Gastrointestinal digestion of olive leaf waste from olive mill production chain in a simulator of the gastrointestinal tract.

BACKGROUND: Currently, olive leaves are considered waste, although their high phenolic content makes them a source of antioxidants that could be used directly. The aim of this work was to study the behavior of phenolic compounds contained in olive leaf from the olive mill production chain during their gastrointestinal digestion. RESULTS: Phenolic compounds in the gastric digestion analyzed by high-performance liquid chromatography-diode array detection-mass spectrometry increased by 58% to 314.5% compared to the meal, while in the intestinal stage they ranged from 1.87 to 9.04 times higher. An increase of between 187% and 903% in bioaccessibility of phenolic compounds in the intestinal phase was observed, except for verbascoside. Furthermore, such compounds were fully bioavailable, except for apigenin-7-O-glucoside, which showed a bioavailability of 56%. CONCLUSION: The current study showed that the concentration of oleuropein, apigegin-7-O-glucoside, hydroxytyrosol hexoside and hydroxytyrosol contained in olive leaf continued to be extracted during the gastrointestinal digestion process. Furthermore, the results obtained with respect to their bioaccessibility and bioavailability suggest a good disposition to pass into the bloodstream where they could exert beneficial effects. Therefore, these results are promising for olive leaf becoming a consumable by-product that could be directly ingested through a simple infusion. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The application of ozonated water rearranges the Vitis vinifera L. leaf and berry transcriptomes eliciting defence and antioxidant responses.

Ozonated water has become an innovative, environmentally friendly tool for controlling the development of fungal diseases in the vineyard or during grape postharvest conservation. However, little information is currently available on the effects of ozonated water sprayings on the grapevine physiology and metabolism. Using the microvine model, we studied the transcriptomic response of leaf and fruit organs to this treatment. The response to ozone was observed to be organ and developmental stage-dependent, with a decrease of the number of DEGs (differentially expressed genes) in the fruit from the onset of ripening to later stages. The most highly up-regulated gene families were heat-shock proteins and chaperones. Other up-regulated genes were involved in oxidative stress homeostasis such as those of the ascorbate-glutathione cycle and glutathione S-transferases. In contrast, genes related to cell wall development and secondary metabolites (carotenoids, terpenoids, phenylpropanoids / flavonoids) were generally down-regulated after ozone treatment, mainly in the early stage of fruit ripening. This down-regulation may indicate a possible carbon competition favouring the re-establishment and maintenance of the redox homeostasis rather than the synthesis of secondary metabolites at the beginning of ripening, the most ozone responsive developmental stage.

Effect of Crocus sativus L. Stigmas Microwave Dehydration on Picrocrocin, Safranal and Crocetin Esters.

The dehydration process is the basis to obtain high quality saffron and to preserve it for a long time. This process modifies saffron's main metabolites that define its quality, and are responsible for the characteristic color, taste, and aroma of the spice. In this work, the effect of microwave dehydration on saffron main metabolites (picrocrocin, safranal and crocetin esters) from Crocus sativus L. stigmas at three determinate powers and different time lapses was evaluated. The results showed that this dehydration process obtained similar or lower crocetin esters content, and after three months of storage, higher concentration was shown in treatments at 440 W for 36 s, 55 s, and 73 s; at 616 W for 90 s; and at 800 W for 20 s. Picrocrocin content was lower and safranal content was higher in all treatments compared to the control both before and after storage. Regarding to commercial quality, microwave dehydration obtained Category I of saffron according to International Standard Organization (ISO) 3632. After three months of storage, treatments at 616 W for 83 s and 800 W for 60 s obtained lower categories. The results obtained suggest that microwave dehydration is a suitable process for obtaining high quality saffron, 800 W with 6 lapses of 20 s being the best conditions studied.

Analysis of Intact Glycosidic Aroma Precursors in Grapes by High-Performance Liquid Chromatography with a Diode Array Detector.

Nowadays, the techniques for the analysis of glycosidic precursors in grapes involve changes in the glycoside structure or it is necessary the use of very expensive analytical techniques. In this study, we describe for the first time an approach to analyse intact glycosidic aroma precursors in grapes by high-performance liquid chromatography with a diode array detector (HPLC-DAD), a simple and cheap analytical technique that could be used in wineries. Briefly, the skin of Muscat of Alexandria grapes was extracted using a microwave and purified using solid-phase extraction combining Oasis MCX and LiChrolut EN cartridges. In total, 20 compounds were selected by HPLC-DAD at 195 nm and taking as a reference the spectrum of phenyl ß-D-glucopyranoside, whose DAD spectrum showed a first shoulder from 190 to 230 nm and a second around 200-360 nm. After that, these glycosidic compounds were identified by High-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-qTOF-MS). Disaccharides hexose pentose were the most abundant group observed with respect to the sugars and monoterpendiols the main aglycones found.

Spraying Ozonated Water on Bobal Grapevines: Effect on Wine Quality.

Ozonated water is being introduced as an alternative phytosanitary treatment to control grapevine diseases in a context in which the reduction of chemical pesticides has become an urgent necessity. In this study, we evaluated the effect of spraying grapevines with ozonated water on the enological, phenolic, and aromatic qualities of Bobal wines during two consecutive growing seasons. In the first season, ozonated water was applied once during the ripening period on grapevines trained on the traditional gobelet system (S1). In the second season, three applications were performed between fruit set and harvest on grapevines grown on a vertical trellis system (S2). The S1 treatment led to a wine with an increased alcoholic degree and a remarkably higher phenolic content, which resulted in preferable chromatic characteristics. The S2 treatment maintained the total phenolic content but significantly enhanced stilbenes and flavanols and also reduced anthocyanins, which negatively affected the wine colour. Regarding aroma, both treatments reduced the content of glycosylated precursors and had different effects on free volatiles, both varietal and fermentative. Thus, the metabolic response of grapevines to the ozonated water stress, and therefore the quality of wines, depended on the ozone dose received by the plants.

Winemaking with vine-shoots. Modulating the composition of wines by using their own resources.

Vine-shoots from two important Vitis vinifera, Airén and Cencibel, have been prepared in two different formats (chip and granule) and added (12 g/L) in their own wines in different winemaking steps. Results have shown significant differences depending in all conditions tested, and wine chemical composition was modulated while in contact with vine-shoots. Compounds such as trans-resveratrol, increased its concentration up to 4 mg/L in Airén white wines. In Cencibel red wines, vanillin was found in a concentration four times above its odor threshold and independently of the vine-shoot variety, format and moment of addition, compounds such as (-)-epicatechin and (+)-catechin increased the concentration with respect to the control wine. When vine-shoots were added after fermentation, ß-ionol appeared for first time in all wines. In case of guaiacol, the higher increment was observed for Airén wines. Therefore, it is possible to elaborate distinctive wines using their own resources.

Toasted vine-shoot chips as enological additive.

Different ways of vine-shoots revalorization have been proposed, but not in wine yet, as for example in the same way as oak chips are being used. In this work, vine-shoot samples were submitted to a thermogravimetric analysis to establish the temperature range for its lignin structure decomposition, resulting between 160 and 180 °C. Then, vine-shoot chips from Airén and Cencibel cultivars, with a particle size around 2.5-3.5 cm, were submitted to six toasting conditions: 160 °C and 180 °C for 45, 60 and 75 min. Their volatile composition was very similar to oak chips, being vanillin the most important compound. Moreover, such vine-shoots have an interesting content of prodelphinidins that together with the stilbenes may contribute to wine antioxidant activity. The toasting conditions at 180 °C/45 min were the most suitable one for releasing the mentioned valuable compounds in order to propose vine-shoots as new enological additive similar to oak chips.

Vine-Shoot Tannins: Effect of Post-pruning Storage and Toasting Treatment.

For the first time vine-shoot tannin composition was carried out by means of HPLC-DAD-ESI-MS/MS. Two vine-shoot cultivars (Airén and Cencibel) with different post-pruning storage times were submitted to a toasting process and assayed. There were no traces of gallotannins nor ellagitannins, but a high proanthocyanidin content and a mean degree of polymerization (mDP) close to 3 were characterized. The higher concentration of proanthocyanidins corresponded to Airén after 6 months post-pruning storage and at 3 months for Cencibel. Procyanidins were the most abundant fraction (70-95%), which decreased with storage, and especially significant was the contribution of B1, B2, and B4 dimers. Prodelphinidins were also found (8-24%), increasing their % with storage time. Toasting produced a considerable reduction of proanthocyanidin content and a loss of a monomer mDP unit, suggesting that if used as oenological tannins, then they may be more bitter and less astringent when compared with the nontoasted vine-shoot samples.

Accumulation of guaiacol glycoconjugates in fruit, leaves and shoots of Vitis vinifera cv. Monastrell following foliar applications of guaiacol or oak extract to grapevines.

Previous studies have shown that volatile compounds present within a vineyard during the growing season can be absorbed by grapevines, assimilated within grapes, and then released during fermentation to influence the final aroma of wine. For example, the accumulation of volatile phenols in glycoconjugate forms following grapevine exposure to bushfire smoke, and their subsequent release during winemaking. This study investigated the accumulation of guaiacol glycoconjugates in the fruit, shoots and leaves of Monastrell grapevines following foliar applications (at veraison) of either an aqueous solution of guaiacol or an aqueous oak extract. Fruit, shoot and leaf samples were then collected at 3 time points between veraison and maturity, and analysed by gas chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry, to quantify guaiacol and its glycoconjugates, respectively. Guaiacol glycoconjugates were observed in fruit and leaves in particular, demonstrating glycosylation occurred after grapevine treatment; however, different glycoconjugate profiles were apparent.

Accumulation of Glycoconjugates of 3-Methyl-4-hydroxyoctanoic Acid in Fruits, Leaves, and Shoots of Vitis vinifera cv. Monastrell following Foliar Applications of Oak Extract or Oak Lactone.

Grapevines are capable of absorbing volatile compounds present in the vineyard during the growing season, and in some cases, volatiles have been found to accumulate in fruits or leaves in glycoconjugate forms, that is, with one or more sugar moieties attached. The presence of oak lactone in wine is usually attributable to oak maturation, but oak lactone has been detected in wines made with fruit from grapevines treated with oak extract or oak lactone. This study investigated the accumulation of glycoconjugates of 3-methyl-4-hydroxyoctanoic acid (i.e., the ring-opened form of oak lactone) in the fruits, leaves, and shoots of Monastrell grapevines following foliar application of either oak extract or oak lactone at approximately 7 days postveraison. Fruits, leaves, and shoots were collected at three different time points, including at maturity. The oak lactone content of fruit was determined by gas chromatography-mass spectrometry, with declining concentrations observed in fruit from grapevines treated with oak lactone with ripening. The concentrations of a ß-d-glucopyranoside of 3-methyl-4-hydroxyoctanoic acid in fruits, leaves, and shoots was determined by liquid chromatography-tandem mass spectrometry, with the highest oak lactone glucoside levels observed in leaves of grapevines treated with oak lactone. A glucose-glucose disaccharide was also tentatively identified. These results demonstrate both ring-opening and glycosylation of oak lactone occurred after experimental treatments were imposed.

Vine-shoot waste aqueous extracts for re-use in agriculture obtained by different extraction techniques: phenolic, volatile, and mineral compounds.

Vine-shoots are an important waste in all viticulture areas that should be re-used with innovative applications. The aim of this work was to produce Airén waste vine-shoot aqueous extracts by four solid-liquid extraction techniques such as conventional solid-liquid extraction (CSLE), solid-liquid dynamic extraction (SLDE-Naviglio), microwave extraction (ME), and pressurized solvent extraction (PSE). Their chemical composition was studied in terms of phenolic, volatile, and mineral compounds. The highest concentrated extracts corresponded to CSLE and SLDE-Naviglio, independent of the conditions tested. The CSLE extracts had the highest flavanols, phenolic acids, and stilbenes contents. The volatile composition, quantified for first time in this work, shows that furanic compounds were the most abundant. All extracts showed an interesting mineral content, which may be assimilated by plants. These results show the agricultural potential of Airén vine-shoot waste aqueous extracts to be used as grape biostimulants and/or foliar fertilizer.

Analysis of red grape glycosidic aroma precursors by glycosyl glucose quantification.

A method has been developed to analyse the glycosidic aroma precursor of non aromatic red grapes in terms of glucose GG. Due to the matrix interferences, an extract free of glycosylated polyphenols (especially anthocyanins and flavonols) and free glucose has been prepared. Such interferences have been eliminated by combining the use of Oasis MCX SPE and Fehling reagent. The glycosyl aroma precursor's fraction was subjected to an acidic hydrolysis (pH 1, 100°C, 1h), where equimolecular proportions of glucose (glucose GG) were released from their respective aglycones and quantified HPLC-IR. Compared with methods that require detailed analysis of the volatile aglycones, this one is able to estimate with good reproducibility the potential aroma of grapes by the only measurement of glucose GG.

Volatile compounds formation in alcoholic fermentation from grapes collected at 2 maturation stages: influence of nitrogen compounds and grape variety.

The aim of this work was to study the influence of nitrogen compounds on the formation of volatile compounds during the alcoholic fermentation carried out with 4 nonaromatic grape varieties collected at 2 different maturation stages. To do this, Monastrell, Merlot, Syrah, and Petit Verdot grapes were collected 1 wk before harvest and at harvest. Then, the musts were inoculated with the same Saccharomyces cerevisiae yeast strain and were fermented in the same winemaking conditions. Amino acids that showed the highest and the lowest concentration in the must were the same, regardless of the grape variety and maturation stage. Moreover, the consumption of amino acids during the fermentation increased with their concentration in the must. The formation of volatile compounds was not nitrogen composition dependent. However, the concentration of amino acids in the must from grapes collected 1 wk before harvest can be used as a parameter to estimate the concentration of esters in wines from grapes collected at harvest and therefore to have more information to know the grape oenological capacity. Application of principal components analysis (PCA) confirmed the possibility to estimate the concentration of esters in the wines with the concentration of nitrogen compounds in the must.

Effect of oak extract application to Verdejo grapevines on grape and wine aroma.

Volatile compounds from a commercial aqueous oak extract application to white Verdejo grapevines at veraison have been studied. Treated grapes under two types of formulation (25% and 100%) have been analyzed at the optimum maturation time, and winemaking was then subsequently carried out. The volatile compounds were analyzed by stir bar sorptive extraction-gas chromatography-mass spectrometry. The results suggest that after the grapevine treatments, grapes store the volatiles in the form of nonvolatile precursors, and some of the volatiles are released during the winemaking process, especially six months after the alcoholic fermentation. The sensory analysis shows that wines maintain the typical aroma properties of Verdejo wines at the end of fermentation; but after six months, the wine color is greener and more astringent, and, in terms of aroma, it has wooden notes as if the wine has been aged in oak barrels.

Study of the evolution of nitrogen compounds during grape ripening. application to differentiate grape varieties and cultivated systems.

The aim of this work was to study the evolution of amino acids and ammonium during grape ripening and to evaluate its application to differentiate grape varieties and cultivated systems (organic and nonorganic). For this purpose, Monastrell, Syrah, Merlot, and Petit Verdot grapes produced using conventional agriculture and Monastrell grape cultivated using organic agriculture, collected during two consecutive harvests at different stages of ripening, were studied. These years of harvest were very different climatic years; even so, the grape varieties presented similar qualitative compositions. Therefore, the percentage of amino acids at harvest moment allowed differentiation of grapes according to variety and cultivated system, regardless of the year. The nitrogen composition could allow estimation of the fermentative aroma potential of grapes. Thus, Syrah was the grape with the greatest aroma potential at harvest. Monastrell nonorganic grape had a concentration of nitrogen compounds superior to that of Monastrell organic grape. In Monastrell, Syrah, and Merlot, traditional varieties in the area, the highest concentration of nitrogen compounds coincided with the highest degrees Baume/total acidity ratio and color index during 2007. Consequently, technological and phenolic maturity of these grape varieties coincided with the maximum composition of nitrogen compounds. However, in 2008, this did not happen because grape ripening was irregular as a consequence of different climatological conditions.

Comparison of stir bar sorptive extraction and solid-phase microextraction to determine halophenols and haloanisoles by gas chromatography-ion trap tandem mass spectrometry.

Solid-phase microextraction by immersion (IS-SPME) and headspace mode (HS-SPME), together with stir bar sorptive extraction (SBSE), have been assayed in combination with gas chromatography-ion trap tandem mass spectrometry (MS/MS) for analysing 2,4,6-trichlorophenol, 2,3,4,6-tetrachlorophenol, 2,4,6-tribromophenol, 2,4,6-trichloroanisole, 2,3,4,6-tetrachloroanisole and 2,4,6-tribromoanisole in different liquid matrices. Once, the optimization of MS/MS fragmentation analysis was carried out, sample enrichment was performed using the three mentioned extraction methods, and comparison through the determination of linearity, and LOD and LOQs were carried out. SBSE and IS-SPME methods described enabled us to determine the target compounds at ng/l levels, concentrations lower than their olfactory threshold, which is not the case of HS-SPME. SBSE showed a higher concentration capability than both SPME techniques, especially when compared to the HS-SPME mode. Thus, SBSE should be the definitive technique to analyse halophenols and haloanisoles in aqueous matrices. SBSE has been also applied to nine aqueous matrices as different as tap water, wines or commercial lemon juice extract.

Generation of saffron volatiles by thermal carotenoid degradation.

Generation of volatiles by thermal treatments has been studied in saffron spice for two reasons: (a) to determine volatile profile changes during simulated aging processes and (b) to study the volatile generation pathway. During the aging process, while the amounts of C10 compounds such as safranal and HTCC increase, the amounts of C9 compounds such as isophorone and 2,6,6-trimethylcyclohexane-1,4-dione decrease. A new compound tentatively identified as 4,5,6,7-tetrahydro-7,7-dimethyl-5-oxo-3H-isobenzofuranone seems to play a very important role in the aging process. The importance of this compound, structurally similar to dihydroactindiolide, was also confirmed when the saffron volatile fraction was analyzed via the degradation of the linear chain of crocetin and crocetin esters and is reported for the first time in this paper. Thermal degradation studies of zeaxanthin, crocetin, and trans and cis crocetin esters isomers allowed us to propose different mechanisms which explain saffron volatile generation depending on the crocetin ester isomer structure.

Influence of oak wood on the aromatic composition and quality of wines with different tannin contents.

The aromatic compounds associated with aging in wood have been studied in Monastrell wines containing different quantities of tannin. Analyses were carried out before and after a six month period in wooden barrels (225 L) of American and French (Allier) medium-toasted oak. Three classes of wine were made: one from a free-run must; one to which enological tannin was added; and one to which was added the wine obtained by pressing the grape pomaces. The aromas were determined by GC-MS, and the quality was evaluated by a group of expert tasters. The aromatic compounds, some of which came from the wines themselves and others which came from the wood, all increased in concentration by the end of the aging process. The ratio between cis- and trans-whiskylactone contents in wines reflected the different types of oak wood used. The note "vanilla" used by the tasters is probably not totally due to the presence of vanillin. The wines of best quality were those with a natural tannin content which had been aged in American oak, whereas the wines to which enological tannin had been added but aged in the same wood were considered the worst. An equalizing effect on the sensorial wines qualities was seen to result from the use of French oak. The furfural, eugenol, and cis-, and in some cases trans-, whiskylactone contents identified the wines from the different classes of wood. The wood from which the barrels were made had a greater effect on wines differentiation than the tannin content.