POLYPHENOLS AND ANTHOCYANINS OF GRAPE POMACE FROM CV. MERLOT
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1 POLYPHENOLS AND ANTHOCYANINS OF GRAPE POMACE FROM CV. MERLOT Branka LEVAJ, Danijela BURSAĆ KOVAČEVIĆ, Marina MARINČIĆ, Verica DRAGOVIĆ-UZELAC Faculty of Food Technology and Biotechnology University of Zagreb, Croatia Laboratory for Technology of Fruits and Vegetables Preservation and Processing
2 Ecological problem Economical problem Grape pomace seeds skin BAS - residue from winemaking
3 The most important in utilization of waste (Galanakis, 2012): To clarify the high added-value ingredients from impurities and toxic compounds, To avoid deterioration and loss of functionality during processing, To ensure the food grade nature of the final product.
4 Utilization of waste obtained in 5 steps (Galanakis, 2012): Pre-treatment to avoid spoilage of waste (grape pomace high proportion of water) (Macro and micro molecules separation) Extraction Isolation and Purification Product formation
5 BAS in red grape pomace skin (Kammemer et al, 2004) Anthocyanins > Phenolic acids > Others: Flavan-3-ols Procyanidins Flavonols Stilbens 39 identified Corrales et al Amount varies upon: cultivar year ripeness (e.g. phenolic acids)
6 Anthocyanins in red grape pomace skin (Kammemer et al. 2004) mg/kg DM (Cabernet Mitos, Germany, 2002), but e.g (Lamberger) malvidin 3-O-glc petunidin 3-O-pcoumaroylglc malvidin 3-O-pcoumaroylglc ** cyanidin 3-O-glc peonidin 3-O-glc petunidin 3-Oacetylglc * malvidin 3-O-acetylglc * delfinidin 3-Oacetylglc * petunidin 3-O-glc cyanidin 3-O-pcoumaroylglc ** delfinidin 3-O-glc peonidin 3-O-pcoumaroylglc ** peonidin 3-O-acetylglc *
7 Phenolic acids in red grape pomace - skin protocatechuic acid caftaric acid gallic acid coutaric acid ferulic acid fertaric acid caffeic acid syringic acid p-coumaric acid from 60.5 (Schwarzriesling, 2002) to mg/kg of dry matter (DM) (Lemberger, 2002) approx. 285mg/kg DM (Cabernet Mitos)
8 Other polyphenolics in red grape pomace - skin Flavan-3-ols catechin epicatechin Procyanidins procyanidin procyanidin B2 epicatechin gallate Flavonols quercetin 3-O-galactoside quercetin 3-O-glucoside quercetin 3-O-glucuronide quercetin 3-O-rhamnoside kaempferol 3-O-glucoside isorhamnetin quercetin kaempferol Stilbens trans-resveratrol trans-polydatin from (Cabernet Mitos, 2002) to mg/kg of DM (Lemberger, 2002)
9 BAS in grape pomace - skin Flavan-3-ols Proantocyanidins Flavonols (Amico et al. 2004) Stilbens Resveratrol
10 BAS from red grape health benefits antioxidants malvidin - protect the cardiovascular system (Quientieri et al. 2013) catechin, epicatechin, quercetin and its glycoside rutin, transresveratrol - protect human low density lipoprotein (LDL) against oxidation (Yilmaz &Toledo 2004) potential anticancer properties (resveratrol - Frémont, 2000) antiinflammatory (resveratrol -Frémont,2000)
11 BAS polyphenolics extraction from red grape pomace skin yield composition purity chemical nature (simple or complex or presence of interfering substances) sample size (liquid-solid ratio, particle size) extraction conditions and technique (solvent±acid, time, temperature, flow) (Naczk & Shahidi, 2004, Pinelo et al. 2005)
12 BAS polyphenolics extraction from red grape pomace skin organic/alcoholic solvent water mixtures for the extraction of vinification solid residues (Bonilla, Mayen, Merida, & Medina, 1999; Negro, Tommasi, & Miceli, 2003): permeability of cell tissue better mass transfer by molecular diffusion (Jayaprakasha, Singh, & Sakariah, 2001) water soluble bioactive compounds.
13 BAS polyphenolics extraction from red grape pomace skin organic/alcoholic ± acid (Sólyom et al. 2014) Solvent type: acetone, ethanol or methanol Different researchers have found different solvent as the best.
14 BAS extraction from grape pomace - skin Modern technique: Ultrasound (González-Centeno et al. 2014) Pulsed electric field (Liu et al. 2011) Supercritical fluid extraction - use of a gas (i.e. CO 2 ) above its critical temperature and pressure, it is between liquid and gas Microwave (Ghafoor et al. 2011) High pressure (Corrales et al. 2009)
15 The aim: to investigate efficiency of extraction from grape pomace - skin on yield of - total phenols - anthocyanins
16 Material: grape pomace Merlot vinary Agrolaguna, 2014, Poreč, Croatia Dried skin Seeds liofilized = freeze dried
17 Methods Extraction: - ethanol/water (1:1) solvent - acidity (addition of HCL: 0, 0.5 and 1 %) - duration of extraction (30, 45 and 60 minutes) - temperature 80 o C Analysis: - total phenols -by Folin-Ciocalteu reagent -anthocyanins -by ph-differential method
18 mg/100g g/100g Results: -higher in case of acidic solvents -less difference between 0.5 and 1.0 %HCl - Time 120, ,000 80,000 60,000 40,000 20,000 Total phenols 0,000 0 % HCl 0.5 % HCl 1.0 % HCl 30 min 28,672 63,825 61, min 25,906 82,906 87, min 30,905 95,931 87,871
19 mg/100g g/100g Results: -higher in case of acidic solvents -less difference between 0.5 and 1.0 %HCl -time 0,45 0,4 0,35 0,3 0,25 0,2 0,15 0,1 0,05 Anthocyanins 0 0 % HCl 0.5 % HCl 1.0 % HCl 30 min 0,1607 0,3066 0, min 0,1392 0,2161 0, min 0,1569 0,2382 0,2435
20 Conclusions yield of total phenols and anthocyanins was higher in case of acidic solvents but with different pattern considering extraction time. yield of total phenols increased with extraction time yield of anthocyanins decreased with time but not remarkable taking into account both classes recommended extraction condition would be ethanol:water (1:1) with 1% HCl/60 min.
21 This research was financed by the funds of the project "Application of innovative technologies in the isolation of bioactive compounds from organic waste in the wine production" The project is co-financed by the European Union from the European Regional Development Fund.
22 References : Galanakis, C.M (2012) Recovery of high added-value components from food wastes Conventional, emerging technologies and commercialized applications.trends in Food Science & Technology 26, Kammerer D., Claus A., Carle R., Schieber A. (2004)Polyphenol Screening of Pomace from Red and White Grape Varieties (Vitis vinifera L.) by HPLC-DAD- MS/MS J. Agric. Food Chem., 52 (14), pp Corrales M., Fernández García A., Butz P., Tauscher B. (2009) Extraction of anthocyanins from grape skins assisted by high hydrostatic pressure. Journal of Food Engineering 90, Amico V., Napoli E.M., Renda A., Ruberto G., Spatafora C., Tringali C. (2004) Constituents of grape pomace from the Sicilian cultivar Nerello Mascalese.Food Chemistry 88, Quintieri AM, Baldino N, Filice E, Seta L, Vitetti A, Tota B, De Cindio B. (2013) Malvidin, a red wine polyphenol, modulates mammalian myocardial and coronary performance and protects the heart against ischemia/reperfusion injury. The Journal of nutritional biochemistry 24 (7) Yilmaz, Y., & Toledo, R. T. (2006). Oxygen radical absorbance capacities of grape/wine industry byproducts and effect of solvent type on extraction of grape seed polyphenols. Journal of Food Composition and Analysis, 19(1), Frémont, L., Biological effects of resveratrol. Life Sciences 66, Naczk, M., & Shahidi, F. (2004). Extraction and analysis of phenolics in food: Review. Journal of Chromatography A, 1054, Pinelo, M., Rubilar, M., Jerez, M., Sineiro, J., & Nunez, M. J. (2005). Effects of solvent, temperature, and solvent-to-solid ratio on the total phenolic content and antiradical acivity of extracts from different compounds of grape pomace. Journal of Agricultural and Food Chemistry, 53, Bonilla, F., Mayen, M., Merida, J., & Medina, M. (1999). Extraction of phenolic compounds from red grape marc for use as food lipid antioxidants. Food Chemistry, 66, Negro, C., Tommasi, L., & Miceli, A. (2003). Phenolic compounds and antioxidant activity from red grape marc extracts. Bioresource Technology, 87, Jayaprakasha, G. K., Singh, R. P., & Sakariah, K. K. (2001). Antioxidant activities of grape seed (Vitis vinifera) extracts on peroxidation models in vitro. Food Chemistry, 73, Ghafoor, K., Park, J., & Choi, Y.-H. (2011). Optimization of supercritical fluid extraction of bioactive compounds from grape (Vitis labrusca B.) peel by using response surface methodology. Innovative Food Science and Emerging Technologies, 11, 485e490. Sólyom K, Solá R, Cocero MJ, Mato RB. (2014) Thermal degradation of grape marc polyphenols. Food Chemistry González-Centeno MR, Knoerzer K, Sabarez H, Simal S, Rosselló C, Femenia A. (2014) Effect of acoustic frequency and power density on the aqueous ultrasonic-assisted extraction of grape pomace (Vitis vinifera L.) - a response surface approach. Ultrason Sonochem. 21(6): Liu, D., Vorobiev, D., Savoire, R., & Lanoiselle, J.-L. (2011). Extraction of polyphenols from grape seeds by unconventional methods and extract concentration through polymeric membrane. In P. S. Taoukis, N. G. Stoforos, V. T. Karathanos, & G. D. Saravacos (Eds.), Food process engineering in a changing world. Proceedings of the 11th International Congress on Engineering and Food, May 22e26, 2011, Athens, Greece (pp. 1939e1940). Athens: Cosmosware.
23 Thank you for your attention
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