The paper presents the results of mMonitoring of qualitative and quantitative composition of polyphenols in wines of trademarks of the Crimea and the Kuban made from Cabernet-Sauvignon, Merlot and Saperavi red grapes. It has been shown that total amount of polyphenols in wine exceeds the European average of 2.5 g/L. Antioxidant activity is also high, exceeding 2.0 g/L in units of the standard antioxidant trolox. Technology and regulatory documentation was developed for obtaining products with a standardized amount of polyphenols from grapes, grape pomace and grape vines. The presents the results of development and practical evaluation of products saturated with grape polyphenols (wine, grape drink, extract) having a high antioxidant activity confirmed by in vitro and in vivo.

The Magarach Institute has been addressing the issue of developing technologies for obtaining functional food products from grapes saturated with grape polyphenols; and of determining main properties and modes of grape polyphenol extraction from potential raw materials (pomace, seeds, crests) for the production of experimental samples of functional products. The aim of the present research was not only to experimentally determine the total stock of polyphenols in pomace, seeds, crests of red and white grape varieties, but also to assess empirically the qualitative and quantitative composition of total polyphenols in pomace, seeds, stems, vines, leaves of red and white grape varieties as functional components of a healthy nutrition. The research has dwelled upon the empiric assessment of the characteristics of grape raw materials (grape must, pomace, seeds, crests) in terms of their technological stock, quantitative and qualitative composition of polyphenols (by HPLC), and the main functional ingredients of the biological activity of grape products. An analysis of the data obtained shows that the aqueous-alcoholic extracts of grape raw materials possess the whole spectrum of polyphenols typical for red grape wines (anthocyanins, flavones, flavan-3-ols, hydroxycinnamic, hydroxybenzoic acids, stilbenes, oligomeric and polymeric procyanidins), however their concentration quantitatively exceeds that found in wines. It has been shown that the total content of water-soluble antioxidants experimentally determined by the amperometric method depends on the content of oligomeric and polymeric procyanidins in the extracts of grape raw materials. Thus, the content of oligomeric procyanidins in the extracts of grape raw materials from the sum of phenolic substances averaged 5.7%; while the content of polymer procyanides went up to 94%. The technological availability of polyphenols of pomace, stalks and grape seeds during water-alcohol extraction in the piled layer without mixing was shown, the equilibrium concentrations of polyphenols transferred into the extract and the potential amounts of polyphenols extracted during single-stage extraction (technological stock) were assessed. The maximum values of technological stock of polyphenols of 29.2–76.02 g/kg dry weight in the stalks of white and red grape varieties were determined.

Обсуждаются результаты экспериментальной оценки потенциала виноградной грозди как источника получения полифенолов винограда, биологически активных функциональных ингредиентов питания. Идентифицированы по качественному и количественному составу полифенолы сусла, выжимок, гребней, семян винограда белых и красных технических сортов Алиготе, Рислинг рейнский, Цитронный Магарача, Каберне-Совиньон, Мускат гамбургский, Пино нуар, а также выжимок, семян, гребней белых и красных сортов винограда из ампелографической коллекции института «Магарач» (Совиньон зеленый, Первенец Магарача, Бияс айбатлы, Мальбек, Ай-Петри, Праздничный Магарача, Голубок, Антей магарачский). Фенольные соединения включают полифенолы как мономерного ряда, так и олигомерные и полимерные проантоцианидины, составляющие около 90% общего количества полифенолов. Мономерная фракция представлена флавоноидами (антоцианы, (+)-D-катехин, (-)-эпикатехин, кверцетин, кверцетин-3-О-глюкозид, кверцетин-3-О-глюкуронид), фенольными кислотами (галловая, сиреневая, кафтаровая, 2-S-глутатионилкафтаровая, коутаровая, кофейная, фертаровая, n-кумаровая, феруловая), а также стильбеноидом транс-ресвератролом. Дана интегральная характеристика биологической активности комплекса полифенолов сусла, выжимок, гребней, семян по суммарной концентрации антиоксидантов; данная величина варьирует в пределах 0,2-65,0 г/дм в пересчете на стандартный антиоксидант Тролокс. Показана технологическая доступность полифенолов выжимок, гребней, семян винограда при водно-этанольной экстракции в насыпном слое без перемешивания, оценены равновесные концентрации полифенолов, переходящих в экстракт, и потенциальные количества извлекаемых при одностадийной экстракции полифенолов (технологический запас). Установлено, что средняя величина извлекаемого технологического запаса фенольных веществ водным раствором этанола с объемной долей 50% варьирует (ТЗФВ) в пределах (г/кг сухой массы): 8-18 в выжимках, 29-38 в семенах, 49-62 в гребнях. Abstact. The results of experimental evaluation of grape bunch potential as a source for obtaining grape polyphenols, biologically active functional food ingredients are discussed. The polyphenols of must, pomace, stalks and seeds of white and red wine grape varieties ‘Aligote’, ‘Rhine Riesling’, ‘Tsitronnyi Magaracha’, ‘Cabernet-Sauvignon’, ‘Muscat d’Hamburg’, ‘Pinot Noir’, as well as pomace, seeds and stalks of white and red wine grape varieties from the Institute Magarach Ampelographic Collection (‘Sauvignon Vert’, ‘Pervenets Magaracha’, ‘Biyas Aibatly’, ‘Malbec’, ‘Ai-Petri’, ‘Prazdnichnyi Magaracha’, ‘Golubok’, Antei Magarachskiy’ were identified. Phenolic compounds include polyphenols both monomeric and oligomeric, as well as polymeric proanthocyanidins, which amount about 90% of the total number of polyphenols. Monomeric fraction is represented by flavonoids (anthocyanins, (+)-D-catechin, (-)-epicatechin, quercetin, quercetin-3-О-glucoside, quercetin-3-О-glucuronide), phenolic acids (gallic, syringic, caftaric, 2-S-glutathionyl caftaric, coutaric, caffeic, fertaric, n-coumaric, ferulic), as well as by stilbenoid trans-resveratrol. An integral characteristic of biological activity of polyphenol complex of must, pomace, stalks and seeds by total concentration of antioxidants is defined; this value ranges from 0.2 to 65.0 g/dm with reference to a standard antioxidant Trolox. Technological availability of polyphenols of grape pomace, stalks and seeds during aqueous-ethanol extraction in the filling layer without mixing was shown; equilibrium concentrations of polyphenols transferring into the extract and potential amounts of polyphenols extracted during single-stage extraction of polyphenols (technological stock) were assessed. It was determined that average quantity of technological stock of phenolic substances extracted by aqueous solution of ethanol with volume fraction of 50% ranges (TSPhS) within the following limits (g/kg of dry matter): 8-18 in pomace, 29-38 in seeds, 49-62 in stalks.

В работе экспериментально оценены этанольные экстракты выжимки винограда красных сортов и спиртовые экстракты из лозы винограда белых и красных сортов по количественному и качественному составу полифенолов - основных функциональных ингредиентов биологической активности продуктов переработки винограда. Анализ полученных данных показывает, что в этанольных экстрактах выжимки винограда сортов Каберне-Совиньон, Пино нуар, Антей магарачский, Мальбек, Саперави, Бастардо магарачский, Мускат гамбургский представлен весь спектр полифенолов, свойственный красным виноградным винам (антоцианы, флавоны, флаван-3-олы, оксикоричные, оксибензойные кислоты, олигомерные и полимерные проантоцианидины), при этом количественное содержание полифенолов кратно превосходит их концентрацию в вине. В этанольных экстрактах лозы винограда сортов Цитронный Магарача, Алиготе, Ркацители, Бастардо магарачский, Каберне-Совиньон обнаружены стильбеновые вещества, которые составляют от 9,2 до 28,1 % от суммы всех фенольных веществ. В спиртовых экстрактах из лозы винограда сорта Пино нуар концентрация стильбеновых веществ достигает 47,2 % от суммы фенольных веществ. Получены пищевые экспериментальные образцы концентратов из выжимки Каберне-Совиньон и лозы винограда сортов Цитронный Магарача, Пино нуар, Каберне-Совиньон. В безалкогольном пищевом концентрате полифенолов из лозы винограда по сравнению с концентратом из виноградной выжимки сорта Каберне-Совиньон отсутствуют антоцианы, оксикоричные кислоты; присутствуют стильбеноиды, которые составляют 2,28 г/дм, или 20,9 % от суммы всех фенольных веществ. Стильбеновые вещества в составе комплекса полифенолов, выделенных из лозы винограда представлены транс-ресвератролом, e-виниферином, транс-пицеидом, транс-пикеатаннолом. Показатель суммарного содержания водорастворимых антиоксидантов зависит от содержания в концентрированных экстрактах виноградной выжимки олигомерных и полимерных проантоцианидинов, а в случае концентрата из лозы винограда - и от концентрации стильбеновых веществ. Abstact. In the work, ethanol extracts of pomace of red grape varieties and alcoholic extracts of vines of white and red grape varieties are experimentally assessed by the quantitative and qualitative composition of polyphenols as the main functional ingredients of biological activity of grape processing products. The obtained data analysis shows that in ethanol extracts of grape pomace from the ‘Cabernet-Sauvignon’, ‘Pinot Noir’, ‘Antei Magarachskiy’, ‘Malbec’, ‘Saperavi’, ‘Bastardo Magarachskiy’, ‘Muscat Hamburg’, the whole range of polyphenols, common to red grape wines (anthocyanins, flavones, flavan-3-ols, oxycinnamic, oxybenzoic acids, oligomeric and polymeric proanthocyanidins), is presented, while quantitative content of polyphenols is many fold higher than their concentration in wine. In ethanol vine extracts of ‘Tsitronnyi Magaracha’, ‘Aligote’, ‘Rkatsiteli’, ‘Bastardo Magarachskiy’, ‘Cabernet-Sauvignon’ grape varieties, stilbene substances, which amount from 9,2 to 28,1% of the sum of all phenolic substances, were found. In alcoholic extracts of the ‘Pinot Noir’ vine, the concentration of stilbene substances reaches 47,2% of the sum of phenolic substances. Experimental food samples of concentrates from the ‘Cabernet-Sauvignon’ pomace and vines of the ‘Tsitronnyi Magaracha’, ‘Pinot Noir’ and ‘Cabernet-Sauvignon’ grape varieties were obtained. In non-alcoholic food concentrate of polyphenols from grape vines, compared to the concentrate from the ‘Cabernet-Sauvignon’ grape pomace, anthocyanins and oxycinnamic acids are absent; stilbenoids, constituting 20.9% of the sum of all phenolic substances and amounting to 2.28 g/dm, are present. Stilbene substances in the composition of polyphenol complex, isolated from grape vines, are represented by trans-resveratrol, e-viniferin, trans-piceid and trans-piceatannol. The index of the total content of water-soluble antioxidants depends on the content of oligomeric and polymeric proanthocyanidins in concentrated extracts of grape pomace; in the case of vine concentrate it also depends on the concentration of stilbene substances.

The present work describes the results of research of a complex of substances obtained from sedimentary wine yeast by extraction with food extractants with the use of ultrasound. The filtrates of alcoholic extracts of yeast and sediments isolated from filtrate were investigated. It has been shown that hydrophobic substances moderately soluble and poorly soluble in alcohol, such as ethyl esters of fatty acids, fatty acids, phospholipids, sterols, squalene, were isolated from sedimentary wine yeast by extraction with alcohol-containing extractants. The mechanical effect of ultrasound provides faster and more complete penetration of the solvent into the intracellular substances. Ultrasound allows one to achieve greater penetration of the solvent into plant tissues and improve mass exchange. Ultrasonic waves, causing cavitation in the liquid, destroy cell walls and promote the release of cell matrix components. Thus, during the US treatment of suspended sedimentary wine yeast a mixture of fatty acid ethyl ester more than 2 times and squalene - 1.5 times, compared to the control, is observed.

Adipose tissue in obesity produces more active oxygen forms and inflammatory cytokines, which results in the development of a systemic inflammatory response syndrome (SIRS) and a metabolic syndrome (MS). Recently, there has been a growing interest in natural products for oxidative stress remodeling. Therefore, it is relevant to study grape polyphenols (GPs) to develop certain techniques for oxidative stress prevention and treatment. The goal of the paper is to examine antioxidant profiles in MS, and the efficacy of its pathogenetic correction with GPs. Materials and Methods. The authors estimated the ratio of peroxidase (PA) activity and thiobarbituric acid active products (TBA-APs), and the ratio of ceruloplasmin (C) and PA. Results. The authors observed stereotypical changes in the systems of antioxidant and antiproteolytic protection in MS. Thus, with MS, there was recorded a decrease in PA/TBA-AP ratio (2.3 compared with 16.6 in norm) and an increase in C/PA ratio (3.17 compared with 0.62 in norm). In the case of “Fenokor” administration for MS remodeling, the level of superoxide dismutase became normal. Moreover, the authors observed the highest level of TBA-active products in experimental animals. MS remodeling with Fenokor was effective: PA/TBA ratio was 2 times higher than in the control, and C/PA ratio was 0.86. Catalase and superoxide dismutase levels were 5 and 1.3 times higher than in norm, respectively. Conclusion. Normalization of PA/TBA-AP and C/PA ratios under “Fenokor” allows us to GPs with a high polyphenol content to correct antioxidant status and reduce systemic inflammatory response syndrome. Keywords: oxidative stress; grape polyphenols. Жировая ткань при ожирении производит большее количество активных форм кислорода и воспалительных цитокинов, что приводит к развитию синдрома системной воспалительной реакции (ССВР) и метаболического синдрома (МС). В последнее время возрастает интерес к использованию продуктов натурального происхождения для коррекции оксидативного стресса. Поэтому представляется актуальным изучение полифенольных продуктов переработки винограда (ПППВ) для разработки методов профилактики и терапии проявлений окислительного стресса. Цель – изучение антиоксидантных профилей при МС, а также эффективности его патогенетической коррекции ПППВ. Материалы и методы. Оценивали отношение активности пероксидазы (ПА) и активных продуктов тиобарбитуровой кислоты (ТБК-АП), а также церулоплазмина (Ц) и ПА. Результаты. Показана определённая стереотипность изменений в системах антиоксидантной и антипротеолитической защиты при МС. Так, при МС регистрировалось снижение ПА/ТБК-АП (2,3 по сравнению с 16,6 в норме) и повышение Ц/ПА (3,17 по сравнению с 0,62 в норме). В случае применения «Фэнокора» для коррекции проявлений МС происходила нормализация содержания супероксиддисмутазы, наблюдался самый высокий среди экспериментальных животных уровень ТБК-активных продуктов. Коррекция МС с помощью «Фенокора» была эффективна: соотношение ПА/ТБА было в 2 раза выше, чем в контроле, а отношение Ц/ПА составляло 0,86. Уровни каталазы и супероксиддисмутазы были в 5 и 1,3 раза выше нормы соответственно. Заключение. Нормализация отношений ПА/ТБК-АП и Ц/ПА при применении препарата «Фэнокор» позволяет использовать ПППВ с высоким содержанием полифенолов для коррекции антиоксидантного статуса и снижения проявлений ССВР. Ключевые слова: оксидативный стресс, полифенолы винограда.

The aim of the work was to study the influence of sparkling wines on the main markers of hypoxia. Material and methods. The experiment was carried out on 25 white male rats weighing 180–200 grams (age 10–12 weeks.). Animals of group 1 were control animals and received drinking water. Animals of the second group received water and were subjected to bloodletting on the 1st week. Animals of the 3rd group were subjected to bloodletting on the 1st week and received diluted sparkling wine as drinking water. Animals of group 4 were not subjected to bloodletting and received diluted sparkling wine as drinking water throughout the experiment. Sparkling wine was given at a dose of 300 ml of wine per 70 kg of body weight. In the blood serum, markers of free radical oxidation and markers of inflammation formed in hypoxic damage were studied. Results: the use of sparkling wines for correction of biochemical disorders in hypoxia led to the strengthening of antioxidant protection systems: the level of superoxide dismutase showed an increase of 77 % (p < 0,05) comparing to the group with simulated hypoxia, the growth of catalase activity by 5 times, the values of trypsin-like activity and elastase-like activity were lower than the hypoxic level by 20 % (p < 0,05) and 59 % (p < 0,01), respectively. Conclusion: the use of sparkling red wine in the correction of free-radical and proteolytic damage, typical for hypoxia, revealed a complex positive effect on a number of important indicators of antioxidant and antiproteolytic protection of the body. This indicates the prospects for further experimental and clinical studies in this direction.

Objective — Biological effects of grape processing products (GPP) rich in polyphenols are still not studied completely. The aim of the investigation was to study co-interactions between peroxidation lipid oxidation (PLO) and nonspecific proteases and their inhibitors activity in the animals with the model of metabolic syndrome (MS) corrected with GPP. Material and Methods — The study was performed on 54 white male rats of the weight of 180-200 g. The animals of experimental groups were fed by the standard menu and were given 10% fructose solution instead the drinking water during 8 weeks, which promoted development of MS. “Enoant”, “Enoant-premium” and “Fenocor” were used as GPP in dosage of 2.5 ml/kg during 4 weeks just after 5th week. The PLO was estimated according to the concentration of thiobarbituric acid active products (TBA-AP), ceruloplasmin (CC), catalase-like activity (CLA), peroxidase-like activity (PLA) and superoxide dismutase activity (SOD). Trypsin-like activity (TLA), elastase-like activity (ELA), alfa-1-antitripsin activity (ATA) and acid-stable inhibitors activity (ASA) were estimated. Results —MS demonstrated rise of TBA-AP by 50.4 % (р<0.01) and TLA by 19.5% (р<0.01), and drop of ATA, SOD, CLA, PLA in comparison with the control. The GPP (especially “Fenocor”) led to the block of pathogenetic link of MS because of suppression of PLO and proteases’ activity and increase of their inhibitors. Conclusion —There is a tight connection between PLO and proteases’ activity in MS. “Fenocor” could be used as MS-modified remedies acting positively on key links of MS.

The chapter presents the results of development and testing of products, rich in polyphenols (wine, drink, extract, food grape concentrate), which has a high antioxidant capacity, and which was confirmed by research both in vitro and in vivo. It was shown that oral intake of innovative products block the development of metabolic syndrome and ischemic myocardial damage in experimental animals. Clinical studies have proved, that patients with coronary heart disease and hypertension can significantly improve their health conditions, if they add innovative products saturated with grape polyphenols into their diet, during treatment at sanatorium or resort. The chapter estimates the vectors of development in viticulture and winemaking industry for further production of healthy food for the population.

The article presents the results of study of the effect of polyphenol-rich materials obtained from grapes on the clinical symptoms, activity of non-specific proteases, and parameters of lipid peroxidation (LPO) of blood of rats with fructose model of metabolic syndrome (MS). White male rats (n=54, with initial body weight of 190-210 g) were randomly divided into five groups: control and 4 experimental groups. Body weight, circumference of the abdomen, blood serum level of glucose, total cholesterol, triglycerides and high density (HDL) lipoproteins were monitored in all animals in dynamics. Also non-specific proteases and their inhibitors were evaluated in rat blood by enzymatic methods, lipid peroxidation profile (malondialdehyde and caeruloplasmin blood serum level, superoxide dismutase blood activity) - by spectrophotometric methods. Modeling of the MS (10% fructose in drinking water) within 8 weeks resulted in statistically significant increase in body mass, abdominal adipose tissue, the activity of elastase-like (ELA) and trypsin-like (TLA) proteinases (20 and 18% respectively), the level of secondary products of LPO (50%), and decrease in activity of superoxide dismutase (15%) compared with the parameters of intact animals. The use of polyphenol-rich materials obtained from grapes with a total content of 1; 1,1 and 4 mg of phenolic compounds (in 0.05 ml) at MS for 4 weeks contributed regression the key clinical signs of MS: significant decrease of glucose and triglyceride levels against the background of increasing the HDL cholesterol were observed. The polyphenol-rich materials obtained from grapes influenced positively on the proteolytic profile (decreased TLA and ELA by 20% at the maximum dosage), contributed to the increase of the level of acid-stable protease inhibitors (by 21%), and reduced the amount of secondary products of LPO (by 34%). The dose-dependent effect of the amount of polyphenol components contained in food concentrates has been revealed.

Experimental data on the antioxidant activity of grape juice, grape concentrates and wine from grapes of Cabernet Sauvignon, Merlot and Saperavi from Crimea and Krasnodar regions was presented. Flavonoids are presented in the form of glycosides of such anthocyanins as delphinidin, malvidin, cyanidin, petunidin, peonidin and also by quercetin and its glycoside, (+)-D-catechin and (-)-epicatechin. Oligomeric procyanidins, which are condensed catechol units (2-6) soluble in water, are presented in significant amounts, and polymeric procyanidins with the amount catechin units greater than 7, insoluble in water, constituted the bulk of polyphenols od wine and concentrates from red grapes (no juice). Among non-flavonoid polyphenols hydroxybenzoic (gallic, syringic) and hydroxycinnamic (caftaric, cautaric) acids are identified, the relative content of which in the amount of polyphenols in the juice is maximum, and minimum is in concentrates. It was found that antioxidant activity for all products in standard Trolox method can be estimated by the equation: Y = 0.53627+0.1395X+0.080439X2-0.00064708X3, with a correlation coefficient r = 0.9952; where: Y--antioxidant activity, g/dm3 by Trolox method; X--mass concentration of phenolic substances on the Folin-Ciocalteu, g/dm3. The equation is valid for Y = 0.76-196.22; X = 1.0-82.67. The results of biological testing of wines Cabernet Sauvignon, Merlot, Saperavi and polyphenol concentrates from grape on the biological model of bioluminescent bacteria Photobacterium leiognathi Sh1 demonstrated the applicability of bioassay to assess the antioxidant activity, which correlates well with the polyphenols content and antioxidant activity by trolox method.