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.

Characterisation of extra virgin olive oils from Galician autochthonous varieties and their co-crushings with Arbequina and Picual cv.

The current trend of the olive oil market is the production of high quality extra from traditional minor olive varieties with peculiar and differentiated characteristics (especially with respect to the aromatic and phenolic composition). In this way, the interest of Galician oil producers (NW Spain) in recovering old autochthonous Local olive fruits has increased substantially in recent years. In order to investigate the potential of the Local olives by either producing high quality monovarietal oils or mixing with the most widespread olives in Galicia (Arbequina and Picual cv.), quality indices, and fatty acid composition as well as volatile and phenolic profiles were determined and compared. All EVOOs studied in this work can be considered as "extra virgin olive oil" due to quality indices fell within the ranges established in legislation. Picual and Local olive oils as well as those resulting from their co-crushing reach values which are required by EU legislation to add the specific health claim on the oil label. Co-crushing Picual:Local (80:20) provided a significant enhancement of grass and apple nuances and a decrease of banana notes with respect to Picual oils. The co-crushing process improved sensory and health properties of Picual extra virgin olive oils. The effect of co-crushing on phenolics, ester volatiles and banana nuances cannot be easily modulated, contrary to quality indices and fatty acid composition, both changing linearly in strict correlation with the fruit mass ratio.

Quality of extra virgin olive oils produced in an emerging olive growing area in north-western Spain.

Systematic studies of physico-chemical and stability-related properties, and chemical composition, of extra virgin olive oils (EVOOs) obtained from drupes cropped in specific regions are of special agricultural interest. This is particularly so with new production areas, where careful selection of the most suitable olive varieties for EVOO production is required. This paper reports the first comprehensive chemical characterisation of EVOOs obtained from three different olive varieties (viz., Picual, Morisca and Manzanilla de Sevilla) grown in a new cultivation area in Galicia (NW Spain). The Morisca variety was that providing the highest industrial oil yield (21%). However, the three types of EVOO exhibited no statistically significant differences in standard quality-related indices other than acidity. Morisca EVOO was that with the lowest content in oleic acid (mean=68%) and highest content in linoleic acid (mean=13%). Also, Morisca EVOO exhibited the highest sterol levels (mean=1,616 mg/kg) and Picual EVOO the lowest (mean=1,160 mg/kg). Picual EVOO contained greater amounts of the phenolic compounds luteolin and pinoresinol than both Morisca and Manzanilla de Sevilla EVOOs. Finally, Manzanilla de Sevilla EVOO exhibited differential attributes, with banana and olive fruit aromatic series prevailing predominantly over bitter-like, pungent-like and leaf series.

Improvements in the malaxation process to enhance the aroma quality of extra virgin olive oils.

The influence of olive paste preparation conditions on the standard quality parameters, as well as volatile profiles of extra virgin olive oils (EVOOs) from Morisca and Manzanilla de Sevilla cultivars produced in an emerging olive growing area in north-western Spain and processed in an oil mill plant were investigated. For this purpose, two malaxation temperatures (20/30 °C), and two malaxation times (30/90 min) selected in accordance with the customs of the area producers were tested. The volatile profile of the oils underwent a substantial change in terms of odorant series when different malaxation parameters were applied.

Ultrasound-assisted emulsification-microextraction for the determination of phenolic compounds in olive oils.

A reliable, sensitive and effective method based on ultrasound-assisted emulsification-microextraction (USAEME) coupled to HPLC-DAD has been developed to identify and quantify several target phenolic compounds from extra virgin olive oils (EVOO). This approach is based on the emulsification of a microvolume of polar organic extractant in a non-polar liquid sample by ultrasound radiation and further separation of both liquid phases by centrifugation. The percentage of methanol/water (v/v) in the extractant, the volume of extractant, and the extraction time as three effective parameters on the extraction were optimised by a central composite design (Box-Behnken response surface) method. The optimised method presented recoveries in EVOO between 91% and 115% for the target analytes (except vanillin with 65%) and a satisfactory precision with relative standard deviations (RSD%) lower than 8.4% for repeatability and reproducibility. The method showed good linearity and limits of detection and quantification were in the range 0.001-0.14 and 0.004-0.47mg/kg, respectively. After method validation, it was successfully applied to the analysis of three EVOO samples. All target compounds were detected in all analysed samples. Tyrosol and hydroxytyrosol were the major phenolic compounds, followed by pinoresinol and luteolin.

Influence of new generation fungicides on Saccharomyces cerevisiae growth, grape must fermentation and aroma biosynthesis.

The influence of ten new generation fungicides (ametoctradin, benthiavalicarb-isopropyl, boscalid, cyazofamid, dimethomorph, fenhexamid, kresoxim-methyl, mepanipyrim, metrafenone, and pyraclostrobin) on the fermentative activity of Saccharomyces cerevisiae yeast was initially evaluated in pasteurised red must. The presence of ametoctradin, dimethomorph and mepanipyrim seemed to affect sugars-to-ethanol yield in the stationary phase. The same fermentation experiments were carried out for these three fungicides in ecological red must from Vitis vinifera cv. Tempranillo. When ecological must was unfiltered, the fermentative activity of yeasts was unaffected by the presence of these selected fungicides. However, when ecological must was filtered beforehand, a slight decrease of biomass and ethanol production (in terms of biomass-to-ethanol yield and sugars-to-ethanol yield, respectively), as well as a decrease in fruity aroma, were registered with respect to the control wine.

Aroma biogenesis and distribution between olive pulps and seeds with identification of aroma trends among cultivars.

The two constitutive parts of four cultivars (Arbequina, Picual, Local and Manzanilla de Sevilla) grown in Spain were separately analysed in order to establish the role of pulp and seed in the biogenesis of extra virgin olive oil (EVOO) aroma through the lipoxygenase (LOX) pathway. C6 and C5 volatile compounds responsible of EVOO aroma were produced by endogenous enzymes in both parts of olive fruits and the differences can be attributed to different enzymes distribution in pulp and seed. According to results, C6 and C5 volatile compounds have mainly their biogenesis in pulp (80-90%) vs. seed (20-10%), independently of the cultivar considered. A linear discriminant analysis was used to establish discriminant aroma compounds between pulp and seed related to the maturity index. A decrease in trans-2-hexen-1-al and an increase in 1-hexanol with ripeness were observed independently of the cultivar considered. Finally, Partial Least Squares (PLS) regression analysis between pulp and seed aroma compounds allowed to establish those volatile compounds that better describe each cultivar.

Evolution of the aromatic profile in Garnacha Tintorera grapes during raisining and comparison with that of the naturally sweet wine obtained.

The postharvest dehydration is one of the most important steps in obtaining a high quality naturally sweet wine and it can play an important role in modulating the production and the release of volatile compounds. However, only a few studies have analysed the changes in the free and bound volatile compounds of grapes throughout the process. In this work, GC-MS was applied to determine the aromatic composition of Garnacha Tintorera grapes subjected to off-vine dehydration or raisining at several points during the process. The total water loss in 83 days was about 62% and the sugar concentration rose from 225 to 464 g/L. Within the free volatile compounds, isoamyl alcohols, benzaldehyde and guaiacol registered the largest increase above the concentration effect due to water loss; while within the bound volatile compounds were isoamyl alcohols, ethyl vanillate and benzoic acid. The aromatic profile of the raisins obtained were mainly caramelised, floral, phenolic and burned.

Floral, spicy and herbaceous active odorants in Gran Negro grapes from shoulders and tips into the cluster, and comparison with Brancellao and Mouratón varieties.

Within the framework of a more and more competitive market, the opportunity to obtain different wines from the same variety cultivated in the same vineyard is becoming of increasing importance. In this study the presence of aroma compounds in Gran Negro (Vitis vinifera L.) grapes was investigated in order to obtain its aroma potential fingerprint taking into consideration the separation of apical (tips) and basal (shoulders) berries of the clusters. In the final stages of maturation, differences were searched in the probable alcohol content, total acidity of the must, as well as in the aromatic composition of skin and flesh from shoulder and cluster tip berries. A GC-MS method was used to determine the aromatic composition. The obtained results showed that there was variability for their aromatic composition. These results are promising for those wine cellars that are considering the separation of berries from tips and shoulders of the clusters for the elaboration of wines with different qualities. For the berries from the tips of the clusters, aromatic alcohols and volatile phenols were mainly found in the flesh (15 and 2 times higher than in the skin, respectively); whereas aldehydes and C6 alcohols were mainly in the skin (4 and 3 times higher than in the flesh, respectively). For this reason, it could be recommended to separate berry skin before enzymatic maceration of the berry flesh must. For the berries from the shoulders of the clusters, the group of volatile phenols showed 2 times more concentration in the skin than in the flesh; it could be recommended to maintain berry skin during enzymatic maceration of the must. Overall, the tips showed a 40% lower level of C6 alcohols (contributing to herbaceous nuances). These results from Gran Negro were compared with those of Brancellao and Mouratón cultivars.

Aroma profile of Garnacha Tintorera-based sweet wines by chromatographic and sensorial analyses.

The aroma profiles obtained of three Garnacha Tintorera-based wines were studied: a base wine, a naturally sweet wine, and a mixture of naturally sweet wine with other sweet wine obtained by fortification with spirits. The aroma fingerprint was traced by GC-MS analysis of volatile compounds and by sensorial analysis of odours and tastes. Within the volatiles compounds, sotolon (73 µg/L) and acetoin (122 µg/L) were the two main compounds found in naturally sweet wine. With regards to the odorant series, those most dominant for Garnacha Tintorera base wine were floral, fruity and spicy. Instead, the most marked odorant series affected by off-vine drying of the grapes were floral, caramelized and vegetal-wood. Finally, odorant series affected by the switch-off of alcoholic fermentation with ethanol 96% (v/v) fit for human consumption followed by oak barrel aging were caramelized and vegetal-wood. A partial least square test (PLS-2) was used to detect correlations between sets of sensory data (those obtained with mouth and nose) with the ultimate aim of improving our current understanding of the flavour of Garnacha Tintorera red wines, both base and sweet. Based on the sensory dataset analysis, the descriptors with the highest weight for separating base and sweet wines from Garnacha Tintorera were sweetness, dried fruit and caramel (for sweet wines) vs. bitterness, astringency and geranium (for base wines).

Aroma potential of Brancellao grapes from different cluster positions.

In this study the presence of aroma compounds in grapes of Brancellao (Vitis vinifera L.) was investigated in order to obtain its aroma potential fingerprint. It is well known that differences exist in aromatic compounds amongst grapevine varieties at ripening stages. Within the framework of an increasingly competitive market, the chance of obtaining different wines from vines of the same variety grown at the same vineyard is becoming of increasing importance. This can be done through the managing of the vineyard, but also some wineries have assayed the separation of the tip and shoulder berries of the clusters of a specific variety with this objective. In this work it is evaluated that, in the final stages of maturation, differences exist in the probable alcoholic degree, total acidity of the must, as well as in the aromatic composition of skin and flesh of berries coming from the tips and shoulders of the clusters. Gas chromatography coupled to mass spectrometry (GC-MS) was used to determine the aromatic composition, in the skin and flesh of each sample, either tip or shoulder berries from the clusters. The obtained results showed that there was not variability for the probable alcoholic degree and total acidity between the shoulders and tips, whereas there was variability for their aromatic composition. For the berries from the tips of the clusters most of volatiles were found in the flesh (except aldehydes) and spicy and floral nuances (with the only exception of ß-ionone) were in higher proportions. For the berries from the shoulders of the clusters, most of volatiles were found in the skin (monoterpenes, norisoprenoids, aldehydes, and C6 alcohols), where the flesh was slightly richer in aromatic alcohols, volatile phenols and pantolactone; ß-ionone and herbaceous nuances were in higher proportions. These results are promising for those wineries that are considering the chance of separating berries from tips and shoulders of the clusters for the elaboration of different quality wines.

A review on the fate of pesticides during the processes within the food-production Chain.

Only the intake of toxicologically-significant amounts can lead to adverse health effects even for a relatively toxic substance. In the case of residues in foods this is based on two major aspects--first, how to determine quantitatively the presence of a pollutant in individual foods and diets, including its fate during the processes within the food production chain; and second, how to determine the consumption patterns of the individual foods containing the relevant pollutants. The techniques used for the evaluation of the fate of pesticides during food processing have been critically reviewed in this paper to determine those areas where improvements are needed or desirable. Options for improvements are being suggested, including, for example, the development of a pan-European food composition database, activities to understand better effects of processing on individual food pesticides, and harmonization of food consumption survey methods with the option of a regular pan-European survey. The ultimate aim is to obtain appropriate estimations for the presence and quantity of a given chemical in a food and in the diet in general. Existing pragmatic approaches are a first crude step to model food pollutant intake. It is recommended to extend, refine, and validate this approach in the near future. This has to result in a cost-effective exposure-assessment system to be used for existing and potential categories of pollutants. This system of knowledge (with information on sensitivities, accuracy, etc.) will guide future data collection.

Relationships between Godello white wine sensory properties and its aromatic fingerprinting obtained by GC-MS.

Godello is a traditional white-grape vine grown widely in the Valdeorras area (Northwest Spain) available information about the sensory and chemical characteristics of which is scant. In this work, exploratory research was conducted with a view to correlate the results of instrumental analyses of the aroma compounds in Godello wine and their sensory perception. Principal Component Analysis (PCA) and Partial Least Squares Regression (PLS) were the multivariate data analysis methods selected. PCA showed the distribution of the Godello wines based on volatile compounds with Odour Activity Values (OAV) higher or near-unity and sensory characteristics, and PLS exposed relationships between sensory descriptors and volatile compounds, in the wines. The compounds that mostly contributed to the flavour of Godello wines were those conferring a fruity (ethyl esters and acetates, accounting 55.1% of total OAV), spicy (fatty acids, 35.3%) or floral aroma (terpenes, 3.1%) based on instrumental analysis. In sensory analysis the descriptors with the highest intensity percent were fruity and floral aromas (floral, apple and citrus), together with herbaceous notes.

Occurrence of soluble organic compounds in thermal waters by ion trap mass detection.

Thermal waters can be regarded as worth developing because of their human health implications and, if thermal tourism is promoted, their importance to the local economy. Galicia, in the North-West of Spain, has numerous sources of thermal waters spread all over the region. They have been used for therapeutic purposes since Roman times. Ourense, with 82 sources (9 spas), is becoming one of the richest Spanish provinces in hydromineral and thermal resources. In this work, the most representative springs of thermal waters from the city of Ourense--Outariz, Chavasqueira, Tinteiro and As Burgas--were analysed in order to identify the presence of several chemical classes of soluble organic compounds responsible for their biological activity. The waters studied are hyperthermal, with temperatures ranged between 45 and 66 degrees C. Different extraction techniques, liquid-liquid extraction (LLE), solid phase extraction (SPE) and solid phase micro-extraction (SPME), combined with gas chromatography-electron impact ion trap mass spectrometry (GC-(EI)ITMS) were used to cover a wide variety of compounds. The most characteristic chemical families of extractable organic compounds detected are thoroughly discussed. Most of the substances belonging to phenol, aldehyde, esther, ketone, and elemental sulphur groups occur in all samples. Large amounts of elemental sulphur were found in Tinteiro spring, providing to this water of numerous beneficial clinical effects.

Photochemical studies of a polybrominated diphenyl ethers (PBDES) technical mixture by solid phase microextraction (SPME).

In this work the photochemical behaviour of a technical mixture of polybrominated diphenyl ethers (PBDEs) (BDE-47, BDE-99, BDE-100, BDE-153 and BDE-154) has been studied. The mixture of BDEs was extracted from aqueous solutions using SPME fibers that were subsequently exposed to different UV irradiation times, procedure so-called Photo-SPME. PBDEs photochemical studies in such medium have been accomplished for the first time. Twenty one different photoproducts, all of them generated by successive bromine atoms losses, have been identified, being their photoformation-photodegradation curves easily determined by the Photo-SPME technique.