ANNALS OF AGRARIAN SCIENCE, vol. 11, no. 2, 2013 ИЗВЕСТИЯ АГРАРНОЙ НАУКИ, Том 11, Ном. 2, 2013
TECHNOLOGY OF PROCESSING OF AGRICULTURAL PRODUCTS ТЕХНОЛОГИЯ ПЕРЕРАБОТКИ СЕЛЬСКОХОЗЯИСТВЕННЫХ ПРОДУКТОВ
OXIDATIVEOXIDATIVE REDUCTIONREDUCTION FORMATION OF BIOLOGICALLY ACTIVE QUERCETIQUERCETINN AND DIHYDROQUERCETIN (TAXIFOLIN) DURING WINE FERMENTATION AND WINE FORMATION
M.M.M.GM. GGG.. Bezhuashvili, * T.MT.M.... Shonia, ** T. SShhhh.. Buachidze ** *Agricultural University of Georgia 13 km, David Aghmashenebeli Ave., Tbilisi, 0131, Georgia; [email protected] **Technical University of Georgia 77, Kostava St., Tbilisi, 0175, Georgia; tekle [email protected]; [email protected] Received: 22.02.13; accepted: 14.04.13
Oxidative reduction transformation of quercetin and dihydroquercetin is studied during alcoholic fermentation of musts made from Rkatsiteli and Tsolikouri wine varieties and also during Kakhetian and Imeretian type wine formation. Alcoholic fermentation was conducted using wine yeasts: Sacch. vini – Kakhuri 42 and Sacch. vini – Tsolikouri 13 . The reduction transformation of quercetin into dihydoquercetin during alcoholic fermentation was observed, which is intensified due to the increase of titrable acidity. During wine formation dihydroquercetin is oxidized into quercetin.
INTRODUCTION Wide spectrum of grape flavonoids procyanidins (oligomeric and polymeric), flavonols, flavanols, flavones, flavonones and others are important chemical substances that are presented in high quality wines. Flavonoids are extracted intensively from the stems and seeds (and other hard parts) of the grapes and are localized in the wine by natural and transformed forms. As flavonoids are localized in the hard parts of the grapes, their content is high in the wines fermented on the must: red wines, Kakhetian and Imeretian type wines. For example total number of one of the groups of flavonoids flavonols in Kakhetian type wines made from Rkatsiteli vine variety is 41, 25 mg/l, and in Imeretian type wines made from Tsolikauri is 23, 7 mg/l [1]. Flavonoids take part in the oxidative reduction transformation during wine maturation and influence greatly the quality of the wines. Besides, due to the medical researches single representatives are biologically active substances and also carry curative prophylactic values towards several diseases. Versus the scientific researches antibacterial [2], antivirus [3] and antioxidant activity of quercetin was determined [4,5]. Cardio protective and antiplatelet activities were also revealed [6,7]. It is determined that rutin and ascorbin acid reveal high therapeutic effect against blood vascular diseases.
Due to the above mentioned the research of the transformation of flavonoids in accordance with antioxidant activity is an interesting subject to be studied. We have studied some flavonoids – quercitrin, rutin, transformation of quercetin and dihydroquercetin in the grape juice during alcoholic fermentation; diagram of transformation was created: for example quercetin and rutin produce quercetin. In the fermented area quercetin stays partly in its natural form or as transformation product [8], by wine yeast samples: Sacch. vini – Kakhuri 42; Sacch. vini. – Rkatsiteli 61; Sacch. vini – Tsolikorui 13; Sacch. chofati da Sacch. oviformisi. During grape juice and model area alcoholic fermentation dihydroquercetin is transformed intensively and one substance among other transformed substances is identical with quercetin transformation product. Intensity of dihydroquercetin by single wine yeast is similar [9]. As we have prolonged the research of flavonoids, at the next stage we have studied: interconversion of quercetin and dihydroquercetin during wine fermentation and wine formation of Kakhetian and Imeretian type of wines
OBJECTIVES AND METHODS The research objects were the wines made from Rkatsiteli vine variety and Imeretian type wines made from Tsolikouri vine variety. The model trial experiment of quercetin transformation was done by fermentation of individual quercetin found in the wines made from Rkatsiteli and Tsolikouri using wine yeasts Sacch. vini – Kakhuri 42 and Sacch. vini – Tsolikorui 13; quantitative analysis of quercetin was conducted using spectrophotometric method and qualitative analysis was conducted by paper chromatography system N buthanol:acetic acid:water (4:1:2). Chromatograms were revealed using vanillin reagent and ALCL 3 solution. We used standard dihydroquercetin and quercetin for the experiment.
RESULTS AND ANALYSIS The accumulation dynamics of quercetin and dihydroquercetin during must fermentation process of Kakhetian type wine made from Rkatsiteli vine variety and Imeretian type wines made from Tsolikouri differ from each other. As a result the content of quercetin in the grape juice made from Rkatsiteli vine variety is 0,35 mg/l and from Tsolikouri vine variety is 0,23 mg/l. Intensive quantitative increase of quercetin during wine must fermentation is a result of extraction of hard parts from grape and also a result of transformation determined by us mainly by division of quercetin and rutin. As a result of intensive fermentation the concentration of quercetin in the fermented must of Rkatsiteli vine variety reaches 7,7mg/l, in Tsolikouri must 1,6 mg/l and then it decreases. Dynamics of dihydroquercetin after 7 8 days of fermentation is changed with increased concentration (Fig.1).
C, mg/l
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Fig 1. Dynamics of accumulation of quercetin and dihydroquercetin Rkatsiteli (1, 3) and Tsolikouri (2,4) in the must during wine fermentation
The titrable acidity of Tsolikouri grape juice was higher than Rkatsiteli grape juice (correspondingly 9,2 g/l and 9, 8 g/l), we decided to study the titrable acidity influence over the creation of dihydroquercetin. We have done the model experiment of alcoholic fermentation of Rkatsiteli and Tsolikouri vine varieties for the simultaneous existence of quercetin and dihydroquercetin where the content of titrable acidity was increased by adding tartaric acidity. The fermentation was conducted using wine yeasts: Sacch. vini – Kakhuri 42 and Sacch. vini – Tsolikouri 13. The data of table show that by increase of titrable acidity the concentration of dihydroquercetin is increased and quercetin decreased. This chractaristic is similar for Rkatsiteli, as well as for Tsolikouri grape fermentation juices. The result of the experiment shows the reduction transformation of quercetin during wine fermentation by creating dihydroquercetin, which is intesified by increase of titrable acidity.
Table. The change of quercetin and dihydroquercetin caused by change of titrable acidity revealed by model trial alcoholic fermentation
Quercetin, mg/l Dihydroquercetin Version of experiment Initial 3 days 7 days 14 days Initial 3 days 7 days 14 days Grape juice of Rkatsiteli – quercetin + dihydroquercetin titrable acidity: g/l 9,2 10 9,8 9,5 9,1 10 10,1 10,3 10,6 9,7 10 9,5 8,8 8,3 10 10,3 10,5 10,9 10,2 10 9,0 8,3 7,7 10 10,4 10,6 11,2 Grape juice of Tsolikouri – quercetin + dihydroquercetin titrable acidity: g/l 9,8 10 9,4 8,8 8,2 10 10,2 10,5 10,9 10,3 10 8,8 8,0 7,5 10 10,5 10,8 11,2 10,8 10 8,5 7,8 7,3 10 10,7 11,0 11,5
To discuss the above mentioned results, the transformation of quercetin and dihydroquercetin during intensive wine fermentation is important (2 7 days). The coincidence of the number of decreased quercetin and increased dihydroquercetin was determined. That is the decreased number of quercetin is not transformed into dihydroquercetin, but the part of it is used for new product formation. Also dihydroquercetin during intensive alcoholic fermentation process is transformed and as a result is increased by small amount. Compared to intensive period of alcoholic fermentation during slow alcoholic fermentation more concentration of dihydroquercetin takes place. It indicates that dihydroquercetin transformation process is weakened, which causes the quantitative increase. During one year formation of Kakhetian and Imeretian type wines the concentration of dihydroquercetin is decreased and quercetin is increased. Due to the analysis of model fermented versions the oxidative transformation of dihydroquercetin into quercetin was determined. The dynamic of this transformation (Fig. 3) shows that during 12 months formation period dihydroquercetin is oxidized intensively 3 months later. These transformations are shown in the scheme below.
Fig. 2. Oxidative transformation of dihydroquercetin to quercetin
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mg/l 7,00 C, 6,00
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Fig 333.3. Quercetin (1) and dihydroquercetin (2) change of Kakhetian type of wine during wine formation
CONCLUSION According to the conducted experiment the interexchange of quercetin and dihydroquercetin (taxifolin) as a result of oxidative reduction process was revealed. During must alcoholic fermentation quercetin is transformed into dihydroquercetin, during wine formation dihydroquercetin is oxidized as quercetin. Reduction of quercetin into dihydroquercetin is intensified in the condition of high titrable acidity. REFERENCES • Shonia T., Bezhuashvili M., Pataraia M . Accumulation Dinamics of the Sum of Flavonoids Found in Kakhetian and Imeretian Type Wines // Viticulture and Winemaking, 4, 2006, pp. 19 20. • Stavric B. Quercetin in our Diet from Potent Mytagen to Probable Antiocarcinogen // Clinical Biochemistry, 27, 1994, pp. 245 248. • Vrijsen R., Everaert L. and Boeje A. Antiviral Activity of Flavones and Potentiation by Ascorbate // J. of Geneology and Virologi, 69, 1998, pp. 1749 1751. • Takahama U. Unhibition of Lipoxygenase Dependent Lipid Peroxidation by Quercetin: Mechanism of Antioxidative Function // Phytochemistry, 24, 1985, pp. 1443 1446. • Bezhuashvili M.G., Meskhi M.U., Bostoghanashvili M.A., Malania M.A. Antioxidant Activity of the Extract Containing Benzo stilben, Determined by “in vitro” // Winemaking and Viticulture, 3, 2005, pp.26 27. • Pace Asciak C. R., Haln S., Diamandis E.P., Soleas G. and Goldberg D. M. The Red Wine Phenolics Trans Resveratrol and Quercetin Block Human Platelet Aggregation and Eicosanoid Synthesis: Implications for Protection Against Coronary Heart Disease // Clinica Chemist Acta, 235, 1995, pp. 207 219. • Conquer Y.A., Maiani G., Azzini E., Raguzzini A. and Hobub B.Y. Supplementation with Quercetin Markedly Increases Plasma Quercetin Concentration Without Effect on Selected Risk Factors for Heart Disease in Healthy Subjects // J. of Nutrition, 128, 1998, pp. 595 597. • Bezhuashvili M.G., Shonia T.M. Influence of Wine Yeast on Different Flavonoids During Alcoholic Fermentation // Winemaking and Viticulture, 5, 2008, pp.22 24. • Shonia T. M., Bezhuashvili M.G., Aplakov A.R. Influence of Wine Yeasts on the Dihydroquercetin During Alcoholic Fermentation // Georgian Engineering News, 1, 2009. pp.193 195.
ОООКИСЛИТЕЛЬНООКИСЛИТЕЛЬНОКИСЛИТЕЛЬНО ВОССТАНОВИТЕЛЬНЫЕВОССТАНОВИТЕЛЬНЫЕ ПРЕВРАЩЕНИЯ БИОЛОГИЧЕСКИ АКТИВНОГО КВЕРЦЕТИНА И ДИГИДРОКВЕРЦЕТИНА (ТАКСИФОЛИННА)А) В ПРОЦЕССАХ АЛКОГОЛЬНОГО БРОЖЕНИЯ И ФОРМИРОВАНИЯ ВИНОМАТЕРИАЛОВ
МММ.М...ГГГГ....БежуашвилиБежуашвилиБежуашвили,, ТТТ.Т...ММММ....ШонияШонияШония,, ТТТ.Т...ШШШШ.. Буачидзе
Исследованы окислительно восстановительные превращения кверцетина и дигидрокверцетина (таксифолина) в алкогольном брожении мезги Ркацители и Цоликоури, а также, соответственно, в период формирования виноматериалов кахетинского и имеретинского типов. Алкогольное брожение проведено штаммами винных дрожжей: Sacch.vini Кахури 42 и Sacch.vini Цоликоури 13 . Установлено восстановительное превращение кверцетина в дигидрокверцетин в процессе алкогольного брожения, которое интенсифицируется увеличением титруемой кислотности. При формировании виноматериалов дигидрокверцетин окисляется в кверцети
სოფოსოფო,, ამის ქვემოთ მინდა მხოლოდ შევარჩიო ნახაზები
C, mg/l 9,00 8,00 7,00 6,00 5,00 4,00 3,00 2,00 1,00 0,00 2 4 6 8 10 12 14 1 2 3 4 τ , Day-Night
FigFigFig.Fig ... 1. Dynamics of accumulation of quercetin and dihydroquercetin Rkatsiteli (1, 3) and Tsolikouri (2,4) in the must during wine fermentation
Fig. 2. Oxidative transformation of dihydroquercetin to quercetin
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mg/l 7,00
C, 6,00 5,00 4,00 3,00 2,00 1,00 0,00 Initial 3 6 9 12 1 2 τ , Months
FigFigFig.Fig ... 333.3. Quercetin (1) and dihydroquercetin (2) change of Kakhetian type of wine during wine formation