Diseño y aplicación de rúbricas a la evaluación de monografías elaboradas en Nutrición y Bromatología mediante el uso de STUDIUM

Memoria ID-0071. Ayudas de la Universidad de Salamanca para la innovación docente, curso 2015-2016

Physico-Chemical and Chromatic Characterization of Malvidin 3-Glucoside-vinylcatechol and Malvidin 3-Glucoside-vinylguaiacol Wine Pigments

[EN] The physicochemical and chromatic features of malvidin 3-glucoside-vinylcatechol (PA1) and malvidin 3-glucoside-vinylguaiacol (PA2) adducts were investigated. Important differences between both pigments were observed. In the investigated pH range (2.0−4.5), our results suggest that PA1 could be considered as a noncovalent dimer of two pyranoflavylium ions (AH)2 which undergo a hydration reaction in two successive steps, with no proton transfer. In contrast, only proton transfer equilibrium between the pyranoflavylium ion and the quinonoid bases was observed for PA2. The hydration and acidity thermodynamic constants of both pyranoanathocyanins were determined by UV−vis spectroscopy. Pigment PA1 was shown to be less sensitive to bisulfite addition than to water addition, and PA2 seems to be largely insensitive to both water and bisulfite additions. The binding constants for the interaction between the pigments and (+)-catechin in aqueous solution and the changes in the CIELAB parameters that it provoked were also determined. The huge increase in the absorption of the pyranoflavylium ion of PA2 when it binds catechin has no equivalent for anthocyanins and nicely outlines the potential of pyranoanthocyanins in the expression of natural colors

Schizanthus grahamii and Schizanthus hookeri. Is there any relationship between their anthocyanin compositions and their different pollination syndromes?

[EN] Three different pollination syndromes (bee, hummingbird and moth-pollination syndromes) have been described for the different species of the genus Schizanthus Ruiz & Pav. (Solanaceae). Two closely related species from a phylogenetic point of view, Schizanthus grahamii and Schizanthus hookeri, show hummingbird and bee-pollination syndromes, respectively. One of the traits used to assign these pollination syndromes is the colour of the petals, which is red in S. grahamii and bluish-pink in S. hookeri. The objective of this work was to establish the differences in the anthocyanin composition of these two Schizanthus species that contribute to the different pollination syndrome. Identification of the pigments was carried out from the chromatographic and spectral data supplied by the HPLC-DAD-MSn analyses of the samples. Alkaline and acid hydrolyses were also performed in the extracts and in some isolated compounds in order to confirm the identities. First difference between these two species was found in the total anthocyanin content, which was about 13-fold higher in S. grahamii than in S. hookeri. Furthermore, the major peak was also different in both cases, thus explaining quantitative and qualitative colour differences between species. Delphinidin 3-O-rutinoside represented ca. 72% of the total area in S. grahamii and petanin (Petunidin 3-O-p-coumaroylrutinoside-5-O-glucoside) accounted for almost 62% of the total area in S. hookeri. The presence of the p-coumaroylrutinosyl moiety in the petanin makes the intramolecular copigmentation possible, thus conferring the bluish-pink hue to the petals of S. hookeri. Delphinidin 3-O-rutinoside is in turn responsible for the red colour exhibited by the petals of S. grahamii and along with other floral traits, responsible for the attraction of hummingbirds to the plant. Pigments structurally related to petanin, which have been often detected in other genus of Solanaceae, were detected in both species and in similar contents, supporting results from previous studies that pointed to a bee-pollinated common ancestor for both species that further specialised acquiring traits attractive to hummingbirds and among them, red colour of the petals which is mainly supplied by delphinidin 3-O-rutinoside

Behaviour and characterisation of the colour during red wine making and maturation

[EN] During winemaking and ageing, the colour of red wine evolves from the initial purple-red hues of young red wines towards more red-orange ones. Simultaneously, a modification in the pigment profile takes place. The stability of pigments isolated from red wine has been usually studied in model solutions. However, studies carried out on wines are scarce. The objective of this work was to analyse the changes in composition, colour and stability in relation to pH and SO2 that take place in a red wine during winemaking and maturation. For this purpose, samples of red wine from Tempranillo grapes were collected periodically during 18 months and submitted to chromatic analysis using the CIELAB and CIELUV colour spaces and high performance liquid chromatography–diode array detection coupled to a mass spectrometer analysis (HPLC–DAD–MS). The results obtained showed the existence of processes capable of causing quantitative and qualitative changes in the colouring material of the wine. These changes, in spite of the relatively short time considered, are sufficient to cause changes of colour that can be perceived by the human eye and lead to the formation of pigments that are more stable against pH and against bleaching by SO2 than the original anthocyanins

Development of a fractionation method for the detection and identification of oak ellagitannins in red wines

[ES] During maturation and ageing in oak barrels wines improve their organoleptic properties. Ellagitannins can be released from wood to the wine and be involved in oxidation reactions and seem to influence the astringency and colour properties of the wine. Nevertheless, the ellagitannins levels are lower than those of other wine constituents and, consequently, they are not easily detected. This study has developed a two-step fractionation method consisting of a solid phase extraction in C-18 Sep-Pak® cartridges followed by size exclusion chromatography in hand-packed Sephadex LH-20 minicolumn for the detection of oak ellagitannins in different types of wines. An HPLC method has also been developed which allows the separation of compounds with the same m/z ratios, facilitating the ellagitannin identification by means of the mass spectrometric analyses. The main oak ellagitannins (grandinin, vescalagin, roburin E and castalagin) were isolated, detected separately and identified in a spiked wine and in three real ones, proving the usefulness of the fractionation method

Changes in the detailed pigment composition of red wine during maturity and ageing

[EN] The qualitative and quantitative changes produced, during maturation and ageing, in the pigment composition of a red wine made from Vitis vinifera cv Tempranillo grapes have been studied. In order to determine the detailed composition of the main pigment families involved in the colour changes, a fractionation of the samples has been carried out. One-hundred and twenty-nine different compounds have been identified and their evolutions with wine age have also been established. The data obtained in the analyses of the fractions by high performance liquid chromatography–diode array detection coupled to mass spectrometry (HPLC–DAD–MS) have been used in pigment identification. In order to confirm the identity of some of these compounds, their syntheses have also been carried out. As far as we know, compounds originated by acylation of the monoglucosides of the anthocyanins with lactic acid as well as 3,7-diglucosides of anthocyanins have, among others, been reported here for the first time. The moments of appearance and disappearance of all the detected compounds have also been established as well as the changes in the levels of the different pigment families and subfamilies originated as a consequence of maturation and ageing of the wine in barrels or in bottles. As wine became older, the percentages of anthocyanins decreased slightly, whereas that of the anthocyanin-derived pigments increased and, above all, compounds providing the wine with orange hues (pyranoanthocyanins). In the last sample, they represented 70% of the anthocyanin-derived pigments. On the contrary, the percentage of compounds providing bluish hues to the wine, direct and acetaldehyde-mediated flavanol–anthocyanin condensation products, decreased. This change in the nature of the anthocyanin-derived pigmentwould explain the colour change observed in the samples, from purple towards orange hues

Separation of pyranoanthocyanins from red wine by column chromatography

[EN] With the aim of monitoring the formation of anthocyanin-derived pigments and contributing to the study of their chromatic properties, stability and relative contribution to the colour of red wines, a method for fractionation of the colouring material was set up. The method was based on the distinct reactivity of the different pigment families towards bisulfite (hydrogen sulfite). The wine, acidified and bleached with NaHSO3, was placed in a Toyopearl® HW-40(s) gel column and submitted to elution with ethanol. Two fractions with different pigment compositions were collected and analysed by liquid chromatographay diode array detection-mass spectrometry. Compounds present in each fraction were identified according to their UV-visible and MSn mass spectra, showing that the first one was mostly constituted of pyranoanthocyanins, whereas the second basically contained anthocyanins and anthocyanin-flavanol condensation products. A large variety of new pigments were detected, some of which had not been previously reported in red wines, as far as we know. Characteristic MS2 and MS3 fragmentation patterns were observed within each family of compounds, which could be further applied for characterisation of unknown pigments in other wines

Determination of phenolic compounds of grape skins during ripening by NIR spectroscopy

[EN] The potential of near infrared spectroscopy (NIRS) to determine the content of phenolic compounds in red grapes has been evaluated. The near infrared spectra of intact grapes and grape skins throughout maturity were recorded using a fibre-optic probe and a transport quartz cup, respectively. Reference values of phenolic compounds were obtained by HPLC-DAD-MS. Modified Partial Least Squares (MPLS) regression was used to develop the quantitative models for flavanols, flavonols, phenolic acids, anthocyanins and total phenolic compounds. The procedure reported here seems to have an excellent potential for fast and reasonable cost analysis. The results of this work show that the models developed using NIRS technology together with chemometric tools allow the quantification of total phenolic compounds and the families of main phenolic compounds in grape skins throughout maturation. The validation of these models showed the best results for the determination of flavonols (differences between HPLC and NIRS of 7.8% using grapes and 10.7% using grape skins) in the external validation procedure. Good results in the external validation were also obtained for the determination of total phenolic compounds (differences of 11.7% using grapes and 14.7% using grape skins). The best results were generally obtained recording the spectra directly in intact grapes

Sensory evaluation of bitterness and astringency sub-qualities of wine phenolic compounds: synergistic effect and modulation by aromas

[EN] Mouthfeel characteristics of wine phenolic compounds (catechin, epicatechin, coumaric acid, caffeic acid, protocatechuic acid and gallic acid) were evaluated by sensory analysis using a Labeled Magnitude Scale for rating the intensity of the perceived sensations. Synergisms on bitterness and on astringent sub-qualities were detected when the phenolic compounds were tested as mixtures in comparison to individual compounds, maintaining the total amount of stimulus constant in all tasted samples. Principal component analysis was applied to observe trends in the data and to indicate relationships between phenolic substances and sensory attributes. It allowed us to easily visualize the synergistic effect discussed above. To our knowledge, this is the first time that astringency synergism between phenolic compounds has been described. This research also shows that wine aromas may modulate the perception of the astringency. In this study, the addition of volatile compounds increased the intensity of the astringent perception of the flavanol solutions and also its persistence

Evaluation of sensory parameters of grapes using near infrared spectroscopy

[EN] Grape seed and skin sensory parameters are important characteristics in making decisions concerning the optimal harvest time and producing high-quality red wines. The potential of near infrared spectroscopy to determine several sensory parameters of seeds and skins was evaluated. Taste (sourness), texture (astringency, tannic intensity, dryness and hardness), visual (colour) and olfactory (intensity and type of aroma) attributes were considered. Calibration models were performed by modified partial least squares regression. The differences between sensory analysis and NIRS analysis in external validation were lower in the case of seeds; they were between 4.5% for hardness and 8.7% for colour. For the skins, differences in external validation were between 9.8% for tannic intensity and 13.7% for astringency. The results obtained show that NIRS technology has considerable potential for predicting the above sensory attributes