Evaluation of the Long-Lasting Flavour Perception after the Consumption of Wines Treated with Different Types of Oenological Additives Considering Individual 6-n-Propylthiouracil Taster Status.

Due to the limited scientific knowledge on the impact of commercial oenological additives on flavour perception, the aim of this work was to evaluate the effect of different types of oenological additives on the long-lasting flavour perception (flavour persistence) during wine tasting, also considering the effect of the individual PROP (6-n-propylthiouracil) taster status (PTS). To do so, white and red wines with two oenotannins (ellagitannin and gallotannin) and a commercial yeast mannoprotein were prepared. A control wine of each type was also made without additives. All the wines were spiked with a mixture of aromatic compounds responsible for the "fruity" and "woody" notes. Retronasal aroma and astringency were evaluated at the same time using time-intensity (TI) methodology and a trained panel (n = 40), including PROP non-tasters (NTs) and tasters (Ts). The results showed a significant effect of PTS on the long-lasting perception of astringency, being Ts who showed higher values than NTs for most TI parameters. However, PTS did not affect aroma persistence. In addition, the three oenological additives had an effect on astringency and retronasal aroma perception. They significantly increased the long-lasting perception of astringency compared to the control, while gallotannin also increased the persistence of the woody aroma.

Understanding If Differences in Salivary Flow Rate and Total Protein Content Triggered by Biological Factors (Sex and Age) Affect Aroma Perception and the Hedonic and Emotional Response of Wine Consumers.

The relationship between oral physiology (e.g., salivary protein content) and aroma perception over wine consumption was investigated in previous work. However, the relationship between oral physiology and the consumer's response to wine is unknown. Additionally, age-gender differences might affect oral physiology and, therefore, inter-individual differences in the consumer's enjoyment of wine. The aim of this work was to study the effect of biological individual factors such as sex and age on salivary flow rate and total protein content, on wine retronasal aroma perception and on the hedonic and self-reported emotional consumer response to wine. Additionally, the relationship between saliva composition, aroma perception and liking was also assessed. Results showed that age and sex influenced saliva composition and aroma perception in wine consumers in the case of red but not white wines. Younger females reported higher aroma intensity of the black pepper descriptor compared to older females. Correlation analysis confirmed the relationship between salivary flow rate and aroma perception and between the salivary protein content and wine acceptability. The interactions between wine polyphenols-saliva-aroma that occurred in the mouth during the oral processing of wine might partially explain these results, although further research will be required to confirm this hypothesis.

Insights on the effect of age and gender on in-mouth volatile release during wine tasting.

This study focus for the first time, in looking for age-gender effects on in vivo volatile release during wine consumption, also considering oral physiological differences (e.g. saliva composition). To do so, the in-mouth Head Space Sorptive Extraction technique was used, which allowed monitoring the oral release of twenty-four different types of volatile compounds from white and red wines. Thirty-two individuals (n = 32) males and females, belonging to two different age groups: young (18-35 y.o) and senior (>55 y.o.) participated in this analytical in vivo study. Results showed differences in volatile release among age-gender groups, which also depended on the volatile compound and wine type. Senior groups (SM, SF) showed a similar release behaviour among them. Contrarily, young males showed a higher release (between 10 and 29%) of alcohols and esters indistinctly of the wine type, while young females showed the lowest oral volatile release among the four age-gender groups. Gender differences in volatile release were more evident in young than in seniors. A higher release of furanic compounds (furfural and 5-methyl furfural) in seniors was likely related to differences in their saliva composition (total protein content, minerals (Mg, Zn) and α-amylase activity).

Temporal changes in salivary composition induced by oral exposure to different wine matrices and the relationship with the behaviour of aroma compounds in the mouth.

The dynamic changes in saliva flow and composition (pH, total protein capacity (TPC), total polyphenol index (TPI) and saliva antioxidant activity (SAOX)) after the exposure of the oral cavity to aromatized wine matrices with different chemical compositions (dealcoholized, alcoholized, and synthetic wines) have been investigated. For this, stimulated saliva from ten volunteers were collected five days per week (from Monday to Friday) during three non-consecutive weeks, before (basal saliva) and after the oral intervention with the wines (5 and 15 minutes later) (n = 450). In order to know the relationship between the changes induced in salivary composition and the amount of aroma retained in the oral cavity, the expectorated wines were also collected (n = 150). Results showed differences in saliva composition (pH, TPI and SAOX) depending on the wine matrix that were only significant in the first five minutes after the oral exposure to the wines. The wines with ethanol produced significantly lower in-mouth aroma retention, while salivary TPI and, to a minor extent, SAOX, were positively related to the aroma retained. These results prove that not only wine aroma composition, but also the physiological changes in saliva induced by the non-volatile chemical composition of the wine play an important role in wine odorant compounds, and likely, in aroma perception.

Bioconversion of Glycosidic Precursors from Sour Guava (Psidium friedrichsthalianum Nied.) Fruit by the Oral Microbiota into Odor-Active Volatile Compounds.

The ability of the human oral microbiota to hydrolyze the glycosidic aroma precursor extract isolated from sour guava (Psidium friedrichsthalianum Nied.) fruits was studied herein. The glycosidic extract (GP) was incubated with a mixture of the oral microbiota isolated from three individuals' saliva to evaluate the hydrolytic capacity of oral bacteria in the generation of odor-active compounds. The oral microbiota was able to release 1-hexanol from GP, under both aerobic and anaerobic conditions. Additionally, the aroma precursor extracts showed a decrease in the growth of harmful oral bacteria (Streptococcus and Actinomyces). This effect can be considered beneficial to human health because these bacteria have been related to different diseases of the bucco-respiratory tract.

Specificity of Saliva Esterases by Wine Carboxylic Esters and Inhibition by Wine Phenolic Compounds Under Simulated Oral Conditions.

The specificity of human esterase activity (EA) from the stimulated (SS) and non-stimulated (NSS) saliva toward different typical wine odorant carboxylic esters and its inhibition by the wine phenolic compounds has been evaluated. For the specificity, six p-nitrophenyl linked esters with different carbon chain lengths (from 2 to 12 carbons) were employed. The five single phenolic compounds (catechin, quercetin, kaempferol, myricetin, and resveratrol) at typical wine concentrations were assayed in the salivary EA inhibition study. Additionally, the inhibition exerted by the mixtures of wine polyphenols was evaluated using four commercial phenolic extracts [a grape seed extract (GSE), the monomers and oligomer fraction of the GSE, and a red wine extract (RWE)]. Finally, the saliva EA under the wine consumption conditions (pH = 5 and 11.3% ethanol) was evaluated. The results showed a higher EA in SS than NSS. It was also shown that the EA was higher toward the smaller than bigger esters regardless of the saliva types (SS or NSS). However, the inhibition exerted on saliva EA by the individual and mixtures of phenolic compounds was proven. Catechin was the phenolic compound that mostly inhibited saliva EA, while resveratrol showed the lowest EA inhibition. This inhibition was mainly related to the concentration of the phenolic compounds, but also with its structure. Finally, under simulated wine consumption, a decrease in EA was produced, which was mainly provoked by the decrease in the salivary pH. Nonetheless, since salivary pH recovers a few seconds after wine consumption, saliva EA might be relevant for the long-lasting perception of wine esters.

Oral Release Behavior of Wine Aroma Compounds by Using In-Mouth Headspace Sorptive Extraction (HSSE) Method.

The oral release behavior of wine aroma compounds was determined by using an in-mouth headspace sorptive extraction (HSSE) procedure. For this, 32 volunteers rinsed their mouths with a red wine. Aroma release was monitored at three time points (immediately, 60 s, and 120 s) after wine expectoration. Twenty-two aroma compounds belonging to different chemical classes were identified in the mouth. Despite the large inter-individual differences, some interesting trends in oral release behavior were observed depending on the chemical family. In general, esters and linear alcohols showed rapid losses in the mouth over the three sampling times and therefore showed a low oral aroma persistence. On the contrary, terpenes, lactones, and C13 norisoprenoids showed lower variations in oral aroma release over time, thus showing a higher oral aroma persistence. Additionally, and despite their low polarity, furanic acids and guaiacol showed the highest oral aroma persistence. This work represents the first large study regarding in-mouth aroma release behavior after wine tasting, using real wines, and it confirmed that oral release behavior does not only depend on the physicochemical properties of aroma compounds but also on other features, such as the molecular structure and probably, on the characteristics and composition of the oral environment.

Evaluation of the Effect of a Grape Seed Tannin Extract on Wine Ester Release and Perception Using In Vitro and In Vivo Instrumental and Sensory Approaches.

This study aimed to systematically evaluate the effect of a commercial grape seed tannin extract (GSE) fully characterized (53% monomers, 47% procyanidins) on wine ester release and perception using a global approach. The behavior of two esters (ethyl hexanoate, ethyl decanoate) was studied in a control wine or in the same wine supplemented with the GSE in preconsumption (in vitro headspace-stir bar sorptive extraction-gas chromatography mass spectrometry (HS-SBSE-GC/MS) and orthonasal perception) and consumption (intraoral-HS-SBSE-GC/MS and dynamic retronasal perception) conditions. For the compound ethyl hexanoate, no significant differences (p > 0.05) among wines were observed in the in vitro analyses while they were observed in the three in vivo experiments (p < 0.05). Thus, the wine supplemented with the GSE showed lower (35%) in vivo release and ortho (36%) and retronasal (16%) perception scores than the control wine. Overall, this suggests that components of the GSE could interact with this compound, directly and/or through complexes with oral components, affecting its release and conditioning its perception. However, perceptual interactions and effects of polyphenols on oral esterases cannot be discarded. On the contrary, the compound ethyl decanoate was not significantly affected by the addition of GSE. In conclusion, the addition of tannin extracts to wines can modulate aroma perception in a compound-dependent manner.

Oral persistence of esters is affected by wine matrix composition.

The present work evaluated for the first time the influence of wine matrix composition on oral ester persistence. To do that, the in mouth behaviour (oral retention and persistence) of six esters was followed in nine individuals after they rinsed their mouths with four rosé wines presenting two levels (low and moderate) of ethanol (0.5% or 10% v/v) and polyphenols (402 ± 10 or 661 ± 33 mg gallic acid/L). Overall, polyphenols and specially, ethanol, affected the oral retention of esters and their subsequent oral persistence, in an individual, compound and concentration dependent manner. The wine with moderate ethanol and low polyphenol content and the wine with low ethanol and moderate polyphenol content presented an increased oral ester persistence respect to the control wine (low ethanol/polyphenol content). However, the wine with moderate ethanol/polyphenol content showed most likely a lower oral ester persistence compared to the rest of the wines. Thus, an interaction between ethanol and polyphenols at specific concentrations could reduce oral ester persistence, and likely the fruity character of wines. The information generated in this research can be used by winemakers to understand how different winemaking techniques, which might alter wine ethanol and polyphenol content, may also affect wine aroma quality.

Interactions Among Odorants, Phenolic Compounds, and Oral Components and Their Effects on Wine Aroma Volatility.

To determine the impact of oral physiology on the volatility of typical wine aroma compounds, mixtures of a synthetic wine with oral components (centrifuged human saliva (HS), artificial saliva with mucin (AS), and buccal epithelial cells (BC)) were prepared. Each wine type was independently spiked with four relevant wine odorants (guaiacol, ß-phenyl ethanol, ethyl hexanoate, and ß-ionone). Additionally, the impact of four types of phenolic compounds (gallic acid, catechin, grape seed extract, and a red wine extract) on aroma volatility in the HS, AS, and BC wines was also assessed. Static headspace was measured at equilibrium by solid phase microextraction-GC/MS analysis. Results showed a significant impact of oral components on the volatility of the four tested odorants. Independently of the type of aroma compound, aroma volatility was in general, higher in wines with BC. Moreover, while guaiacol and ethyl hexanoate volatility was significantly lower in wines with HS compared to wines with AS, ß-ionone showed the opposite behavior, which might be related to metabolism and retention of mucin, respectively. Phenolic compounds also showed a different effect on aroma volatility depending on the type of compound and wine. Gallic acid had little effect on polar compounds but it enhanced the volatility of the most hydrophobic ones (ethyl hexanoate and ß-ionone). In general, flavonoid type polyphenols significantly reduced the volatility of both polar (guaiacol and ß-phenyl ethanol) and hydrophobic compounds (ß-ionone in HS and BC wines), but through different mechanisms (e.g., π-π interactions and hydrophobic binding for polar and apolar odorants respectively). On the contrary, flavonoids enhanced the volatility of ethyl hexanoate, which might be due to the inhibition exerted on some salivary enzymes (e.g., carboxyl esterase) involved in the metabolism of this odorant molecule.

Effect of saliva composition and flow on inter-individual differences in the temporal perception of retronasal aroma during wine tasting.

In order to determine if inter-individual differences in saliva composition and flow influence the perception of specific aromatic stimuli elicited by esters after wine consumption, ten individuals were selected and instructed in the recognition of four aromatic stimuli elicited by four ester compounds, which were added to a rosé wine. The whole panel was firstly characterised by their salivary flow, composition (pH, total protein content, macro- and micro- minerals) and rheological properties (viscosity), and secondly, the panellists were trained in a dynamic sensory method for the evaluation of retronasal aroma intensity at discrete time intervals (5, 60, 120, and 180 s) after wine expectoration. Significant inter-individual differences (p < 0.05) in saliva composition were found in most salivary parameters. Differences in the intensity ratings among individuals were also found for the four aroma attributes. Spearman correlation analysis between saliva parameters and aroma intensity over time showed a strong positive correlation between salivary flow and aroma perception for some aroma and time points. This correlation was higher in the immediate than in the long lasting perception and greater for aroma attributes elicited by short chain length esters (isoamyl acetate, ethyl butanoate and ethyl hexanoate) (than for attributes elicited by larger esters (ethyl decanoate).

Effects of Ethanol Concentration on Oral Aroma Release After Wine Consumption.

This paper evaluates, for the first time, the effects of ethanol concentration on the dynamics of oral (immediate and prolonged) aroma release after wine consumption. To do this, the intraoral aroma release of 10 panelists was monitored at two sampling points (0 and 4 min) after they rinsed their mouths with three rosé wines with different ethanol content (0.5% v/v, 5% v/v and 10% v/v) that were aromatized with six fruity esters (ethyl butanoate, isoamyl acetate, ethyl pentanoate, ethyl hexanoate, ethyl octanoate and ethyl decanoate). Overall, the results indicated that the extent of the effects of ethanol content on the oral aroma release were influenced by the subject, the ethanolconcentration and the type of aroma compound. This effect was also different in the immediate than in the prolonged aroma release. In the first in-mouth aroma monitoring, an increase in the ethanol content provoked a higher release of the more polar and volatile esters (ethyl butanoate, ethyl pentanoate), but a lower release for the more apolar and less volatile esters (ethyl octanoate, ethyl decanoate). Regarding the prolonged oral aroma release, an increase of ethanol content in wine increased the oral aroma release of the six esters, which might also increase the fruity aroma persistence in the wines. Future works with a higher number of individuals will be needed to understand the mechanisms behind this phenomenon.

Development of an in-mouth headspace sorptive extraction method (HSSE) for oral aroma monitoring and application to wines of different chemical composition.

An in-mouth headspace sorptive extraction (HSSE) procedure for the in-mouth volatile sampling of wine aroma compounds during wine tasting has been developed. The procedure is based on the application of a PDMS (polydimethylsiloxane) twister contained inside a tailored made glass tube placed into the headspace of the mouth after rinsing and spitting-off the wine, which is then followed by gas chromatography mass spectrometry analysis (GCMS). Various parameters that might affect the performance of the method (extraction time, aroma concentration) were firstly investigated. Despite the short selected in-mouth extraction time (30 s); the application of the in-mouth HSSE procedure using real wines allowed the detection of >30 volatile compounds from different chemical families in the oral cavity in a single run, and which are also present in the wine at very low concentrations. Additionally, the in-mouth HSSE profile allowed us to distinguish between wines types (with different non-volatile and volatile composition) in a similar way to that when using data from the headspace of the wine (wine-HSSE-GCMS). The simplicity, sensitivity, good repeatability and the easy automatization of this procedure, makes this technique a reliable and feasible tool to determine the chemical and biochemical changes of these compounds in the mouth in real physiological conditions providing useful -in vivo analytical data to better correlate with sensory studies.

Assessment Wine Aroma Persistence by Using an in Vivo PTR-ToF-MS Approach and Its Relationship with Salivary Parameters.

To better understand wine aroma persistence, the nasal cavity of nine volunteers was monitored by Proton Transfer Reaction-Time of Flight-Mass Spectrometry (PTR-ToF-MS) after they rinsed their mouths with three rosé wines (one control and the same wine supplemented with two tannin extracts) during four minutes. Wines were aromatised with a mixture of five target aroma compounds. Results showed that wine aroma persistence was highly compound-dependent: while esters disappeared very fast, other compounds such as linalool remained in the oral cavity for longer times after wine expectoration. A low effect of tannins (at 50 mg/L) on nasal cavity parameters was observed, with the exception for the compound ethyl decanoate that was significantly higher released in the presence of tannins. Strong interindividual differences on aroma persistence were also found. Significant positive correlations with the salivary total protein content and negative with the salivary flow were observed for specific compounds. This work has studied for the first time in vivo wine aroma persistence in real time from an analytical perspective.

Individual differences and effect of phenolic compounds in the immediate and prolonged in-mouth aroma release and retronasal aroma intensity during wine tasting.

To explore the role of phenolic compounds in oral aroma release during wine tasting, four rosé wines supplemented with three types of commercial phenolic extracts and a control wine were evaluated. Wines were aromatized with a mixture of six target aroma compounds. -In vivo oral aroma release was monitored in six volunteers at two different times after wine rinsing, just after spitting of the wine (immediate release), and four minutes later (prolonged release). To check the sensory meaning of these changes, descriptive analysis using a trained panel (n = 10) was also performed. Results showed a strong individual effect on total oral aroma release at the two sampling points. After the oral exposure to wines with phenolic extracts, a lower release of most target aroma compounds was also determined. Lower intensity scores for some aroma attributes in wines with phenolic extracts were found, showing a good agreement between the two scientific approaches.

Effect of saliva esterase activity on ester solutions and possible consequences for the in-mouth ester release during wine intake.

The aim of this study was to investigate the role of saliva esterase activity on carboxylic esters typically associated with pleasant and fruity aromas in wine. For this, ex vivo experiments using the same fresh and inactivated (without enzymatic activity) human saliva with a mixture of carboxylic esters with different aliphatic chain length (ethyl butanoate, ethyl pentanoate, ethyl hexanoate, ethyl octanoate, ethyl decanoate, and isoamyl acetate) were prepared. Liquid-liquid extraction with dichloromethane and GC-MS analysis were applied to the saliva systems in order to determine the reduction in ester content and the formation of their corresponding metabolic products (carboxylic acids) in the saliva systems before and after incubation at 37C. In addition, to check if there was a relationship between the susceptibility of esters to saliva hydrolysis and the amount of in-mouth ester release during wine intake, the remaining oral amount of each ester was determined by comparing the intraoral amount immediately after spitting out the wine and 4 min later. Ex vivo experiments showed ester degradation by saliva esterase enzymes mainly acted on long chain esters (ethyl octanoate and ethyl decanoate), which gave rise to the formation of their corresponding carboxylic acids. Nonetheless, in spite of their higher susceptibility to saliva enzymes, in vivo experiments showed that long chain carboxylic esters remained in the oral cavity long after swallowing. This confirmed that ester hydrophobicity is closely related to the in-mouth temporal release of these odorants and, therefore, behind wine aroma persistence. In wines, esters represent a group of aromatic compounds of great interest since they are linked to pleasant fruity aroma nuances. Today wine consumers are demanding fresh and long persistent fruity aromatic wines. The present research contributes to better understanding the relationship between ester content in the wine and oral aroma release experienced during wine tasting, considering the changes in these compounds during oral processing. This is a necessary step when trying to unravel the factors involved in wine aroma perception and in consumer preferences, and it represents a necessary knowledge in promoting winemaking practices (e.g., the use of selected microorganisms) for improving the type and amount of these aroma compounds in the wine.

Changes in the volatile profile of Pinot noir wines caused by Patagonian Lactobacillus plantarum and Oenococcus oeni strains.

The ability of Patagonian L. plantarum and O. oeni strains to change the volatile profile of a sterile Pinot noir wine was studied through fermentation assays, at laboratory scale. Two strains of each LAB species were selected based on data regarding to their ability to survive in wine and to consume l-malic acid. Both O. oeni strains but only one L. plantarum (UNQLp 11) strain were able to remain viable, consuming l-malic acid through the fermentation assay with a concomitant increase of l-lactic acid. The volatile profile of Pinot noir wine, before and after LAB inoculation, was measured by using HS-SPME gas chromatography technique. This analysis showed that alcohols were the main volatile compounds after alcoholic fermentation and that after fermentation with the selected LAB strains, a decrease in the volatile alcohols concentration and an increase in the volatile esters concentration could be observed. The O. oeni UNQOe 73.2 strain produced the most notable change in the volatile profile, with the production of some important odorant esters at higher concentration, compared to O. oeni UNQOe 31b strain. Although, L. plantarum UNQLp 11 strain showed a better performance in the consumption of l-malic acid, this strain had a low capacity to modify the volatile compounds profile after incubation in red wine. The results found in the present work showed that different strains selected as potential malolactic starters could have different behavior when are incubated in real wine. Although L. plantarum UNQLp 11 strain showed a good consumption of l-malic acid, the O. oeni UNQOe 73.2 strain exhibited superior capacity to improve the flavor of wine due to its esterase activity that produce an increase of fruity and creamy odorants.

Aroma release in the oral cavity after wine intake is influenced by wine matrix composition.

The aim of this study has been to investigate if wine matrix composition might influence the interaction between odorants and oral mucosa in the oral cavity during a "wine intake-like" situation. Aroma released after exposing the oral cavity of three individuals to different wines (n=12) previously spiked with six target aromas was followed by an -in vivo intra-oral SPME approach. Results showed a significant effect of wine matrix composition on the intra-oral aroma release of certain odorants. Among the wine matrix parameters, phenolic compounds showed the largest impact. This effect was dependent on their chemical structure. Some phenolic acids (e.g. hippuric, caffeic) were associated to an increase in the intra-oral release of certain odorants (e.g. linalool, ß-ionone), while flavonoids showed the opposite effect, decreasing the intra-oral release of aliphatic esters (ethyl hexanoate). This work shows for the first time, the impact of wine composition on oral-mucosa interactions under physiological conditions.

Targeted Metabolomic Analysis of Polyphenols with Antioxidant Activity in Sour Guava (Psidium friedrichsthalianum Nied.) Fruit.

Psidium is a genus of tropical bushes belonging to the Myrtaceae family distributed in Central and South America. The polar extract of Psidium friedrichsthalianum Nied. was partitioned with ethyl ether, ethyl acetate, and n-butanol, and the total phenolic content and antioxidant activity were measured by Folin-Ciocalteu and ABTS assays, respectively. The ethyl acetate fraction exhibited both the highest phenolic content and antioxidant activity. Due to the complexity of this fraction, an analytical method for the comprehensive profiling of phenolic compounds was done by UPLC-ESI/QqQ in MRM (multiple reaction monitoring) mode. In this targeted analysis, 22 phenolic compounds were identified, among which several hydroxybenzoic, phenylacetic, and hydroxycinnamic acid derivatives were found. This is the first time that (+)-catechin, procyanidin B1, procyanidin B2, and (-)-epicatechin have been reported as constituents of sour guava. A fractionation by exclusion size, C18-column chromatography, and preparative RRLC (rapid resolution liquid chromatography) allowed us to confirm the presence of ellagic acid and isomeric procyanidins B, well-known bioactive compounds. The content of phenolic compounds in this fruit shows its potential for the development of functional foods.

Intra-oral adsorption and release of aroma compounds following in-mouth wine exposure.

Wine "after-odour" defined as the long lasting aroma perception that remains after wine swallowing is an outstanding characteristic in terms of wine quality but a relatively unstudied phenomenon. Among the different parameters that might affect wine after-odour, the adsorption of odorants by the oral mucosa could be important but has been little explored. In this work, the impact of the chemical characteristics of aroma compounds on intra-oral adsorption was assessed by an in vivo approach that determined the amounts of odorants remaining in expectorated wine samples. In addition, the subsequent aroma release after in-mouth wine exposure was studied by means of intra-oral SPME/GC-MS using three different panellists. Oral adsorption of the aroma compounds added to the wines ranged from 6% to 43%, depending on their physicochemical characteristics. A progressive intra-oral aroma decrease at different decay rates depending on compound type and panellist was also found. The strength of the aroma-oral mucosa interactions seems to explain these results more than the amount of compound adsorbed by the oral mucosa.