Procyanidin Fractions Research Articles

Procyanidins from Cranberry Press Residues-Extraction Optimization, Purification and Characterization.

Procyanidins are a polyphenolic group that can be found in a variety of foods such as chocolate, tea, cranberries and others. Type A procyanidins can be found in a handful of sources and one of the richest sources are American cranberries. These compounds possess antioxidative, anticancer and anti-inflammatory activities and are most widely used as prevention for urinary tract infections. Cranberries are utilized for jam and juice production, and the latter produces industrial food waste press residues. Press residues contain free and bound procyanidins which can be extracted for use as nutraceuticals. In this study, the extraction of cranberry press residues has been optimized using RSM and the resulting extracts have been purified and fractionated. The obtained procyanidin fractions have been investigated for their antioxidative potential and analyzed using LC-ESI-FTICR-HRMS to determine individual procyanidins. The optimization showed that the optimal extraction can be conducted using acetone in a concentration of 53% without the addition of an acidifying agent. Strong correlation was observed for procyanidin contents and their antioxidative activity using DPPH, ABTS and FRAP methods. The purified fractions contained 78 individual (65 Type A) procyanidins with the degree of polymerization of up to 9.

Pre-Treatment with Grape Seed Extract Reduces Inflammatory Response and Oxidative Stress Induced by Helicobacter pylori Infection in Human Gastric Epithelial Cells.

Helicobacter pylori (H. pylori) is a pathogenic bacteria identified as a potential risk factor for gastritis, gastric ulcers and gastric cancer. During the stomach colonization, H. pylori triggers a strong inflammatory response and subsequent oxidative stress, which are associated with tissue damage. For this reason, it is of particular interest to develop alternative natural tools that enable modulation of the associated damaging immune response. With this purpose, we obtained grape seed extract (GSE) from sweet (not fermented) food grade seeds. The aim of our study was to investigate the effect of GSE and its two enriched procyanidins fractions (OPC and PPC) on the inflammatory process and oxidative stress produced by different H. pylori strains in human gastric epithelial cells (AGS). Anti-inflammatory activity was evaluated by measuring the level of interleukin-8 (IL-8) secretion. IL-8 production was significantly reduced in H. pylori-infected human gastric epithelial cells pre-treated with GSE or its enriched fractions when compared with non-pre-treated infected cells (from 21.6% to 87.8%). Pre-treatment with GSE or its fractions significantly decreased intracellular reactive oxygen species (ROS) production in AGS cells after infection, depending on the H. pylori strain. Our results also showed that GSE and its fractions demonstrate antibacterial activity against all strains of H. pylori used in the study. This work demonstrates the effectiveness of GSE enriched in procyanidins against the main events associated with H. pylori infection.

Exploring interactions between pectins and procyanidins: Structure-function relationships

During fruit and vegetable processing, procyanidins interact with cell walls and form complexes, which further impact their potential health effects. Among cell wall polysaccharides, pectins have the highest affinity for procyanidins. Binding of two procyanidin fractions with twelve pectins of different linearity and size was investigated by ITC, UV–Visible spectrophotometry and HPSEC-MALLS. Pectins interacted preferentially with highly polymerized procyanidins except beet pectins probably because of steric hindrance due to abundant feruloylated arabinans. Linear pectins had higher affinity for procyanidins: this was verified both for comparison between botanical origins (kiwifruit > apple > beet pectins) and between extraction conditions. Debranched pectins, extracted at pH 2.0, had higher affinity and aggregation capacities with procyanidins than those extracted at other pHs. However, the factors affecting pectins of different origins seemed to be different. High molar mass, intrinsic viscosity and hydrodynamic radius contributed more to increased adsorption of procyanidins by apple and beet pectins. Highly linear kiwifruit pectins, with high homogalacturonan content and lower branching ratio bound preferentially to procyanidins. The enthalpy/entropy proportion of the interaction between kiwifruit pectins and procyanidins was higher than that of apple and beet pectins, which suggested more hydrogen bonding. Predominance of homogalacturonan regions and high degree of methylation thus appeared key structural features of pectins for high affinity for procyanidins, while high degree of branching was detrimental. These findings provide the structural foundation for selectivity of interactions in molecular-level.

Comparison of anti-hyperglycemic activities between low- and high-degree of polymerization procyanidin fractions from cacao liquor extract

Procyanidins are thought to be effective for prevention of hyperglycemia. To clarify the degree of polymerization (DP) of procyanidins on anti-hyperglycemic effects, we prepared low-DP (DP ≤ 3) and high-DP (DP ≥ 4) procyanidin fractions from cacao liquor, and examined whether these procyanidin fractions prevent hyperglycemia in vitro. We found that both fractions promoted glucose uptake in L6 myotubes in a dose-dependent manner accompanied by translocation of GLUT4 to the plasma membrane. Moreover, both fractions stimulate phosphorylation of AMP-activated protein kinase (AMPK). In these beneficial effects, low-DP fraction was more effective than high-DP one. On the other hand, high-DP fraction showed greater inhibitory effect of intestinal α-glucosidase activity than low-DP fraction in a cell free-incubation system. In conclusion, cacao liquor procyanidins have anti-hyperglycemic activities and low-DP procyanidins mainly contribute to stimulate glucose uptake by glucose transporter 4 translocation through AMPK-dependent pathway in skeletal muscle while high-DP procyanidins mainly contributes to inhibit the α-glucosidase activity in small intestine.

Green Synthesis of Gold Nanoparticles Capped with Procyanidins from Leucosidea sericea as Potential Antidiabetic and Antioxidant Agents

In this study, procyanidins fractions of dimers and trimers (F1–F2) from the Leucosidea sericea total extract (LSTE) were investigated for their chemical constituents. The total extract and the procyanidins were employed in the synthesis of gold nanoparticles (Au NPs) and fully characterized. Au NPs of 6, 24 and 21 nm were obtained using LSTE, F1 and F2 respectively. Zeta potential and in vitro stability studies confirmed the stability of the particles. The enzymatic activity of LSTE, F1, F2 and their corresponding Au NPs showed strong inhibitory alpha-amylase activity where F1 Au NPs demonstrated the highest with IC50 of 1.88 µg/mL. On the other hand, F2 Au NPs displayed the strongest alpha-glucosidase activity at 4.5 µg/mL. F2 and F2 Au NPs also demonstrated the highest antioxidant activity, 1834.0 ± 4.7 μM AAE/g and 1521.9 ± 3.0 μM TE/g respectively. The study revealed not only the ability of procyanidins dimers (F1 and F2) in forming biostable and bioactive Au NPs but also, a significant enhancement of the natural products activities, which could improve the smart delivery in future biomedical applications.

ABTS On-Line Antioxidant, α-Amylase, α-Glucosidase, Pancreatic Lipase, Acetyl- and Butyrylcholinesterase Inhibition Activity of Chaenomeles Fruits Determined by Polyphenols and other Chemical Compounds.

This study aimed to identify and quantify the chemical composition and polyphenolic profile of 19 cultivars of Chaenomeles × superba, Chaenomeles japonica, and Chaenomeles speciosa by liquid chromatography coupled with photodiode array detector and quadrupole time-of-flight electrospray ionization mass spectrometry (LC-PDA-QTOF-ESI-MS). Antioxidant (ABTS on-line, ABTS, FRAP, and ORAC), as well as in vitro biological activities, i.e., the ability to inhibit α-amylase, α-glucosidase, pancreatic lipase, acetylcholinesterase (AChE), butyrylcholinesterase (BuChE), and 15-lipoxygenase (15-LOX) were determined. Most of the Chaenomeles species and cultivars analyzed in this study have not been examined in this respect until now. Fruits contained 30.26 to 195.05 mg of vitamin C, 0.65 to 1.69 g of pectin, 0.32 to 0.64 g of ash, 0.60 to 3.98 g of sugars, and 41.64 to 110.31 g of organic acids in 100 g fresh weight. The lowest content of total polyphenols showed C. speciosa ‘Rubra’ (57.84 g/kg dry weight, dw) while C. × superba ’Nicoline’ (170.38 g/kg dw) exhibited the highest concentration of those compounds. In the phenolic compounds, polymeric procyanidin fraction predominated (65%) with procyanidin B2, C1, and (−)-epicatechin the most abundant. The antioxidant capacity measured by ABTS assay was mainly formed by polymeric procyanidins and flavan-3-ols, which was confirmed by ABTS on-line profiling. Chaenomeles fruits showed high potential for inhibition of α-glucosidase and pancreatic lipase. The analyzed cultivars displayed greater potential for acetylcholinesterase (AChE) inhibition than for butyrylcholinesterase (BuChE). The data indicate that Chaenomeles fruits could be regarded as a promising source of bioactive functional food.

Development of a New Cell-Based Oral Model To Study the Interaction of Oral Constituents with Food Polyphenols.

Some polyphenols have unpleasant taste properties such as astringency, which could interfere with consumers' choices. The knowledge on astringency mechanisms points that astringency is a complex phenomenon probably related to more than one physical-chemical mechanism. Thus, this work aims to develop a new and more realistic cell-based model containing human saliva, mucosa pellicle, and an oral cell line (HSC-3) to understand the oral molecular events that could contribute to the overall astringency perception. This model was then used to study the interactions with a food procyanidin fraction (PF) by HPLC. In general, the results revealed higher interaction (synergism) for the model with all the referred oral constituents (mucosa pellicle, salivary proteins, and HSC-3 cell line, HSCMuSp) when compared to the interaction with individual constituents, the PF + cells or PF + saliva. Regarding the procyanidins, a significant interaction was observed for the procyanidin monomer EcG, procyanidin dimers B7 and B2G, and trimer C1.

African baobab (Adansonia digitata) fruit as promising source of procyanidins

Baobab is an endemic African plant rich in health-promoting compounds and still not extensively studied. Characterization of bioactive molecules in baobab fruit by means of LC system coupled with a photodiode array and a triple quadrupole mass spectrometer detector is here reported. This work provides a qualitative and quantitative profile of polyphenols in baobab pulp, shell, kernel and seeds. Baobab pulp is a rich source of ascorbic acid, (−)-epicatechin and procyanidin B2 (135.6, 104.8, and 100.1 mg/100 g). Moreover, a HPLC preparative method to isolate procyanidin fraction from pulp was developed, revealing how the antioxidant activity of baobab pulp not only depend on the ascorbic acid content but also on the presence of procyanidins. The use of baobab fruit pulp as promising healthy food or food supplement is also discussed.

Cacao liquor procyanidins prevent postprandial hyperglycaemia by increasing glucagon-like peptide-1 activity and AMP-activated protein kinase in mice.

Procyanidins have been reported to possess potential for the prevention of hyperglycaemia. However, there are very few data for procyanidins about the difference the degree of polymerisation (DP) has on anti-hyperglycaemic effects. Moreover, the underlying molecular mechanisms by which procyanidins suppress hyperglycaemia are not yet fully understood. In the present study, we prepared procyanidin fractions with different DP, namely low-DP (DP≤3) and high-DP (DP≥4) fractions, from a cacao liquor procyanidin-rich extract (CLPr). These fractions were administered orally to Institute of Cancer Research (ICR) mice and their anti-hyperglycaemic effects were examined. We found that CLPr and its fractions prevent postprandial hyperglycaemia accompanied by an increase in the plasma glucagon-like peptide-1 (GLP-1) level with or without glucose load. In the absence of glucose load, both fractions increased the plasma insulin level and activated its downstream signalling pathway in skeletal muscle, resulting in promotion of the translocation of GLUT4. Phosphorylation of AMP-activated protein kinase (AMPK) was also involved in the promotion of GLUT4 translocation. High- and low-DP fractions showed a similar activation of insulin and AMPK pathways. In conclusion, cacao liquor procyanidins prevent hyperglycaemia by promoting GLUT4 translocation in skeletal muscle, and both the GLP-1-activated insulin pathway and the AMPK pathway are involved in the underlying molecular mechanism.

In vitro evaluation of the anti-digestion and antioxidant effects of grape seed procyanidins according to their degrees of polymerization

Few studies have evaluated the anti-digestion effects of grape seed procyanidins (GSPs) compared to their strong anti-oxidant activities according to different degrees of polymerization (DPs). Additionally, the effects of GSPs with different DPs on the binding sites of digestive enzymes are unexplored. In this work, the anti-digestive and anti-oxidant activity of GSPs with different DPs were verified and compared. Anti-digestive activity assays were conducted on key digestive enzymes. Although all procyanidins fractions possessed antioxidant activity as expected, there was no significant difference in the antioxidant activity among procyanidins with different DPs. In contrast, the anti-digestive activity of the fractions increased significantly as the DP increased. Meanwhile, molecular docking provided a putative mechanism of anti-digestion, and GSPs can block the enzyme sites by hydrogen interactions, hydrophobic interactions and electrostatic interactions. These results indicated that the higher-molecular-weight procyanidins had equal or lower antioxidant activity and a greater anti-digestion effect than lower-molecular-weight procyanidins.

Effect of Proanthocyanidin-rich Extracts from Pinus radiata Bark on Termite Feeding Deterrence

Antioxidants are known to affect the feeding habits of termites and a good source is pine bark which contains high levels of antioxidants which can be extracted with neutral solvents. In this study procyanidins (PCs)-rich MeOH-extract and a hot-water extract (HWE) were prepared from Pinus radiata bark. HWE was further separated into crude PCs (CPCs), polymeric PCs (PPCs), and low-molecular-weight PCs (LMWPCs) fractions. The MeOH extract and these fractions were examined for termite (Reticulitermes flavipes Kollar) antifeedant activities using a no-choice test. Catechin was used as a positive control. The LMWPCs was found to be the most effective deterrent to termite feeding. The comparative efficiency of the compounds tested were LMWPCs > PPCs > CPCs > MeOH extract, with the latter being more dependent on molecular weight than on its antioxidant activity.

Metabolite Profile and Antiproliferative Effects in HaCaT Cells of a Salix reticulata Extract.

Phenolic constituents of Salix reticulata (Salicaceae) and antiproliferative activity of an extract and individual compounds were investigated in immortalized human non-tumorigenic keratinocytes (HaCaT). A MeOH extract from aerial parts afforded several flavonoids, including luteolin and apigenin glycosides (2-5 and 9) and catechin (1), two procyanidin fractions, and the phenolic glucosides picein (6), triandrin (7), and salicortin (8). In an adenosine triphosphate assay, the MeOH extract reduced cell viability by approximately 60 % at a concentration of 100 µg/mL. Cell proliferation was assessed with a BrdU incorporation ELISA assay. The extract inhibited proliferation of HaCaT cells in a concentration-dependent manner, with approximately 50 % inhibition at 100 µg/mL. In time-lapse assays, the extract showed distinct inhibitory effects on cell migration at concentrations of 12.5, 25, and 50 µg/mL. The activity of selected constituents was also determined. Luteolin-7-O-β-glucuronide (3) significantly inhibited cell proliferation at concentrations of 10 and 50 µM. In contrast, luteolin-7-O-β-glucopyranoside (2) and a procyanidin fraction (P1) had only weak effects, while picein (6) and salicortin (8) did not affect cell proliferation. Luteolin-7-O-β-glucuronide (10 µM) and, to a lesser extent, the procyanidin fraction (10 µg/mL) also inhibited cell migration.

Monomeric cocoa procyanidins enhances β‐cell function by increasing mitochondrial respiration

A hallmark of type 2 diabetes (T2D) is β‐cell dysfunction and the eventual loss of functional β‐cell mass. Therefore, mechanisms that can improve β‐cell function could be used as a treatment for the millions of individuals with T2D. Recent studies have shown that monomeric, oligomeric and polymeric cocoa derived procyanidins have vastly different effects on obesity, insulin resistance and glucose tolerance during high fat feeding. Based on these previously published data, we hypothesized that cocoa derived procyanidins may have beneficial effects on improving β‐cell function. 832/13 INS‐1 derived β‐cells and primary rat islets cultured with monomeric cocoa procyanidins demonstrated enhanced glucose stimulated insulin secretion (GSIS), while cells cultured with total cocoa extract, oligomeric procyanidins or polymeric procyanidins demonstrated no improvement. The increased GSIS observed with culture in the presence of monomeric procyanidin fractions corresponded with enhanced mitochondrial respiration, suggesting improvements in β‐cell fuel utilization. We demonstrated upregulation of electron transport chain components Complex III, IV and V after culture with monomeric procyanidins. Monomeric procyanidins also resulted in an increased cellular redox state and increased glutathione concentration, which corresponds with Nrf2 nuclear localization. We propose a model by which cocoa derived monomeric procyanidins enhance the cellular redox state, allowing Nrf2 to migrate to the nucleus and enhance expression of key mitochondrial genes that results in enhanced GSIS and improved β‐cell function. These results suggest that cocoa derived monomeric procyanidins could be leveraged as a treatment modality for patients with T2D.Support or Funding InformationBYU MEG Grant to JST, Diabetes Action Research and Education Grant to JST, BYU ORCA Grant to JDR and BFB

Molecular study of mucin-procyanidin interaction by fluorescence quenching and Saturation Transfer Difference (STD)-NMR.

Astringency is closely related to the interaction between procyanidins and salivary proteins (SP). The aim of this work was to study the interaction between mucin, a SP responsible for saliva lubricating properties, with different procyanidins (B4 dimer, tetramer (TT) and fractions of oligomeric procyanidins), and the influence of several conditions [pH, ionic strength, procyanidins' mean degree of polymerization (mDP) and different solvents (ethanol or dimethylsulfoxide)] on this interaction by fluorescence quenching and Saturation Transfer Difference (STD)-NMR. For fractions of oligomeric procyanidins, the mucin-procyanidin interaction increased with mDP; however, for pure compounds, procyanidin TT has lower affinity than dimer B4 which could be due to a lower structural flexibility imposed by its complex structure. Furthermore, EtOH and DMSO can disrupt the main driving forces of these interactions, hydrophobic interactions and hydrogen bonds, respectively, lowering significantly the binding constants.

Protein-Tannin Interactions of Tryptic Digests of α-Lactalbumin and Procyanidins.

Protein-tannin interactions on a molecular level were investigated by using a model system containing peptides of α-lactalbumin and berry tannins (procyanidins). Oxidation of isolated tryptic peptide LDQWLCEK (m/z 1034) with procyanidin B2 or procyanidin fraction (PF) isolated from aronia juice was monitored by LC-ESI-MS. Procyanidin B2 and PF showed radical scavenging activities toward oxidation of the peptide with the peptide also preventing procyanidin B2 from degradation. Oxidation enhanced the cleavage of peptide between tryptophan and glutamine. Interaction products arising from WLCEK or WLCE residue and degradation product of procyanidin B2 were also identified using both size exclusion chromatography and LC-MS. Tryptophan and lysine were the amino acids most prone to interact with procyanidin B2. The study shows that protein-tannin interaction takes place during oxidation leading to both degradation of the parent compounds and formation of interaction products. This may in turn affect the quality of protein and tannin containing food.

Contribution of Human Oral Cells to Astringency by Binding Salivary Protein/Tannin Complexes

The most widely accepted mechanism to explain astringency is the interaction and precipitation of salivary proteins by food tannins, in particular proline-rich proteins. However, other mechanisms have been arising to explain astringency, such as binding of tannins to oral cells. In this work, an experimental method was adapted to study the possible contribution of both salivary proteins and oral cells to astringency induced by grape seed procyanidin fractions. Overall, in the absence of salivary proteins, the extent of procyanidin complexation with oral cells increased with increasing procyanidin degree of polymerization (mDP). Procyanidin fractions rich in monomers were the ones with the lowest ability to bind to oral cells. In the presence of salivary proteins and for procyanidins with mDP 2 the highest concentrations (1.5 and 2.0 mM) resulted in an increased binding of procyanidins to oral cells. This was even more evident for fractions III and IV at 1.0 mM and upper concentrations. Regarding the salivary proteins affected, it was possible to observe a decrease of P-B peptide and aPRP proteins for fractions II and III. This decrease is greater as the procyanidins' mDP increases. In fact, for fraction IV an almost total depletion of all salivary proteins was observed. This decrease is due to the formation of insoluble salivary protein/procyanidin complexes. Altogether, these data suggest that some procyanidins are able to bind to oral cells and that the salivary proteins interact with procyanidins forming salivary protein/procyanidin complexes that are also able to link to oral cells. The procyanidins that remain unbound to oral cells are able to bind to salivary proteins forming a large network of salivary protein/procyanidin complexes. Overall, the results presented herein provide one more step to understand food oral astringency onset.

Preparative separation of cacao bean procyanidins by high-speed counter-current chromatography.

In this work, an efficient method for preparative separation of procyanidins from raw cacao bean extract by high-speed counter-current chromatography (HSCCC) was developed. Under the optimized solvent system of n-hexane-ethyl acetate-water (1:50:50, v/v/v) with a combination of head-tail and tail-head elution modes, various procyanidins fractions with different polymerization degrees were successfully separated. UPLC, QTOF-MS and 1H NMR analysis verified that these fractions contained monomer up to pentamer respectively. Dimeric procyanidin B2 (purity>86%) could be isolated by HSCCC in a single run. Other individual procyanidins in these fractions could be further isolated and purified by preparative HPLC. The developed HSCCC together with preparative HPLC techniques appeared to be a useful tool for large preparation of different procyanidins from cacao beans. Furthermore, by antioxidant activity assays, it was proved that both fractions and individual procyanidins possessed greater antioxidant activities compared to standard trolox. The antioxidant activities of procyanidins increase as the increase of their polymerization degree.

Principal component analysis of metabolites in urine from rats treated with procyanidin A2 and two oligomeric procyanidin fractions from cranberry

Principal component analysis of metabolites in urine from rats treated with procyanidin A2 and two oligomeric procyanidin fractions from cranberry

Cocoa procyanidins with different degrees of polymerization possess distinct activities in models of colonic inflammation

Procyanidins are available in the diet from sources such as cocoa and grapes. Procyanidins are unique in that they are comprised of repeating monomeric units and can exist in various degrees of polymerization. The degree of polymerization plays a role in determining the biological activities of procyanidins. However, generalizations cannot be made regarding the correlation between procyanidin structure and bioactivity because the size–activity relationship appears to be system dependent. Our aim was to screen fractions of procyanidins with differing degrees of polymerization in vitro for anti-inflammatory activities in models of colonic inflammation. Monomeric, oligomeric and polymeric cocoa procyanidin fractions were screened using cell models of disrupted membrane integrity and inflammation in human colon cells. High-molecular-weight polymeric procyanidins were the most effective at preserving membrane integrity and reducing secretion of interleukin-8 in response to inflammatory stimuli. Conversely, oligomeric procyanidins appeared to be the least effective. These results suggest that polymeric cocoa procyanidins may be the most effective for preventing loss of gut barrier function and epithelial inflammation, which are critical steps in the pathogenesis of metabolic endotoxemia, inflammatory bowel disease and colon cancer. Therefore, further investigations of the potential health-protective benefits of cocoa procyanidins with distinct degrees of polymerization, particularly high-molecular-weight procyanidins, are warranted.

Chemical characterization of procyanidins from Spatholobus suberectus and their antioxidative and anticancer activities

Spatholobus suberectus (SS) is an excellent functional and medicinal food source of dietary procyanidins, which have numerous health benefits. Ten procyanidin fractions with different polymerization degrees were prepared from SS. For the first time, trimers to 11-mers were detected in SS extracts, and their compositions were elucidated through thiolysis, ESI-MS and MALDI-TOF-MS analyses. The results determined that the mean polymerization degree (mDP) of the crude SS procyanidins was 5.2 and that the mass distribution of the monomer and the oligomers (mDP of 1.0–5.7) was 70.4%, whereas that of the polymers (mDP of 6.2–11.8) was 25.6%. Catechin and epicatechin were the main monomeric units with minor quantities of epigallocatechin and gallocatechin. The observed interflavan linkages were both A-type (minor type) and B-type (major type). The fractions enriched in the monomer and oligomers showed the highest activities, including antioxidant activity and inhibitory activities on both the proliferation of MCF-7 breast cancer cells and lactate dehydrogenase A (LDH-A). These results suggested that the monomer- and oligomer-enriched procyanidin fractions from SS could undergo further development to become a beneficial source of antioxidants, LDH-A inhibitors, and potential inhibitors of breast cancer.