin flavanols. Two separate trials were carried out in thermohygrometric-controlled chambers at 20 °C and 70% relative humidity. The first trial was conducted under an ozone-enriched atmosphere at 30 μL/L, whereas the second trial was performed under an air atmosphere as a control. Two red wine grape varieties were studied, Barbera and Nebbiolo (Vitis vinifera L.), for their different phenolic composition. Berry skin flavanol composition was determined by high-performance liquid chromatography after phloroglucinolysis and size-exclusion chromatography. The results showed that dehydration and ozone effects were variety-dependent. In Barbera skins, being characterized by lower proanthocyanidin contents, the two effects were significant and their combination showed interesting advantages related to lower proanthocyanidin loss as well as higher prodelphinidin and lower galloylation percentages. In Nebbiolo, skin flavanol composition was barely affected.

Proanthocyanidins are plant specialized metabolites which are beneficial to animal nutrition and health. This study determined how proanthocyanidin profiles of sainfoin (Onobrychis viciifolia) and birdsfoot trefoil (Lotus corniculatus) change during the forage conservation process and along the digestive tract of lamb. We determined soluble, protein- and fiber-bound proanthocyanidins by spectrophotometric methods and soluble proanthocyanidin profiles by UPLC-MS/MS. During the conservation process, the total proanthocyanidin contents reduced in both forages and the relative proportion of insoluble proanthocyanidins increased, especially in sainfoin. The soluble proanthocyanidins, their mean degree of polymerization and the relative prodelphinidin share declined in both feed species. In the abomasum of lambs fed sainfoin silage, most of the proanthocyanidins were in insoluble form bound to proteins and fibers, but in the small and large intestines, the proportion of soluble proanthocyanidins increased again. For lambs fed birdsfoot trefoil, the trend was not so clear as proanthocyanidins were already mainly soluble in the abomasum. Nevertheless, a large part of soluble proanthocyanidins was recovered in the digestive tract but could not be detected by the UPLC-MS/MS method used. This study suggests that proanthocyanidins have probably been metabolized in the digestive tract by the resident microbiota.

Proanthocyanidins (PAs) are highly bioactive plant specialized metabolites. One of their most characteristic features is their ability to precipitate proteins. In this study, eleven plant species were used to study the structure&ndash, activity patterns between PAs and their protein precipitation capacity (PPC) with bovine serum albumin. To obtain a comprehensive selection of PAs with highly variable procyanidin to prodelphinidin ratios and mean degree of polymerizations, nearly 350 subfractions were produced from the eleven plant species by semi-preparative liquid chromatography. Their PA composition was defined by tandem mass spectrometry and high-resolution mass spectrometry, and their PPC was measured with a turbidimetry-based well-plate reader assay. The distribution of the PPC within plant species varied significantly. The mean degree of polymerization of the PAs had a strong correlation with the PPC (r = 0.79). The other structural features were significant from the PPC point of view as well, but they contributed to the PPC in different ways in different plant species. Retention time, prodelphinidin proportion, and mean degree of polymerization explained 64% of the measured variance of the PPC.

Berries of genus Vaccinium are rich in flavonoids and proanthocyanidins (PAs). We studied the PA composition and biosynthesis in bilberry (Vaccinium myrtillus L.) tissues and during fruit development. Soluble PAs, analyzed by UHPLC–MS/MS, were most abundant in stem and rhizome with the mean PA polymerization level varying between 4 and 6 in all tissues. Both A- and B-type PAs were present in all tissues. Procyanidin subunits were more common than prodelphinidin subunits in PAs. During fruit ripening, the amount of procyanidin subunits decreased while prodelphinidin subunits and F3′5′H expression increased, indicating a shift in biosynthesis toward the delphinidin branch of the flavonoid pathway. Epicatechin was the most abundant flavan-3-ol in all tissues. Expression of ANR and three isolated LAR genes, analyzed by qRT-PCR, showed connection to accumulation of PAs and flavan-3-ols biosynthesized from different flavonoid branches. Insoluble PAs accumulated during berry development, suggesting that PAs are not recycled after biosynthesis.

Isoetes sinensis Palmer is a critically endangered, first-class protected plant in China. Until now, researchers have primarily focused on the ultrastructure, phylogeny, and transcriptomes of the plant. However, flavonoid profiles and bioactivity of I. sinensis have not been extensively investigated. To develop the endangered I. sinensis for edible and medicinal purposes, flavonoid content, chemical constitution, and antioxidant activities were investigated in this study. Results revealed the following. 1) The total flavonoid content was determined as 10.74 ± 0.25 mg/g., 2) Antioxidant activities were stronger than most ferns, especially ABTS free radical scavenging activities. 3) Four flavones, containing apigenin, apigenin-7-glucuronide, acacetin-7-O-glcopyranoside, and homoplantageninisoetin; four flavonols, namely, isoetin, kaempferol-3-O-glucoside, quercetin-3-O-[6”-O-(3-hydroxy-3-methylglutaryl)-β-D-glucopyranoside], and limocitrin-Neo; one prodelphinidin (procyanidins;) and one nothofagin (dihydrochalcone) were tentatively identified in the mass spectrometry-DAD (254nm) chromatograms. This study was the first to report on flavonoid content and antioxidant activities of I. sinensis. Stronger antioxidant activity and flavonoid content suggests that the endangered I. sinensis is an important and potentially edible and medicinal plant.

Proanthocyanidins (PAC) were isolated from pecan kernel and shell; one fraction was obtained from kernel (KR I ) and 3 fractions from shell (SH I , SH II and SH III ). The average degree of polymerization (ADP) of these fractions was estimated by phloroglucinolysis and the coupling of butanolysis and vanillin reactions. ADP decreased in the order SH I > SH II > SH III > KR I . Prodelphinidin in monomer distribution decreased in the same order. Fractions were tested as inhibitors of carbohydrate, lipid and protein hydrolysis in a simulated human digestive system. Fraction KR I was the best inhibitor. Fraction SH III , inhibited carbohydrate and lipid hydrolysis but was a poor inhibitor of proteolysis. This suggests that PAC oligomers from pecan kernel and shell could help to slow dietary carbohydrate and fat absorption, and that structural features like monomer composition and ADP are responsible from their different inhibitory potency.

Astringency is a parameter of great importance for wine quality. During wine maturation in wooden barrels it can be enhanced by tannins extracted by the wood. However, their concentration is not directly correlated with astringency. Mean Degree of Polymerization (mDP) is often employed to characterize the structural characteristics of tannins in wines, providing also information on their proanthocyanidin content. The prodelphinidin content (%P), as well as the percentage of epigallocatechin (EGC) in extension and terminal units, have been negatively linked with astringency. Since the intensity of astringency is a decisive factor for the quality of a barrel-aged wine in this experiment, the structural characteristics of wine proanthocyanidins were determined in four different Greek wines, two white (Vilana and Dafni) and two red (Kotsifali and Mandilari). All wines matured for nine months in different types of containers (stainless steel tanks and barrels made of different woods). The prodelphinidin (terminal or extension) content was determined by means of phloroglucinolysis, followed by HPLC-UV-MS analysis, while astringency was assessed by a panel of trained tasters. Multivariate analysis showed that only white wines that matured in Acacia barrels contained high %P values independently of the variety used and were characterized as less astringent by the panel. Moreover, the determination of %extension and terminal EGC units revealed that the tannins of the wines maturing in Acacia barrels, contain higher amounts of EGC as terminal units. These findings could be used by winemakers in order to select the appropriate type of barrel according to the type of wine. Keywords: Astringency, proanthocyanidin, tannin, barrel

This data article contains raw and processed data related to research published in Teixeira et al. (2016) [1]. Here we introduce data acquired from the synthesis of a prodelphinidin dimer gallate. All synthesis steps are described and a dataset for the removal of the protecting on prodelphinidin synthesis is presented. With hydrogenolysis in situ with triethylsilane the hydrogen required is produced and used at the same time, making the reaction possible without resorting to bottled hydrogen. Full NMR and HPLC-ESI-MS analysis data is also provided. Keywords: Prodelphinidins, Synthesis, Hydrogenation

Our study investigated the effects of condensed tannins (CT) on rumen in vitro methane (CH4) production and fermentation characteristics by incubating lucerne in buffered rumen fluid in combination with different CT extracts at 0 (control), 40, 80 and 120 g CT/kg of substrate DM. Condensed tannins were extracted from four sainfoin accessions: Rees ‘A’, CPI63763, Cotswold Common and CPI63767. Gas production (GP) was measured using a fully automated GP apparatus with CH4 measured at distinct time points. Condensed tannins differed substantially in terms of polymer size and varied from 13 (Rees ‘A’) to 73 (CPI63767) mean degree of polymerization, but had relatively similar characteristics in terms of CT content, procyanidin: prodelphinidin (PC: PD) and cis:trans ratios. Compared to control, addition of CT from CPI63767 and CPI63763 at 80 and 120 g CT/kg of substrate DM reduced CH4 by 43% and 65%, and by 23% and 57%, respectively, after 24-h incubation. Similarly, CT from Rees ‘A’ and Cotswold Common reduced CH4 by 26% and 46%, and by 28% and 46% respectively. Addition of increasing level of CT linearly reduced the maximum rates of GP and CH4 production, and the estimated in vitro organic matter digestibility. There was a negative linear and quadratic (p < 0.01) relation between CT concentration and total volatile fatty acid (VFA) production. Inclusion of 80 and 120 g CT/kg of substrate DM reduced (p < 0.001) branched-chain VFA production and acetate: propionate ratio and was lowest for CPI63767. A decrease in proteolytic activity as indirectly shown by a change in VFA composition favouring a shift towards propionate and reduction in branched-chain VFA production varied with type of CT and was highest for CPI63767. In conclusion, these results suggest that tannin polymer size is an important factor affecting in vitro CH4 production which may be linked to the CT interaction with dietary substrate or microbial cells.

The objective of this work was to characterize condensed tannins (CT) from six tropical forage legumes and to determine their biological activity. The monomers propelargonidin, prodelphinidin and procyanidin were analyzed, as well as extractable condensed tannin (ECT), protein-bound CT (PBCT) and fiber-bound CT (FBCT), molecular weight, degree of polymerization, polydispersity index, and biological activity by protein precipitate by phenols (PPP) of leaves of the legumes Cajanus cajan, Gliricidia sepium, Stylosanthes capitata x Stylosanthes macrocephala (stylo), Flemingia macrophylla, Cratylia argentea, and Mimosa caesalpiniifolia, and of the bark of this latter species. Differences were observed in the concentrations of ECT, PBCT, PPP, and total condensed tannin among species, but not in that of FBCT. The highest value of PBCT occurred in F. macrophylla. Total CT varied from nondetected concentration in C. argentea to the highest concentration in M. caesalpiniifolia leaves that contain the greatest levels of PPP. No differences were observed for polymerization degree in stylo, F. macrophylla, and M. caesalpiniifolia. Leaves of stylo, C. cajan, and G. sepium, all containing between 20 and 50 g kg-1 total CT, should be beneficial CT sources, if offered as sole feeds in ruminant diets. The ratio of prodelphinidin:procyanidin varies from 10:80 (stylo) to 65:35 (F. machrophylla), and propelargonidin is only determined in C. argentea. Resumo: O objetivo deste trabalho foi caracterizar taninos condensados de seis leguminosas forrageiras e determinar sua atividade biológica. Os monômeros propelargonidina, prodelfinidina e procianidina foram analisados, assim como taninos condensados extraíveis (TCE), taninos condensados ligados a proteínas (TCLP) e ligados a fibras (TCLF), massa molecular, grau de polimerização, índice de polidispersão e atividade biológica, por meio de proteínas precipitadas por fenóis (PPF), em folhas das leguminosas Cajanus cajan, Gliricidia sepium, Stylosanthes capitata x Stylosanthes macrocephala (estilosantes), Flemingia macrophylla, Cratylia argentea e Mimosa caesalpiniifolia, e na casca desta última espécie. Foram observadas diferenças nas concentrações de TCE, TCLP, PPF e total de taninos condensados entre as espécies, mas não na de TCLF. O maior valor de TCLP ocorreu em F. macrophylla. O total de TC variou de não detectável em C. argentea à maior concentração em folhas de M. caesalpiniifolia, que contêm os maiores níveis de PPF. Não se observaram diferenças quanto ao grau de polimerização em estilosantes, F. macrophylla e M. caesalpiniifolia. Folhas de estilosantes, C. cajan e G. sepium, todas com total de TC entre 20 e 50 g kg-1, poderiam ser fontes benéficas de TC, se oferecidas como única fonte de alimentos em dietas de ruminantes. A proporção prodelfinidina:procianidina variou de 10:80 (stylo) a 65:35 (F. machrophylla), e a propelargonidina é determinada somente em C. argentea.

Salal ( Gaultheria shallon Pursh.) is a wild perennial shrub of the Ericaceae and common in coastal forests of western North America, and its berries were an important traditional food for First Nations in British Columbia. Salal berries were investigated for phytochemical content and antioxidant capacity over the course of fruit development. The proanthocyanidin content was extremely high in young berries (280.7 mg/g dry wt) but dropped during development to 52.8 mg/g dry wt. By contrast, anthocyanins accumulated only at the late berry stages. Total antioxidant capacity, as measured by the 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) method, reflected both proanthocyanidin and anthocyanin content, and in mature berries reached 36 mmol Trolox equivalents/100 g dry wt. More detailed phytochemical analysis determined that delphinidin 3- O -galactoside is the dominant anthocyanin, and that the berries are also rich in procyanidins, including procyanidin A2 which has been implicated in anti-adhesion activity for uropathogenic E. coli . Proanthocyanidins were 60% prodelphinidin, and overall concentrations were higher than reported for many Vaccinium species including blueberry, lingonberry, and cranberry. Overall, the phenolic profile of salal berries indicates that these fruit contain a diversity of health-promoting phenolics.

The objective of this study is to investigate the bioactive compounds and antioxidant activity of coffee and tea mangrove (locally known in Indonesia) produced from the fruit of Rhizophora stylosa. Furthermore, three raw materials of coffee mangrove were also investigated to clarify their potencies. The crude extracts of five samples were subjected to antioxidant assay using DPPH. The results show that the extract of tea mangrove has the strongest activity; then, it was successfully fractionated using different polarity of solvents and yielded acetone and methanol fractions that had high antioxidant activity. The acetone fraction was purified and gave fractions A1, A2, A3, A4, A5, and A6, but only A2 and A3 indicated antioxidant activity and, therefore, they were subjected to further purification. Fractions A3 (caffeine) and A2 gave AS1 (N,N-dimethyl-L-alanine), AS2 (quercetin-3-O-galactopyranoside), AS3 (dodecanoic acid), and AS4 that had the similar 1H-NMR spectrometric results with AS2, while the methanol fraction did not exhibit clear peaks on the chromatogram by HPLC. Therefore, the precipitation method was conducted to purify this fraction, and the precipitate was analyzed by NMR spectra. The results from 1H and 13C NMR indicate that this fraction is a typical polymer of condensed tannins, containing procyanidin and prodelphinidin units.

International audience; The variable anthelmintic efficacy of condensed tannins (CT) against gastrointestinal nematodes may depend on CT concentration, composition, or fate along the digestive tract. We analyzed CT concentration and composition by acetone-HCl-butanol and thiolysis coupled to HPLC-MS in digesta and feces of lambs. Lambs had been infected with Haemonchus contortus and Trichostrongylus colubriformis and received sainfoin pellets and hazelnut skins of contrasting prodelphinidin/procyanidin ratios. The digesta and feces had lower CT concentrations than the original feeds but similar concentration patterns across the digestive compartments. The changes in assayable CT concentrations between rumen, abomasum, and small intestine may be due to complex formation between CT and other dietary components. However, the large CT disappearance (61-85%) from feed to feces could also indicate that CT may have been structurally modified, degraded, or absorbed during digestion. Interestingly, there were no changes in the structural features of assayable CT in the digesta.

Free, esterified, and insoluble-bound phenolics were extracted from selected lentil cultivars. Total phenolics, flavonoids and proanthocyanidin contents were determined. Catechin, epicatechin and procyanidins B were predominant in all fractions of all tested samples as evaluated using HPLC-DAD-ESI-MSn. Methyl vanillate, procyanidin dimer A, and prodelphinidin dimer A were identified and quantified in lentils for the first time. Procyanidin dimer A was detected only in the insoluble-bound form, methyl vanillate was present in the esterified and insoluble-bound one and prodelphinidin dimer A was found in all forms. The presence of unrevealed compounds present only in the insoluble-bound form shows that ignoring the presence of phenolics linked to the cell wall of lentils may underestimate their potential health benefits. The antioxidant activities and the inhibition of cupric ion-induced human low-density lipoprotein peroxidation and peroxyl radical induced supercoiled plasmid DNA strand scission demonstrated that lentil seeds may be considered as a functional food.

The phenolic composition of leaves from Phyllanthus acuminatus L., a plant commonly used in Costa Rica as traditional medicine, was studied using UPLC-ESI-MS on an enriched phenolic extract. A total of 20 phenolic compounds were identified, comprising eight flavonoids (two flavanones—pinocembrin isomers and six derivatives from apigenin, chrysin, quercetin, and kaempferol); seven ellagitannins, two flavan-3-ols (prodelphinidin B dimer and (epi)gallocatechin); and three phenolic acids (ellagic acid, trimethylellagic acid, and ferulic acid). All of these compounds are reported for the first time in P. acuminatus, while previously reported in the genus Phyllanthus. Antioxidant evaluation was performed for P. acuminatus phenolic extract obtaining DPPH results with a remarkably low IC50 value of 0.15 µg/mL. Also, cytotoxicity on gastric AGS and colon SW20 adenocarcinoma cell lines was evaluated, and highly promising results were obtained, with IC50 values of 11.3 µg/mL and 10.5 µg/mL, respectively. Furthermore, selectivity index values obtained when comparing cytotoxicity on normal Vero cells was SI > 20 for both cancer cell lines, indicating a particularly high selectivity. Additionally, Justicidin B, a metabolite extensively studied for its antitumoral activity, was isolated from a non-polar extract of P. acuminatus, and comparatively evaluated for both bioactivities. The DPPH value obtained for Justicidin B was moderate (IC50 = 14.28 µg/mL), while cytotoxicity values for both AGS (IC50 = 19.5 µg/mL) and SW620 (IC50 = 24.8 µg/mL) cell lines, as well as selectivity when compared with normal Vero cells (SI = 5.4 and 4.2 respectively), was good, but lower than P. acuminatus extract. These preliminary results suggest that P. acuminatus enriched phenolic extract containing flavonoids, ellagitannins, flavan-3-ols, and phenolic acids, reported for the first time in this plant, could be of interest for further cancer cytotoxicity studies to elucidate structure–bioactivity relationships, and the molecular mechanisms and pathways. Consejo Nacional de Rectores/[Ref 115B0653]/CONARE/Costa Rica Universidad de Costa Rica/[]/UCR/Costa Rica Instituto Tecnológico de Costa Rica/[]/TEC/Costa Rica UCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de Química UCR::Vicerrectoría de Docencia::Salud::Facultad de Medicina::Escuela de Medicina UCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de Biología

The structural chemistry of proanthocyanidin molecules has been investigated in temperate zone plants, but few studies have been done with plants of the Amazonian lowland tropical wet forests where herbivore pressure is more extensive and diverse. Using MALDI-TOF mass spectrometry, we report unique properties of the proanthocyanidin structural chemistry in two neotropical Cecropia species, C. polystachya, a myrmecophyte with mutualistic ants, and C. sciadophylla, a non-myrmecophyte lacking mutualistic ants. Our preliminary data suggests the presence of reportedly uncommon propelargonidin subunits in a majority of proanthocyanidin oligomers. The presence of 3-O-gallate proanthocyanidin monomers was also detected in the mass spectra of both species. Unlike other studies that have examined species growing at higher latitudes, oligomers composed of procyanidin, propelargonidin, and their 3-O-gallates were present in both Cecropia species while the presence of oligomers containing prodelphinidin units were absent or at lower levels. These distinctive features may suggest that proanthocyanidins in some tropical plant species may be an untapped source of proanthocyanidin structural complexity that warrants further investigation. Several differences between spectra of the two Cecropia species could also point to the presence of anti-herbivore defense tradeoffs between chemical defense quality and biotic defense between the two species.

Total synthesis of prodelphinidin B1, B2, and B4 has been accomplished. The key step is Lewis acid-mediated equimolar condensations between an epigallocatechin and/or a gallocatechin nucleophile and an epigallocatechin and/or a gallocatechin electrophile. The antitumor effects of synthetic prodelphinidin B1-B4 against human PC-3 prostate cancer cell lines have been investigated. These compounds showed significant antitumor effects. Their activity seemed to be little bit stronger than EGCG and prodelphinidin B3, known antitumor agent., Article, TETRAHEDRON LETTERS. 54(52):7188-7192 (2013)

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 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.

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.

Parasitic gastrointestinal nematodes (GIN) of livestock are increasingly developing resistance to synthetic nematocidal drugs. Moreover, the use of nematocides can induce ecotoxicity by affecting free-living nematodes. Condensed tannins (CT) are a structurally diverse group of bioactive plant compounds possessing anthelmintic activity against GIN. We investigated the relationship between the chemical structure of contrasting, purified CT and nematocidal effects using Caenorhabditis elegans. We also explored whether the nematocidal activity of CT could synergize with trans-cinnamaldehyde (CIN). A non-significant correlation was evident between the ability of CT fractions to inhibit C. elegans motility and the molar proportion of prodelphinidin subunits in purified CT samples. Synergistic inhibition of motility was achieved by combinations of CT and CIN. Galloylation of procyanidins was also a key factor for synergy. To increase the nematocidal effect of CT, plant sources containing CT with specific structural features could be selected and combined with compounds acting in synergy.

This study investigated the separate and combined anthelmintic (AH) effects of different phenolic compounds, including condensed tannins and flavonoids, all of which are known to occur in willow leaves, a potentially valuable dry season feed. A range of contrasting model tannins, which span the whole range of willow tannins, were isolated from tilia flowers, goat willow leaves, black currant leaves and red currant leaves. All together, the tested compounds represented the major tannin types (procyanidins and prodelphinidins) and flavonoid types (flavonols, flavones and flavanones). The larval exsheathment inhibition assay (LEIA) was used to assess their in vitro effects on Haemonchus contortus third stage larvae. Arbutin, vanillic acid, and taxifolin proved to be ineffective whereas naringenin, quercetin and luteolin were highly effective at 250 μM concentrations. Procyanidin (PC) tannins tended to be less active than prodelphinidin tannins (PD). Experiments with combinations of tannins and quercetin or luteolin revealed for the first time the existence of synergistic AH effects between tannins and flavonoid monomers. They also provided evidence that synergistic effects appear to occur at slightly lower concentrations of PC than PD. This suggests that the AH activity of condensed tannins can be significantly enhanced by the addition of quercetin or luteolin. This information may prove useful for plant breeding or selection and for designing optimal feed mixtures., Graphical abstract, Highlights • Tannins and flavonoids showed anti-parasitic effects against Haemonchus contortus. • This is the first report of their synergistic effects on larval exsheathment. • Flavonoids enhanced especially the anthelmintic activities of procyanidin tannins.

Ephedra sinica, an important plant in Chinese traditional medicine, contains a complex mixture of proanthocyanidin oligomers as major constituents; however, only the minor components have been chemically characterized. In this study, oligomers with relatively large molecular weights, which form the main body of the proanthocyanidin fractions, were separated by adsorption and size-exclusion chromatography. Acid-catalyzed degradation in the presence of mercaptoethanol or phloroglucinol led to the isolation of 18 fragments, the structures of which were elucidated from their experimental and TDDFT-calculated ECD spectra. The results indicated that (?)-epigallocatechin was the main extension unit, while catechin, the A-type epigallocatechin?gallocatechin dimer, and the A-type epigallocatechin homodimer, were identified as the terminal units. Among the degradation products, thioethers of gallocatechin with 3,4-cis configurations, a B-type prodelphinidin dimer, a prodelphinidin trimer with both A- and B-type linkages, and a prodelphinidin dimer with an α-substituted A-type linkage were new compounds. In addition, a phloroglucinol adduct of an A-type prodelphinidin dimer, a doubly-linked phloroglucinol adduct of epigallocatechin, and a unique product with a flavan-3-ol skeleton generated by the rearrangement of the aromatic rings were also isolated., Molecules, 22(8), 1308; 2017

The phenolic composition of Tunisian Aleppo pine barks was studied. Colorimetric assays, Fourier transformed infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR) were used to examine Aleppo pine tannins. Analyzes showed that Aleppo pine barks are rich in condensed tannins, especially in procyanidin and prodelphinidin tannins. The reactivity to formaldehyde test (Stiasny number) showed the possible use of Aleppo pine tannins in wood adhesive formulation. Thermomechanical analysis (TMA) and strength analysis of Aleppo pine tannin/hexamin based resin showed important bonding properties.

International audience; Matrix assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry was used to examine the oligomer types and their distribution of jubjube (Zyziphus jujuba Mill.) root bark tannin extract. The flavonoid oligomers, up to hexamers, of lower degree of polymerization than that of commercial tannins, are very frequently covalently linked to carbohydrate monomers and dimers. The site of carbohydrate linkage to the flavonoid has been identified by 13C NMR spectroscopy. Fisetinidin, robinetinidin, catechin and gallocatechin flavonoid units are all present in this tannin, all entering to form mixed oligomers based on these four units. This forbids the classification of this extract as just a simple procyanidin, prorobinetinidin, profisetinidin or prodelphinidin.

Proanthocyanidins in Chinese bayberry leaves (PCBLs) were qualitatively analyzed. NMR data suggest that PCBLs are mostly composed of (epi)gallocatechin gallate units. Matrix-assisted laser desorption time-of-flight MS data indicate 95 possible prodelphinidin structures, ranging from dimers to tridecamers. Preparative normal-phase HPLC and further analysis by reverse-phase HPLC together with electrospray ionization MS enabled detection of 20 compounds, including seven newly identified compounds in Chinese bayberry leaves. The antioxidant capacity of PCBLs was evaluated by (1,1-diphenyl-2-picryl-hydrazyl), ferric-reducing antioxidant power, and oxygen radical absorption capacity assays. The EC50 of DPPH radical scavenging activities (as 50% decrease in the initial DPPH concentration) were 7.60 µg. The FRAP and ORAC values were 8859.33 ± 978.39 and 12991.61 ± 1553.34 µmol Trolox equivalents per gram, respectively. The results indicate the high antioxidant potency of PCBLs.

International audience; The flavan-3-ol and proanthocyanidin composition of Aglianico seeds and skins were for the first time determined by HPLC-MS in comparison with the international grapes Merlot and Cabernet Sauvignon. Monomers [(+)-catechin C, (-)-epicatechin EC, (-)-epicatechin-3-O-gallate, ECG] and oligomers [B1, B2, B3, B4 dimers and trimer C1] were identified and quantified in grape extracts. In order to evaluate the reactivity towards salivary proteins of model wine solutions of seeds and skins monomeric/oligomeric and polymeric fractions, the Saliva Precipitation Index (SPI) was carried out. Fractions were also analyzed for their mean degree of polymerization (mDP), percentage of galloylation (%G) and of prodelphinidin (%P) by phloroglucinolysis. Aglianico was the most effective in precipitating proteins than Merlot and Cabernet Sauvignon, mainly for the high percentage of galloylation of grape fractions. The mDP and the percentage of ECG in terminal units resulted to significantly contribute to the precipitation of salivary proteins by grape proanthocyanidins.

International audience; MALDI-TOF, HPLC-ESI-TOF and 13C-NMR techniques were used to analyse the structure of non-purified aqueous chestnut shell tannin extracts. In addition, the influence of the extraction agent (water or aqueous solutions of Na2SO3 and/or NaOH) on tannin structure was analysed by MALDI-TOF in order to select the extract with the best properties for wood adhesives. Using HPLC-ESI-TOF, catechin/epicatechin, gallocatechin/epigallocatechin, dicatechin structures, dicatechin structures without a hydroxyl group, galloyl-glucoses and ellagic acid were identified as the main monomeric components in the aqueous extract. 13C-NMR and MALDI-TOF spectra revealed that extracts are mostly composed of procyanidin and prodelphinidin structures although prorobinetidins might be also present. MALDI-TOF spectrometry was used to identify the extract oligomeric components. Extractions with Na2SO3 and/or NaOH produced changes in the predominant structures in the extracts and on the amount of sugar units linked to the flavonoid structures, which decreased in the presence of Na2SO3. Extract obtained using low Na2SO3 and NaOH concentrations (1.5 and 0.75 %, respectively) was selected as more suitable for wood adhesive preparation.

Condensed tannins were extracted from grape pomace using a water medium in presence of three different bases as catalyst (NaOH, Na 2 CO 3 or NaHCO 3 ) and were characterized by FT-IR and MALDI-TOF spectrometry. It was demonstrated that the tannin extracts contained oligomers up to 6 repeating flavonoid units, with dominant procyanidin with a minor amount of prodelphinidin units. The structure of grape pomace tannin extracted did not present noticeable discrepancy exception of the ratio of the opening pyran ring which was different as a function of the catalyst used. Two different formulations (nonfortified tannin adhesive and fortified with addition of 20% of polymeric 4,4′-diphenyl methane diisocyanate (pMDI)) were used to press one layer particle board. It has been proved that the extraction processes largely affect the adhesive properties of the extracts obtained from the same raw material. Especially the particleboards prepared from tannins extracted using Na 2 CO 3 as catalyst and without pMDI, give a high internal bonding value (IB = 0.45 MPa), which was good enough to pass relevant international standard specifications for interior grade panels. Formaldehyde emission of these panels is very low, well under the European Norm requirements (≤6.5 mg/100 g panel).

One approach to reducing enteric production of greenhouse gases (GHG; i.e. , nitrous oxide and methane) is to feed tanniniferous forage plants such as sainfoin. The objective of this study was to investigate, in vitro , effects of the content and structural characteristics of condensed tannins (CT) in the whole plant, leaves and stems of sainfoin ( Onobrychis viciifolia ) on the digestive process in the rumen. Sainfoin, studied during the first vegetation cycle at the end of flowering, and during the second vegetation cycle at the start of flowering, was incubated in serum bottles containing buffered rumen fluid for 3.5 and 24 h. To assess effects of CT, incubations were with and without added polyethylene glycol (PEG), to neutralise CT effects. Biological activity and prodelphinidin (PD) content were higher (P cis value was lower (P 4 values for stems and leaves at two vegetation cycles (P 2 :CH 4 ratio was lower (P 4 production, or the CO 2 :CH 4 ratio, at either 3.5 or 24 h of incubation. At both incubation periods, in vitro true organic matter digestibility (IVTOMD) was higher for leaves than stems (P 3 –N for leaves and stems in both vegetation cycles. For the whole plant, PEG resulted in higher NH 3 –N content (P 4 production without altering its organic matter digestibility and N value, but research on the nutritional impact of CT must not only be based on CT content in plant tissue, but include structure-activity considerations of CT.

In this paper, a survey of our studies on almond polyphenols including their chemical characterization and further bioavailability in humans is reported. Combination of analytical techniques (LC-DAD/fluorescence, LC/ESI-MS and MALDI-TOF-MS) allowed us, for the first time, the identification of A- and B-type procyanidin, propelargonidin and prodelphinidin polymers in almond skins. Glucuronide, O-methyl glucuronide, sulfate and O-methyl sulfate derivatives of (epi)catechin, as well as the glucuronide conjugates of naringenin and isorhamnetin, and sulfate conjugates of isorhamnetin, together with conjugates of hydroxyphenylvalerolactones were detected in plasma and urine samples after the intake of almond skin polyphenols. In addition, numerous microbial-derived metabolites, including hydroxyphenylpropionic, hydroxyphenylacetic, hydroxycinnamic, hydroxybenzoic and hydroxyhippuric acids were also identified. Depending of the type of metabolite, maximum urinary excretion was attained at different time in comparison to the control group in the course of the 24-h period of urine excretion, allowing us to establish the onset of microbial metabolism., This work has received financial support from the CONSOLIDER INGENIO 2010 Programme: FUN-C-FOOD CSD2007- 063 from the Spanish Ministry of Science and Innovation (MICINN). I.G. was the recipient of a fellowship from the I3P Program funding by the European Social Fund. M.U.-S. thanks the FPI fellowship from MICINN. M.M. thanks the ‘‘Ramón y Cajal” post-doctoral program. R.L.L. thanks the ‘‘Fondo de Investigación Sanitaria” postdoctoral program (F.I.S. CD06/00161) from MICINN.

This EU funded 'HealthyHay'project stablished a sainfoin (Onobrychis vicifolia) germplasm bank at NIAB, Cambridge, with 306 accessions from around the world. A screening method was developed to characterise tannins by thiolytic degradation [1] directly in green plants for the first time. the method was validated by separate analysis of unextractable, extractable and purified tannins using thiolysis, HPLC-GPC and MALDI-TOF MS. Most tannins (58 to 73% of the total) could be recovered after Toyopearl HW50 fractionation with water, aqueous methanol and acetone. the greatest losses during purification occurred amongst larger molecular weight tannins with mean degree of polymerisation (mDP) > 18. The composition of water-,aqueous methanol- and acetone-soluble tannins differed considerably in their mDP and trans/cis ratios, but not in their prodelphinidin/orocyanidin (PD/PC) ratios.

E-mail: brillouet@cirad.fr Publication Inra prise en compte dans l'analyse bibliométrique des publications scientifiques mondiales sur les Fruits, les Légumes et la Pomme de terre. Période 2000-2012. http://prodinra.inra.fr/record/256699; International audience; Tannins were extracted by acetone/water 60:40 from skin and flesh of four cashew apple genotypes from Brazil and Benin (West Africa), and separated from monomeric phenols. Tannins were submitted to acid-catalysed degradation in the presence of phloroglucinol and the products were analysed by HPLC-DAD/ESI-MS. Both skin and flesh tannins contained high percentages of (-)-epigallocatechin and (-)epigallocatechin-O-gallate, followed by minor quantities of (-)-epicatechin and (-)-epicatechin-3-O-gallate; 100% of the compounds were the 23-cis configuration. Skin tannins were half as galloylated (similar to 20%) than flesh tannins (similar to 40%). Their weight-average molecular weight (M-w) was high

Using acid-catalyzed degradation in the presence of cysteamine, the condensed tannins from Lithocarpus glaber leaves were characterized, following thiolysis, by means of reversed-phase HPLC, 13C-NMR and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) analyses. The thiolysis reaction products showed the presence of the procyanidin (PC) and prodelphinidin (PD) structures. The 13C-NMR spectrum revealed that the condensed tannins were comprised of PD (72.4%) and PC (27.6%), and with a greater content of cis configuration rather than the trans configuration of C2–C3. The MALDI-TOF MS analysis proved the presence of PD units, and the maximum degree of polymerization (DP) was an undecamer. The antioxidant activity of condensed tannins from L. glaber leaves was evaluated by using a free radical scavenging activity assay.

Tannin-rich forages offer an alternative to anthelmintic chemicals to control gastrointestinal nematodes. However, the mode of action of such bioactive plants still needs to be assessed. Previous studies have shown that extracts of tannin-rich plants interfere with the first phase of host invasion, i.e., the exsheathment of infective larvae (L3s). In the current study, we examined the hypothesis that exposure to tannins could also affect the second phase of larval establishment, i.e., the tissue association/penetration of the exsheathed L3s into the digestive mucosae. An in vitro direct challenge technique using fundic explants was applied in this study. The main parasite model was Haemonchus contortus. The objectives were to verify: (i) whether a modification of the association/penetration of L3s with the mucosae occurred after contact with sainfoin extract; (ii) whether this is a dose-dependent phenomenon; (iii) whether tannins were responsible for these effects; (iv) whether these effects were dependent on the parasite species; and (v) how the biochemical structure of tannins might influence these effects. Following 3h contact with sainfoin extract at 1,200 microg/ml, the penetration of exsheathed L3s of H. contortus and Teladorsagia circumcincta into fundic explants was significantly reduced. Moreover, a dose-response relationship was found for H. contortus. For both nematodes, the changes were totally alleviated after addition of polyvinyl polypyrrolidone, an inhibitor of tannins, to the sainfoin extract, suggesting that tannins play a major role in the observed effects. Comparison of results obtained with different monomers of condensed tannins confirms a relationship between structure and activity, the prodelphinidin monomers and galloyl-derivatives being more effective than the procyanidin monomers. Combined with the delay or the inhibition of larval exsheathment previously shown, these effects could explain how tanniniferous plants reduce the establishment of infective larvae in small ruminants.