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Impact of wine technology on the variability of resveratrol and piceids in Saperavi (Vitis vinifera L.)
Annals of Agrarian Science xxx (2016) 1e4
Contents lists available at ScienceDirect
Annals of Agrarian Science journal homepage: http://www.journals.elsevier.com/annals-of-agrarianscience
Impact of wine technology on the variability of resveratrol and piceids in Saperavi (Vitis vinifera L.) M.A. Surguladze*, M.G. Bezhuashvili Agricultural University of Georgia, Institute of Viticulture and Oenology, 240, David Aghmashenebeli Alley, Tbilisi, 0159, Georgia
a r t i c l e i n f o
a b s t r a c t
Article history: Received 18 October 2016 Accepted 18 October 2016 Available online xxx
The biologically active stilbenoids-resveratrol and its glucosides were identified in the dry bulk wines of different types made with red-grape vintage variety of Saperavi (Vitis vinifera L.): cis-resveratrol, transresveratrol, cis-piceid and trans-piceid. Red and pink wines were made by different technology: I e dry, pink, of a European type; alcoholic fermentation with natural microflora; II e dry, pink, of a European type; alcoholic fermentation with dry yeast “B2000”. III e red, dry; alcoholic fermentation with no-stem pomace and aging on it for 5 months; IV e dry, red, of a Kakhetian type; alcoholic fermentation with stem pomace with natural microflora and aging on it for 5 months; V e dry, red, of a Kakhetian type, with preliminary fermentation of cluster stems, then, alcoholic fermentation with natural microflora and aging on it for 5 months. The impact of the wine-making technology on the variability of the concentrations of these substances is proved. Kakhetian bulk wines (IV and V) also differ from one another. Bulk wine-V made by the fermentation of grappa fermented in advance contains little concentration of study stilbenoids as a result of the oxidation transformations caused by the preliminary treatment. The role of grape juice, stem and seed in the localization of resveratrols and piceids in the bulk wines is identified. © 2016 Agricultural University of Georgia. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Keyword: Antitumor
Introduction The chemical content of red grapes and wine presents a rich spectrum of the components of different classes. Among them, the diversified profile of phenol compounds with flavonoid (procyanidins e oligomeric and polymeric, flavanols, flavonols, anthocyanins, flavonons, etc.) and non-flavonoid (stilbenoids, phenolic acids, phenol aldehydes, etc.) compounds is noteworthy. Intense research of stilbenoids in red wines is associated with the “French paradox” phenomenon. The studies proved the high biological activity of the said compounds [1]. Resveratrol, a monomer representative of stilbenoids, in red wines is fixed as two isomers: cis-resveratrol and trans-resveratrol [2]. These compounds form glycosidic and other derivatives and are localized in wines as cis-piceid and trans-piceid, dimers: pallidol, delta- and epsilon-viniferins, trimer: alphaviniferin, tetramer: hopeaphenol. In addition, red wines contain astringin: piceatannol glucoside and astringinin: piceatannol, etc. [3,4].
* Corresponding author. E-mail addresses: [email protected] (M.A. Surguladze), [email protected] (M.G. Bezhuashvili). Peer review under responsibility of Journal Annals of Agrarian Science.
m.
The biological activity of stilbenoids in various fields is proved with experiments in vitro and in vivo. For instance, trans-resveratrol is characterized by antioxidant [5] and inhibitory [6] activities, cardio-protective [7], antitumor (skin and gastrointestinal cancers) [8], antidiabetic [9], skin protecting [10] and other effects. The biological activity of epsilon-viniferin is presented by antioxidant [11], anti-inflammatory [12], antitumor [13] and neuroprotective [6] effects, as well as Sirt.1 activating and 5-alphareductase inhibiting properties [14]. Hopeaphenol has antitumor [15] and antiinflammatory [16] effects. The study of vine stilbenoids in Georgia was initiated by us in Institute of Horticulture, Viticulture and Winemaking of Georgia in 1991. Trans-resveratrol [17], ε-viniferin [18] and two tetramer stilbens, including hopeaphenol [19] were isolated and identified in one-year-old vine canes. Trans-resveratrol, ε-viniferin, trans-piceid and cis-piceid [20e26] were isolated and quantified in red-grape vintage vines growing in Georgia and wines. Trans-resveratrol was found to have a full inhibiting power of Agrobacterium tumefaciens causing vine cancer [27]. Trans-resveratrol stimulates wine yeasts: Sacch. vini- Kahuri 42 and Sacch. chodati- Teliani 77 in the process of alcoholic fermentation what is evidenced by an intensified growth and development of yeasts [28]. The antioxidant action of stilbenoids isolated from the vine evidenced by the
http://dx.doi.org/10.1016/j.aasci.2016.10.002 1512-1887/© 2016 Agricultural University of Georgia. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/).
Please cite this article in press as: M.A. Surguladze, M.G. Bezhuashvili, Impact of wine technology on the variability of resveratrol and piceids in Saperavi (Vitis vinifera L.), Annals of Agrarian Science (2016), http://dx.doi.org/10.1016/j.aasci.2016.10.002
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M.A. Surguladze, M.G. Bezhuashvili / Annals of Agrarian Science xxx (2016) 1e4
inhibition degree of malondialdehyde formation in human blood serum with “in vitro” experiments, occurs in the following order: trans-resveratrol (105%) < ε-viniferin (118%) < tetrameric stilben (169%) < hopeaphenol (178%) [29]. Trans-resveratrol and ε-viniferin influence the process of malic-and-lactic acid fermentation in Saperavi bulk wine evidenced by the inhibition of the formation of the intermediate product [30]. Resveratrol isomers and stilbene derivatives are characterized by different actions depending on the environment they occur. They limit the enzymes causing the inflammation processes, obstruct the origination of thrombocytes and development of atherosclerosis. These conclusions were made as a result of the research of the wines produced in some European countries, and the wine of a Kakhetian type from Georgia was among the wines being studied [31]. The concentration of stilbenoids in red wines is influenced by the methods of technological treatment of the bulk wine. The concentrations of trans-resveratrol and ε-viniferin in the bulk wines treated with gelatin, fish glue, bentonite or cold, are reduced [32]. Out of the following bulk wines made of Saperavi variety: dry, naturally sweet and fortified ones, the concentration of transresveratrol is naturally highest in the fortified wines [33]. Following the above-mentioned, the study of vine and wine stilbenoids, the biologically active substances, is topical. By continuing the study of stilbenoids in Saperavi variety, this time, the goal of the study is to determine the variability of cis- and transresveratrol and their glucosides: cis-piceid and trans-piceid depending on the wine technology. Objectives and methods
Saperavi variety: I e dry, pink, of a European type; alcoholic fermentation with natural microflora; II e dry, pink, of a European type; alcoholic fermentation with dry yeast “B2000”. III e red, dry; alcoholic fermentation with natural microflora and with no-stem pomace, aging on it for 5 months; IV e dry, red, of a Kakhetian type; alcoholic fermentation with stem pomace with natural microflora and aging on it for 5 months; V e dry, red, of a Kakhetian type, with preliminary fermentation of cluster stems, then, alcoholic fermentation with natural microflora and aging on it for 5 months. In experiment was used grapes 2014 vintage from lowland vineyard. Wine analyses were done in June of 2015 year. We also used hard grape parts, such as stems and seeds, as the study objects. We isolated the stilbenoid-containing fractions from the bulk wines and hard parts of Saperavi grapes with relevant treatment, as per plan 1. HPLC analysis We identified the stilbenoids by using the method of highpressure liquid chromatography with the following conditions: chromatograph “Varian”, column Supelcosil™ LC18, 250X4, 6 (mm); solvent A, 0.025%, trifluoroacetic acid; solvent B, acetonitrile (ACN)/A, 80/20, gradient regime 0e35 min. 20e50% B; 35e40 min. 50e100% B; 41e46 min. 100% B; 46e48 min. 100e20% B; 48e53 min. 20% B. Eluent transfer velocity 1 ml/min, wavelength 306 nm and 286 nm [4]. The samples for chromatography were filtered with a membrane filter (0.45 mm) in advance. We should note that we carried out the analysis on both 306 nm and 286 nm waves twice for each, and presented their mean values.
The objects of the study were various bulk wines made of
Please cite this article in press as: M.A. Surguladze, M.G. Bezhuashvili, Impact of wine technology on the variability of resveratrol and piceids in Saperavi (Vitis vinifera L.), Annals of Agrarian Science (2016), http://dx.doi.org/10.1016/j.aasci.2016.10.002
M.A. Surguladze, M.G. Bezhuashvili / Annals of Agrarian Science xxx (2016) 1e4
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Table 1 Contents of resveratrol and piceids (mg/l) in Saperavi bulk wines. Wines
Trans-resveratrol
Cis-resveratrol
Total resveratrol
Trans-piceid
Cis-piceid
Total piceid
I II III IV V
0.076 0.176 0.35 0.24 0.65
0500 0920 1.28 2.38 0.84
0.576 1096 1.63 2.62 1.49
4.9 4.3 5.9 5.1 4.8
12.2 11.5 14.7 14.1 13.7
17.1 15.8 20.6 19.2 18.5
Results and discussion The impact of the wine-making technology on the concentrations of cis- and trans-resveratols and piceids is clear (Table 1). The impact of dry yeasts on the piceids in the grape must of the European-type bulk wines is clearly seen during the alcoholic fermentation. The cis- and trans-resveratols formed as a result of glucosidal bond splitting are localized in the European-type bulk wine. We proved this fact by the alcoholic fermentation of the grape must in Saperavi, as well as red-grape vintage varieties, such as Aladasturi and Mujuretuli (2013 vintage) [24]. An increase of the total amount of resveratrol is followed by the decrease of the total amount of piceids. The concentration of cis-resveratrol exceeds that of trans-resveratrol. Piceids contain bulk of cis-isomers. In this connection, the results of our past experiments are worthwhile. In particular, more quantity of trans-piceid was found in Saperavi grape juice than cis-piceid, while the quantity of cis-piceid in the grape skin was much more than that of trans-piceid Consequently, the great quantity of cis-isomers in the study bulk wines is the result of the transformation of the trans-forms into the cis-form in the process of the bulk wine formation. The same variability of cisand trans-resveratrol and piceids was fixed in bulk wine-III made by the fermentation of Saperavi no-stem pomace fermentation.
The data of Kakhetian bulk wines differ from those of the European-type bulk wines. Bulk wine-IV is distinguished for high concentration of cis-resveratrol and a big quantitative difference from trans-resveratrol (2.38 mg/l and 0.24 mg/l). It should be noted that Kakhetian bulk wine is made by the alcoholic fermentation of stem pomace (grape juice þ stem þ skin þ pip) and each of them enriches the bulk wine with relevant extract substances, including stilbenoids. We identified trans-resveratrol in the stems of redgrape vintage vine varieties growing in Georgia. These varieties are: Saperavi, Cabernet Sauvignon, Otskhanuri Sapere, Aleksandrouli, Mujuretuli, Shavkapito, Tavkveri, Aladasturi, Dzelshavi and Ojaleshi [25]. The contents of resveratrol and piceids in the hard grape parts fixed by the given experiment are given in Table 2. The hard grape parts are different sources of resveratrol and piceid accumulation in Kakhetian bulk wines. Skin is the richest structure what is proved by relevant studies. Dominant in a stem is trans-resveratrol, while only traces of cis-resveratrol are seen in it. Stems contain cis- and trans-piceids with the amount of the former much exceeding that of the latter. Pips contain little transresveratrol and only traces of cis-resveratrol. The amount of cispiceid exceeds that of trans-piceid. Kakhetian bulk wines (IV and V) differ from each other. Bulk wine-V made by the fermentation of grapes fermented in advance contains less than detection limit of study stilbenoids as a result of the oxidation transformations caused by the preliminary treatment. Conclusion So, the experiment evidenced the impact of wine-making technology on the total concentration of resveratrol and piceids. Unlike the European-type wines, in Saperavi bulk wines made with Kakhetian technology, the stilbenoids coming from the grape skin, stem and pips are localized. One should note that the liquid chromatograms also showed other known and non-identified stilbenoids, and their study is the subject of the future studies. Maintenance of the biologically active stilbenoids in high-quality Saperavi wines without impairing the organoleptic properties of the wines is important for their functional purposes. Considering the gained results is an important precondition for reaching this goal. References
Table 2 Content of resveratrol and piceids in hard grape parts (mg/100 g dry wt.). Objects
Trans-resveratrol
Cis-resveratrol
Trans-piceid
Cis-piceid
Stems Seed
24.18 1.24
A little amount A little amount
2.35 0.75
5.23 2.31
[1] S. Renaud, M. De Lorgeril, Wine, alcohol, platelets, and the French paradox for coronary heart disease, Lancet 339 (8808) (1992) 1523e1526. rez, R.M. Lamuela-Raventos, A.L. Waterhouse, M.C. de la Torre[2] A.I. Romero-Pe Boronat, Levels on cis- and trans-resveratrol and their glucosides in white and rose' Vitis vinifera wines from Spain, J. Agric. Food Chem. 44 (1996) 2124e2128.
Please cite this article in press as: M.A. Surguladze, M.G. Bezhuashvili, Impact of wine technology on the variability of resveratrol and piceids in Saperavi (Vitis vinifera L.), Annals of Agrarian Science (2016), http://dx.doi.org/10.1016/j.aasci.2016.10.002
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[19] M. Bezhuashvili, L. Mujiri, A. Shashkov, O. Chzhov, A. Stomakhin, Stilben tetramers of one-year vine shoots, Bioorg. Chem. 23 (1997) 979e987 (in Russian). [20] M. Kokhtashvili, M. Bezhuashvili, Identification of trans-resveratrol in some red grape species, Georgian Eng. News 4 (1998) 104e106 (in Russian). [21] M. Kokhtashvili, M. Bezhuashvili, M. Pataraia, Study of Trans-resveratrol in Dry Table Red Wines, vol. 19, Collection of Works of Georgian Agrarian University, Tbilisi,, 2002, pp. 79e86 (in Georgian). [22] M. Bezhuashvili, N. Vepkhishvili, T. Kobaidze, L. Shubladze, D. Okruashvili, Content of the biologically active trans-resveratrol and ε-viniferin in color vine varieties growing in Georgia, Bull. Georgian Natl. Acad. Sci. 5 (2011) 61e64. [23] M. Bezhuashvili, L. Shubladze, D. Okruashvili, Trans-piceid stilbenoid in the juice and skin of the red grape(Vitis vinifera L.) varieties growing in Georgia, Bull. Georgian Natl. Acad. Sci. 7 (2013) 74e79. [24] M.G. Bezhuashvili, M.A. Surguladze, P.N. Vashakidze, Biologically active stilbenoids of saperavi grape (Vitis vinifera L.), in: 3rd International Conference on Pharmaceutical Sciences. “looking towards the Future, Honoring the Past” Tbilisi, May 29-31, 2015, pp. 66e67. Georgia. Abstract Book. [25] M.G. Bezhuashvili, P.N. Vashakidze, N.G. Vepkhishvili, L.D. Elanidze, Transformation of biologicaly active stilbenoids from grapevine to red wine, Food Suppl. Beverage Ann. Agrar. Sci. 12 (2014) 63e70. [26] M.A. Surguladze, M.G. Bezhuashvili, Change correlation of saperavi (Vitis vinifera L.) stilbenoides to wine type, in: 7th Imternational Conference on Polyphenols and Health, Congress Center Tours, France, 2015. October 27e30. [27] M. Bezhuashvili, M. Kokhtashvili, Z. Lomtatidze, K. Mamulashvili, Impact of trans-resveratrol on the growth and development of some microorganisms, Collect. Sci. Works Postgrad. Stud. Seek. Deg. IV (1999) 210e213 (in Georgian). [28] M. Bezhuashvili, T. Kobaidze, M. Kokhtashvili, M. Meskhi, Z. Sturua, Some biological activities of vine stilbens and new georgian grape products with a functional designation, in: 33 rd World Congress of Vine and Wine, Tbilisi, 2010, 20e25 June. [29] M. Bezhuashvili, M. Меskhi, M. Bostoganashvili, M. Malania, Anti-oxidant activity of stilben-containing extract in experiments “in vitro”, Winemak. Vitic. 3 (2005) 26e27. [30] M. Bezhuashvili, N. Vepkhishvili, D. Okruashvili, Influence of phenolic compounds over the malolactic fermentation in red wines, Biodiversity and Biotechnology, in: Conference of Georgian Agrarian University, 2011. December 5e6. [31] Z. Kutil, V. Temml, D. Maghradze, M. Pribylova, M. Dvorakova, D. Schuster, T. Vanek, P. Landa, Impact of wines and wine constituents on Cyclooxygenase1, Cyclooxygenase-2, and 5-lipoxygenase catalytic activity, Mediat. Inflamm. 2014 (2014). ID 178931 8. [32] M. Мeskhi, M. Bezhuashvili, Effects of the Technological treatment in wines on the contents of the stilbenoids compounds, Magarach Vitic. Winemak. 3 (2003) 22e24 (in Russian). [33] M. Kokhtashvili, Studying of Resveratrol in Some Standard Red Vine Grapes and its Wines, Doctoral Thesis, Tbilisi, 2006, p. 25 (in Georgian).
Please cite this article in press as: M.A. Surguladze, M.G. Bezhuashvili, Impact of wine technology on the variability of resveratrol and piceids in Saperavi (Vitis vinifera L.), Annals of Agrarian Science (2016), http://dx.doi.org/10.1016/j.aasci.2016.10.002
Contents lists available at ScienceDirect
Annals of Agrarian Science journal homepage: http://www.journals.elsevier.com/annals-of-agrarianscience
Impact of wine technology on the variability of resveratrol and piceids in Saperavi (Vitis vinifera L.) M.A. Surguladze*, M.G. Bezhuashvili Agricultural University of Georgia, Institute of Viticulture and Oenology, 240, David Aghmashenebeli Alley, Tbilisi, 0159, Georgia
a r t i c l e i n f o
a b s t r a c t
Article history: Received 18 October 2016 Accepted 18 October 2016 Available online xxx
The biologically active stilbenoids-resveratrol and its glucosides were identified in the dry bulk wines of different types made with red-grape vintage variety of Saperavi (Vitis vinifera L.): cis-resveratrol, transresveratrol, cis-piceid and trans-piceid. Red and pink wines were made by different technology: I e dry, pink, of a European type; alcoholic fermentation with natural microflora; II e dry, pink, of a European type; alcoholic fermentation with dry yeast “B2000”. III e red, dry; alcoholic fermentation with no-stem pomace and aging on it for 5 months; IV e dry, red, of a Kakhetian type; alcoholic fermentation with stem pomace with natural microflora and aging on it for 5 months; V e dry, red, of a Kakhetian type, with preliminary fermentation of cluster stems, then, alcoholic fermentation with natural microflora and aging on it for 5 months. The impact of the wine-making technology on the variability of the concentrations of these substances is proved. Kakhetian bulk wines (IV and V) also differ from one another. Bulk wine-V made by the fermentation of grappa fermented in advance contains little concentration of study stilbenoids as a result of the oxidation transformations caused by the preliminary treatment. The role of grape juice, stem and seed in the localization of resveratrols and piceids in the bulk wines is identified. © 2016 Agricultural University of Georgia. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Keyword: Antitumor
Introduction The chemical content of red grapes and wine presents a rich spectrum of the components of different classes. Among them, the diversified profile of phenol compounds with flavonoid (procyanidins e oligomeric and polymeric, flavanols, flavonols, anthocyanins, flavonons, etc.) and non-flavonoid (stilbenoids, phenolic acids, phenol aldehydes, etc.) compounds is noteworthy. Intense research of stilbenoids in red wines is associated with the “French paradox” phenomenon. The studies proved the high biological activity of the said compounds [1]. Resveratrol, a monomer representative of stilbenoids, in red wines is fixed as two isomers: cis-resveratrol and trans-resveratrol [2]. These compounds form glycosidic and other derivatives and are localized in wines as cis-piceid and trans-piceid, dimers: pallidol, delta- and epsilon-viniferins, trimer: alphaviniferin, tetramer: hopeaphenol. In addition, red wines contain astringin: piceatannol glucoside and astringinin: piceatannol, etc. [3,4].
* Corresponding author. E-mail addresses: [email protected] (M.A. Surguladze), [email protected] (M.G. Bezhuashvili). Peer review under responsibility of Journal Annals of Agrarian Science.
m.
The biological activity of stilbenoids in various fields is proved with experiments in vitro and in vivo. For instance, trans-resveratrol is characterized by antioxidant [5] and inhibitory [6] activities, cardio-protective [7], antitumor (skin and gastrointestinal cancers) [8], antidiabetic [9], skin protecting [10] and other effects. The biological activity of epsilon-viniferin is presented by antioxidant [11], anti-inflammatory [12], antitumor [13] and neuroprotective [6] effects, as well as Sirt.1 activating and 5-alphareductase inhibiting properties [14]. Hopeaphenol has antitumor [15] and antiinflammatory [16] effects. The study of vine stilbenoids in Georgia was initiated by us in Institute of Horticulture, Viticulture and Winemaking of Georgia in 1991. Trans-resveratrol [17], ε-viniferin [18] and two tetramer stilbens, including hopeaphenol [19] were isolated and identified in one-year-old vine canes. Trans-resveratrol, ε-viniferin, trans-piceid and cis-piceid [20e26] were isolated and quantified in red-grape vintage vines growing in Georgia and wines. Trans-resveratrol was found to have a full inhibiting power of Agrobacterium tumefaciens causing vine cancer [27]. Trans-resveratrol stimulates wine yeasts: Sacch. vini- Kahuri 42 and Sacch. chodati- Teliani 77 in the process of alcoholic fermentation what is evidenced by an intensified growth and development of yeasts [28]. The antioxidant action of stilbenoids isolated from the vine evidenced by the
http://dx.doi.org/10.1016/j.aasci.2016.10.002 1512-1887/© 2016 Agricultural University of Georgia. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/).
Please cite this article in press as: M.A. Surguladze, M.G. Bezhuashvili, Impact of wine technology on the variability of resveratrol and piceids in Saperavi (Vitis vinifera L.), Annals of Agrarian Science (2016), http://dx.doi.org/10.1016/j.aasci.2016.10.002
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M.A. Surguladze, M.G. Bezhuashvili / Annals of Agrarian Science xxx (2016) 1e4
inhibition degree of malondialdehyde formation in human blood serum with “in vitro” experiments, occurs in the following order: trans-resveratrol (105%) < ε-viniferin (118%) < tetrameric stilben (169%) < hopeaphenol (178%) [29]. Trans-resveratrol and ε-viniferin influence the process of malic-and-lactic acid fermentation in Saperavi bulk wine evidenced by the inhibition of the formation of the intermediate product [30]. Resveratrol isomers and stilbene derivatives are characterized by different actions depending on the environment they occur. They limit the enzymes causing the inflammation processes, obstruct the origination of thrombocytes and development of atherosclerosis. These conclusions were made as a result of the research of the wines produced in some European countries, and the wine of a Kakhetian type from Georgia was among the wines being studied [31]. The concentration of stilbenoids in red wines is influenced by the methods of technological treatment of the bulk wine. The concentrations of trans-resveratrol and ε-viniferin in the bulk wines treated with gelatin, fish glue, bentonite or cold, are reduced [32]. Out of the following bulk wines made of Saperavi variety: dry, naturally sweet and fortified ones, the concentration of transresveratrol is naturally highest in the fortified wines [33]. Following the above-mentioned, the study of vine and wine stilbenoids, the biologically active substances, is topical. By continuing the study of stilbenoids in Saperavi variety, this time, the goal of the study is to determine the variability of cis- and transresveratrol and their glucosides: cis-piceid and trans-piceid depending on the wine technology. Objectives and methods
Saperavi variety: I e dry, pink, of a European type; alcoholic fermentation with natural microflora; II e dry, pink, of a European type; alcoholic fermentation with dry yeast “B2000”. III e red, dry; alcoholic fermentation with natural microflora and with no-stem pomace, aging on it for 5 months; IV e dry, red, of a Kakhetian type; alcoholic fermentation with stem pomace with natural microflora and aging on it for 5 months; V e dry, red, of a Kakhetian type, with preliminary fermentation of cluster stems, then, alcoholic fermentation with natural microflora and aging on it for 5 months. In experiment was used grapes 2014 vintage from lowland vineyard. Wine analyses were done in June of 2015 year. We also used hard grape parts, such as stems and seeds, as the study objects. We isolated the stilbenoid-containing fractions from the bulk wines and hard parts of Saperavi grapes with relevant treatment, as per plan 1. HPLC analysis We identified the stilbenoids by using the method of highpressure liquid chromatography with the following conditions: chromatograph “Varian”, column Supelcosil™ LC18, 250X4, 6 (mm); solvent A, 0.025%, trifluoroacetic acid; solvent B, acetonitrile (ACN)/A, 80/20, gradient regime 0e35 min. 20e50% B; 35e40 min. 50e100% B; 41e46 min. 100% B; 46e48 min. 100e20% B; 48e53 min. 20% B. Eluent transfer velocity 1 ml/min, wavelength 306 nm and 286 nm [4]. The samples for chromatography were filtered with a membrane filter (0.45 mm) in advance. We should note that we carried out the analysis on both 306 nm and 286 nm waves twice for each, and presented their mean values.
The objects of the study were various bulk wines made of
Please cite this article in press as: M.A. Surguladze, M.G. Bezhuashvili, Impact of wine technology on the variability of resveratrol and piceids in Saperavi (Vitis vinifera L.), Annals of Agrarian Science (2016), http://dx.doi.org/10.1016/j.aasci.2016.10.002
M.A. Surguladze, M.G. Bezhuashvili / Annals of Agrarian Science xxx (2016) 1e4
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Table 1 Contents of resveratrol and piceids (mg/l) in Saperavi bulk wines. Wines
Trans-resveratrol
Cis-resveratrol
Total resveratrol
Trans-piceid
Cis-piceid
Total piceid
I II III IV V
0.076 0.176 0.35 0.24 0.65
0500 0920 1.28 2.38 0.84
0.576 1096 1.63 2.62 1.49
4.9 4.3 5.9 5.1 4.8
12.2 11.5 14.7 14.1 13.7
17.1 15.8 20.6 19.2 18.5
Results and discussion The impact of the wine-making technology on the concentrations of cis- and trans-resveratols and piceids is clear (Table 1). The impact of dry yeasts on the piceids in the grape must of the European-type bulk wines is clearly seen during the alcoholic fermentation. The cis- and trans-resveratols formed as a result of glucosidal bond splitting are localized in the European-type bulk wine. We proved this fact by the alcoholic fermentation of the grape must in Saperavi, as well as red-grape vintage varieties, such as Aladasturi and Mujuretuli (2013 vintage) [24]. An increase of the total amount of resveratrol is followed by the decrease of the total amount of piceids. The concentration of cis-resveratrol exceeds that of trans-resveratrol. Piceids contain bulk of cis-isomers. In this connection, the results of our past experiments are worthwhile. In particular, more quantity of trans-piceid was found in Saperavi grape juice than cis-piceid, while the quantity of cis-piceid in the grape skin was much more than that of trans-piceid Consequently, the great quantity of cis-isomers in the study bulk wines is the result of the transformation of the trans-forms into the cis-form in the process of the bulk wine formation. The same variability of cisand trans-resveratrol and piceids was fixed in bulk wine-III made by the fermentation of Saperavi no-stem pomace fermentation.
The data of Kakhetian bulk wines differ from those of the European-type bulk wines. Bulk wine-IV is distinguished for high concentration of cis-resveratrol and a big quantitative difference from trans-resveratrol (2.38 mg/l and 0.24 mg/l). It should be noted that Kakhetian bulk wine is made by the alcoholic fermentation of stem pomace (grape juice þ stem þ skin þ pip) and each of them enriches the bulk wine with relevant extract substances, including stilbenoids. We identified trans-resveratrol in the stems of redgrape vintage vine varieties growing in Georgia. These varieties are: Saperavi, Cabernet Sauvignon, Otskhanuri Sapere, Aleksandrouli, Mujuretuli, Shavkapito, Tavkveri, Aladasturi, Dzelshavi and Ojaleshi [25]. The contents of resveratrol and piceids in the hard grape parts fixed by the given experiment are given in Table 2. The hard grape parts are different sources of resveratrol and piceid accumulation in Kakhetian bulk wines. Skin is the richest structure what is proved by relevant studies. Dominant in a stem is trans-resveratrol, while only traces of cis-resveratrol are seen in it. Stems contain cis- and trans-piceids with the amount of the former much exceeding that of the latter. Pips contain little transresveratrol and only traces of cis-resveratrol. The amount of cispiceid exceeds that of trans-piceid. Kakhetian bulk wines (IV and V) differ from each other. Bulk wine-V made by the fermentation of grapes fermented in advance contains less than detection limit of study stilbenoids as a result of the oxidation transformations caused by the preliminary treatment. Conclusion So, the experiment evidenced the impact of wine-making technology on the total concentration of resveratrol and piceids. Unlike the European-type wines, in Saperavi bulk wines made with Kakhetian technology, the stilbenoids coming from the grape skin, stem and pips are localized. One should note that the liquid chromatograms also showed other known and non-identified stilbenoids, and their study is the subject of the future studies. Maintenance of the biologically active stilbenoids in high-quality Saperavi wines without impairing the organoleptic properties of the wines is important for their functional purposes. Considering the gained results is an important precondition for reaching this goal. References
Table 2 Content of resveratrol and piceids in hard grape parts (mg/100 g dry wt.). Objects
Trans-resveratrol
Cis-resveratrol
Trans-piceid
Cis-piceid
Stems Seed
24.18 1.24
A little amount A little amount
2.35 0.75
5.23 2.31
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M.A. Surguladze, M.G. Bezhuashvili / Annals of Agrarian Science xxx (2016) 1e4
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Please cite this article in press as: M.A. Surguladze, M.G. Bezhuashvili, Impact of wine technology on the variability of resveratrol and piceids in Saperavi (Vitis vinifera L.), Annals of Agrarian Science (2016), http://dx.doi.org/10.1016/j.aasci.2016.10.002