The influence of acid on astringency of alum and phenolic compounds

Astringency of aqueous solutions of phenolic compounds (grape seed tannins, tannic acid, catechin and gallic acid) increased upon addition of citric acid, whereas the astringency of alum was reduced. Astringency of alum was decreased equivalently by addition of equi-sour levels of lactic acid, citric acid or hydrochloric acid. The difference between alum and the phenolic compounds is speculated to result from chemical modifications affecting binding of the astringents with oral proteins rather than cognitive differences. Chelation of the aluminum ion in alum by acids reduces its availability for interacting with salivary proteins or epithelial proteins. In contrast, the increased astringency produced upon acidification of phenolic compounds is speculated to result from the pH driven increase in the affinity of the phenols for binding with proteins. These results suggest that alum cannot be used interchangeably with phenolic astringents in psychophysical studies.      

INTERACTION OF ASTRINGENCY AND TASTE CHARACTERISTICS1

The perception of astringency and basic taste in mixtures and their interaction effects were investigated by two procedures. In Experiment 1, focused and nonfocused testing procedures were compared using mixtures of low and high concentrations of alum and basic taste solutions. Both procedures yielded taste and astringency intensities that were modality‐dependent. Nonfocused testing was used in Experiment 2 to investigate the interactions of astringent phenolic (tannic acid) and nonphenolic (alum) compounds with each basic taste. Sweetness of sucrose increased with increased concentration with or without alum or tannin present. Changes in salty, bitter, and sour taste intensities were modality‐dependent. Astringency either remained unchanged or decreased with the addition of sucrose, sodium chloride, citric acid, or caffeine depending upon the taste concentration. Bitterness of tannin and alum at high concentrations was suppressed by the addition of sucrose, sodium chloride, or citric acid; sourness also decreased in the presence of sucrose or sodium chloride as well as a high level of caffeine.     

SELECTION OF AN ASTRINGENCY REFERENCE STANDARD FOR THE SENSORY EVALUATION OF BLACK TEA

Astringent and bitter sensations are characteristic sensory qualities of black tea. Three different classes of potential astringent reference standards (two concentrations each of alum and tannic acid and three fruit juices) were evaluated in this study. The perceived astringency, bitterness and sourness of each were profiled using computerized time-intensity and compared with the astringent intensity of a standardized brew of black tea. The differences in temporal profiles of potential reference standards across taste attributes were evident and intensity ratings were found to be dependent upon the stimulus and its concentration. Both concentrations of tannic acid were evaluated as the highest in perceived bitterness. For the juices, a strong sour taste was perceived in addition to astringency. It was concluded that the best reference standard for the astringency of black tea is a solution of 0.7 g/L alum as it is low in perceived bitterness and sourness.     

Psychophysical evidence that oral astringency is a tactile sensation

Aluminum potassium sulphate [AIK(SO₄)₂ 12H₂O—‘alum’]was tested for its ability to elicit oral astringency independentlyof its ability to elicit taste. First, the astringency producedby alum (10 g/l; 21.1 mM) on the non-gustatory surfaces betweenthe gum and the upper lip was measured. Subjects reported thatalum elicited sensations of astringency when the upper lip wasmoved laterally against the gum. Second, subjects dipped thetongue into two solutions—alum or a mixture solution thatapproximated the taste of alum—and attempted to determinewhich solution was more astringent. A slight tendency was foundfor subjects to identify the mixture as more astringent thanthe alum. However, the same subjects reported the alum to bemore astringent than the mixture when the same solutions wereapplied under the upper lip. These two experiments support thehypothesis that tactile stimulation is important for producingastringency, whereas taste stimulation of the anterior tongueis not. Third, after the application of alum, lubricating rinses(water, sucrose, Salivart®, corn oil, and the subjects'own saliva) were compared for their ability to decrease astringency.The lubricants reliably decreased original astringency, butto varying degrees depending upon their lubricating properties.All three experiments suggest that tactile sensations causedby increased friction (decreases,in salivary lubrication) betweenoral membranes are the primary basis of astringent sensations.     

Comparison of the effects of concentration, pH and anion species on astringency and sourness of organic acids

The separate effects of concentration, pH and anion species on intensity of sourness and astringency of organic acids were evaluated. Judges rated sourness and astringency intensity of lactic, malic, tartaric and citric acid solutions at three levels of constant pH varying in normality and at three levels of constant concentration varying in pH. To assess the comparative sourness and astringency of the organic acid anions of study, binary acid solutions matched in pH and titratable acidity were also rated. As pH was decreased in equinormal solutions, both sourness and astringency increased significantly (P < 0.001). By contrast, as the normality of the equi-pH solutions was increased, only sourness demonstrated significant increases (P < 0.001) while astringency remained constant or decreased slightly. At the lowest normality tested, all solutions were more astringent than sour (P < 0.05). Although lactic acid was found to be significantly more sour than citric acid (P < 0.05), no other sourness or astringency differences among the organic acid anions were noted. This study demonstrates for the first time that astringency elicited by acids is a function of pH and not concentration or anion species, and confirms that sourness is independently influenced by concentration, pH and anion species of the acid.      

Time-course of astringent sensations

Qualitative and quantitative perceptual reactions to astringentmaterials were examined for three diverse chemical substances(alum, tannic acid and tartaric acid) at several concentrationsproducing moderate to strong levels of perceived sensation.Group discussions were held to determine language appropriateto describe the sensations arising from solutions of the threecompounds and a composite ballot of six rating scales (astringency,mouth drying, puckery feeling, mouth roughing, bitterness andsourness) was developed. For both experiments, two concentrationsof each compound were rated on the six attributes for five tosix minutes, a discrete-point time-intensity scaling procedure.All ratings showed roughly exponential decays over time. Theintensity ratings for each attribute were found to depend onboth the particular astringent substance and concentration tested.The results from experiment 2 suggested that the four tactileattributes of drying, puckery feeling, roughing, and overallastringency may not be totally interchangeable and that theremay be multiple sub-qualities in the sensory reactions groupedas astringency. It is recommended that future structure-activitystudies make use of time-intensity procedures with multiplerated attributes, using 1 g/1 alum as a reference material,since it is relatively low in perceived bitterness and sourness,but produces pronounced drying, roughing, puckery/drawing sensations.     

Intensity and hedonic judgments of taste mixtures: an information integration analysis

Anderson's (1981) information integration approach was usedto examine taste mixture integration for intensity and hedonicjudgments of sucrose/sodium chloride and sucrose/citnc acidsolutions. In Experiment I, total intensity and hedonic ratingswere made for factorial combinations of sucrose and sodium chlorideor citric acid. The total intensity judgments produced an integrationpattern characterized by extreme subadditivity at high soluteconcentrations. (Subadditivity refers to the tendency for totalmixture intensity to be rated as less than the sum of the unmixedcomponent intensities.) The intensity judgment integration patternswere essentially identical for the two mixture types. However,the pattern of integration for the sucrose/sodium chloride andsucrose/citric acid mixtures differed for the hedonic ratings.Sucrose tended to eliminate the unpleasantness associated withincreasing concentrations of citric acid, while it only modulatedthis trend for sodium chloride. In Experiment II, subjects ratedthe individual sweet, salty and sour components of the mixturesto determine whether mixture suppression of the component tastescould account for the subadditivity of the total intensity judgmentsand/or the pattern of results for the hedonic ratings. It wasfound that sucrose suppressed the sour component of the sucrose/citricacid mixtures more than the salty component of the sucrose/sodiumchloride solutions This difference in component suppressionseemed to account for the hedonic integration patterns of thetwo mixture types which suggests that mixture suppression isan important factor to consider when predicting the pleasantnessof simple taste mixtures.     

Turbidity as a measure of salivary protein reactions with astringent substances

Binding of tannins to proline-rich proteins has been proposed as an initial step in the development of astringent sensations. In beer and fruit juices, formation of tannin-protein complexes leads to the well-known effect of haze development or turbidity. Two experiments examined the development of turbidity in human saliva when mixed with tannins as a potential in vitro correlate of astringent sensations. In the first study, haze was measured in filtered human saliva mixed with a range of tannic acid concentrations known to produce supra-threshold psychophysical responses. The second study examined relationships among individual differences in haze development and the magnitude of astringency ratings. Mostly negative correlations were found, consistent with the notion that high levels of salivary proteins protect oral tissues from the drying effects of tannic acid.     

Astringent Subqualities in Acids

Astringency, astringent subqualities (drying, roughing and puckering)and sourness were compared among six acids: hydrochloric, lactic,citric, acetic, fumaric and malic acids. The attribute profilesof organic acids were similar to each other but different fromhydrochloric acid, the only inorganic acid, which was the mostastringent and the least sour. In a second experiment, two inorganicacids (hydrochloric and phosphoric) and two organic acids (citricand malic) were tested at three concentration levels. At approximatelyequal levels of overall sensory impact, the inorganic acidswere alike in astringency and sourness, receiving higher ratingsfor roughing and drying, and lower ratings for sourness thanthe organic acids. Interactions with concentration (differencesin psychophysical functions) for the subquality of drying werenoted, in addition to the differences in the astringent subqualitiesof roughing and drying seen across acids in both experiments.The higher level of astringency for inorganic acids suggeststhat the current model for tannin binding to salivary proteinsas an explanation of astringency needs to be extended to includea direct pH-dependent effect. Chem. Senses 20: 593–600,1995     

Amiloride-sensitive and amiloride-insensitive responses to NaCl + acid mixtures in hamster chorda tympani nerve

Component signaling in taste mixtures containing both beneficial and dangerous chemicals depends on peripheral processing. Unidirectional mixture suppression of chorda tympani (CT) nerve responses to sucrose by quinine and acid is documented for golden hamsters (Mesocricetus auratus). To investigate mixtures of NaCl and acids, we recorded multifiber responses to 50 mM NaCl, 1 and 3 mM citric acid and acetic acid, 250 μM citric acid, 20 mM acetic acid, and all binary combinations of each acid with NaCl (with and without 30 μM amiloride added). By blocking epithelial Na(+) channels, amiloride treatment separated amiloride-sensitive NaCl-specific responses from amiloride-insensitive electrolyte-generalist responses, which encompass all of the CT response to the acids as well as responses to NaCl. Like CT sucrose responses, the amiloride-sensitive NaCl responses were suppressed by as much as 50% by citric acid (P = 0.001). The amiloride-insensitive electrolyte-generalist responses to NaCl + acid mixtures approximated the sum of NaCl and acid component responses. Thus, although NaCl-specific responses to NaCl were weakened in NaCl-acid mixtures, electrolyte-generalist responses to acid and NaCl, which tastes KCl-like, were transmitted undiminished in intensity to the central nervous system. The 2 distinct CT pathways are consistent with known rodent behavioral discriminations.     

Cytotoxic activity of hydrolyzable tannins against human oral tumor cell lines--a possible mechanism.

Hydrolyzable tannins showed higher cytotoxic activity against human oral squamous cell carcinoma and salivary gland tumor cell lines than against normal human gingival fibroblasts, whereas gallic acid, a component unit of tannins, showed much weaker selective cytotoxicity. The cytotoxic activity of dimeric compounds was generally higher than that of monomeric compounds. Macrocyclic ellagitannin oligomers, such as oenothein B, woodfordin C and woodfordin D showed the greatest cytotoxic activity, and their activity (per given number of molecules) was one order higher than those of gallic acid and epigallocatechin gallate, a major component of green tea. These compounds induced apoptotic cell death characterized by DNA fragmentation (as demonstrated by the TUNEL method) and cleavage of cytokeratin 18 by activated caspase(s) (as demonstrated by M30 monoclonal antibody). ESR spectroscopy revealed that these macrocyclic compounds at higher concentrations produced their own radicals and significantly enhanced the radical intensity of sodium ascorbate, possibly by their prooxidant actions. Catalase failed to eliminate their apoptosis-inducing activity, reducing the possibility of the involvement of hydrogen peroxide production in the extracellular fraction. These observations suggested that the antitumor activity of macrocyclic ellagitannin oligomers reported previously might be explained by their apoptosis-inducing activity.     

EFFECTIVENESS OF RINSES IN ALLEVIATING BITTERNESS AND ASTRINGENCY RESIDUALS IN MODEL SOLUTIONS1,2

A two‐part study determined the effectiveness of gum‐based rinses with or without oil for alleviating residuals of a bitter (0.8 g/L caffeine solution) and an astringent (1g/L alum solution) stimuli in serial responses using a sip and spit method. In Experiment 1, rinsing with deionized water was compared to rinsing with 0.3% xanthan gum in water alone or with 5% corn oil and 0.55% carboxymethyl cellulose (CMC) in water alone or with 5% or 10% corn oil. The 0.3% xanthan gum and 5% corn oil mixture resulted in the highest stimulus intensity difference before and after rinsing, whereas 0.55% CMC had a value of nearly zero for residual effect. Results of a two‐alternative forced choice test used in Experiment 2 indicated that both were equally effective for reducing bitterness residuals, but 0.55% CMC solution resulted in the lowest (p=0.007) residual effect for astringency. Therefore, the 0.55% CMC rinse was considered an effective interstimulus rinse to use for assessing both bitterness and astringency in model solutions.     

Volatile compounds associated to the loss of astringency in persimmon fruit revealed by untargeted GC–MS analysis

High resolution volatile profiling (67 compounds identified) of fruits from 12 persimmon cultivars was established and used to characterize the different astringency types of persimmon fruit before and after deastringency treatment. Analysis of the volatile profile of fruit enables us to differentiate between cultivars that at the moment of harvest produced non-astringent fruit (Pollination Constant Non Astringent—PCNA-type) from astringent ones (non-PCNA-type). Fruit failing to accumulate astringent compounds naturally (PCNA fruit) showed high levels of 3(2H)-benzofuranone, while this compound was not detected in any astringent type fruit (non-PCNA). In addition to this, PCNA cultivars also showed at harvest higher accumulation of benzeneacetaldehyde and lipid-derived aldehydes (hexanal, heptanal, octanal and decanal) than non-PCNA fruit. The application of postharvest deastringency treatment to all non-PCNA cultivars resulted on an important insolubilization of tannins. In general the CO₂-treatment enhanced the levels of acetaldehyde, however those cultivars showing high levels of dihydrobenzofuran at harvest did not present an increment of acetaldehyde. In contrast, all non-PCNA cultivars exhibited an important accumulation of lipid-derived aldehydes due to CO₂-treatment. Therefore, we propose that lipid-derived aldehydes (mainly decanal, octanal and heptanal) may be playing a role in the astringency loss. Our results suggest that 3(2H)-benzofuranone, benzeneacetaldehyde and lipid-derived aldehydes could be used as markers for both natural and artificial loss of astringency.     

Responses of lipid membranes of taste sensor to astringent and pungent substances

Astringent substances and pungent substances were studied usinga multichannel taste sensor with lipid membranes. The electric-potentialpattern constructed of eight outputs from the membranes hasinformation of taste quality and intensity. Pungent substances,such as capsaicin, pipeline and allyl isothiocyanate, had noeffect on the membrane potentials of the lipid membranes. Onthe other hand, astringent substances such as tannic acid, catechin,gallic acid and chlorogenic acid changed the potentials remarkably.A principal component analysis of the patterns in electric potentialchanges caused by tbe taste substances revealed the astringencyis located between bitterness and sourness.     

Expression of ethylene response genes during persimmon fruit astringency removal

Thirteen ethylene signaling related genes were isolated and studied during ripening of non-astringent ‘Yangfeng’ and astringent ‘Mopan’ persimmon fruit. Some of these genes were characterized as ethylene responsive. Treatments, including ethylene and CO₂, had different effects on persimmon ripening, but overlapping roles in astringency removal, such as increasing the reduction in levels of soluble tannins. DkERS1, DkETR2, and DkERF8, may participate in persimmon fruit ripening and softening. The expression patterns of DkETR2, DkERF4, and DkERF5 had significant correlations with decreases in soluble tannins in ‘Mopan’ persimmon fruit, suggesting that these genes might be key components in persimmon fruit astringency removal and be the linkage between different treatments, while DkERF1 and DkERF6 may be specifically involved in CO₂ induced astringency removal. The possible roles of ethylene signaling genes in persimmon fruit astringency removal are discussed.     

<Emphasis Type="Italic">ADH</Emphasis> and <Emphasis Type="Italic">PDC</Emphasis> genes involved in tannins coagulation leading to natural de-astringency in Chinese pollination constant and non-astringency persimmon (<Emphasis Type="Italic">Diospyros kaki</Emphasis> Thunb.)

Pollination constant non-astringency (PCNA)-type persimmons are the most desirable cultivar because the fruit loses astringency naturally and does not require any treatments for edibility. The mechanism of natural astringency loss in Chinese PCNA (C-PCNA)-type persimmon is probably related to the coagulation of soluble tannins into insoluble tannins, which is quite different from that in the Japanese PCNA (J-PCNA) type. In this work, three types of persimmon cultivars were sampled: ‘Luotian-tianshi’ (C-PCNA), ‘Maekawa-jirou’ (J-PCNA), and ‘Mopanshi’ (pollination constant astringent (PCA)) were sampled. Three DkADH and four DkPDC genes were isolated from C-PCNA plants. Three candidate genes for soluble tannins coagulation identified in C-PCNA fruit (DkADH1, DkPDC1, and DkPDC2) were characterized through combined analysis of spatiotemporal expression patterns and tannin and acetaldehyde contents during fruit development. Transient over-expression in persimmon leaves showed that DkADH1 and DkPDC2 led to a significant decrease in the levels of soluble tannins in infiltrated leaves. These results indicated that DkADH and DkPDC genes should be considered key genes for natural astringency loss in C-PCNA types.     

Binary taste mixture interactions in prop non-tasters, medium-tasters and super-tasters

It is generally assumed that the mutual, but asymmetric, suppression of the components in binary taste mixtures is an invariant property of the human psychophysical response to such mixtures. However, taste intensities have been shown to vary as a function of individual differences in sensitivity, indexed by the perceived bitterness of 6-n-propylthiouracil (PROP). To determine if these variations in taste perception influence taste mixture interactions, groups of PROP super-, medium- and non-tasters assessed four binary taste mixtures: sweet-bitter [sucrose/quinine hydrochloride (QHCl)], sweet-sour (sucrose/citric acid), salty-bitter (NaCl/QHCl) and salty-sour (NaCl/citric acid). In each experiment, subjects received factorial combinations of four levels of each of two tastants and rated individual taste intensities and overall mixture intensity. For each taste quality, super-tasters typically gave higher ratings than either medium- or non-tasters, who tended not to differ. There were also group differences in the interactions of the mixtures' components. Super-tasters rated the overall intensity of the mixtures, most likely reflecting integration of the taste components, as greater than medium- and non-tasters, who again showed few differences. In sweet-bitter mixtures, non-tasters failed to show the suppression of sweetness intensity by the highest QHCl concentration that was evident in super- and medium-tasters. These data show that the perception of both tastes and binary taste mixture interactions varies as a function of PROP taster status, but that this may only be evident when three taster groups are clearly distinguished from one another.     

EVALUATION OF BITTERNESS AND ASTRINGENCY OF (+)-CATECHIN AND (-)-EPICATECHIN IN RED WINE AND IN MODEL SOLUTION

Two stereoisomeric phenolic compounds, (+)-catechin and (-)-epicatechin, were rated for perceived intensity of oral astringency and bitterness by trained judges using the scalar method. Mouth drying and mouth roughening were also assessed, since they are often associated with astringency. Amounts of 375, 750 and 1500 mg/L of each compound were tasted in red wine, and in a model system, similar in composition to a dry table wine. Preliminary tests showed that these concentrations were above the threshold level but within the range found in wine. A control sample (model solution or wine without the addition of the above phenolic compounds) was also evaluated. The results showed that the two compounds were both bitter and astringent. The high (-)-epicatechin concentration was significantly more bitter and astringent than the equal concentration of (+)-catechin in the model solution. Mouth drying and roughening ratings showed a similar increasing pattern with the ratings of astringency particularly at the higher concentrations. However, these attributes were rated differently from astringency suggesting that although they contribute to astringency, they are not subsumed by it.     

Effect of condensed tannins addition on the astringency of red wines

Astringency has been defined as a group of sensations involving dryness, tightening, and shrinking of the oral surface. It has been accepted that astringency is due to the tannin-induced interaction and/or precipitation of the salivary proline-rich proteins (PRPs) in the oral cavity, as a result of the ingestion of food products rich in tannins, for example, red wine. The sensory evaluation of astringency is difficult, and the existence of fast and reliable methods to its study in vitro is scarce. So, in this work, the astringency of red wine supplemented with oligomeric procyanidins (condensed tannins), and the salivary proteins (SP) involved in its development were evaluated by high-performance liquid chromatography analysis of human saliva after its interaction with red wine and by sensorial evaluation. The results show that for low concentration of tannins, the decrease of acidic PRPs and statherin is correlated with astringency intensity, with these families having a high relative complexation and precipitation toward condensed tannins comparatively to the other SP. However, for higher concentrations of tannins, the relative astringency between wines seems to correlate's to the glycosylated PRPs changes. This work shows for the first time that the several families of SP could be involved in different stages of the astringency development.