Application of the INFOGEST Standardized Method to Assess the Digestive Stability and Bioaccessibility of Phenolic Compounds from Galician Extra-Virgin Olive Oil.

The INFOGEST standardized method was applied to assess the potential bioaccessibility and bioaccessibility of the phenolic compounds from a Galician extra-virgin olive oil (EVOO). The in vitro digestion model involves three steps and generates two fractions after each one: an aqueous fraction (namely, water phase (Wp)) and an oily fraction (namely, oily phase (Op)). The results showed that secoiridoids were the most abundant family in the Galician EVOO polar fraction, representing 98% of the total phenolic compounds. After oral digestion, phenolic acids and simple phenols were mainly detected in Wp, while lignans and flavonoids were mostly found in Op. After gastric digestion, extensive hydrolysis of secoiridoids was observed to generate free tyrosol, hydroxytyrosol, and hydroxytyrosol acetate. The instability of secoiridoids after intestinal digestion was again responsible for the release of simple phenols, which were mainly recovered in Wp together with flavonoids. In contrast, lignans were stable to duodenal conditions and remained in Op.

Effect of olive ripening degree on the antidiabetic potential of biophenols-rich extracts of Brava Gallega virgin olive oils.

The diet management is imperative to anticipate risk factors that favour the development of diseases; indeed, the intake of virgin olive oil could be an alternative natural source of α-glucosidase enzyme inhibitors, which delay the digestion rate of carbohydrates. Consequently, the impact of diabetes mellitus (DM) could be diminished. Extra Virgin Olive Oils (EVOO) were elaborated from Galician autochthonous variety 'Brava Gallega' with olives selected at three different degree of ripeness (ripening index, RI: 1.4, 3.0, 5.5) in order to assess the effect of maturation on overall chemical composition, sensory quality, and enzyme inhibition. The phenolic profile of the EVOOs determined by LC-ESI-IT-MS exhibited quantitative differences as ripening advanced; for example oleocanthal, tyrosol, luteolin and apigenin concentrations were higher in the overripe olive oil (RI 5.5). Anyway, the phenolic extracts (from every tested RI) were more active than acarbose. In particular, those obtained from the most mature olives displayed the most powerful inhibitory activity (IC50 value of 143 µg of dry extract/mL). In addition, the significant effect of these compounds (i.e. luteolin, apigenin, tyrosol and oleocanthal) on the inhibitory activity of the olive oil extracts was demonstrated. Our results suggest that, regardless of RI, the inhibitory activity of 'Brava Gallega' olive oils could represent a valuable strategy for reinforcing the health claim of olive oil for phenolic compounds.

Tetraconazole alters the methionine and ergosterol biosynthesis pathways in Saccharomyces yeasts promoting changes on volatile derived compounds.

The influence of antifungal tetraconazole residues (either as an active substance or as a commercial formulation product) on the fermentative activity of Saccharomyces cerevisiae yeast was evaluated in pasteurized Garnacha red must by using laboratory-scale fermentation assays. The presence of this fungicide promoted a slight decrease in glucose consumption. Volatile fermentative-derived compounds were evaluated in deep. Statistically significant changes were found in methionol (with a mean decrease of around 24%), fatty acids (with increments ranged from 23% to 66%), and ethyl esters (with increases ranged from 23% to 145%) contents when grape musts were enriched with the commercial formulation at both contamination levels assayed. Based on protein mass fingerprinting analysis, it was possible to relate these variations on volatiles content with changes in the activity of several enzymes (Met3p, Met14p, Adh2p, Hmg1p, Erg5p, Erg6p, Erg11p, and Erg20p) involved in the secondary metabolism of yeasts.

Mepanipyrim residues on pasteurized red must influence the volatile derived compounds from Saccharomyces cerevisiae metabolism.

The impact of mepanipyrim (Mep) and its corresponding commercial formulation (Mep Form) on Saccharomyces cerevisiae metabolites was assessed, separately, by using laboratory-scale wine fermentation assays on pasteurized red must. The presence of Mep did not alter the fermentation course. With regard to volatiles formed at the intracellular level by fermenting yeast cells, Mep residues affected mainly the acetate and ethyl ester biochemical pathways. In particular, the target acetates showed a notorious increment, >90%, in presence of commercial Mep Form at the higher dose assayed. The addition of Mep and Mep Form, at both tested levels, highly increased ethyl caprylate (between 42 and 63%) and ethyl caprate (between 36 and 60%) contents as the same as their respective fatty acid precursors. No important effects were observed on colour and non-volatile pyranoanthocyanins, probably due to the low anthocyanin content characteristic of pasteurized musts.

The involvement of phenolic-rich extracts from Galician autochthonous extra-virgin olive oils against the α-glucosidase and α-amylase inhibition.

'Brava' and 'Mansa de Figueiredo' extra-virgin olive oils (EVOOs) are two varieties identified from north-western Spain. A systematic phenolic characterization of the studied oils was undertaken by LC-ESI-IT-MS. In addition, the role of dietary polyphenols from these EVOOs has been evaluated against the inhibition of key enzymes (α-glucosidase and α-amylase) in the management of diabetes mellitus (DM). Oleuropein and ligstroside derivatives comprised 83% and 67% of the total phenolic compounds in 'Brava' and 'Mansa de Figueiredo' EVOOs, respectively. The main secoiridoids from oleuropein were DOA (3,4-DHPEA-EDA, 59 and 22 mg kg-1, respectively) and the main isomer of OlAgl (3,4-DHPEA-EA, 74 and 23 mg kg-1). The main secoiridoids from ligstroside were D-LigAgl (p-HPEA-EDA or oleocanthal, 23 and 167 mg kg-1) and the main isomer of LigAgl (p-HPEA-EA, 214 and 114 mg kg-1). For α-glucosidase, both EVOO extracts displayed stronger inhibitory activity (IC50 values of 60 ±â€¯8 and 118 ±â€¯9 µg mL-1, respectively) than the commercial inhibitor acarbose (IC50 = 356 ±â€¯21 µg mL-1). Nevertheless, for α-amylase, only 'Brava' extracts showed anti-α-amylase capacity. A daily VOO intake lower than the requirements of EFSA seem to be enough to reach both 50% for α-glucosidase and 25% for α-amylase inhibition. These findings support the potential health benefits derived from Galician EVOOs that might be probably linked to the outstanding high concentration levels of phenolic acids and flavonoids.

Evaluation of the neuroprotective and antidiabetic potential of phenol-rich extracts from virgin olive oils by in vitro assays.

In this work, phenol-rich extracts from 'Cornicabra' and 'Picual' virgin-olive oils (EVOOs) were examined, for the first time, to establish their capacity to inhibit key enzymes involved in Alzheimer's disease (AD) (acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and 5-lipoxygenase (LOX)), major depressive disorder (MDD) and Parkinson's disease (PD) (monoamine oxidases: hMAO-A and hMAO-B respectively), and diabetes mellitus (DM) (α-glucosidase and α-amylase). 'Cornicabra' displayed the best inhibitory activity against all enzymes, when compared to 'Picual': BuChE (IC50 = 156 ±â€¯4 and 308 ±â€¯33 mg mL-1), LOX (IC50 = 26 ±â€¯0.5 and 37 ±â€¯3 mg mL-1), hMAO-A (IC50 = 20 ±â€¯2 and 37 ±â€¯0.2 mg mL-1), hMAO-B (IC50 = 131 ±â€¯7 and 215 ±â€¯13 mg mL-1) and α-glucosidase (IC50 = 154 ±â€¯17 and 251 ±â€¯31 mg mL-1), respectively. The behaviour observed can be associated with the higher content of secoiridoids, lignans and phenolic acids in 'Cornicabra' EVOO.

The phenolic chemistry and spectrochemistry of red sweet wine-making and oak-aging.

A natural sweet wine (NSW) was made with dried grapes from Vitis vinifera L. cv Garnacha Tintorera. A fortified sweet wine (FSW) was also obtained: the maceration-alcoholic fermentation of Garnacha Tintorera must was stopped by addition of ethanol 96% (v/v). UV/Vis spectrophotometry and HPLC/DAD-ESI/MS were applied to determine, respectively, the evolution of colour and phenolic compounds in Garnacha Tintorera based-sweet wines during aging. In sweet wines, aging decreased a(∗) (red/green), colour saturation and lightness and increased b(∗) (yellow/blue), and hue angle. Most of the phenolic compounds determined, such as anthocyanins, esters of hydroxycinnamic acids, flavan-3-ols monomers, oligomers and polymers decreased in both sweet wines during aging. On the contrary, hydroxybenzoic and hydroxycinnamic acids and vitisins increased after one year of aging. Despite that both terminal and extension subunit compositions show very small changes, mean degree of polymerisation of proanthocyanidins decline slightly as aging progressed in both sweet wines.

Garnacha Tintorera-based sweet wines: detailed phenolic composition by HPLC/DAD-ESI/MS analysis.

Sweet wines are traditionally elaborated in Galicia (the N.W. corner of Spain). The denomination of origin (DO) Valdeorras, one of the five DOs in Galicia, wants to promote the production and marketing of new sweets wines. The first one is made with dried red grapes Vitis vinifera L. Garnacha Tintorera (GNSW); this cultivar is a teinturier cultivar which has excellent potential to produce wines from raisined grapes. The second one, a fortified sweet wine aged in oak barrels (GFSW). Additionally a dry young wine (GBW) was produced from the same variety. Their aroma profiles and chromatic characteristics (determined by simple spectrophotometric methods) have been previously established. Now, proanthocyanidins, flava-3-ol monomers, anthocyanins, phenolic acids, flavonols and resveratrol were determined by HPLC, for the same three wines. The highest concentration of total proanthocyanidins (PAs) was evaluated in the GBW (525mgL(-1)), which was about 2-fold the concentration in the GNSW (236mgL(-1)) and about more 10-fold the concentration in the GFSW (44mgL(-1)). No apparent difference in the aDP (mean degree of polymerisation) was observed for the GBW (1.9) and the GNSW (2.1), whereas a slightly lower value was obtained for the GFSW (1.5). Total anthocyanin concentration was described as follow as GBW: 390mgL(-1)≫GNSW: 57mgL(-1)>GFSW: 25mgL(-1), which indicates that sweet wines were polymerised in great extent. Only vitisin A and B were found the main concentration in GFSW when compared to GBW by the ageing process. In sweet wines, phenolic acids (hydroxybenzoic and hydroxycinnamic acids) and flavonols were lowest when compared to GBW and resveratrol not was found in sweet wines.

Evolution of colour and phenolic compounds during Garnacha Tintorera grape raisining.

The off-vine drying is one of the most important steps in the production of a high quality naturally sweet wine. However, only few studies have analyzed the changes in colour and phenolic compounds of the grapes throughout the process. In this work, UV/Vis spectrophotometry and HPLC/DAD-ESI/MS were applied to determine, respectively, the evolution of colour and phenolic compounds in the red grapes of Garnacha Tintorera during the raisining process. The total water loss in 83 days was about 62% and the sugar concentration rose from 225 to 464 g L(-1). Browning and low-medium molecular weight compounds increase during the dehydration process. In CIELab coordinates, raisining decreases a(*) (red/green) and lightness and increases b(∗) (yellow/blue), colour saturation and hue angle. In general, most of the phenolic compounds determined (anthocyanins, flavonols, esters of hydroxycinnamic acids, flavan-3-ol monomers and proanthocyanidins) increase but to a lesser extent than expected because of the water loss of the grapes during drying. The lower increase was observed for esters of hydroxicinnamic acids suggesting these compounds could undergo strong enzymatic degradation. Despite this, both the terminal and the extension subunit compositions show little changes, while the mean degree of polymerization of proanthocyanidins decreases as raisining progresses.

Garnacha Tintorera-based sweet wines: chromatic properties and global phenolic composition by means of UV-Vis spectrophotometry.

Valdeorras (the N.W. corner of Spain) wants to promote the production and marketing of new sweet wines. The present work represents the first study on sweet wines manufactured with red grapes Vitis vinifera L. Garnacha Tintorera, a teinturier cultivar. Two different red sweet wines were elaborated: the first one was made with dried grapes; Vitis vinifera L. Garnacha Tintorera has excellent potential to produce wines from raisined grapes; the second one, a fortified sweet wine aged in oak barrels. Different red Garnacha Tintorera-based wines (a dry base wine, GBW; a naturally sweet wine, GNSW; and a fortified sweet wine, GFSW) were characterized. Chromatic characteristics and phenolic compounds were established by spectrophotometric methods in order to assess the technology of Garnacha Tintorera-based sweet wines. High molecular weight brown polymers, produced during the grape drying process and isolated from sweet wines by the dialysis process, were responsible for the brown colour of sweet wines. As a consequence, yellowness of sweet wines was also higher which was confirmed by colorimetric indexes. With respect to phenolic content, GFSW presented the lowest content because the maceration-alcoholic fermentation was stopped through the addition of alcohol before the diffusion of red pigments from skins to must was complete. GNSW presented the highest phenolic content due to the concentration effect resulting from evaporation of water from the grapes. Anthocyanins of sweet wines were polymerised in great extent. The percentage of polymerised tannins was sufficient to guarantee the aging process of sweet wines.

Evolution of flavonoids in Mouratón berries taken from both bunch halves.

Galicia (N.W. Spain) is a Spanish region with several old-traditional winegrowing areas. There are autochthonous grapevine varieties, such as Vitis vinifera L. cv. Mouratón, considered a biodiversity resource in viticulture and an opportunity for Galician sustainable wine production. Therefore, it is necessary to assess the potential of traditional cultivars to produce quality red wines. In this work, anthocyanin and flavonol evolution was followed in red berries from V. vinifera L. cv. Mouratón. The novelty of this study is that grapes were separately collected from two different positions (tips and shoulders) within the cluster, over ripening to examine the effects of berry position within the fruit cluster on the flavonoid compounds. Derivatives of five anthocyanins (malvidin, peonidin, petunidin, delphinidin and cyanidin) and derivatives of six flavonols (quercetin, myricetin, kaempherol, laricitrin isorhamnetin and syringetin) were detected in both positions within the cluster. Dynamic of anthocyanins (from 819 mg/kg to 1206 mg/kg in tips; and from 786 mg/kg to 1077 mg/kg in shoulders) and dynamic of flavonols (from 25mg/kg to 41 mg/kg in tips; and from 18 mg/kg to 21 mg/kg in shoulders) confirmed their upward trends over ripening. Grapes located inside the shoulder bunch receive less sunlight radiation than those located inside the tip bunch and this fact could explain the different accumulation observed for both positions. These results can be useful for winemakers in order to obtain different final red wine quality.