By: Denise M. Gardner
It’s that time of year again: bottling time! The past year’s vintage is slowly starting to take up too much room in the cellar and now is the time for decision making in terms of preparing for the pending vintage. Finalizing a good bottling schedule before harvest starts is an essential good winemaking practice, but bottling comes with its own set of challenges.
It is not uncommon for winemakers to express feelings of “not being able to sleep at night” when wines get bottled, as they are worried about possible re-fermentation issues. As wine naturally changes through its maturity, it is easy to feel insecure about bottling wines, especially those wines that may have had challenges associated with it throughout production.
However, there are several analytical tests that winemakers can add to their record books every year to ensure they are bottling a sound product. The following briefly describes a series of analytical tests that provide information to the winemaker about stability and potential risks associated with the product when it goes in bottle.
Basic Wine Analysis Pre-Bottling:
This first list is the bare minimum data that should be measured and recorded for each wine getting bottled, regardless of the wine’s variety or style. Keeping accurate records of these chemistries is also helpful in case something goes wrong while the bottle is in storage or after it is purchased by a customer.
pH is essential to know as it gives an indication for the wine’s stability in relation to many chemical factors including sulfur dioxide, color, and tannin. For example, high pH (>3.70) wines provide an indication that more free sulfur dioxide is needed to obtain a 0.85 ppm molecular free sulfur dioxide content. At the 0.85 ppm molecular level, growth of any residual yeast and bacteria in the wine should be adequately inhibited.
High pH wines tend to have issues with color stability. At this point, color stability can be addressed by blending or with use of color concentrates (e.g., Mega Purple). Keep in mind that if the wine is blended with another wine, all chemical analyses, including pH, should be completed on the blend (as opposed to average individual parts) prior to bottling.
Free and Total Sulfur Dioxide Concentration
In the United States, total sulfur dioxide is regulated and must fall under 350 mg/L for all table wines (CFR: https://www.ecfr.gov/cgi-bin/text-idx?SID=eddaa2648775eb9b2423247641bf5758&mc=true&node=pt27.1.24&rgn=div5#sp27.1.24.a).
However, the free sulfur dioxide concentration provides an indication to the winemaker regarding antioxidant strength and perceived antimicrobial protection. To inhibit growth of yeast and bacteria during bottle storage, a 0.85 ppm molecular free sulfur dioxide concentration must be obtained. The free sulfur dioxide concentration required to meet the molecular level is dependent on pH. Therefore, free sulfur dioxide additions should be altered and based on a wine’s pH for optimal antimicrobial protection.
Analytically, it can be daunting to measure free sulfur dioxide as the wet chemistry set up looks intimidating. However, many small commercial wineries have benefited from the integration of a modified aeration-oxidation (AO) system, and with a little practice, have been relatively successful at monitoring free sulfur dioxide concentrations. A few wineries have worked to validate use of Vinmetrica’s analyzer (https://vinmetrica.com/), and found results comparable to those obtained by use of the AO system.
Residual (or Added) Sugar
Any remaining sugar in the bottle, whether through an arrested fermentation or direct addition, can pose a risk for re-fermentation post-bottling. This is especially true if the winery lacks good cleaning and sanitation practices. Nonetheless, it is a good idea to assess the sugar content pre-bottling to record a baseline value of the sugar concentration going into bottle. If bottles were to start re-fermenting, a sugar concentration could be analyzed and used to compare against the baseline value in order to assess the potential of yeast re-fermentation.
For wineries with minimal residual sugar concentrations, a glucose-fructose analysis (often abbreviated glu-fru) is often used to help determine accurate sugar content. For wines with added sugar an inverted glucose-fructose analysis may be required.
If you are concerned about potential risk for Brettanomyces (Brett) bloom post-bottling, it is usually encouraged to reduce the sugar content in the finished wine below 1% (<10 g/L sugar) in the bottle.
Malic Acid Concentration
While using paper chromatography to monitor malolactic fermentation (MLF) is useful, it does not give an accurate reflection of residual malic acid concentration. In fact, some winemakers find that a paper chromatogram may show a MLF has been “completed,” but would prefer to have lower residual malic acid concentrations remaining in the wine.
During my time at an analytical company, 0.3 g/L of malic acid and below was considered “dry.” This is typically a safe level of residual malic acid to avoid post-bottling MLF.
Volatile acidity (VA) is federally regulated, and levels are indicated in the Code of Federal Regulations (CFR: https://www.ecfr.gov/cgi-bin/text-idx?SID=eddaa2648775eb9b2423247641bf5758&mc=true&node=pt27.1.24&rgn=div5#sp27.1.24.a). For most states, with California as an exception, the maximum allowable VA for red wines is 1.40 g/L acetic acid (0.14 g/100 mL acetic acid) and for white wines is 1.20 g/L acetic acid (0.12 g/100 mL acetic acid).
Monitoring VA through production is a good indicator of acetic acid bacteria spoilage. At minimum, wineries should record VA
- immediately post-primary fermentation,
- periodically through storage (e.g., every 2-3 months) and
Whiling monitoring VA, sharp increases in VA should alarm the winemaker of some sort of contamination. Typically, these increases are caused by acetic acid bacteria, which can only grow with available oxygen.
As a general rule of thumb, knowing the final alcohol concentration is a good idea. Alcohol content helps determine a tax class for the wine and is required for the label.
Titratable Acidity (TA)
All wines are acidic in nature as they fall under the pH 7.00. However, titratable acidity (TA) acts as an indicator for the sour sensory perception associated with a given wine. For example, two wines, Wines 1 and 2, with a pH of 3.40 may have different TAs. If Wine 1 has a TA of 8.03 g/L tartaric acid while Wine 2 has a TA of 6.89 g/L tartaric acid, Wine 1 would likely taste more acidic (assuming all other variables are the same).
Cold stability tests are often recommended to ensure the wine is cold stable, and will, therefore, not pose a threat of precipitating tartrate crystals during its time in bottle. Not all wines require a cold stability process (e.g., seeding and chilling). Cold stability testing can be done prior to a cold stabilization step in order to avoid extraneous processing operations, saving time and money.
For more information on cold stability processes and testing, please visit Penn State Extension’s website: http://extension.psu.edu/food/enology/analytical-services/cold-stabilization-options-for-wineries
Additionally, haze formation is a potential risk post-bottling. While hazes do not typically offer any safety threat to wine consumers, they often look unappealing. Protein hazes tend to make the wine look cloudy. Some varieties are more prone to protein hazes then others, and running a protein stability trial could minimize the risk for a protein haze in-bottle.
It is important to remember that due to the fact protein stability is influenced by pH, cold stability production steps should take place before analyzing the wine for protein stability and before going through any necessary production steps to make the wine protein stable. This is due to the fact that cold stability processes ultimately alter the wine’s pH, and the chemical properties of proteins are influenced by the pH.
Analysis for Those that May Consider Bottling Unfiltered:
Yeast and Bacteria Cultures (Brett, Yeast, Lactic Acid Bacteria, Acetic Acid Bacteria)
Having a microscope in the winery can be a great reference point in terms of scanning for potential microbiological problems. However, if the winery does not have a microscope, but knows that some microbiological issues or risks may exist in a wine, having a lab set test the wine on culture plates is a good indicator for potential growth risks during the wine’s storage.
If the wine is going to be bottled using a sterile filtration step, keep in mind that wines are not bottled sterile. Assuming the absolute filtration method is working properly, the wine has potential to become re-contaminated with yeasts and bacteria from the point of which it exits the filter. In fact, it is not uncommon for wines to pick up yeast or bacteria contamination during the bottling process.
Managing free sulfur dioxide concentrations can help inhibit any potential growth from contamination microorganisms if the proper antimicrobial levels (0.85 ppm molecular) are obtained at that wine’s pH and retained during the bottle’s storage.
4-EP and 4-EG Concentrations for Reds
For wines that may have had a Brettanomyces (Brett) bloom, knowing the concentrations of 4-EP and 4-EG in the wine going into bottle is a good result to keep on file. If a Brett bloom occurs later in the bottle, it is likely (although, not guaranteed) that the volatile concentration of 4-EP and/or 4-EG may increase and confirm the problem.
Furthermore, evaluating a wine for 4-EP and 4-EG concentrations can also help isolate a possibility of Brett existence, especially if their concentrations are below threshold. However, it should be noted that both compounds can also exist in wines that are stored in wood, even without a Brett contamination.
Double Check: PCR for Reds
Brett can be a tricky yeast to isolate and identify. It is usually recommended to run multiple analytical tests related to Brett in order to confirm its existence or removal from a wine. While culture plating identifies living populations of microorganisms, PCR cannot typically differentiate between live and dead cells as it is measuring the presence of DNA. A microorganism’s DNA can get into a wine after yeast death and through autolysis. Therefore, a positive PCR result for Brettanomyces is hard to confirm if the result includes live cells, dead cells, or a combination of both.
Culture plating can help confirm the presence of active, live cells, but the success rate of growing Brettanomyces in culture plates is variable.
Nonetheless, scanning wines by PCR for Brett can help winemakers isolate a general presence and risk of Brett in their wines.
Still Worried About Your Wine Post-Bottling?
Bottle sterility testing is helpful, especially when a winemaker wants to ensure wines have been bottled cleanly. For this type of testing, it is best to sample a few bottles
- at the beginning of a bottling run,
- immediately before any breaks,
- immediately after any breaks, and
- at the end of a bottling run.
Bottles can, again, be evaluated under a microscope and evaluated for the presence of microorganisms. Bottles can also be sent to a lab for culture plating. The growth of yeasts or bacteria from culture plates at this stage indicates a failure of the sterile filtration system or contamination of the wine post-filtration. Clean wines, obviously, should help put a winemaker’s mind at ease as it matures in bottle.
Ensuring a wine’s stability post-bottling is a challenge. However, with proper cleaning and sanitation methods coupled with the right analytical records, winemakers can reduce their worry. For information on any of these topics, please visit:
- An Overview of Winery Sanitation by Patricia Howe: https://www.umpqua.edu/images/areas-of-study/career-technical/viticulture-enology/downloads/conferences/technical-symposia/2011-march-wine-flaws/2011-ts-howe-winery-sanitation.pdf
- Making Cleaning and Sanitation Practical for the Small Commercial Winery by Denise M. Gardner: http://bit.ly/PracticalWinerySanitation
- Minimizing Spoilage of Wines in Barrel by Denise M. Gardner: http://bit.ly/WineBarrelSanitation
- Bottling Line Cleaning Protocol by Scott Labs: http://www.scottlab.com/uploads/documents/technical-documents/1191/Bottling%20Line%20Cleaning%20Protocol.pdf
- Preparing Wines for Bottling by Enartis Vinquiry: http://www.enartis.com/upload/images/03_2016/160311011309.pdf
- Starting a Lab in a Small Commercial Winery: http://extension.psu.edu/food/enology/analytical-services/setting-up-your-winerys-lab
- Wine Analytical Labs: How Your Winery Can Use Them: http://extension.psu.edu/food/enology/analytical-services/wine-analytical-labs-how-your-winery-can-use-them
By: Denise M. Gardner
The long months post-harvest require regular attention by cellar staff and winemakers to ensure that wine quality is upheld through storage conditions. Wine stability, while somewhat nebulous, is essential to obtain in order to ensure the wine’s quality will be upheld post-sale. Below is a list of cellar maintenance practices that are recommended in preparation before the growing (and bottling) season.
Monitor Sulfur Dioxide Concentrations
Now (i.e., the winter and spring months) is a good time to regularly check sulfur dioxide concentrations of wines sitting in tanks and barrels waiting to get bottled. At minimum, wines should be checked once a month for free sulfur dioxide concentrations. Some winemakers opt to check barreled wines every other month in order to minimize frequently opening the barrel.
Proper sanitation and sampling is required for best analytical results:
- Use clean sampling bottles when taking wine samples
- Make sure that you sanitize any valves or sampling ports before and after releasing a sample from a tank. At the very least, you can use a food-grade alcohol solution spray or a citric acid-sulfur dioxide mix as a sanitizing agent.
- Properly clean and sanitize wine thieves or other sampling devices each time you use it to take a sample from a barrel or the top of tank. Warm water is not enough to sanitize a wine thief. We recommend using a citric acid-sulfur dioxide mix for quick dipping in between barrel sampling.
For wines that have completed primary fermentation and/or malolactic fermentation, maintaining a molecular free sulfur dioxide concentration is helpful to reduce the risk of yeast and bacterial spoilage. For a review on sulfur dioxide and making sulfur dioxide additions, please refer to this Penn State Wine Made Easy fact sheet.
Cold (Tartrate) Stabilization
Cold stabilization is often utilized to avoid the precipitation of tartrate crystals, which is common in instable wines at cooler temperatures.
In 2012, Virginia (Smith) Mitchell, now head winemaker at Galer Estate Winery, wrote a primer on cold stabilization techniques available for wine producers: http://extension.psu.edu/food/enology/analytical-services/assessment-of-cold-stabilization This primer covered everything from how to analyze for cold stability to the use of carboxymethylcellulose (CMC) to avoid tartaric acid crystallization in wine.
Prior to putting a wine through cold stabilization, it is worth the time and effort to analyze the wine for cold stability. Not all wines end up having cold stabilization problems. For those wines that do not, going through the cold stabilization process can actually minimize wine quality by stripping out delicate aromas and flavors, or altering taste or mouthfeel attributes of the wine. This doesn’t touch upon the amount of wasted time and effort to cold stabilize wines that are otherwise cold stable.
The above report recommends several testing procedures to ensure tartrate stability of a wine.
With the relatively warmer 2015-2016 winter, many winemakers may need to turn to artificial chilling in order to cold stabilize their wines properly. Again, this could be used as an argument to test wines prior to cold stabilization to minimize the use of electricity and to better manage the flow of wines in and out of the cold stabilization tank.
Wines that do undergo cold stabilization will likely have changes in pH and titratable acidity (TA) that can ultimately affect other parameters of the wine: protein (heat) stability, color, sulfur dioxide concentrations, and volatile acidity. It is prudent to check these components analytically following the cold stabilization process.
Protein (Heat) Stabilization
Proteins in wine can elicit hazes in wines post-bottling that may be off-putting to some consumers. While the proteins cause no effect on wine quality, they do cause an alteration in the appearance of the wine. Some varieties, like Gruner Veltliner, have naturally high concentrations of proteins, and, therefore, require a more aggressive approach to protein fining. Other varietals, however, may not require protein fining with bentonite at all.
Wines should undergo protein (heat) stability after they are cold stabilized due to the fact that cold stabilization will affect the acidity (pH and TA) of the wine, and therefore, alter protein stability properties of the wine. Again, winemakers are encouraged to check the wine for protein stability prior to treating a wine with bentonite.
Bentonite is a fining agent used to bind any proteins in a wine that would otherwise be considered unstable. However, if the addition of bentonite is unnecessary (i.e., the wine is protein stable and does not provide a component for bentonite to bind to, bentonite can bind to other components in the wine, most specifically: aroma and flavor active compounds. While this has been shown in the research literature, it is unclear how detrimental the loss of aromatic compounds is to the wine (Marchal and Waters 2010). Additionally, bentonite additions have been noted to strip color out of rosé and red wines (Butzke 2010).
A summary from UC Davis on heat stability testing can useful to understand the positive points and limitations of protein stability testing. Protocols for heat stability tests can be found here from Dr. Bruce Zoecklein. Additionally, ETS Labs has provided a small summary of how to interpret heat stability results, which can be helpful for wineries that are not used to reading analytical results on this test.
Additionally, wineries can submit wines to ISO-accredited labs for a bentonite trial in which the lab pinpoints the exact concentration of bentonite needed to heat stabilize the wine. This may be helpful to avoid making too little or too much bentonite additions, which costs time and labor in the winery.
Finally, if wineries are conducting their own bench trials, they are encouraged to use the same lot of bentonite in both the trials and the commercial application (Marchal and Waters 2010). This is due to the natural variability associated with most bentonite products. Finally, unless otherwise stated by the supplier, bentonite should always be blended in chlorine-free, hot (60°C, 140°F) water (Butzke 2010), and allowed to cool to room temperature so that the bentonite can swell. Allowing the slurry to cool will ensure that the wine is not exposed to a hot slurry.
Butzke, C. 2010. “What Should I use: sodium or calcium bentonite?” In: Winemaking Problems Solved. Christian E. Butzke, Ed. Woodhead Publishing Limited and CRC Press, Boca Raton, FL. ISBN: 978-1-4398-3416-9
Marchal, R. and Waters, E.J. 2010. “New directions in stabilization, clarification and fining of white wines.” In: Managing wine quality, volume 2. Andrew G. Reynolds, Ed. Woodhead Publishing Limited, Great Abington, UK. ISBN: 978-1-84569-798-3
Iland, P., N. Bruer, A. Ewart, A. Markids, and J. Sitters. 2012. Monitoring the winemaking process from grapes to wine: techniques and concepts, 2nd edition. Patrick Iland Wine Promotions Pty. Ltd., Adelaide, Australia. ISBN: 978-0-9581605-6-8.
Penn State Extension Wine Made Easy: Sulfur Dioxide Management: http://extension.psu.edu/publications/ee0093
Penn State Extension: Assessment on Cold Stabilization: http://extension.psu.edu/food/enology/analytical-services/assessment-of-cold-stabilization
UC Davis: Heat Stability Testing: http://wineserver.ucdavis.edu/pdf/attachment/88%20stability%20tests%20and%20haze%20formation%20.pdf
Virginia Tech: Protein Stability Determination in Juice and Wine (1991): http://www.apps.fst.vt.edu/extension/enology/downloads/ProteinS.pdf
ETS Labs: Interpreting Heat Stability Tests: https://www.etslabs.com/assets/PTB011-Interpretation%20of%20Heat%20Stability%20Results%20and%20Turbidity%20Readings.pdf
By: Denise M. Gardner
Wine protein chemistry is a daunting subject to tackle for a wine novice, and can also be a challenge for those who have been in the industry for years.
Proteins exist in wine in a state of charge: positive, negative, or neutral. The stability, and charge, of proteins is determined by the protein’s isoelectric point (or pI), in which the net charge of the protein is 0. Proteins are most unstable at their pI. Isoelectric points are based on pH, and alterations of pH in the wine will ultimately affect the charge of proteins in the wine. Alterations to wine chemistry (i.e. cold stabilization, acidification or deacidification, blending, and use of some fining agents) may alter protein stability of a given wine.
[Production Note: Due to the fact that protein stability is based on pH, winemakers should allow a wine to go through cold stabilization before testing and treating for protein stability. This is because alterations to the wine’s pH and titratable acidity (TA) will occur as a result of cold stabilization.]
Factors that affect protein stability
In general, lower pH wines have less protein stability problems than higher pH wines. This is a consideration for those producing lower pH (<3.4) wines. It is possible that the wines may not need bentonite additions.
The only way to evaluate if a wine is a protein stable is through a heat/protein stability test. For a protocol on protein stability, please refer to Bruce Zoecklein’s “Wine Analysis and Production” book (page 469-473) or via this online protocol. Most experts recommend following an ethanol-based test for better accuracy affiliated with protein stability. Additionally, wineries can contract certified wine labs to run protein stability tests for them, in addition to determining a rate for bentonite additions.
Tannin content can also affect the stability of wine proteins. As some tannins bind with proteins, having a higher concentration of tannins in the wine may make the wine more protein stable. This is one why some wine grape varieties, like Pinot Noir or some hybrids, have more protein instability problems than others. Additionally, white and rosé wines are more likely to carry protein instability problems as they are typically lower in tannins available to bind to proteins, but other factors contribute to this generality.
Some wine grape varieties may also be high in wine proteins, which dictate a need for more aggressive protein fining treatments. Gruner Veltliner and native varieties like Concord and Niagara anecdotally tend to have more consistent issues related to protein stability than other varieties. For Gruner Veltliner, juice treatment with bentonite additions is often recommended. Native varieties tend to require a treatment with Sparkolloid.
Be forewarned that protein stability can alter for a given grape variety each vintage year; a wine harvested in one vintage year may be protein stable for that particular year, but not so for a second, consecutive year. Protein stability varies with vintage season, and should be evaluated on each wine annually.
Why is bentonite used to treat protein instabilities?
Bentonite is a clay-based fining agent used in winemaking to bind and remove some wine proteins through precipitation or acts as a clarifying agent. Winemakers should note that there is a lot of variation between bentonite products and suppliers, and some products may be more effective compared to others. Winemakers should also ensure that the bentonite they are using is strictly for wine additions.
Bentonite is negatively charged, and will, therefore, bind with positively charged proteins in wine. It is possible to treat a wine with bentonite and still have protein stability problems by:
- A concentration effect in which case all of the proteins in the treated wine have not been bound by bentonite.
- A charge effect in which negatively charged proteins have not been bound and, therefore, remain in the wine.
In contrast, the fining agent Sparkolloid offers alternatives to winemakers as it is primarily positively charged and will bind to negatively charged proteins. Winemakers are always recommended to do bench trials before treating a wine with any fining agent.
Like many fining agents, bentonite may have some alternative side effects including aroma/flavor and color stripping. While it is sometimes preferred to add bentonite without testing a wine for protein stability, winemakers run several risks towards the wine including:
- No knowledge towards the protein stability of the wine even after treatment of bentonite. (Keep in mind that some wines require 10+ pounds per 1,000 gallon additions of bentonite.)
- Unnecessary stripping of aroma/flavor composition or wine color when there is no protein instability issue in the wine. Depending on the wine, this can come as a detriment to wine quality.
- Potential off-flavor development in aroma-neutral varieties.
- Economic losses for the winery if unnecessary additions of bentonite are made to a wine without protein instabilities (i.e. waste of product). Or a waste of resources if the wine hazes in the bottle even after a bentonite addition was made during production. If no analytical evaluation to reassure the winery that the addition was successful at stabilizing the wine, the time and cost affiliated with un-bottling, treating, and re-bottling wine can be significant. [Note: Protein stability tests are not a 100% guarantee, but provide a winemaker with more information than guessing a wine’s protein stability]
- Unnecessary use of employee time for fining if the wine does not require a bentonite addition.
Practical Directions for Wineries
Wineries should consider routine analysis for protein stability to avoid guesswork affiliated with bentonite additions. Some wineries may find they are making unnecessary bentonite additions or find a need for making larger bentonite additions. After a fining treatment, wines should be re-evaluated for protein stability to ensure that the treatment has worked properly.
When adding bentonite to any wine, it is recommended that the fining agent be hydrated in warm, chlorine-free water. Bentonite additions should be thoroughly mixed into a wine for approximately 15 minutes, and racked after a week’s settling time. Most wine suppliers will provide preparation and use instructions for wineries unfamiliar with bentonite treatments.
More Practical Information
This article does not touch upon a lot of the theory, chemistry, preparation of bentonite, and wine treatment for protein stability issues. For more information available online, please consider the following resources:
“Fining Agents for Use in Wine” from The Wine Lab
Wine Analysis and Production by Bruce W. Zoecklein et al. (ISBN: 0-8342-1701-5)
Winemaking Problems Solved, edited by Christian E. Butzke (ISBN: 978-1-84569-475-3 or 978-1-4398-3416)