Effects of different oenological technologies on the evolution of anthocyanin derivatives determined by LC-MS during red wine fermentations. Encapsulation of phenolic grape pomace extracts for cosmetic applications

167 p.This PhD thesis is focused on to two topics related to wine-making: (i) the study of the influence of novel oenological techniques on the phenolic composition of wine during its elaboration, and (ii) the valorization of grape pomace waste resulting from wine elaboration.Phenolic compounds occur naturally in plants and fruits. Red grapes are one of the fruits with the highest content of phenolics, which account for their color, flavor and antioxidant and anti-inflammatory properties. Wine is a popular and appreciated beverage elaborated through a fermentation process from grape must and grapes. In Spain, red wine is of great importance, especially in the area of La Rioja, where red wine has a Protected Designation of Origin (PDO). Red wine is rich in phenolic compounds, mainly anthocyanins and anthocyanin derivatives. Pedoclimatic conditions of gravevine areas affect the chemical composition of grape berries, and thus of the wine produced from these grapes. Wine elaboration techniques also influence the composition of wine and its evolution during the wine-making process and its maturation in the long term.In this work, different oenological technologies were evaluated to study their influence on the phenolic composition of wine during its elaboration process. For this purpose, wine elaborations were performed in 2018 and 2019 in Bodegas Faustino, under controlled conditions at pilot scale, employing the main grape cultivar under the PDO "Rioja" named Tempranillo. The oenological technologies studied were: (i) microoxygenation during malolactic fermentation (MLF); (ii) ionic exchange technologies to lower the pH of the grape must at the beginning of alcoholic fermentation (AF); (iii) tannin addition at the beginning of AF; and (iv) the use of a mixture of must from different grape cultivars. An experimental design including all the variables of the oenological technologies was carried out, and the wine elaborations were monitored and sampled from the start of AF until the end of MLF.In the must and wine samples, 68 phenolic compounds (13 anthocyanins, 27 anthocyanin derivatives and 28 tannins) were determined during the wine-making process by HPLC-DAD and HPLC-DAD-ESI(+)-MS/MS. The concentration phenolic profiles of 8 anthocyanins and anthocyanin derivatives, selected because of their relevance in wine characteristics, were treated mathematically in order to study their evolution kinetics, i.e. extraction from grape, formation and degradation. The kinetic constants were statistically compared (i) between different compounds under the same oenological conditions, in order to study their different stability and evolution kinetics; and (ii) between different oenological conditions employed during wine elaboration, to observe the influence of the oenological techniques on the evolution kinetics.Results of the concentration measurements and the consequent kinetic study showed that the main anthocyanin derivatives present in must and wine are pyranoanthocyanins (i.e. Vitisin A, Vitisin B and derivatives with vinylphenol), followed by anthocyanin-flavan-3-ol condensation products, both direct (F- A+ or A+-F) or indirect, linked by an ethyl bridge (A+-ethyl-F). Vitisin B reaches higher concentration levels than the other compounds during wine elaboration; however, its stability is lower in the long term, leading to a higher degradation of the compound and a lower concentration in wine at the end of fermentations. In contrast, Vitisin A shows a higher stability during fermentations, leading to a more important contribution of this compound to wine color at the end of the elaboration. Malvidin-3-O-glucoside-vinylphenol showed a similar stability as Vitisin A, and thus, it also contributes to wine color. Among the anthocyanin-flavan-3-ol condensation derivatives, the order of stability found is F-A+ > A+-ethyl-F > A+-F; catechin-malvidin-3-O-glucoside (F-A+) being the most important regarding wine color contribution at the end of fermentations.Related to the studied oenological variables, the use of a secondary grape cultivar mixed with the main grape cultivar Tempranillo compared to the use of the single main grape cultivar, led to an increase in the formation constants of pyranoanthocyanins and a decrease in those of anthocyanin-flavan-3-ol condensation products. The application of ionic exchange resulted in a decrease in the concentrations of pyranoanthocyanins and an increase in the concentrations of the anthocyanin-flavan-3-ol condensation products at the end of the fermentations. The addition of tannins and the use of microoxygenation during MLF did not produce significant changes in the concentration of the studied compounds or their kinetic constants.Grape pomace residues from the wine elaborations were employed in order to obtain extracts rich in phenolic compounds with potential cosmetic applications. These extracts were analyzed by HPLC-DAD and HPLC-DAD-ESI(+)-QToF/MS: 13 anthocyanins were identified and quantified, and 24 minor phenolics (flavan-3-ols, flavonols, hydroxycinnamic and hydroxybenzoic acids) were identified. The extracts were formulated in liposomal vesicles, i.e. liposomes and transfersomes, with the aim of enhancing their health beneficial properties and making them suitable for application on skin. The nanoformulations prepared were characterized in terms of size, shape and stability by transmission electron microscopy (TEM), dynamic light scattering (DLS) and zeta potential, and their antioxidant activities and total phenolic contents were assayed by means of the DPPH and FRAP and Folin-Ciocalteu assays respectively. Furthermore, the cytocompatibility of the nanoformulations containing the extracts, as well as their inhibition capacity of intracellular Reactive Oxygen Species (ROS) generation were evaluated. Nanoformulation of the extracts obtained from the grape pomace residues of wine-making process yielded stable, safe, effective and usable formulations for the treatment of skin under oxidative stress conditions. The in vitro studies highlighted the ability of the vesicle formulations to counteract ROS overproduction, enhancing the effect of the extracts in the cells

Tempranillo grape extract in transfersomes: A nanoproduct with antioxidant activity

Polyphenols are gaining increasing interest due to their beneficial properties to human health. Grape pomace, the by-product of wine production, is a source of these bioactive compounds. An extract from Tempranillo grape pomace was obtained and characterized qualitatively and quantitatively. The major components found were anthocyanins, flavan-3-ols, and flavonols. To improve the bioavailability of these compounds, the extract was formulated in phospholipid vesicles, namely transfersomes. Spherical unilamellar vesicles around 100 nm each were obtained. The antioxidant activity of both the extract and the transfersomes was evaluated by using colorimetric assays (i.e., DPPH, FRAP, and Folin–Ciocalteu). The cells’ viability and the antioxidant activity were assessed in keratinocytes. The results showed that the extract and the transfersomes had no cytotoxic effects and exerted remarkable antioxidant activity, which was more evident in a vesicle formulation. These findings highlighted the potential of the Tempranillo grape pomace extract and the efficacy of the incorporation into phospholipid vesicles.Fil: Asensio Regalado, Carlos. Universidad del País Vasco; EspañaFil: Alonso Salces, Rosa Maria. Universidad Nacional de Mar del Plata. Instituto de Investigaciones en Producción, Sanidad y Ambiente - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones en Producción, Sanidad y Ambiente; ArgentinaFil: Gallo, Blanca. Universidad del País Vasco; EspañaFil: Berrueta, Luis A.. Universidad del País Vasco; EspañaFil: Era, Benedetta. Università Degli Studi Di Cagliari.; ItaliaFil: Pintus, Francesca. Università Degli Studi Di Cagliari.; ItaliaFil: Caddeo, Carla. Università Degli Studi Di Cagliari.; Itali

Stepwise strategy based on 1H-NMR fingerprinting in combination with chemometrics to determine the content of vegetable oils in olive oil mixtures

1H NMR fingerprinting of edible oils and a set of multivariate classification and regression models organised in a decision tree is proposed as a stepwise strategy to assure the authenticity and traceability of olive oils and their declared blends with other vegetable oils (VOs). Oils of the 'virgin olive oil' and 'olive oil' categories and their mixtures with the most common VOs, i.e. sunflower, high oleic sunflower, hazelnut, avocado, soybean, corn, refined palm olein and desterolized high oleic sunflower oils, were studied. Partial least squares (PLS) discriminant analysis provided stable and robust binary classification models to identify the olive oil type and the VO in the blend. PLS regression afforded models with excellent precisions and acceptable accuracies to determine the percentage of VO in the mixture. The satisfactory performance of this approach, tested with blind samples, confirm its potential to support regulations and control bodies. Keywords: Adulteration; Authentication; Decision tree; Multivariate data analysis; Nuclear magnetic resonance; Olive oil

Determination of Vegetable Oils in Olive Oil Mixtures Using a Stepwise Strategy Based on H-1-Nmr Fingerprinting and Pattern Recognition

Final version is available in open access in publisher’s site.-- This abstract is based on the published article: Rosa María Alonso-Salces, Luis Ángel Berrueta, Beatriz Quintanilla-Casas, Stefania Vichi, Alba Tres, María Isabel Collado, Carlos Asensio-Regalado, Gabriela Elena Viacava, Aimará Ayelen Poliero, Enrico Valli, Alessandra Bendini, Tullia Gallina Toschi, José Manuel Martínez-Rivas, Wenceslao Moreda, Blanca Gallo. Stepwise strategy based on 1H-NMR fingerprinting in combination with chemometrics to determine the content of vegetable oils in olive oil mixtures. Food Chemistry, Volume 366, 2022, 130588, https://doi.org/10.1016/j.foodchem.2021.130588.--http://hdl.handle.net/10261/261907Peer reviewe

Tempranillo Grape Extract in Transfersomes: A Nanoproduct with Antioxidant Activity

Polyphenols are gaining increasing interest due to their beneficial properties to human health. Grape pomace, the by-product of wine production, is a source of these bioactive compounds. An extract from Tempranillo grape pomace was obtained and characterized qualitatively and quantitatively. The major components found were anthocyanins, flavan-3-ols, and flavonols. To improve the bioavailability of these compounds, the extract was formulated in phospholipid vesicles, namely transfersomes. Spherical unilamellar vesicles around 100 nm each were obtained. The antioxidant activity of both the extract and the transfersomes was evaluated by using colorimetric assays (i.e., DPPH, FRAP, and Folin–Ciocalteu). The cells’ viability and the antioxidant activity were assessed in keratinocytes. The results showed that the extract and the transfersomes had no cytotoxic effects and exerted remarkable antioxidant activity, which was more evident in a vesicle formulation. These findings highlighted the potential of the Tempranillo grape pomace extract and the efficacy of the incorporation into phospholipid vesicles

Reactivity of trivalent metals towards the Keggin-type [SbW9O33]9- trilacunary polyoxoanion

Systematic studies on the reactivity of trivalent metals (Sc, Cr, Mn, Fe, Co, Al, Ga) towards the Keggin-type [SbW9O33]9- trilacunary anion have been carried out. The usual methods for the chemical and structural characterization of polyoxometalates have been applied to the obtained novel metal-substituted polyoxoanions

Productos Naturales en alimentos, bebidas y plantas

The chemical analysis of food and beverages has a huge interest in food science in order to develop deep knowledge of natural products, which can lead to an improvement of nutritional quality and manufacturing methods, as well as the detection of geographical origins, bad practices, adulterations and frauds.Polyphenols are a relevant group of natural products which are widely found in the Plant Kingdom. Due to their antioxidant, antimicrobial and antiparasitic properties these compounds have attracted attention for their many benefits for human health (e.g. cardiovascular diseases, cancer...), and have also been associated with the development of biotic resistance in plants. In addition, they are a key element in explaining some food properties, such as flavour, bitterness, astringency, aroma and colour.Los polifenoles son un grupo muy importante de productos naturales debido a su ubicuidad en el reino vegetal y su interés para explicar muchas de las propiedades de los alimentos, tales como el amargor, la astringencia, el aroma y el color, además de sus muchos efectos beneficiosos para la salud humana, debidos sobre todo a su gran potencial antioxidante.Fil: Gallo, Blanca. Universidad del País Vasco; EspañaFil: Berrueta, Luis Ángel. Universidad del País Vasco; EspañaFil: Sasía Arriba, Andrea. Universidad del País Vasco; EspañaFil: Asensio Regalado, Carlos. Universidad del País Vasco; EspañaFil: Alonso Salces, Rosa Maria. Universidad Nacional de Mar del Plata. Instituto de Investigaciones en Producción, Sanidad y Ambiente - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones en Producción, Sanidad y Ambiente; ArgentinaZientzia eta Teknologia Fakultateko VII. Ikerkuntza Jardunaldiak; VII Jornadas de Investigación de la Facultad de Ciencia y TecnologíaEspañaUniversidad del País VascoEuskal Herriko Unibertsitate

Effect of climate change on the polyphenolic composition of the main varieties of grape from La Rioja (Spain) and new oenological strategies to correct these effects on the quality of red wine

Even though grapes grow all around the planet those aimed for high quality wine production occur in areas with very specific climate conditions. Moreover, each individual variety of grape requires conditions that are even more particular. Thus, any change on the climate or meteorological patterns can affect the chemistry of the grape and the final quality of wine. Recent studies have explored the impact of climate change on the worldwide production of grape, and some highlight an advance on the vintage date associated with a temperature increase, this being for instance, 18-21 days in the South of France from 1940 to 2000. Regarding the chemical composition of wine, anthocyanins and condensed tannins are the main type of polyphenolic compounds found in red wine. Anthocyanins are the main pigments of red grapes whereas tannins play an important role on the astringency and bitterness of wine. During wine making process, both types of compounds undergo chemical transformations leading to more stable polymeric pigments. The mechanisms of these reactions and the structures of these derivative pigments have been a recurrent topic of study. With the advances on the analytical techniques, some of these structures have been elucidated, discovering products of the direct and indirect condensation of anthocyanins and tannins. Additionally, anthocyanins have been found to react with other minor components present in wine, yielding a new class of compounds called pyranoanthocyanins and producing changes in the hue of the wine [3]. A better knowledge of the formation kinetics and evolution of these pigments during wine production will allow the producer to adapt the oenological practice to control and optimise the final quality of red wine. This work is aimed towards the study of climate influence on the polyphenolic composition of must and wine from Tempranillo grape cultivar, the main variety of grape in La Rioja (Spain), and its evolution during wine production.Fil: Asensio-Regalado, Carlos. Universidad del País Vasco; EspañaFil: Sasía-Arriba, Andrea. Universidad del País Vasco; EspañaFil: Poliero, Aimará Ayelen. Universidad del País Vasco; EspañaFil: Alonso Salces, Rosa Maria. Universidad Nacional de Mar del Plata. Instituto de Investigaciones en Producción, Sanidad y Ambiente - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones en Producción, Sanidad y Ambiente; Argentina. Universidad Nacional de Mar del Plata, Facultad de Ciencias Exactas y Naturales, Centro de Investigación en Abejas Sociales (CIAS); ArgentinaFil: Gallo, Blanca. Universidad del País Vasco; EspañaFil: Berrueta, Luis Ángel. Universidad del País Vasco; EspañaXXXth International Conference on PolyphenolsTurkuFinlandiaGroupe Polyphénols and University of Turk

A Liposomal Formulation to Exploit the Bioactive Potential of an Extract from Graciano Grape Pomace

Antioxidant compounds with health benefits can be found in food processing residues, such as grape pomace. In this study, antioxidants were identified and quantified in an extract obtained from Graciano red grape pomace via a green process. The antioxidant activity of the extract was assessed by the DPPH and FRAP tests, and the phenolic content by the Folin–Ciocalteu test. Furthermore, nanotechnologies were employed to produce a safe and effective formulation that would exploit the antioxidant potential of the extract for skin applications. Anthocyanins, flavan-3-ols and flavanols were the main constituents of the grape pomace extract. Phospholipid vesicles, namely liposomes, were prepared and characterized. Cryo-TEM images showed that the extract-loaded liposomes were predominantly spherical/elongated, small, unilamellar vesicles. Light scattering results revealed that the liposomes were small (~100 nm), homogeneously dispersed, and stable during storage. The non-toxicity of the liposomal formulation was demonstrated in vitro in skin cells, suggesting its possible safe use. These findings indicate that an extract with antioxidant properties can be obtained from food processing residues, and a liposomal formulation can be developed to exploit its bioactive value, resulting in a promising healthy product

Untargeted Metabolomic Liquid Chromatography High-Resolution Mass Spectrometry Fingerprinting of Apple Cultivars for the Identification of Biomarkers Related to Resistance to Rosy Apple Aphid

Liquid chromatography high-resolution mass spectrometry fingerprinting together with pattern recognition techniques was used to determine the metabolites involved in the susceptibility of apple cultivars to rosy apple aphid (RAA). Preprocessing of ultra-high-performance liquid chromatography coupled to electrospray ionization and quadrupole time-of-flight mass spectrometry raw data of resistant and susceptible apple cultivars was carried out with XCMS and CAMERA packages. Univariate statistical tools and multivariate data analysis highlighted significant different profiles of the apple metabolomes according to their tolerance to RAA. Optimized and cross-validated Partial least squares discriminant analysis and orthogonal projections to latent structures discriminant analysis models confirmed trans-4-caffeoylquinic acid and 4-p-coumaroylquinic acid as biomarkers for the identification of resistant and susceptible apple cultivars to RAA and disclosed that only hydroxycinnamic acids are involved in the disease susceptibility of cultivars. In this sense, the final steps of the biosynthesis of caffeoylquinic acid (CQA) and p-coumaroylquinic acid (p-CoQA) become decisive because the isomerization of 5-CQA to 4-CQA is favored in resistant cultivars, whereas the isomerization of 5-p-CoQA to 4-p-CoQA is favored in susceptible cultivars.Fil: Alonso Salces, Rosa Maria. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones en Sanidad Producción y Ambiente. - Comisión de Investigaciones Científicas de la Provincia de Buenos Aires. Instituto de Investigaciones en Sanidad Producción y Ambiente; ArgentinaFil: Berrueta, Luis A.. Universidad del País Vasco; EspañaFil: Abad García, Beatriz. Universidad del País Vasco; EspañaFil: Sasía Arriba, Andrea. Universidad del País Vasco; EspañaFil: Asensio Regalado, Carlos. Universidad del País Vasco; EspañaFil: Dapena, Enrique. No especifíca;Fil: Gallo, Blanca. Universidad del País Vasco; Españ