Determination of organic acid and polyphenol content in grape seeds obtained with pressurized water exraction

Abstract

In this study, using an eco–friendly extraction system, it is aimed to obtained extract has high polyphenol and organic acid content from black grape seeds which are one of the most important industrial waste for wine and grape juice. For this purpose grape seed extract was obtained by using pressurized water extractor on pilot scale at 50 bar, 50°C and 90 min and its polyphenol and organic acid content was determined by chromatographic and spectroscopic methods. Total phenolic content was found as 49.77% (w/w) in terms of gallic acid equivalent and antioxidant activity that was determined by DPPH method was 90.90% as inhibition, 98.13% as antiradical activity. As a result of high performance liquid chromatography (HPLC) analysis, tartaric, oxalic, L–malic acid and fumaric acid were found and the presence of hydroxytyrosol and (+)–catechin as polyphenol were revealed in extract. Grape seed extract is a commercially available product in the market. The extract obtained by the pressurized water extraction method, has a strong aroma and desirable stability with organic acids in the structure as well as the rich of polyphenols content. Thus, grape seed extract which is a higher quality and more prominent health effects will be possible to put on the market with the support of eco–friendly extraction method used.

Keywords

• pressurized water exraction,organic acid,polyphenols,grape seed

Basınçlı su ekstraksiyonuyla elde edilen üzüm çekirdeği ekstraktının polifenol ve organik asit içeriğinin belirlenmesi

Abstract

Bu çalışmada doğa dostu bir ekstraksiyon sistemi kullanılarak, şarap ve üzüm suyu sanayilerinin önemli atıklarından biri olan siyah üzüm çekirdeklerinden polifenol ve organik asit içeriği zengin bir ekstraktın elde edilmesi hedeflenmiştir. Bu amaçla pilot ölçekte basınçlı su ekstraksiyonu sistemi yardımıyla 50 bar basınç altında, 50°C sıcaklıkta, 90 dk’da elde edilen üzüm çekirdeği ekstraktının polifenol ve organik asit içeriği, kromatografik ve spektroskopik yöntemlerle belirlenmiştir. Üzüm çekirdeği ekstraktının toplam fenolik madde miktarı gallik asit eş değeri cinsinden %49.77 (w/w) ve DPPH yöntemiyle belirlenen antioksidan aktivitesi % inhibisyon cinsinden 90.90, antiradikal etkinliği ise %98.13 olarak tespit edilmiştir. Yüksek performans sıvı kromatografisi (HPLC) analizi sonucunda organik asit olarak ekstrakt içerisinde sırasıyla tartarik, okzalik, L–malik ve fumarik asitlerin bulunduğu ve polifenol olarak hidroksitirosol ve (+)–kateşin’in olduğu tespit edilmilştir. Üzüm çekirdeği ekstraktı, piyasada ticari olarak satışı gerçekleşen bir üründür. Basınçlı su ekstraksiyonu yöntemiyle elde edilen ekstrakt, zengin polifenol içeriğinin yanısıra yapısındaki organik asitlerin varlığıyla güçlü bir aromaya ve arzu edilen stabiliteye sahiptir. Böylece bu çalışmada kullanılan basınçlı su ekstraksiyon yönteminin de desteğiyle, daha kaliteli ve sağlık etkileri daha baskın üzüm çekirdeği ekstraktının piyasaya sunulmasının mümkün olacağı düşünülmektedir.

Keywords

• basınçlı su ekstraksiyonu,organik asitler,polifenoller,üzüm çekirdeği

References

1. Dönmez, A., 2015. Denizli Bölgesinde Yetiştirilen Bazı Üzüm Çeşitlerinin Resveratrol ve Suda Çözünen Vitaminlerinin Kuruma Kinetiği (Yüksek Lisans Tezi). Pamukkale Üniversitesi Fen Bilimleri Enstitüsü Gıda Mühendisliği Anabilim Dalı, Denizli, 105s.
2. Otağ, M. R., 2015. Denizli Çal Yöresinde Yetişen Bazı Üzüm Çeşitlerinin Farklı Olgunlaşma Evreleri ve Kurutulması Sonrasında Bazı Özellikleri ile Resveratrol İçeriğinin Belirlenmesi (Doktora Tezi). Pamukkale Üniversitesi Fen Bilimleri Enstitüsü Gıda Mühendisliği Anabilim Dalı, Denizli, 165s.
3. Perumalla, A. V. S. and N. S. Hettiarachchy, 2011. Green Tea and Grape Seed Extracts Potential Applications in Food Safety and Quality. Food Research International 44:827–839.
4. Lau, D. W. and A. J. King, 2003. Pre and Post–Mortem Use of Grape Seed Extract in Dark Poultry Meat to Inhibit Development of Thiobarbituric Acid Reactive Substances. Journal of Agricultural and Food Chemistry 51:1602−1607.
5. Rein, D., T. G., Paglieroni, D. A., Pearson, T., Wun, H. H., Schmitz, R., Gosselin and C. L., Keen, 2000, Cocoa and Wine Polyphenols Modulate Platelet Activation and Function. The Journal of Nutrition 130:2120−2126.
6. Shanmuganayagam, D., M. R., Beahm, H. E., Osman, C. G., Krueger, J. D., Reed and J. D. Folts, 2002, Grape Seed and Grape Skin Extracts Elicit a Greater Antiplatelet Effect when Used in Combination than when Used Individually in Dogs and Humans. The Journal of Nutrition 132(12):3592−3598.
7. Clouatre, D. L. and C. Kandaswami, 2005. Grape Seed Extract. In Encyclopedia of Dietary Supplements (Eds. P. Coates, M. Blackman and G. Cragg). New York, NY: Marcel Dekker, ABD. pp: 309−325.
8. Bentivegna, S. S. and K. M. Whitney, 2002, Subchronic 3–Month Oral Toxicity Study of Grape Seed and Grape Skin Extracts. Food and Chemical Toxicology 40(12):1731−1743.

9. Monteleone, E., N. Condelli, C. Dinnella and M. Bertuccioli, 2004, Prediction of Perceived Astringency Induced by Phenolic Compounds. Food Quality and Preference 15:761−769.
10. Weber, H. A., A. E. Hodges, J. R. Guthrie, B. M. O’Brien, D. Robaugh, A. P. Clark, R. K. Harris, J. W. Algaier and C. S. Smith, 2007. Comparison of Proanthocyanidins in Commercial Antioxidants: Grape Seed and Pine Bark Extracts. Journal of Agricultural and Food Chemistry 55(1):148−156.
11. Mahmoudi, M., K. Charradi, F. Limam and E. Aouania, 2016. Grape Seed and Skin Extract as Anadjunct to Xenical Therapy Reducesobesity, Brain Lipotoxicity and Oxidativestress in High Fat Diet Fed Rats. Obesity Research & Clinical Practice DOI: 10.1016/j.orcp.2016.04.006.
12. Ferruzzi, M. G., J. K. Lobo, E. M. Janle, B. Cooper, J. E. Simon, Q. L. Wu, C. Welch, L. Ho, C. Weaver and G. M. Pasinetti, 2009. Bioavailability of Gallic Acid and Catechins from Grapeseed Polyphenol Extract is improved by Repeated Dosing Inrats: Implications for Treatment in Alzheimer’s disease. Journal of Alzheimer’s Disease 18(1):113—124.
13. Du Y., H., Guo and H., Lou, 2007. Grape Seed Polyphenols Protect Cardiaccells from Apoptosis via Induction of Endogenous Antioxidant Enzymes. Journal of Agricultural and Food Chemistry 55(5):1695–1701.
14. Bagchi, D., M., Bagchi, S.J., Stohs, D.K., Das, S.D., Ray, C.A., Kuszynski, S.S., Joshi and H.G., Pruess, 2000. Free Radicals and Grape Seed Proanthocyanidin Extract: Importance in Human Health and Diseaseprevention. Toxicology 148(2–3):187–197.
15. Khazri, O., K., Charradi, F., Limam, M. V., El Mayc and E., Aouania, 2016. Grape Seed and Skin Extract Protects Against Bleomycin–Induced Oxidative Stress in Rat Lung. Biomedicine&Pharmacotherapy 81:242–249.
16. Khanal, R.C., L.R. Howard and R.L. Prior, 2009. Procyanidin Content of Grape Seed and Pomace, and Total Anthocyanin Content of Grape Pomace as Affected by Extrusion Processing. Journal of Food Science 74:174–182.
17. Casazza, A.A., B. Aliakbarian and P. Perego, 2011. Recovery of Phenolic Compounds from Grape Seeds: Effect of Extraction Time and Solid–Liquid Ratio. Natural Product Research 25:1751–1761.
18. Negro, C., L., Tommasi and A., Miceli, 2003. Phenolic Compounds and Antioxidant Activity from Red Grape Marc Extracts. Bioresource Technology 87(1):41−44.
19. Renaud, S., M. de Lorgeril, 1992. Wine, Alcohol, Platelets, and the French Paradox for Coronary Heart Disease. Lancet 339:1523–1526.
20. Kaur, M., R., Mandair, R., Agarwal and C., Agarwal, 2008, Grape Seed Extract Induces Cell Cycle Arrest and Apoptosis in Human Colon Carcinoma Cells. Nutrition and Cancer 60:2–11.
21. Silva, L.R. and M. Queiroz, 2016, Bioactive Compounds of Red Grapes from Dão Region (Portugal): Evaluation of Phenolic and Organic Profile. Asian Pacific Journal of Tropical Biomedicine 6(4):315–321.
22. Dopico–García, M.S., A., Fique, L., Guerra, J.M., Afonso, O., Pereira, P., Valentão, P.B., Andrade and R.M., Seabra, 2008, Principal Components of Phenolics to Characterize Red Vinho Verde Grapes: Anthocyanins or Non–Coloured Compounds? Talanta 75:1190–202.
23. Mato, I., S., Suárez–Luque and J.F., Huidobro, 2007, Simple Determination of Main Organic Acids in Grape Juice and Wine by Using Capillary Zone Electrophoresis with Direct UV Detection. Food Chemistry 102:104–12.
24. Fennema, O.R., 1985. Food Chemistry. Marcel Dekker Inc., New York, ABD. pp:991.
25. Nakilcioğlu–Taş, E., 2016, Zeytin Çekirdeğinin Pilot Ölçekte Basınçlı Su Ekstraktörü Kullanılarak Su Ekstraktının Eldesi ve Mikroenkapsülasyonu (Doktora Tezi). Ege Üniversitesi Fen Bilimleri Enstitüsü Gıda Mühendisliği Anabilim Dalı, İzmir, 242s.
26. Singleton, V. L. and J. A., Rossi, 1965, Colorimetry of Total Phenolics with Phosphomolybdic–Phosphotungstic Acid Reagents. American Journal of Enology and Viticulture 16:144–153.
27. Li, Y., C., Guo, J., Yang, J., Wei, J., Xu and S., Cheng, 2006, Evaluation of Antioxidant Properties of Pomegranate Peel Extract in Comparision with Pomegranate Pulp Extract. Food Chemistry 96:254–260.
28. Singh, R.P., K.N.C., Murthy and G.K., Jayaprakasha, 2002, Studies of the Antioxidant Activity of Pomegranate (Punica granatum) Peel and Seed Extracts Using in vitro Models. Journal of Agricultural and Food Chemistry 50:81–86.
29. Alper, N., P., Onsekizoğlu and J., Acar, 2011, Effects of Various clarification Treatments on Phenolic Compounds and Organic Acid Compositions of Pomegranate (Punıca granatum L.) Juice. Journal of Food Processing and Preservation 35:313–319.
30. Mandić, A. I., S. M. Djilas, J. M. Čanadanović–Brunet, G. S. Ćetković and J. J. Vulić, 2009. Antioxidant Activity of White Grape Seed Extracts on DPPH Radicals. Acta Periodica Technologica 40:53–61.
31. Jayaprakasha, G. K., T. Selvi and K. K. Sakariah, 2003. Antibacterial and Antioxidant Activities of Grape (Vitis vinifera) Seed Extracts. Food Research International 36:117–122.
32. Shiyaji, J., 2007. Determination of Antioxidant Activity of Polyphenol Extract from Grape Seeds (Master Thesis). San Jose State University The Faculty of the Department of Nutrition and Food Science, California, pp:49.
33. Baydar, N. G., Z. Babalık, F. H. Türk and E. S. Çetin, 2011. Phenolic Composition and Antioxidant Activities of Wines and Extracts of Some Grape Varieties Grown in Turkey. Journal of Agricultural Sciences 17:67–76.
34. Farhadi, K., F. Esmaeilzadeh, M. Hatami, M. Forough and R. Molaie, 2016. Determination of Phenolic Compounds Content and Antioxidant Activity in Skin, Pulp, Seed, Cane and Leaf of Five Native Grape Cultivars in West Azerbaijan Province, Iran. Food Chemistry 199:847–855.
35. Curko, N., K. Kovacevic–Ganic, L. Gracin, M. Ðapic, M. Jourdes and P. L. Teissedre, 2014. Characterization of Seed and Skin Polyphenolic Extracts of Two Red Grape Cultivars Grown in Croatia and Their Sensory Perception in a Wine Model Medium. Food Chemistry 145:15–22.
36. Rockenbach, I. I., E. Jungfer, C. Ritter, B. Santiago–Schübel, B. Thiele, R. Fett and R. Galensa, Characterization of Flavan–3–ols in Seeds of Grape Pomace by CE, HPLC–DAD–MSn and LC–ESI–FTICR–MS. Food Research International 48:848–855.