This diploma thesis deals with application of pressurized liquid extraction (PFE) on grape skins of Vitis vinifera cultivars Alibernet and Svatovavřinecké to gain extraction of anthocyanin pigments in the form of 3-monoglucosides. This thesis deals with comparison efficiency pressurized liquid extraction (PFE) for the range temperatures 40–120 °C and the Soxhlet extraction with four extraction solvents. The identification of the anthocyanin pigments by high-performance liquid chromatography with diode array detection (HPLC/DAD) based on Synergi C12 column separation (Phenomenex) was performed. The anthocyanin pigments were identified at wavelength 520 nm. The complete amount of anthocyanin pigments grape skins was depended on the extraction solvent and temperature in range 0,38–244,06 mg/g for cultivar Alibernet and 0,28–105,01 mg/g for cultivar Svatovavřinecké. The contents of anthocyanin pigments was determined in the samples of wine both cultivars in the range 3,83–2836,84 mg/l too. PFE followed by HPLC/DAD employing Synergi C12 column provides relatively fast, quantitative and reproducible determination of anthocyanins in grape skins and wines.

Thesis (PhD)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: South African red wine is often acknowledged world wide as being full bodied and deep in colour. This is often the result of high temperatures that is experienced during the important growth stages of grapes especially post véraison. In the Robertson area in South Africa however, temperatures often exceeds the range for optimal anthocyanin development during these growth stages. The distinction between grapes being technologically ripe and being ripe on a phenolic level is also accepted as an important determining factor for the perfect time to pick grapes. In co-operative wineries such as Robertson Winery (RW) where grapes are delivered from a large area and different producers, it is difficult to individualise grape blocks when it comes to ripeness level in terms of sugar or phenolic ripeness. In most circumstances a generalised set of parameters for deeming grapes ripe or acceptable for delivery is the best substitute. The levels of these parameters are based on research literature that is available for the area as well as data collected through years of maintaining the vineyards of that area. The grape parameters that are currently being used by RW for ripeness and quality are pH, titratable acidity (TA) and sugar level. In recent years RW in conjunction with the Department of Viticulture and Oenology, Stellenbosch University, decided to investigate more parameters to determine the quality of grapes at the time of harvest. Most importantly for the grape growers this quality is connected to a price point and therefore compensation. Two important quality parameters of red wine are the red colour and mouth feel of wine. Anthocyanin and tannins are respectively connected to these two quality attributes and are both widely accepted as quality indicators. Wine with high anthocyanin and tannin content often originates from grapes with a high colour and phenolic profile. The existence of a correlation between grape and wine anthocyanin and tannin content is therefore the basis of attempting to use these parameters in the grape to predict end wine’s colour and phenolic quantity. Determination of anthocyanin and tannin content of grapes has already become part of some private owned wineries’ standard set of determinations. However, sample preparations, extractions and consumables needed are all factors that need to be reduced to make the measurement and therefore the use of these parameters more viable in a co-operative cellar laboratory, where large volumes of grapes are received during harvest. The first objective of this work was to determine the levels of anthocyanin and tannin in red grapes from different vineyard blocks from the producers of RW from three successive vintages. This would give insight as to what can be seen as a low and high anthocyanin and tannin content for grapes received at the cellar. For this purpose, blocks of the most important red wine cultivars for RW was selected and analysed for these compounds. The ranges and average levels of anthocyanin and tannin content were determined using measurement techniques that could be used by any winery. The average mono flavanol and total colour level of the grapes were found to be lower than those often reported in literature, with total grape flavanols being higher. However, a wide range of values for these compounds were found that correlated with those found in other studies. The possible reasons for differences in levels of occurrence of these compounds were discussed and mostly pertain to differences in cultivar, micro climatic and season. The second objective was to determine the correlation between levels of colour and phenolic compounds in grapes and their corresponding wines. Such correlations will form the foundation for the use of phenolic content to predict the colour and phenolic potential of the wine and possibly wine quality as well. When the grape and wine colour and phenolic data were correlated for all seasons and cultivars inclusive it was found that grape and wine colour showed better correlations than for instance total phenols and tannins. This was especially true for total colour pigments in red grapes, measured with HPLC, when correlated with certain spectrophotometric analysis of wine colour. Cultivar and season as well as the synergism between the two were further investigated for its role in affecting correlations. When these relationships were further differentiated by season and by cultivar the resulting correlations varied. This work contributed a great deal of information to support the use of grape colour and phenolic compounds for the prediction of end wine colour and phenolic composition. The third objective was to investigate near infrared spectroscopy (FT-NIR) as a viable option to rapidly measured anthocyanins, tannins and total phenolics in red grapes. If proven successfully, this could be employed by a large cellar such as RW. FT-NIR has been used with success on grape extracts and in this instance the focus was to establish a calibration on the grape homogenate itself. Preliminary results showed that FT-NIR could be applied for the use of determination of anthocyanin and tannin levels in red grapes originating from RW. The prediction of total phenols was not found to be as accurate, but this could also be due to the reference method that was used. This work brought some interesting, practical information not only of importance for RW, but all wineries that are concerned with improving the basis on which grape quality is determined. The use of aerial data mapping for indicating areas regarding important grape colour and phenolic parameters was used in this study and is a very visual way of showing the distribution of certain ripeness parameters over a large area. Correlations between the grape and wines of such a large amount of red grape blocks for a specific area have not also been reported in South Africa before. The use of FT-NIR to determine anthocyanins and tannin concentrations in grape homogenates is also novel for its use in South African wineries. This work may assist grape and wine producers as well as analysts on the phenolic and colour profile of grapes and wines from RW. / AFRIKAANSE OPSOMMING: Suid-Afrikaanse rooiwyn word wêreld-wyd geken aan ‘n dieprooi kleur en vol struktuur. Die grootste rede vir hierdie verskynsel is hoë temperature wat ervaar word tydens rypwording en veral na véraison. In die Robertson wynstreek is temperature egter tydens rypwording dikwels vêr bo dit was as optimaal vir antosianien ontwikkeling beskou word. Die gepaste tyd om druiwe te pluk word nie net gedryf deur die tegnologiese rypheidsvlak nie, maar ook deur fenoliese rypheid. In ‘n koöperatiewe kelder omgewing soos Robertson Wynkelder (RW) word ‘n hoë lading druiwe elke dag ontvant vanaf verskillende produsente oor ‘n breë streek. Dit maak dit moeilik om te bepaal watter druiwe werklik beide tegnologies en fenolies ryp is. Die beste manier om hiervoor te vergoed is om ‘n standaard te stel vir ‘n reeks voorafbepaalde parameters. Die vlakke van die gekose parameters is, word bepaal deur navorsinguitsette sowel as die geskiedkunde data wat ingesamel is vanaf elkeen van die bepaalde blokke. Die parameters wat tans in gebruik is by RW om oesdatum en kwaliteit by inname te bepaal is pH, titreerbare suur (TA) en suiker vlak. Die tekortkoming hier is dat kwaliteit van druiwe beswaarlik met slegs hierdie informasie kan bepaal word, maar dat dit die betaling van die produsent by aflewering wesenlik kan beïnvloed. Dit het RW genoop om in samewerking met die Departement van Wingerd en Wynkunde, Universiteit van Stellenbosch nog parameters te ondersoek wat hierdie rypheid- en kwaliteitsbepaling by inname sou kon versterk. Twee belangrike faktore wat kwaliteit van rooiwyn bepaal is die kleur en struktuur. Antosianiene en tanniene is onderskeidelik verantwoordelik vir hierdie kwaliteits eienskappe van wyn. Wyn wat bestempel word as hoog in kleur en tannien inhoud word dikwels verbind met druiwe wat hoog is in hierdie faktore. Die moontlike korrelasie tussen die antosianien en tannien inhoud van druiwe en die wyn wat daarvan berei word is dus die basis waarop die potensiële toepassing van hierdie parameters berus. Die bepaling van antosianien en tannien vlakke word reeds in sommige laboratoriums gedoen. Die monster voorbereidings tyd, ekstraksies, toerusting en verbruikbare items nodig om hierdie tipe analieses te doen is egter hoog. Die analiese van hierdie komponente is meer lewensvatbaar in groot laboratoriums (soos in ‘n koöperatiewe kelder) waar groter volume druiwe ingeneem word gedurende parstyd. Die eerste doelwit van hierdie studie was om te bepaal teen watter vlakke antosianiene en tanniene in druiwe voorkom, spesifiek van die Robertson area. Die het behels ‘n wye verskeidenheid van blokke, verspreid oor die hele streek wat oor 3 seisoene gemonitor is in terme van veral kleur en tanniene maar ook ander belangrike parameters. Die idee hier is om insig te kry rakende watter vlakke bestempel kan word as laag en hoog in terme van antosianien en tanniene vir die Robertson streek. Daarvoor is slegs die mees aangeplantste rooi kultivars gebruik. Die verspreiding en gemiddelde vlakke waarteen antosianien en tanniene voorkom was bepaal deur gebruik te maak van metodes wat as relatief algemeen in laboratoria gebruik word. Die gemiddelde mono-flavonoïed en totale kleur pigment inhoud van die druiwe was laer as van die vlakke wat in die literatuur beskikbaar is, met totale flavanole wat hoër was. Die wyer verspreiding van die waardes het egter beter gekorreleer met die waardes soos beskryf in die literatuur. Die moontlike redes vir die verskillende vlakke word in die studie bespreek en word waarskynlik bepaal deur verskille in kultivar, mikro-klimaat en seisoen. Die tweede doelwit was om te bepaal of daar ‘n korrelasie te vinde is tussen die kleur en tannien inhoud van die druiwe en ooreenstemmende wyne. Sulke tipe korrelasies sal die basis vorm om antosianien en tannien inhoud van wyn reeds in die druiwe fase te kan voorspel. Nadat die ingesamelde druif en wyn data as ‘n geheel beskou was, was dit sigbaar dat die wynkleur parameters beter korrelasies bied as meeste tannien en totale fenole. Dit was veral waar in die geval van totale kleur pigmente soos gemeet met die HPLC teenoor die wynkleur parameters gemeet met spektrofotometriese metodes. Verdere ondersoeke in terme van die impak wat die kultivar en seisoenale kan hê het tot variërende korrelasies gelei.. Hierdie werk het ‘n groot bydrae gelewer om voorspellings van wyn kleur en fenoliese inhoud reeds met sukses vanaf die druif te bepaal. Derdens het die werk fourier transformasie naby infrarooi skandering (FT-NIR) ondersoek as ‘n lewensvatbare metode vir die bepaling van antosianien, tannien en totale fenoliese inhoud van druiwe en wyn. FT-NIR word reeds oor ‘n wye reeks wyne en druiwe ekstraksiemonsters toegepas en die doelwit hier was om druiwe homogenaat as matriks te kalibreer. Voorlopige resultate het bevind dat antosianien en tannien vlakke in druiwe van RW gemeet kan word met die FT-NIR, maar dat die kalibrasie vir totale fenole nog verbeter kan word. Hierdie werk het ‘n wye reeks interessante en prakties bruikbare informasie na vore gebring wat van onskatbare belang is vir RW en ander kelders wat besorgd is oor die verbetering van algemene druifkwaliteit. Geografiese kaarte wat belangrike druifkleur en fenoliese parameters aandui is in hierdie studie gebruik en wys hoe data visueel voorgestel kan word om die geheelindruk van gekose parameters oor ‘n groot area te vergelyk. Korrelasies tussen druiwe en wyn van so ‘n groot hoeveelheid druiwe blokke is nog nooit voorheen in Suid-Afrika getoon nie. Dieselfde geld vir die gebruik van FT-NIR vir die meet van kleur en fenoliese parameters in druiwe homogenate. Hierdie werk kan druiwe- en wynproduseerders sowel as analiste assisteer in terme van die kleur en fenoliese profiel van druiwe en wyn van RW. / Robertson Winery with Mr Bowen Botha as well as THRIP for funding this project

Os compostos fenólicos presentes em uvas e vinhos apresentam atividades antioxidantes benéficas à saúde humana, além de indicarem a qualidade desses produtos. Assim, esta tese teve como objetivo desenvolver, validar e aplicar métodos para a análise dos compostos fenólicos e avaliar a atividade antioxidante de uvas e vinhos tintos finos provenientes de uma nova região vitivinícola brasileira (Campanha Gaúcha - RS, Brasil). Ao total, foram utilizadas 103 amostras de vinhos finos tintos obtidos a partir de diferentes processos de vinificação (experimentais e comerciais), variedades (10), safras (2004 a 2015) e regiões (97 das microrregiões Ocidental, Central e Meridional da Campanha Gaúcha e seis de outras regiões do RS). Além disso, foram utilizadas oito amostras de uvas finas tintas da Campanha Gaúcha, das safras 2014 e 2015. O primeiro método, por cromatografia líquida de alta eficiência com detector de arranjo de diodos (HPLC-DAD), com propósito de dosar dois marcadores de compostos fenólicos, trans-resveratrol e quercetina, mostrou-se adequado para diferenciar 12 vinhos tintos do RS. O segundo método, por HPLC-DAD e espectrometria de massas de alta resolução, possibilitou o doseamento de seis fenóis bioativos (trans-resveratrol, quercetina, miricetina, quempferol, trans-Ɛ-viniferina e ácido trans-cinâmico) em 87 amostras da Campanha Gaúcha. Os teores mais elevados de compostos fenólicos foram encontrados em: variedades Arinarnoa, Marselan, Rebo, Syrah, Tempranillo e Teroldego; microrregião da Campanha Central; safra de 2013. Os vinhos foram classificados mediante análises de agrupamento hierárquico (HCA) e de componentes principais (PCA), conforme seus respectivos processos de vinificação, variedades e microrregiões. O terceiro e último método abordado foi com o intuito de analisar um maior número de compostos fenólicos, o que foi possível através da técnica de extração em fase sólida por troca catiônica forte. Esse método foi utilizado para análises de uvas e seus vinhos experimentais correspondentes da Campanha Gaúcha, por HPLC-DAD e por cromatografia líquida de ultra eficiência acoplado a espectrômetro de massas (UPLC-MS). Uvas com alto teor de galato de (-)-epicatequina originaram vinhos com elevada concentração de fenóis biativos. Tais amostras também foram analisadas quanto ao índice de fenóis totais e quanto à atividade antioxidante in vitro, demonstrando uma correlação positiva entre uvas e vinhos das mesmas variedades. Nesse mesmo estudo, foi possível atribuir atividades antioxidantes in vivo (modelo Caenorhabditis elegans) superiores nas amostras com teores mais elevados de astilbina e flavanóis, como a (+)-catequina, a (-)-epicatequina e a (-)-epigalocatequina. Assim, esses compostos, principalmente a astilbina, poderão ser utilizados como modelos para modificações moleculares na área de química medicinal. Além disso, o estudo dos compostos fenólicos pode contribuir para a melhoria da qualidade de vinhos da Campanha Gaúcha, trazendo benefícios diretos à vitivinicultura nacional e indiretos a toda a sociedade. / Phenolic compounds found in grapes and wines show antioxidant activities with health benefits for humans, and also indicate the quality of these products. Therefore, this thesis aimed to develop, validate and investigate methods for phenolic compound analysis, and evaluate the antioxidant activity of grapes and wines from a new Brazilian viticulture region (Campanha Gaúcha, Southern Brazil). In total, 103 red wine samples were used. These samples were obtained from different winemaking processes (experimental and commercial), varieties (10), vintages (2004 to 2015), and regions (97 from occidental, central e meridional from Campanha Gaúcha and six from other Southern Brazilian regions). Eight grape samples, 2014 and 2015 vintages, from Campanha Gaúcha were also used. The first method, using high-performance liquid chromatography with a diode array detector (HPLC-DAD) was used to quantify two phenolic compound markers, trans-resveratrol, and quercetin, and was adequate for differentiating 12 red wines from Southern Brazil. The second method, using HPLC-DAD and high-resolution mass spectrometry, enabled the quantification of six bioactive phenols (trans-resveratrol, quercetin, myricetin, kaempferol, trans-Ɛ-viniferin, and trans-cinnamic acid) in 87 samples from Campanha Gaúcha. The highest levels of phenolic compounds were found in the Arinarnoa, Marselan, Rebo, Syrah, Tempranillo, and Teroldego varieties; central Campanha Gaúcha micro-region varieties; and the 2013 vintage. The wines were classified according to their winemaking processes, varieties, and micro-regions using hierarchical cluster and principal component analyses. The third and final method aimed to analyze a large number of phenolic compounds using a solid phase extraction technique employing strong cation exchange. This method was used to analyze grapes and their corresponding wines from Campanha Gaúcha using HPLC-DAD and ultra-performance liquid chromatography coupled with mass spectrometry. Grapes with high levels of (-)-epicatechin galate corresponded to wines more concentrated in bioactive phenols. The samples were also analyzed using the total phenol content index and in vitro antioxidant activity, showing a positive correlation between grapes and wines of the same varieties. In the same study, it was possible to attribute the in vivo antioxidant activities (Caenorhabditis elegans model) to samples with higher levels of astilbin and flavanols such as (+)-catechin, (-)-epicatechin, and (-)-epigalocatechin. Therefore, such compounds, mainly astilbin, may be used for a molecular modification model in medicinal chemistry. Moreover, the study of phenolic compounds may contribute to the improvement of the quality of wines from Campanha Gaúcha, and can bring several direct benefits for national viticulture and indirect benefits to all society.

Os compostos fenólicos presentes em uvas e vinhos apresentam atividades antioxidantes benéficas à saúde humana, além de indicarem a qualidade desses produtos. Assim, esta tese teve como objetivo desenvolver, validar e aplicar métodos para a análise dos compostos fenólicos e avaliar a atividade antioxidante de uvas e vinhos tintos finos provenientes de uma nova região vitivinícola brasileira (Campanha Gaúcha - RS, Brasil). Ao total, foram utilizadas 103 amostras de vinhos finos tintos obtidos a partir de diferentes processos de vinificação (experimentais e comerciais), variedades (10), safras (2004 a 2015) e regiões (97 das microrregiões Ocidental, Central e Meridional da Campanha Gaúcha e seis de outras regiões do RS). Além disso, foram utilizadas oito amostras de uvas finas tintas da Campanha Gaúcha, das safras 2014 e 2015. O primeiro método, por cromatografia líquida de alta eficiência com detector de arranjo de diodos (HPLC-DAD), com propósito de dosar dois marcadores de compostos fenólicos, trans-resveratrol e quercetina, mostrou-se adequado para diferenciar 12 vinhos tintos do RS. O segundo método, por HPLC-DAD e espectrometria de massas de alta resolução, possibilitou o doseamento de seis fenóis bioativos (trans-resveratrol, quercetina, miricetina, quempferol, trans-Ɛ-viniferina e ácido trans-cinâmico) em 87 amostras da Campanha Gaúcha. Os teores mais elevados de compostos fenólicos foram encontrados em: variedades Arinarnoa, Marselan, Rebo, Syrah, Tempranillo e Teroldego; microrregião da Campanha Central; safra de 2013. Os vinhos foram classificados mediante análises de agrupamento hierárquico (HCA) e de componentes principais (PCA), conforme seus respectivos processos de vinificação, variedades e microrregiões. O terceiro e último método abordado foi com o intuito de analisar um maior número de compostos fenólicos, o que foi possível através da técnica de extração em fase sólida por troca catiônica forte. Esse método foi utilizado para análises de uvas e seus vinhos experimentais correspondentes da Campanha Gaúcha, por HPLC-DAD e por cromatografia líquida de ultra eficiência acoplado a espectrômetro de massas (UPLC-MS). Uvas com alto teor de galato de (-)-epicatequina originaram vinhos com elevada concentração de fenóis biativos. Tais amostras também foram analisadas quanto ao índice de fenóis totais e quanto à atividade antioxidante in vitro, demonstrando uma correlação positiva entre uvas e vinhos das mesmas variedades. Nesse mesmo estudo, foi possível atribuir atividades antioxidantes in vivo (modelo Caenorhabditis elegans) superiores nas amostras com teores mais elevados de astilbina e flavanóis, como a (+)-catequina, a (-)-epicatequina e a (-)-epigalocatequina. Assim, esses compostos, principalmente a astilbina, poderão ser utilizados como modelos para modificações moleculares na área de química medicinal. Além disso, o estudo dos compostos fenólicos pode contribuir para a melhoria da qualidade de vinhos da Campanha Gaúcha, trazendo benefícios diretos à vitivinicultura nacional e indiretos a toda a sociedade. / Phenolic compounds found in grapes and wines show antioxidant activities with health benefits for humans, and also indicate the quality of these products. Therefore, this thesis aimed to develop, validate and investigate methods for phenolic compound analysis, and evaluate the antioxidant activity of grapes and wines from a new Brazilian viticulture region (Campanha Gaúcha, Southern Brazil). In total, 103 red wine samples were used. These samples were obtained from different winemaking processes (experimental and commercial), varieties (10), vintages (2004 to 2015), and regions (97 from occidental, central e meridional from Campanha Gaúcha and six from other Southern Brazilian regions). Eight grape samples, 2014 and 2015 vintages, from Campanha Gaúcha were also used. The first method, using high-performance liquid chromatography with a diode array detector (HPLC-DAD) was used to quantify two phenolic compound markers, trans-resveratrol, and quercetin, and was adequate for differentiating 12 red wines from Southern Brazil. The second method, using HPLC-DAD and high-resolution mass spectrometry, enabled the quantification of six bioactive phenols (trans-resveratrol, quercetin, myricetin, kaempferol, trans-Ɛ-viniferin, and trans-cinnamic acid) in 87 samples from Campanha Gaúcha. The highest levels of phenolic compounds were found in the Arinarnoa, Marselan, Rebo, Syrah, Tempranillo, and Teroldego varieties; central Campanha Gaúcha micro-region varieties; and the 2013 vintage. The wines were classified according to their winemaking processes, varieties, and micro-regions using hierarchical cluster and principal component analyses. The third and final method aimed to analyze a large number of phenolic compounds using a solid phase extraction technique employing strong cation exchange. This method was used to analyze grapes and their corresponding wines from Campanha Gaúcha using HPLC-DAD and ultra-performance liquid chromatography coupled with mass spectrometry. Grapes with high levels of (-)-epicatechin galate corresponded to wines more concentrated in bioactive phenols. The samples were also analyzed using the total phenol content index and in vitro antioxidant activity, showing a positive correlation between grapes and wines of the same varieties. In the same study, it was possible to attribute the in vivo antioxidant activities (Caenorhabditis elegans model) to samples with higher levels of astilbin and flavanols such as (+)-catechin, (-)-epicatechin, and (-)-epigalocatechin. Therefore, such compounds, mainly astilbin, may be used for a molecular modification model in medicinal chemistry. Moreover, the study of phenolic compounds may contribute to the improvement of the quality of wines from Campanha Gaúcha, and can bring several direct benefits for national viticulture and indirect benefits to all society.

Os compostos fenólicos presentes em uvas e vinhos apresentam atividades antioxidantes benéficas à saúde humana, além de indicarem a qualidade desses produtos. Assim, esta tese teve como objetivo desenvolver, validar e aplicar métodos para a análise dos compostos fenólicos e avaliar a atividade antioxidante de uvas e vinhos tintos finos provenientes de uma nova região vitivinícola brasileira (Campanha Gaúcha - RS, Brasil). Ao total, foram utilizadas 103 amostras de vinhos finos tintos obtidos a partir de diferentes processos de vinificação (experimentais e comerciais), variedades (10), safras (2004 a 2015) e regiões (97 das microrregiões Ocidental, Central e Meridional da Campanha Gaúcha e seis de outras regiões do RS). Além disso, foram utilizadas oito amostras de uvas finas tintas da Campanha Gaúcha, das safras 2014 e 2015. O primeiro método, por cromatografia líquida de alta eficiência com detector de arranjo de diodos (HPLC-DAD), com propósito de dosar dois marcadores de compostos fenólicos, trans-resveratrol e quercetina, mostrou-se adequado para diferenciar 12 vinhos tintos do RS. O segundo método, por HPLC-DAD e espectrometria de massas de alta resolução, possibilitou o doseamento de seis fenóis bioativos (trans-resveratrol, quercetina, miricetina, quempferol, trans-Ɛ-viniferina e ácido trans-cinâmico) em 87 amostras da Campanha Gaúcha. Os teores mais elevados de compostos fenólicos foram encontrados em: variedades Arinarnoa, Marselan, Rebo, Syrah, Tempranillo e Teroldego; microrregião da Campanha Central; safra de 2013. Os vinhos foram classificados mediante análises de agrupamento hierárquico (HCA) e de componentes principais (PCA), conforme seus respectivos processos de vinificação, variedades e microrregiões. O terceiro e último método abordado foi com o intuito de analisar um maior número de compostos fenólicos, o que foi possível através da técnica de extração em fase sólida por troca catiônica forte. Esse método foi utilizado para análises de uvas e seus vinhos experimentais correspondentes da Campanha Gaúcha, por HPLC-DAD e por cromatografia líquida de ultra eficiência acoplado a espectrômetro de massas (UPLC-MS). Uvas com alto teor de galato de (-)-epicatequina originaram vinhos com elevada concentração de fenóis biativos. Tais amostras também foram analisadas quanto ao índice de fenóis totais e quanto à atividade antioxidante in vitro, demonstrando uma correlação positiva entre uvas e vinhos das mesmas variedades. Nesse mesmo estudo, foi possível atribuir atividades antioxidantes in vivo (modelo Caenorhabditis elegans) superiores nas amostras com teores mais elevados de astilbina e flavanóis, como a (+)-catequina, a (-)-epicatequina e a (-)-epigalocatequina. Assim, esses compostos, principalmente a astilbina, poderão ser utilizados como modelos para modificações moleculares na área de química medicinal. Além disso, o estudo dos compostos fenólicos pode contribuir para a melhoria da qualidade de vinhos da Campanha Gaúcha, trazendo benefícios diretos à vitivinicultura nacional e indiretos a toda a sociedade. / Phenolic compounds found in grapes and wines show antioxidant activities with health benefits for humans, and also indicate the quality of these products. Therefore, this thesis aimed to develop, validate and investigate methods for phenolic compound analysis, and evaluate the antioxidant activity of grapes and wines from a new Brazilian viticulture region (Campanha Gaúcha, Southern Brazil). In total, 103 red wine samples were used. These samples were obtained from different winemaking processes (experimental and commercial), varieties (10), vintages (2004 to 2015), and regions (97 from occidental, central e meridional from Campanha Gaúcha and six from other Southern Brazilian regions). Eight grape samples, 2014 and 2015 vintages, from Campanha Gaúcha were also used. The first method, using high-performance liquid chromatography with a diode array detector (HPLC-DAD) was used to quantify two phenolic compound markers, trans-resveratrol, and quercetin, and was adequate for differentiating 12 red wines from Southern Brazil. The second method, using HPLC-DAD and high-resolution mass spectrometry, enabled the quantification of six bioactive phenols (trans-resveratrol, quercetin, myricetin, kaempferol, trans-Ɛ-viniferin, and trans-cinnamic acid) in 87 samples from Campanha Gaúcha. The highest levels of phenolic compounds were found in the Arinarnoa, Marselan, Rebo, Syrah, Tempranillo, and Teroldego varieties; central Campanha Gaúcha micro-region varieties; and the 2013 vintage. The wines were classified according to their winemaking processes, varieties, and micro-regions using hierarchical cluster and principal component analyses. The third and final method aimed to analyze a large number of phenolic compounds using a solid phase extraction technique employing strong cation exchange. This method was used to analyze grapes and their corresponding wines from Campanha Gaúcha using HPLC-DAD and ultra-performance liquid chromatography coupled with mass spectrometry. Grapes with high levels of (-)-epicatechin galate corresponded to wines more concentrated in bioactive phenols. The samples were also analyzed using the total phenol content index and in vitro antioxidant activity, showing a positive correlation between grapes and wines of the same varieties. In the same study, it was possible to attribute the in vivo antioxidant activities (Caenorhabditis elegans model) to samples with higher levels of astilbin and flavanols such as (+)-catechin, (-)-epicatechin, and (-)-epigalocatechin. Therefore, such compounds, mainly astilbin, may be used for a molecular modification model in medicinal chemistry. Moreover, the study of phenolic compounds may contribute to the improvement of the quality of wines from Campanha Gaúcha, and can bring several direct benefits for national viticulture and indirect benefits to all society.

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