357 Analysis of Complex Technologies for Obtaining Wine Products with Increased Antioxidant Properties* Yurij V. Grishin1, Anatoliy N. Kazak2 [0000-0001-7678-9210], Nikolay N. Oleinikov2 [0000-0002-9348-9153], Nadezhda I. Gallini2[0000-0001-8176-6419], Petr V. Chetyrbok2[0000-0002-0115-9158] 1Magarach All-Russia National Research Institute for Viticulture and Wine-Making, Yalta, Russia 2V.I. Vernadsky Crimean Federal University, Simferopol, Russia kazak_a@mail.ru Abstract. This paper presents the results of studies of the phenolic composition and antioxidant activity of table wine materials from grapes of white technical European varieties grown in the soil and climatic conditions of the southern re- gions of Russia, depending on the method of its processing and using the solid components of the grape bunch. The study showed that the use of tegumental maceration provides an increase in the mass concentration of monomeric forms of phenolic compounds by 1.8 times, antioxidant activity by 1.3 times; partial (1/3 of grape sugars) fermentation of pulp with ridges allows increasing the mass concentration of monomeric forms of phenolic compounds by 2.8 times, and. and antioxidant activity 1.8 times; complete fermentation of sugars in pulp with ridges leads to an increase in the mass concentration of monomeric forms of phe- nolic compounds in products by 5.0 times and antioxidant activity by 2.1 times compared to the traditional "white" grape processing method, without contacting the must with the solid parts of the grapes. The developed complex technology of processing grapes of white technical European varieties with the use of wine- making wastes will make it possible to obtain high-quality products with in- creased antioxidant properties. Keywords: Antioxidant Activity, Tegumental Maceration, Biologically Active Substances, High-Performance Liquid Chromatography (HPLC), Wine Mate- rial, Grape, Grape Crush, Componential Composition, Stems, Phenolic Com- pounds, Monomeric Forms of Phenolic Compounds, Procyanidins. 1 Introduction The key to longevity and full life is the use of enriched foods in nutrition, as is custom- ary, for example, in Japan, where the introduction of a healthy (functional) nutrition program has made it possible to increase life expectancy by 8 years throughout the * Copyright 2021 for this paper by its authors. Use permitted under Creative Commons License Attribution 4.0 International (CC BY 4.0). 358 country [1-4]. According to the US Department of Health, the minimum daily intake of antioxidants required by the human body is 3000-5000 units. ORAC (Oxygen Radical Absorption Capacity) is a unit for measuring the number of antioxidants, expressed in micromoles of Trolox per unit mass (μTE / 100g) [5-7]. It is known from the literature that when processing 1 thousand tons of grapes for must and wine, about 120 tons of pomace, 4 tons of seeds, 5 tons of ridges, and other secondary products of winemaking are formed. The characteristics of the ridges separated during crushing of grapes as a waste of winemaking have the following characteristics: mass concentration of sugars - 1.5-2.0 g / dm3, mass fraction of phenolic substances 3-6%, mineral substances up to 2.5%, tartaric acid up to 0, sixteen]. In the scientific literature, data on the study of antioxidant activity (AOA) of fruit crops, grape ridges, phenolic compounds, and AOA of white sparkling wines from various producing countries, quality and safety of wines obtained by secondary fermentation are also known. According to J. Tauchen's research [7], still, white wines made from white technical grapes (Sauvignon Blanc, Chardonnay, and Rkatsiteli) according to the technology gen- erally accepted in Europe, which involves the rapid separation of the must from the hard parts of the grapes, have a low mass concentration of phenolic substances. varying in the range from 0.2 to 0.29 g / dm3. At the same time, the wine made from Rkatsiteli grapes using the Kakhetian technology (fermentation of pulp with ridges) was distin- guished by a higher mass concentration of phenolic substances, which amounted to 0.4 g / dm3. The key to longevity and full life is the use of enriched foods in nutrition, as is customary, for example, in Japan, where the introduction of a healthy (functional) nutrition program has made it possible to increase life expectancy by 8 years throughout the country [8-11]. According to the US Department of Health, the minimum daily intake of antioxi- dants required by the human body is 3000-5000 units. ORAC (Oxygen Radical Ab- sorption Capacity) is a unit for measuring the number of antioxidants, expressed in micromoles of Trolox per unit mass (μTE / 100g) [11-13]. It is known from the litera- ture that when processing 1 thousand tons of grapes for must and wine, about 120 tons of pomace, 4 tons of seeds, 5 tons of ridges, and other secondary products of winemak- ing are formed. The characteristics of the ridges separated during crushing of grapes as a waste of winemaking have the following characteristics: mass concentration of sugars - 1.5-2.0 g / dm3, mass fraction of phenolic substances 3-6%, mineral substances up to 2.5%, tartaric acid up to 0, sixteen]. In the scientific literature, data on the study of antioxidant activity (AOA) of fruit crops, grape ridges, phenolic compounds, and AOA of white sparkling wines from various producing countries, quality and safety of wines obtained by secondary fermentation are also known. According to J. Tauchen's research [7], still, white wines made from white technical grapes (Sauvignon Blanc, Chardonnay, and Rkatsiteli) according to the technology gen- erally accepted in Europe, which involves the rapid separation of the must from the hard parts of the grapes, have a low mass concentration of phenolic substances. varying in the range from 0.2 to 0.29 g / dm3. At the same time, the wine made from Rkatsiteli 359 grapes using the Kakhetian technology (fermentation of pulp with ridges) was distin- guished by a higher mass concentration of phenolic substances, which amounted to 0.4 g / dm3 [14-18]. The purpose of the article is to analyze the phenolic composition and antioxidant activity of table wine materials from grapes of white technical European varieties grown in the soil and climatic conditions of the southern regions of Russia using ma- chine analysis technologies. 2 Materials and Methods The objects of research were samples of wine materials from grapes of white technical varieties: Rkatsiteli, Rhine Riesling, and Chardonnay, grown in the soil and climatic conditions of the southern regions of Russia. The samples under study were obtained by micro-winemaking using the traditional technological method "in white" (control) and using complex technologies for processing grapes with hard parts of the grape bunch (ridges, skin, and seeds). A sampling of the test samples was carried out follow- ing GOST 31730-2012 [17], sample preparation - by GOST 26671-2014. The main physical and chemical indicators of samples of table white wine materials were deter- mined by standardized and accepted in winemaking methods of analysis [18-21]. Anti- oxidant activity was determined by the chemiluminescence method using a Photochem photochemiluminometer (Analytik Jena AG, USA). The mass concentration of phenolic substances was determined by the photocolori- metric method. The qualitative and quantitative composition of phenolic substances in the objects of the study was determined by HPLC using an Agilent Technologies chro- matographic system (model 1100, United States) with a diode array detector. To sepa- rate substances of polyphenolic nature, we used a Zorbax SB-C18 chromatographic column with a size of 2.1 × 150 mm, filled with silica gel with a grafted octadecylsilyl phase with a sorbent particle size of 3.5 μm. Chromatography was performed in a gra- dient mode. A 0.6% aqueous solution of trifluoroacetic acid and methanol was added to the elu- ent. The flow rate of the eluent was 0.25 ml/min, the volume of the injected sample was 1 μl. Components were identified by their retention time. Calculation of the quantitative content of individual components was carried out using calibration plots of the depend- ence of the peak area on the concentration of a substance, built from solutions of indi- vidual substances. Gallic acid, caffeic acid, (+) - D-catechin, quercetin dihydrate (Fluka Chemie AG, Switzerland) and (-) - epicatechin, lilac acid (Sigma-Aldrich, Switzerland) were used as standards. All determinations were carried out in triplicate. The research results were processed by standard methods of mathematical statistics. The standard deviation of the measurement results is no more than 5%. 3 Results and Discussion The main chemical and technological indicators of the samples of table white wine materials prepared by various technological schemes corresponded to GOST 32030- 360 2013. The article introduces the concept of tegumental maceration. Tegumental (lat. Tegumentum "protective layer, shell") maceration is a technological method, which consists in the fact that immediately after crushing grapes and obtaining pulp (must, skin, seeds), an estimated amount of 75 mg/dm3 of sulfurous anhydride (antiseptic and reducing agent) and it is cooled to 10-12 ° C, followed by 24-hour extraction at a given temperature. As a result of the use of tegumental maceration, more complete extraction of aro- matic and biologically active compounds occurs, and the introduction of sulfurous an- hydride into the resulting pulp (in a stream) provides fast, within the first 5 minutes, the binding of sulfurous acid with phenolic compounds, aldehydes, sugars, and some other substances, which ensures their reliable protection against oxidation and the premature start of fermentation. Analysis of the data obtained by the HPLC method (Tables 1 and 2) showed that the identified component composition of the phenolic compounds of table wine mate- rials from grapes of white technical European varieties obtained using various complex processing technologies consists of monomeric forms - flavones, flavan-3-ols, hy- droxybenzoic and hydroxycinnamic acids, as well as from polymeric forms - oligo- meric and polymeric procyanidins. Table 1. Phenolic component composition of samples of table wine materials from white tech- nical European grape varieties obtained by various processing technologies Mass concentration, mg / dm3 № Name of wine material Oxyben- Fla- Flavan-3- Oxyric zoic vones ols acids acids Chardonnay "in white" 1 0,29 8,01 1,80 18,90 way (control) Chardonnay tegumental 2 12,35 10,70 6,15 23,60 maceration Chardonnay, 1/3 sugar 3 4,15 56,95 13,00 28,70 fermentation with combs Chardonnay, complete 4 37,40 119,56 15,69 42,26 fermentation of sugars with combs Rkatsiteli "in a white" way 5 0,20 14,70 1,30 25,00 (control) Rkatsiteli, 1/3 sugar fermentation 6 3,60 62,40 8,20 29,60 with combs Rkatsiteli full of sugars 7 4,70 111,60 17,80 34,40 fermentation with ridges Rhine Riesling, "white" way 8 - 12,65 3,01 31,13 (control) Rhine Riesling, 1/3 sugar 9 7,34 65,24 12,49 36,96 fermentation with combs Rhine Riesling, complete 10 16,20 115,01 19,92 48,00 fermentation of sugars with ridges 361 Table 2. Phenolic component composition and AOA of samples of table wine materials from white technical European grape varieties obtained by various processing methods Mass concentration, mg / dm3 AOA, № Name of wine material Oligomeric Polymeric g / dm3 procya- procya- nidins nidins 1 Chardonnay "in white" way (control) 90,00 104,00 0,79 2 Chardonnay tegumental maceration 408,00 496,00 1,00 3 Chardonnay, 1/3 sugar fermentation 163,00 433,00 1,32 with combs 4 Chardonnay, complete fermentation of 291,00 770,00 1.65 sugars with combs 5 Rkatsiteli "in a white" way (control) 259,00 375,00 0,87 6 Rkatsiteli, 1/3 sugar fermentation with 626,00 1328,00 1,42 combs 7 Rkatsiteli full of sugars fermentation 786,00 2122,00 1,74 with ridges 8 Rhine Riesling, "white" way (control) 106,00 146,00 0,81 9 Rhine Riesling, 1/3 sugar fermentation 209,00 662,00 1,50 with combs 10 Rhine Riesling, complete fermentation 276,00 840,00 1,68 of sugars with ridges The use of tegumental maceration (Fig. 1) makes it possible to obtain in the wine ma- terial 1.8 times higher content of monomeric forms of phenolic compounds and 1.3 times higher value of antioxidant activity than the traditional method of processing grapes "in white", without contacting the must with the hard parts of the grapes (con- trol). In the course of the research, it was found that the values of the mass fraction of individual components of the monomeric phenolic composition of wine materials ob- tained according to the generally accepted technology for obtaining table white wines (control) are arranged in the following sequence: hydroxycinnamic acids (64.3%)> fla- van-3-ols (29, 9%)> hydroxybenzoic acids (5.2%)> flavones (0.6%). Also, as a result of research, it was found that the values of the mass fraction of individual components of the monomeric phenolic composition of wine materials ob- tained using complex technologies for processing grapes are located in the following sequence: tegumental maceration method - oxycinnamic acids (44.7%)> flavones (23.4%) > flavan-3-ols (20.3%)> hydroxybenzoic acids (11.6%); fermentation of 1/3 of the sugars of the pulp with ridges - flavan-3-ols (56.2%)> hydroxycinnamic acids (29.0%)> hydroxybenzoic acids (10.2%)> flavones (4.6%); complete fermentation of 362 sugars with ridges - flavan-3-ols (59.6%)> hydroxycinnamic acids (21.4%)> flavones (10.0%)> hydroxybenzoic acids (9.0%). Fig. 1. Influence of complex grape processing technologies on the monomeric phenolic compo- sition of white table wine materials. 4 Conclusions The identified component composition of phenolic compounds of table wine materials from Riesling Rhine, Rkatsiteli, Chardonnay grapes obtained using various processing methods (the traditional "white" method and using solid parts of grapes) consists of monomeric forms - flavones, flavan-3-ols, hydroxybenzoic and hydroxycinnamic ac- ids, and polymeric - oligomeric and polymeric procyanidins. The quantitative compo- sition of phenolic compounds of wine materials and antioxidant activity were also es- tablished. 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