Chemistry, Mouthfeel, and Consumer: Understanding Red Wine Astringency via Sensory and Analytical Approaches (Year 1). AVF Project #2025-2954.

AVF Proj. ID: 20252954 / /Cat.: , ,
Primary Investigator(s): Misha T. Kwasniewski, PhD, Associate Research Professor, Department of Food Science, Pennsylvania State University, 326 Erickson Food Science Building, University Park, PA 16802; Phone: 814-865-6842; Email: mtk5407@psu.eduby

Summary of Progress:

Wine astringency is a major driver of red wine style and consumer experience, yet it remains hard to predict when wines appear similar by routine tests.  Phenolic compounds are chemically diverse, and their effects depend on molecular structure and interactions with the wine matrix and saliva, so wines with similar pH, ethanol, and conventional total phenolic measurements can still feel meaningfully different.

During the first 6 to 7 months of Year 1, we advanced and applied our structure-informed phenolic fingerprinting approach to a targeted set of commercial California Cabernet Sauvignon wines selected to be chemically narrow by traditional metrics.  Key components of the analytical workflow have progressed rapidly, resulting in two peer-reviewed publications (Lin et al., 2025; Lin et al., 2026).  We also completed the first round of mouthfeel-focused sensory mapping with semi-trained wine consumers (n = 17) and collected paired saliva samples before and after wine consumption for planned Year 2 proteomics.

Early analyses indicate the structure-informed fingerprints explain the sensory space better than basic composition or conventional spectrophotometric phenolic assays.  In multivariate models, basic chemistry provided little predictive value, bulk phenolic assays predicted the primary sensory dimension but performed poorly for the second, and TFF provided the strongest overall prediction (Q2 approximately 0.71 for Dimension 1 and 0.37 for Dimension 2).  Within this chemically narrow wine set, Dimension 1 is most associated with condensed tannins and oxidation-derived features, while Dimension 2 is more associated with hydrolysable tannins and stilbenes.  We also observe clear person-to-person variability that is not explained by wine chemistry alone, motivating the planned Year 2 focus on saliva characterization.

A second mapping study to validate predictive performance with an independent wine set has been approved and is scheduled to begin in late February, and results will feed directly into Year 2 saliva proteomics and targeted follow-up testing.

This work is advancing practical, structure-informed tools to better explain and predict astringency and mouthfeel differences among wines that appear similar by standard chemistry.  Improved prediction in this chemically narrow space can reduce trial-and-error decision-making and help wineries more consistently achieve intended mouthfeel styles.  By quantifying when person-to-person biology (saliva) contributes to sensory variability, the project also supports future translation to consumer preference and segmentation, strengthening the ability of growers and wineries to align production choices with desired product profiles.

Managing Tannins in the Winery. AVF Project #2025-2978.

AVF Proj. ID: 20252978 / /Cat.: ,
Primary Investigator(s): James F. Harbertson, Washington State University, Viticulture and Enology Department, 359 University Drive, 509-372-7506, jfharbertson@wsu.eduby

Summary of Progress:

To further investigate the structural changes to grape tannins and the impact on grape tannin concentration by glutathione additions, an experimental design was carried out with red wine made to two separate maceration lengths and glutathione added at multiple time points throughout the winemaking process.  Cabernet Sauvignon grapes were harvested from Cold Creek Vineyard on October 7, 2025, and the fruit processed at the Wine Science Center.  For the control maceration wines, pressing occurred 14 days after crush, pharmaceutical-grade reduced glutathione was added on day 15 and day 45 post-crush to three 60 L kegs of wine at each addition.  For extended maceration wines, tanks with wine still on skins and seeds were combined for extended storage on day 13 after crush, reduced glutathione added on day 15 to three of the 210L tanks, sealed on day 16, and stored blanketed with inert gas.  Extended maceration wines were then pressed 44 days after crush, and reduced glutathione added at day 45 to three additional extended maceration wines in 60L kegs.  Glutathione was added to each tank or keg at a concentration of 2mM.  Thus far phenolic measurements using traditional assays show early additions of glutathione have minimal impact on protein precipitable tannin concentration.  In our previous experiments glutathione was added post-fermentation so this result was anticipated, and we have planned future additional treatments.  Overall, the current experiment has highlighted the potential importance of addition timing on the effectiveness of glutathione modification of tannins and has created a clear investigative route for lab scale experiments with regards to tannin modification and reduction.

Disappointingly we have been hampered with operational setbacks that have prevented us from doing our lab scale experiments and utilizing more sophisticated analysis on the winemaking treatments such as mass spectrometry.  We expect to carry out these experiments in early 2026 to remain on track for our objectives.

Eighteen Cabernet Sauvignon wines were made during the 2025 vintage varying the length of maceration with glutathione additions made during stages of maceration and wine development to evaluate its impact.  Thus far we have seen limited impacts using traditional assay methods.  Overall, the current experiment has highlighted the potential importance of addition timing on the effectiveness of glutathione modification and has created a clear investigative route for evaluating glutathione at lab scale with regards to tannin modification and reduction.

Improvement of Wine Quality: Tannin and Polymeric Pigment Chemistry

AVF Proj. ID: 1388 / /Cat.: ,
Primary Investigator(s): Andrew Waterhouseby

Summary

As we were breaking new ground from several perspectives, the project took some unexpected turns. At this point, we have completed several goals. First, we have created a table of expected wine pigments based on known reactions between anthocyanins and proanthocyanidins, goals 2a and 2b. These results were published in two papers, the first describing how to enumerate all possible proanthocyanidins that could be distinguished by mass spectrometry, up to a degree of polymerization (DP) of 10 (Cave and Waterhouse 2019), and the second, taking that list and “reacting” those tannins with the pigment to create a list of all possible wine pigments. This was a list of over 1 million structures. (Cave et al. 2019)

Having that list, we then compared the masses of those proposed structures with mass spectral data from two sources, QToF and ICR. In the case of the QToF, we could match about 10% of the several hundred observable compounds (Cave et al. 2020). However, the ICR is much more sensitive and precise, and here we could observe about 18,000 signals, and of those could match over 20% of the signals to entries in our table (Cave et al. 2020). Unfortunately, many signals could be represented by more than one structure, so additional study is needed to determine which of the options is present. These results addressed goals 1a, 1b and 2c. The data we now have will require further mass spectral analysis to discriminate between a number of redundant structures.

Anthocyanin-Cell Wall Interactions Effect on Tannin Extraction

AVF Proj. ID: 2219 / /Cat.:
Primary Investigator(s): Andrew L. Waterhouseby

The objectives of this proposal have been to do the following:

1) Perform extractions on white grape skins at 5 time points (13,17,20,22, and 25 °Brix) throughout maturity with varying levels of added anthocyanins.

• Extracted tannin will be analyzed for concentration, average molecular size, subunit composition, activity, and fraction extracted.
• Measure the levels of pectin methylation in the white grape skins

2) Conduct red-styled fermentations on Sauvignon blanc and Cabernet Sauvignon at 300 pounds per fermentation, with and without an anthocyanin addition. Measurements are the same as Objective 1.

The overall purpose of this proposal is to determine anthocyanins role on the extraction of procyanidin material due to interactions with cell wall material of grape skins. Activity to date has been the isolation and purification of color along with the sampling and extraction of skins with varying concentration of anthocyanins added. Furthermore, 300lb fermentations were conducted, in triplicate, under standard red wine making conditions with both Sauvignon Blanc and Cabernet Sauvignon with a 1.4% (v/v) addition of color concentrate.

Significance of Oak Ellagitannin Chemical Structure to Wine Oxidation

AVF Proj. ID: 2219 / /Cat.: ,
Primary Investigator(s): Andrew L. Waterhouseby

The pathway of wine oxidation, as currently understood, encompasses the cascade of reactions incited by the oxidation of phenols in the presence of oxygen, eventually coming to the conversion of ethanol into acetaldehyde. While it is evident that wines consume oxygen over time and acetaldehyde becomes increasingly apparent with age, the rate of oxidation can vary unpredictably among different wines, and it was hypothesized that different phenolic structural features, particularly those of oak ellagitannins, are the reason for such variability in oxidation. It was originally proposed that ellagitannins and other phenols with distinct functionalities be studied for their effects on oxygen consumption and acetaldehyde production. However, in light of recent studies conducted by our laboratory demonstrating that the input of oxygen does not guarantee the output of acetaldehyde, it was decided that attempting to study the pathway of oxidation in its entirety, from oxygen to acetaldehyde, would not be an effective approach.

Given the complexity of wine oxidation, a more sensible strategy would be to study the effects of phenolic structure on individual reactions rather than the pathway as a whole. In the first step of wine oxidation, the oxidation of phenols is coupled to the reduction of oxygen by iron, which acts as a shuttle for electrons between phenols and oxygen. A more specific hypothesis now is phenolic structure affects their reactivity with iron, subsequently affecting oxygen consumption and the remainder of the wine oxidation pathway. The initial reactions of wine oxidation may be characterized by the redox cycling of iron between its two oxidation states: Fe(II) and Fe(III). The addition of electrons to oxygen occurs with the oxidation of Fe(II) to Fe(III), and in the opposite direction, the loss of electrons from phenols takes place with the reduction of Fe(III) to Fe(II). The ratio of Fe(II) to Fe(III) should thus depend on the relative reaction rates of Fe(II) with oxygen and that of Fe(III) with phenols.

A quick and simple spectrophotometric method for iron speciation, employing the complexing agent Br-PADAP, is currently being optimized and validated, to be used not only to assess differential rates of iron reduction by structurally diverse phenols, but also by the industry to more generally measure the “redox status” of their wines. Our laboratory’s modified version of the Br-PADAP assay is simple and inexpensive, requiring a sample volume of only 200 μL and a reaction time of 10 min, and is done directly in a cuvette. Validation of the assay is currently underway; difficulty lies in the fact that there does not exist a standard method for iron speciation to compare, thus alternative methods of validation are being considered. This research would not only improve management of oxidation, but also furnish a more complete understanding of phenolic oxidation, with the ultimate goal being the prediction of wine aging based on phenolic content and composition.

Red Wine Tannin Interaction with Polysaccharides

AVF Proj. ID: 1850 / /Cat.: ,
Primary Investigator(s): James Kennedyby

The objectives of this proposal are to do the following:

  1. Investigate polysaccharide interaction with tannins in wine.
  2. Determine how intermolecular interactions between macromolecules in wine influence wine mouthfeel.

Following the first two years of activities, excellent progress has been made. Ten wineries were recruited and ten wines were provided with all different winemaking processes. Laboratory activities have made significant progress, with polysaccharides and tannins extraction from wine and chemical characterization. Interactions between tannins from red wines and polysaccharides have been characterized by dynamic light scattering. The effect of pH and ethanol content on the interactions between tannin and polysaccharide has been investigated.

Tannin Structure-Activity Relation to Red Wine Astringency

AVF Proj. ID: 1691 / /Cat.: ,
Primary Investigator(s): James Kennedyby

The objectives of this proposal have been to do the following:

In a companion project being submitted to the Agricultural Research Institute (California State University research initiative)

II. Conduct extended sensory studies with the University of California, Davis, on a subset of the above wines to determine the relationship between sensory and instrumental analysis of red wine mouthfeel.

These objectives are consistent with the highest priority research objective as outlined by the American Vineyard Foundation.

The overall purpose of this proposal is to determine the role of grape and wine production practices on tannin structure and perception.  In combination, tannin activity will be monitored by high performance liquid chromatography (HPLC) so that a comparison can be made between human perception and HPLC measurement.

Following the second year of activities, excellent progress has been made.  The major activity during the second year of this study has been to monitor the development of berries in approximately 75 Napa Valley blocks.  Monitoring the tannin activity in these blocks is expected to provide information on the role that grape production practices can have in overall grape and corresponding wine tannin.

Red Wine Tannin Interaction with Polysaccharides

AVF Proj. ID: 1850 / /Cat.: ,
Primary Investigator(s): James Kennedyby

The objectives of this proposal are to do the following:

I. Determine variation in tannin activity as a function of polysaccharides structure from yeast and/or grape in red wine.

II. Relate the polysaccharide-tannin interaction variation to the wine mouthfeel.

In a companion project funded by the Agricultural Research Institute (California State University research initiative)

III. Improve the understanding of tannin-polysaccharide interactions consequences on red wine stability.

These objectives are consistent with the highest priority research objective as outlined by the American Vineyard Foundation.
The overall purpose of this proposal is to determine the role of polysaccharides from yeast and/or grape in finished red wine on tannin structure and activity. In combination, competitive interactions between tannins and proteins or polysaccharides will be elucidated by spectrophotometry after variation of matrix parameters, so that an elaboration of a functioning interactions model can be made.

Improvement of Wine Quality: Tannin and Polymeric Pigment Chemistry

AVF Proj. ID: 1850 / /Cat.: ,
Primary Investigator(s): James Kennedyby

Seven vintages of UC Davis Oakville Cabernet Sauvignon spanning 35 years was selected for analysis by a complimentary suite of mass spectrometric techniques. The vintages from 1974 to 2009 selected by informal sensory evaluation and basic wine chemistry to be of a representative vertical style were: 1974, 1981, 1988, 1994, 2001, 2003 and 2009. All wines were produced at UC Davis for research purposes and stored in the UC Davis cellar together. This sample was determined to be the most controlled and uniform sample of wines varying by vintage only as could be obtained for the project. Al basic wine chemistry was obtained, pH, ethanol concentration etc. as were various assays for comparison to mass spectrometric data.

The following objective were submitted for the grant proposal under which this work was peformed.

1) Observe the evolution of pigmented tannin throughout aging

a. Employ our method of complimentary mass spectrometric techniques (ICR, QTOF, QTrap) for comprehensive identification of wine matrix compounds.

b. Observe the changes in relative abundance, depletion and accumulation in pigmented tannin composition (35 Year Vertical)

c. Correlate pigmented tannin structural analysis with well-known molecular characteristics (mDP, mass recovery) and newly developed sensory representative analyses (“grippiness”) to better convey the impact of these discoveries

2) Apply existing synthetic technologies.

a. Development of a library of standards for quantitation and calibration.

b. Postulate wine pigment precursors for examination of mechanistic pathways.

c. Employ the standards to quantitate the classes of polymeric pigment in wine.

Wine Research Assays

Phloroglucinolysis was performed on the wines used for the study (Table 1) as well as their extracts (Table 2). The mass recovery demonstrates typical value dropping off well below 80%after 7 years. Molecular mass after 50%elution also follows expected patterns of increase with age. This indicates overall larger tannin in the older wines as supported by McRae et al 2012. The molecular mass is a measure of intact tannins, as opposed to the mDP which measures only those tannins which could be cleaved by phloroglucinol. The inconsistency between these two values is reconciled by the mass recovery, indicating that more complexity of the tannin structure is a factor in the older wines, for instance additional interflavan bonds preventing phloroglucinol attack.

Tannin Structure-Activity Relation to Red Wine Astringency

AVF Proj. ID: 590 / /Cat.: ,
Primary Investigator(s): James A. Kennedyby

Optimizing wine quality with regard to mouth feel is a quest for wineries that are trying to fine tune the astringency of their wines to a desirable level. Consistent with that, various wineries had agreed and participated in the extended maceration project in order to understand the driving force behind astringency. With that in mind, this project was born to comprise different analytical techniques, which are necessary to construct a conclusive understanding of tannin’s activity. During the 2014 vintage, maceration trials were conducted in five Napa Valley wineries. Winery selection was based upon winemaker interest. For each winery, fermentations were set up according the the specific cooperators protocol. At various times during the course of fermentation/maceration, samples were collected following pumpover operations. Following collection, samples were transported and stored at 7 degrees Centigrade until processed. For tannin isolation, each sample was filtered using a 9.0cm Whatman filter paper (20-25 μm pore size), diluted with milli-Q water (50:50 v/v) and then loaded onto a preconditioned column containing Toyopearl chromatography resin (HW 40C). Purification of tannins was conducted according to Aron et al.1 Briefly, and following application to the column, the column was washed with water followed by 50%v/v aqueous methanol and then tannins were eluted with 66%v/v acetone in water. The acetone was removed by rotary evaporation and the aqueous portion containing the tannin was lyophilized to a powder. Gravimetric yields were determined and the tannin powders were stored in glass, air tight, vials after being sparged with N2 and stored under -20°C. Tannin isolates underwent analysis using a variety of analytical techniques including gel permeation chromatography, phloroglucinolysis and stickiness. The analysis of tannins’ stickiness has recently been introduced by Barak et al.2 and further developed by Revelette et al.3. Samples were prepared according to the above methods and the enthalpy of interaction was determined for tannin polymers absorbing at 280 and 520 nm. Secondly, tannin powders underwent acid catalyzed degradation in the presence of excess phloroglucinol (phloroglucinolysis) to determine the subunit composition, average degree of polymerization and conversion yield following the method of Kennedy and Jones.

Following phloroglucinolysis, tannins were analyzed by gel permeation chromatography which provided information on size distribution5. Across all experiments, a total of 104 tannin isolates were prepared. Given that these isolates represent tannins collected at various points during fermentation and maceration and across different blocks and varieties, it is believed that a realistic picture of commercial structure variation for Napa Valley has been achieved. Results to Date are as follows: Winery 1 investigated the effect of extended maceration on tannin composition and activity. One Cabernet sauvignon block was harvested and equal portions of fruit were contained in two identical fermentation vessels. The must underwent a prefermentation soak for four days before inoculation. Samples for the control tank were collected on a semi-daily basis until press day (total soak time was 15 days). On the last day (day 15), the first extended maceration sample was collected. Similarly, extended maceration samples were taken on a semi-daily basis until press day (total soak time was 26 days).

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