Phenolic Fractions from Muscadine Grape “Noble” Pomace can Inhibit Breast Cancer Cell MDA‐MB‐231 Better than those from European Grape “Cabernet Sauvignon” and Induce S‐Phase Arrest and Apoptosis

Tons of grape pomace which still contained a rich amount of plant polyphenols, is discarded after winemaking. Plant polyphenols have multi‐functional activities for human body. In this study, polyphenols of pomaces from Muscadinia rotundifolia “Noble” and Vitis vinifera “Cabernet Sauvignon” were extracted and fractionated, and then they were analyzed with LC‐MS and the inhibitory effects on breast cancer cells were compared. The inhibition on MDA‐MB‐231 cells of fractions from “Noble” was further evaluated. The results showed that polyphenols from 2 grape pomaces could be separated into 3 fractions, and ellagic acid and/or ellagitannins were only detected in fractions from “Noble” pomace. All 3 fractions from “Noble” pomace inhibited MDA‐MB‐231 better than MCF‐7. But fraction 2 from “Cabernet Sauvignon” inhibited MCF‐7 better while fraction 1 and fraction 3 inhibited both 2 cells similarly. Moreover, the fractions from “Noble” pomace rather than “Cabernet Sauvignon” can inhibit MDA‐MB‐231 better. Finally, fractions from “Noble” pomace can induce S‐phase arrest and apoptosis on MDA‐MB‐231. These findings suggested the extracts from grape pomace especially those from “Noble,” are potential to be utilized as health beneficial products or even anti‐breast cancer agents.     

Applications of Wine Pomace in the Food Industry: Approaches and Functions

Winemaking generates large amounts of wine pomace, also called grape pomace. This by‐product has attracted the attention of food scientists and the food industry, due to its high content in nutrients and bioactive compounds. This review mainly focuses on the different published approaches to the use of wine pomace and its functions in the food industry. Traditionally, wine pomace has been used to obtain wine alcohol, food colorings, and grape seed oil. More recently, research has focused in the production of other value‐added products, such as extracts of bioactive compounds, mainly phenols, recovery of tartaric acid, and the making of flours. The most common functions associated with wine pomace products are their use as antioxidants, followed by their use as fortifying, coloring, and antimicrobial agents. These products have mainly been applied to the preparation of meat and fish products and to, a lesser extent, cereal products.     

Evaluation of the bioactive compounds and the antioxidant capacity of grape pomace

Grape pomace is an agro‐industrial residue produced worldwide and mainly employed for animal feed or as a fertiliser. Several studies have shown that grape pomace is a rich source of bioactive compounds, such as phenolic compounds, polysaccharides, fatty acids and others. Three varieties of grape pomace were evaluated in this study. Antioxidant activity was determined by three different methods, namely, DPPH˙, ABTS and ferrous ion assays. Fatty acids and phenolic compounds were identified, respectively, by gas and liquid chromatography. Results showed that grape pomace is a rich source of polyunsaturated fatty acids and phenolic compounds. Cabernet Sauvignon pomace revealed higher values of total phenolic compounds, total flavonoids and total monomeric anthocyanin, and presented the lowest value of EC₅₀, and thus a higher antioxidant activity among the samples analysed.     

Screening grape seeds recovered from winemaking by‐products as sources of reducing agents and mammalian α‐glucosidase and α‐amylase inhibitors

Grape seeds collected from vinification of various grape varieties were extracted by supercritical CO₂ for oil recovery. The defatted residues thus obtained were considered as a re‐utilisable co‐product and assessed for phenolic content, reducing capacity and inhibitory activities against mammalian α‐amylase and α‐glucosidase enzymes. Supercritical CO₂ treatment led to higher recovery of anthocyanins. Reducing capacity of phenolic extracts reached up to ~2200 mmolFe(II) kg^?1, much higher than that of various natural phenolic sources. The anthocyanin‐rich extracts showed the highest inhibitory effectiveness towards α‐glucosidase (I₅₀ value equal to ~40 μg gallic acid equivalents (GAE)/mL ~ half than acarbose). Inhibitory effectiveness towards α‐amylase activity was similar among grape varieties, with I₅₀ values comparable to that of acarbose and correlated with proanthocyanidin contents. These results could pave the way for an efficient processing of grapes, including cascade processes, namely: winemaking, oil extraction from recovered grape seeds and phenolic extraction from defatted grape seeds as potential cost‐effective nutraceuticals.     

The evaluation of grape pomace distillates from selected red grape varieties

This report describes the composition of the volatiles from grape pomace distillated that are commercially available in Greece. These spirits, which are a valuable by‐product of the winemaking industry, were obtained by distillation of grape pomace after fermentation under anaerobic conditions. The volatiles were distilled twice. Each distillate was separated in to 4 equal (by volume) fractions and the changes in volatile concentration during distillation were measured. Assays were done by gas/liquid chromatography (for alcohols, carboxylic acids, esters and aldehydes). The average values for volatile concentrations obtained for Greek grape pomace distillates were compared with the corresponding values for grape pomace manufactured in other Mediterranean countries.     

Comparison of fining red wines with purified grape pomace versus commercial fining agents: effect on wine chromatic characteristics and phenolic content

The large quantity of grape pomace (seeds, skin, and peduncle) produced during the winemaking process can be a problem and the search for procedures which could permit their valorisation is considered an important issue. This study investigates the use of purified grape pomace as a fining agent for reducing the level of some wine phenolic compounds, especially tannins. For this, purified grape pomace was applied to three young red wines of different phenolic composition and the results were compared with the effect of some common commercial fining agents. The results demonstrated that grape pomace can be used to decrease the wine tannin content with similar results to those obtained with casein, while also reducing anthocyanin content although to a similar extent to when bentonite is used. The effect of purified Monastrell grapes pomace is similar in the three different studied wines although it affected the most to wines with high monomeric anthocyanin content.     

Short communication: Compositional characteristics and aromatic profile of caciotta cheese obtained from Friesian cows fed with a dietary supplementation of dried grape pomace

The aim of the present work was to explore the chemical-sensorial characteristics and aromatic profile of caciotta cheese obtained from Friesian cows fed a diet enriched with grape pomace obtained from red grape (Vitis vinifera L.). Dietary enrichment with grape pomace influenced the production of caciotta cheeses in interesting ways from a compositional point of view, as cheese samples were rich in polyphenols, giving a high antioxidant potential. From a biochemical standpoint, we noted a slight decrease of proteolysis during ripening, whereas, according to the analysis of volatile compounds, lipolysis was the most relevant phenomenon in samples. The presence of bioactive compounds also modified the fatty acid profile of milk and cheese, leading to an increase in concentration of linoleic, vaccenic, and rumenic acids. No significant variations were found in the sensory profile. These results showed the potential of dietary grape pomace intake to influence the chemical-nutritional and nutraceutical properties of cow milk and cheeses, whose introduction to the market could be attractive to consumers, providing interesting implications for the dairy industry. Finally, our results identified of a valid use of an agro-industrial by-product, grape pomace, whose disposal generally presents economic and environmental problems.     

Influence of ethanol/water ratio in ultrasound and high‐pressure/high‐temperature phenolic compound extraction from agri‐food waste

The valorisation and management of agri‐food waste are currently hot investigation topics which probe the recovery of valuable compounds, such as polyphenols. In this study, high‐pressure/high‐temperature extraction (HPTE) and ultrasound‐assisted extraction (UAE) have been used to study the recovery of phenolic compounds from grape marc and olive pomace in hydroalcoholic solutions. The main phenolic compounds in both extracts were identified by HPLC‐DAD. Besides extraction yield (total polyphenol and flavonoid content) and the antiradical power, polyphenol degradation under HPTE and UAE has also been studied. HPTE with ethanol 75% gave higher phenolic extraction yields: 73.8 ± 1.4 mg of gallic acid equivalents per gram of dried matter and 60.0 mg of caffeic acid equivalents per gram of dried matter for grape marc and olive pomace, respectively. In this study, the efficient combination of ethanol/water mixture with HPTE or UAE has been used to enhance the recovery of phenolic compounds from grape marc and olive pomace. HPLC‐DAD showed that UAE prevents phenolic species degradation damage because of its milder operative conditions.     

Anthocyanin dyes extracted from grape pomace for the purpose of textile dyeing

BACKGROUND: Pomace from different grape varieties was studied with regard to the content of extractable anthocyanins. RESULTS: Total anthocyanin concentrations of 24.5–126 mg dm^?3 were found in the extracts. The amount of extractable pigment was dependent on the variety of grape. The extracts were applied in textile dyeing operations using cotton fabric pre‐mordanted with tannin as textile substrate. Red/violet shades were obtained that showed good water‐fastness; however, limited light‐fastness indicated need for improvement to fulfil requirements for textile application. CONCLUSION: Colour measurement by means of CIE‐Lab coordinates and Kubelka–Munk value showed a direct correlation between extractable anthocyanin concentration and colour depth of the dyed material. Thus the analytically determined concentration of anthocyanins in the aqueous extract can serve as a measure of quality of grape pomace as raw material for textile dyeing application. Copyright © 2007 Society of Chemical Industry     

Procyanidin Content of Grape Seed and Pomace, and Total Anthocyanin Content of Grape Pomace as Affected by Extrusion Processing

ABSTRACT: Grape juice processing by‐products, grape seed and pomace are a rich source of procyanidins, compounds that may afford protection against chronic disease. This study was undertaken to identify optimal extrusion conditions to enhance the contents of monomers and dimers at the expense of large molecular weight procyanidin oligomers and polymers in grape seed and pomace. Extrusion variables, temperature (160, 170, and 180 °C in grape seed, and 160, 170, 180, and 190 °C in pomace) and screw speed (100, 150, and 200 rpm in both) were tested using mixtures of grape seed as well as pomace with decorticated white sorghum flour at a ratio of 30 : 70 and moisture content of 45%. Samples of grape seed and pomace were analyzed for procyanidin composition before and after extrusion, and total anthocyanins were determined in pomace. Additionally, chromatograms from diol and normal phase high‐performance liquid chromatography were compared for the separation of procyanidins. Extrusion of both grape by‐products increased the biologically important monomer and dimers considerably across all temperature and screw speeds. Highest monomer content resulted when extruded at a temperature of 170 °C and screw speed of 200 rpm, which were 120% and 80% higher than the unextruded grape seed and pomace, respectively. Increases in monomer and dimer contents were apparently the result of reduced polymer contents, which declined by 27% to 54%, or enhanced extraction facilitated by disruption of the food matrix during extrusion. Extrusion processing reduced total anthocyanins in pomace by 18% to 53%. Practical Applications: Extrusion processing can be used to increase procyanidin monomer and dimer contents in grape seed and pomace. Procyanidins in grape by‐products have many health benefits, but most are present as large molecular weight compounds, which are poorly absorbed. Extrusion processing appears to be a promising technology to increase levels of the bioactive low molecular weight procyanidins.     

The inhibitory effects of wine phenolics on lysozyme activity against lactic acid bacteria

The lysozyme of hen's egg white is used in winemaking to control spontaneous lactic acid bacteria (LAB). A total of eight LAB strains, isolated from grape must and wine, were used to assess the inhibitory effects of wine phenolics on lysozyme activity. The presence of phenolics, extracted from grape pomace, in growth medium reduced the mortality rate due to the lysozyme activity. This effect was especially clear in the case of strains belonging to Lactobacillus uvarum, Pediococcus parvulus and Oenococccus oeni, which are more sensitive to lysozyme than L. plantarum and L. hilgardii strains. Cell lysis assays carried out on four strains sensitive to lysozyme and Micrococcus lysodeikticus ATCC 4698, used as a reference strain, confirmed the inhibition of grape pomace phenolics on the muramidase. There was no interference from non-flavonoids, flavanols and flavonol compounds, when they were tested individually, on the lysozyme activity against the strains. Anthocyanins extracted from grape skins slightly inhibited the activity only against M. lysodeikticus. However, proanthocyanidins extracted from seed berries, strongly inhibited the lysozyme. In this extract, dimers were the predominant oligomers of flavan-3-ol. The study demonstrated that the effectiveness of lysozyme against LAB in red winemaking is related to the amount of low molecular weight proanthocyanidins that are released when the grapes are macerating.     

Antioxidant activities of organic grape, pomace, juice, must, wine and their correlation with phenolic content

The highest values of antioxidant activity (AOA), inhibition of low‐density lipoproteins (LDL‐diene, LDL‐MDA) and total phenols were determined in pomace (82.30 and 82.60%), grapes (68.91%) and must [2750 mg L^?1 gallic acid equivalents (GAE)], respectively. For all parameters (AOA, iLDL‐diene, iLDL‐MDA, total phenols) the highest values were obtained in Cabernet Sauvignon and Merlot grape varieties. Positive correlations were determined between AOA and iLDL‐diene (r = 0.809); AOA and total phenols (r = 0.528); total phenols and iLDL‐diene (r = 0.451) with significance of P < 0.005. The results of principal component analysis demonstrated that the location of iLDL‐diene and AOA is the same for pomace, must and red wines. The total phenols are found in the same place in red wines, red grapes, pomace and must. The results emphasize the importance of must, pomace and red wine for inhibiting LDL‐oxidation.     

High‐pressure recovery of anthocyanins from grape skin pomace (Vitis vinifera cv. Teran) at moderate temperature

The objective was to evaluate the influences of the high hydrostatic pressure extraction parameters on the recovery of anthocyanins from the grape skin pomace extracts (Vitis vinifera cv. Teran) under moderate temperatures. Studied parameters were: solvents (methanol and ethanol); solvent concentrations (30, 50, and 70%); pressures (300, 400, and 500 MPa); times (3, 6.5, 10 min); and temperatures (22, 26, 30 °C). Predominant anthocyanins in all extracts were malvidins (malvidin‐3‐glucoside as the main compound) representing 55.77% of overall anthocyanin content. The type of solvent did not significantly influence anthocyanin extraction yield, while decreased solvent concentration (increased solvent‐to‐water ratio) significantly improved extraction of anthocyanins. Increase of pressure enhanced extraction yield of the anthocyanins but temperature showed stronger impact on the anthocyanins recovery. This investigation evidenced that the best conditions for HHPE of anthocyanins from grape pomace were extraction time 3.39 min, extraction temperature 29.48 °C, pressure 268.44 MPa and solvent concentration 70%.

Practical applications

Due to increased interest for the use of cheap winery byproducts as a source of expensive polyphenols for functional food production, skins from grape pomace became valuable raw material. Therefore, there is a need to evaluate the influence of HHPE innovative extraction technology on its nutritive value during processing. Optimizing parameters for polyphenolic recovery from grape pomace is directly related with nutritional value and economics of food engineering during industrial processing. Obtained results showed that the HHPE under lower temperatures is suitable for the extraction process of anthocyanins from grape skin pomace, but more research is needed to identify other food‐grade solvents with their corresponding concentrations that are useful for the extraction assisted with high hydrostatic pressure.     

Screening of bioactive components in grape and apple vinegars: Antioxidant and antimicrobial potential

Vinegars contain several bioactive compounds that are characterized according to the type of the raw material, such as grape vinegars and apple vinegars. Liquid chromatography coupled to diode array detection and electrospray ionization tandem mass spectrometry was used for identification and quantification of phenolic compounds. Antioxidant properties of vinegars were determined by 2,2‐diphenyl‐1‐picrylhydrazyl and 2,2′‐azino‐bis‐3‐ethylbenzthiazoline‐6‐sulphonic acid assays. Antimicrobial activities of vinegars were examined with an agar disc diffusion method with Staphylococcus aureus , Escherichia coli and Pseudomonas aeruginosa . Gallic acid and chlorogenic acid were found to be the major phenolic acids accounting for the largest proportion of the total phenolic acid contents in grape vinegars and apple vinegars. Within the flavonols, quercetin‐3‐O‐galactoside and quercetin were detected as the major compounds in grape vinegars. Apple vinegars were characterized by phloridzin, phloretin and high chlorogenic acid content. Antimicrobial activity results indicated that grape vinegars exhibited higher antimicrobial activity against tested bacterial strains correlated with their higher antioxidant capacity. Copyright © 2017 The Institute of Brewing & Distilling     

Recovery and fractionation of major apple and grape polyphenols from model solutions and crude plant extracts using ion exchange and adsorbent resins

Model systems composed of phenolic compounds, amino acids and mixtures thereof mimicking apple pomace and grape skin extracts were used to examine adsorption and ion exchange behaviour of one adsorbent as well as four anion and four cation exchange resins in a column system at pH 1, 5 and 7. Furthermore, apple pomace and grape skin extracts were loaded onto the resins to evaluate the transferability of the results obtained in the model experiments. In addition, different eluents were applied for the successive recovery of phenolic compounds from the resins. The results provide valuable information on highly complex adsorption and exchange systems. The data indicate both enhanced and decreased polyphenol binding rates as a result of amino acid addition depending on the composition of the solutions applied to the resins. The application of appropriate process parameters during adsorption and desorption was shown to allow selective recovery of compound classes or even individual compounds. As the model experiments did not allow complete prediction of polyphenol binding behaviour, further extract compounds not investigated in the present study presumably also interfere with polyphenol recovery. The results help to systematically optimise industrial processes, thus improving cost‐efficiency and selectivity of polyphenol recovery.     

Value Adding to Red Grape Pomace Exploiting Eco-friendly FT-NIR Spectroscopy Technique

Agro-food industries require sustainable and profitable alternatives to disposal of their by-products. Grape pomace is a winemaking residue that represents a low-cost natural source of phenolic compounds with recognized antioxidant properties. In this work, Fourier-transform near-infrared (FT-NIR) spectroscopy and chemometric analysis were exploited to the characterization of red grape pomace composition (content of seeds and skins) and chemical properties as total phenolic content (TPC) and total antioxidant capacity (TAC). Raw (n?=?96) and milled (n?=?96) samples were evaluated by NIR spectroscopy and by classical batchwise assays, Folin-Ciocalteu, and ABTS for TPC and TAC, respectively, after different storage times (1 week to 2 months). Grape seeds had higher levels of TPC and TAC per sample dry weight when compared to grape skins. FT-NIR spectra of raw and milled samples were calibrated against content (%) of skins and seeds, TPC, and TAC using partial least squares (PLS) modeling. Spectral wavelength selection and latent variables were optimized for the lowest root-mean-square errors. PLS models’ results showed higher linearity for milled samples (0.936?>?R ²?>?0.914) when compared to raw samples (0.885?>?R ²?>?0.928). The range error ratio (RER) was between 10 and 14 for raw samples, while for milled grape pomace, it ranged from 15 to 18. Results confirmed that NIR spectroscopy can be applied to winemaking residues with virtually no sample processing needed to estimate the content of grape seeds and skins, the total phenolics, and total antioxidant capacity. Therefore, FT-NIR technique represents a non-destructive and eco-friendly technique to foster added value of grape pomace residues before time-consuming extraction steps are performed.     

Grape by‐product extracts against microbial proliferation and lipid oxidation: a review

The wine industry is responsible for the production of million tons of waste, such as grape skin, stalk, sludge and seeds, which can be considered inexpensive sources of phenolic compound owing to incomplete extraction during wine production. Phenolic compounds, also called polyphenols, comprise the most abundant bioactive compounds in grape and are recognized by their antioxidant and antimicrobial potential. Because of their functional properties, extracts obtained from grape wastes, which are rich in phenolic compounds, can be employed in the development of many products, ranging from medical to food applications, decreasing the growth of spoilage and pathogenic microorganisms and inhibiting lipid oxidation. These characteristics are motivating the research for alternative sources of natural antioxidant and antimicrobial agents, aimed at decreasing the use of artificial additives, which have been associated with some toxic effects. This article provides a review of the use of grape by‐product extracts and their bioactive compounds as natural antioxidant and antimicrobial agents in food products. © 2016 Society of Chemical Industry     

Effects of time of grape pomace fermentation and distillation cuts on the chemical composition of grape marcs

 The effects of fermentation time and distillation cuts on the composition of distillates in terms of ethanal, 1,1-diethoxyethane, methanol, 1-butanol, 2-butanol, 1-propanol, 2-methyl-1-propanol, 2-methyl-1-butanol, 3-methyl-1-butanol, ethyl acetate, ethyl 2-hydroxypropanoate, 3-methylbutyl acetate, hexyl acetate, 2-phenylethyl acetate, hexanol, trans-2-hexenol, trans-3-hexenol, cis-3-hexenol, 2-phenylethanol, ethyl butyrate, ethyl hexanoate, ethyl octanoate, ethyl decanoate, ethyl dodecanoate, 2-methylpropanoic acid, 3-methylbutanoic acid, hexanoic acid, octanoic acid, decanoic acid, and dodecanoic acid were assessed through data generated according to a factorial design using analysis of variance and principal component analysis. Four times of storage of pomace obtained following winemaking of two grape varieties of white Verde wine (Alvarinho and Loureiro) and three distillation cuts were considered; volatile compounds in the 24 samples generated were analyzed directly, and indirectly after extraction and concentration, by capillary gas chromatography. The results generated have suggested clear differences (P<0.05) between distillate cuts obtained throughout fermentation times for each grape variety. The major differences between the different distillate fractions analyzed were accounted for by the contents of diethyl butanoate, ethyl 2-hydroxypropanoate, and the sum of 3-methylbutanoic and 2-methylpropanoic acids for Loureiro, whereas the main differences were accounted for by the contents of diethyl butanoate and the sum of the carboxylic acids for Alvarinho.     

Supercritical CO2 extraction of oleanolic acid from grape pomace

In this work, we investigated the possibility to extract oleanolic acid through supercritical fluid extraction from waste materials such as grape pomace, which can represent a yet unexploited source of highly added‐value products. Preliminary supercritical fluid extractions were performed on dried grape skin samples and compared with solid–liquid extraction. After optimising extraction parameters such as temperature, pressure and extraction time, the ratio between supercritical and solid–liquid extraction yields was approximately 1/5. Supercritical CO₂ extractions were also carried out on dried grape pomace samples. In this case, the ratio between supercritical and solid–liquid extraction yields was increased up to 1/3, thanks to a combination of static and dynamic extraction and the use of ethanol as modifier. This extraction technique proved successful in extracting oleanolic acid from grape pomace samples with extraction yields that were comparable with those obtained with well‐established extraction techniques such as solid–liquid extraction.     

Phenolic compounds and antioxidant activity of seed and skin extracts of red grape (Vitis vinifera and Vitis labrusca) pomace from Brazilian winemaking

The phenolic compounds content and antioxidant activity of seed and skin of pomace from the vinification of grape varieties widely produced in Brazil were investigated with a view to their exploitation as a potential source of natural antioxidants. There was a greater concentration of phenolic compounds in the seeds (2128 to 16,518 mg of catechin equivalents (CE)/100 g) than in the skins (660 to 1839 mg CE/100 g). The highest antioxidant activity values determined as DPPH radical-scavenging ability and ferric reducing-antioxidant power (FRAP) were found for the seeds of the Pinot Noir variety (16,925 ??mol Trolox equivalents (TE)/100 g and 21,492 ??mol Fe²⁺/100 g, respectively) and in the skin extracts of the Isabel variety (3640 ??mol TE/100 g and 4362 ??mol Fe²⁺/100 g, respectively). The skin of Cabernet Sauvignon and Primitivo varieties had the highest contents of anthocyanins (935 and 832 mg/100 g, respectively). The grape seed extracts were rich in oligomeric and polymeric flavanols. The data suggested that grape seed and skin extracts may be exploited as antioxidant agents.