Harvest Guide 2021

HARVEST GUIDE E T S L A B O R AT O R I E S | 2 0 2 1

pages 04-05 Maturity Monitoring The ETS Grape Maturity Monitoring Panels Panels provide sets of analyses requested to monitor fruit maturity.

CONTENT

pages 06-17

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ETS Juice Panel Get the complete picture for informed winemaking with the harvest Juice Panel.

page 18 Measuring Proteins in Juice ETS has developed Enzyme-Linked Immunosorbent Assays (ELISA) to monitor proteins associated with heat instability.

pages 19-20 Scorpions Find out what microbes are coming in on your grapes using Scorpion diagnostics.

page 21 Non-Saccharomyces Yeasts Diagnostics The PCR-based diagnostics can be used to determine implant success with commercial strains of non-Saccharomyces yeasts

pages 22-23 Volatile Acidity Recognize the microbes and conditions that lead to VA formation to develop an effective monitoring and prevention program.

page 24 Potential Alcohol Get a closer look at using glucose+fructose analysis to estimate potential alcohol.

page 25 Sugar Analysis Sugar can mean a number of things. See what's behind your "Residual Sugar" numbers.

pages 26-29

Phenolics Program ETS offers a full suite of advanced HPLC-based analytical tools to evaluate phenolic compounds in grapes and wines.

pages 30-31 Aromas Detect and prevent common (and uncommon) sensory flaws.

pages 32-39 The Impact of Wildfires ETS offers an extended panel for volatile smoke markers and a glycosylated markers panel this harvest.

pages 40-43 Wildfire Impact FAQ Your most asked questions, answered.

pages 44-45 Wine Authenticity at ETS Nuclear Magnetic Resonance (NMR)

pages 46-51 Harvest Toolkit This short guide will give you the highlights of our most requested Harvest testing.

page 53 Harvest Satellite Analysis Use this quick reference to see which Juice Analyses are offered at your satellite lab.

page 54 Newberg Courier Service We know you're busy, so let us come to you for your sample pickup and supplies drop off needs.

page 55 The Vintage Portal Identify vintage trends in juice chemistry and grape phenolic development in real time.

pages 56-57 Sampling and Shipping Make sure you get the most out of your results using these sampling guidelines.

pages 58-63 Our Locations p. 59- St. Helena

p. 62- Newberg

p. 60- Healdsburg

p. 63- Walla Walla

p. 61- Paso Robles

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MATURITY

MONITORING THE ETS GRAPE MATURITY MONITORING PANELS AND SUGAR PER BERRY PANELS

WHAT A R E T H E S E PA N E LS ?

The ETS Grape Maturity Monitoring Panels Panels provide sets of analyses requested to monitor fruit maturity.

They include the traditional measurements of juice solids (Brix) and acidity (Titratable Acidity and pH), enhanced by the addition of a more accurate determination of fermentable sugars (glucose + fructose) and malic acid, as the degradation of this organic acid is a well-known marker of ripening.

They also include berry size parameters (volume and weight), as well as a less familiar measurement: sugar per berry. Sugar per berry is a calculation based on the average berry volume as measured by Dyostem®, the tool of choice to determine sugar on a per berry basis. The Dyostem® also provides an assessment of

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berry volume variability expressed as a Coefficient of Variation (%) and a histogram of the berry volume distribution is provided in our reports.

WHY M ON I TORI N G S U GAR P ER BERRY? Monitoring Sugar per Berry, starting shortly after veraison, allows growers to determine the duration and rate of sugar loading. During this phase, vines synthesize and actively transport sugar into berries. At the end of this phase, Brix usually keeps increasing due to berry dehydration, however, and simply monitoring Brix cannot determine when the sugar loading phase stops. Conversely, when this point is reached, the actual quantity of sugar accumulated in each berry remains unchanged (see Fig 1).

ACTIVE SUGAR LOADING PERIOD

VERAISON

NO MORE ACTIVE SUGAR LOADING

DAY «0 »

(PHYSIOLOGICAL RIPENESS)

The duration of the sugar loading phase, the time at which this phase ends, as well as the Brix achieved at that time, are all important indicators of wine growing conditions.

Excessive or insufficient vigor, water availability and resistance to heat stress all have an impact. Vines under often desired moderate hydric stress conditions typically reach between 21 and 23 Brix at the end of sugar loading.

In most cases vines with excessive vigor see their sugar loading stop before these levels are reached, as vegetative growth may compete with sugar synthesis and accumulation. Such vines also often poorly resist heat waves and tend to “shut down” more easily, which can prompt a sudden end of sugar accumulation in berries.

At the other end of the spectrum, vines under excessive water stress, subject to nutrient deficiencies or diseased, also have difficulty reaching typical Brix levels at the end of sugar loading (see Fig 2).

Furthermore, the end of sugar loading triggers a variety of maturity events influencing the development of grape aroma compounds and phenolics. It is often used as an indication of when to start grape phenolics measurements (see Vineyard Decisions – Grape Phenolic Panel p. 27).

P OT E N T I AL D EGR E E AT S U GAR LOAD I N G S TO P ( % VO L . )

SUGAR QUANTITY PER BERRY (MG)

FIG 1: THE SUGAR LOADING CONCEPT

FIG 2: SUGAR LOADING & VINE WATER STATUS MERLOT 2012, PAUILLAC 14

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M O D E R AT E WAT E R RESTRICTION

12

N O WAT E R RESTRICTION

11

NUTRITION DEFICIENCIES ( N , K)

WATE R S TRE SS

H I GH YI E L D 10

0

5 1 2 3 4 6 7 8 P R E DAW N L E AF WAT E R P OT E N T I AL D U RIN G RIPEN IN G ( B AR , AB S O LU T E VALU E )

Working Together: ETS is proud to be par tnered with Fruition Sciences and Vivelys (D yostem ®), to help you better characterize vineyards, closely monitor grape ripening, and make the most informed har vest decisions.

IN THE LABORATORY: A LOOK AT OUR DYOSTEM I N DUST RIA L N UM E RIC CAM E RA P C + TO U CH S CR EEN

L ED L I G H TS

A NA LYSIS P L AT E 5

ETS JUICE

P A N E L

Winemakers rely on juice chemistry analysis for a more complete picture of must composition at harvest that goes beyond traditional TA, pH, and °Brix. This is critical as juice chemistry can be different from vintage to vintage. Combining modern tools gives vital insights to make informed vineyard management decisions, choose harvest dates, predi ct/adjust wine composition and facilitate fermentations.

A PPLY ING JUICE C H E M I S T RY TO WI NE M A K I N G A thorough analytical picture gives winemakers the ability to better predict their wine composition, and plan appropriate winemaking strategies in response to changing must compositions. Whether you analyze your free-run juice, monitor your mid-fermentation chemistry, or do both, it is important to understand the analytical results in context within the fermentation process stage.

Analyzing juice pre-fermentation provides data that gives winemakers the opportunity to identify anything unusual about the current vintage, and to compare differences in composition from vintage to vintage. These insights affect winemaking decisions including acid adjustments and fermentation strategy.

U SING JUICE ANALYS I S TO P RE D I C T W I N E COM POSITION Clients sometimes ask why they see a difference in the concentration of acids, or potential alcohol vs. final alcohol, when comparing their juice samples to mid- to end-stage fermentation samples.

It is not unusual to observe differences between the levels of acids, potassium and sugar/potential alcohol in different samples from the same vineyard. Variations can occur depending on the vineyard sampling strategy and how representative the samples are of the vineyard.

How juice samples are prepared matters, e.g. how thoroughly each grape sample is crushed and mixed. In white winemaking, the differences in composition between free run juices and the different press fractions are well known.

In red winemaking, free-run juices obtained after filling tanks may not reflect the actual content of the tank, since components such as acids and potassium can initially be sequestered at high levels in grape tissue next to the skin. As the grape tissue breaks down during cold soak, fermentation, and maceration, the resulting extraction of acids and potassium from the tissue into the juice can contribute to the observed differences.

Likewise, the sugar in raisins or shriveled grapes may take a long time to release during red winemaking, causing an underestimate of fermentable sugar and therefore potential alcohol. Mid-point analysis on the fermenting wine (analyzing glucose+fructose and ethanol) may give a more accurate picture of fermentable sugar and potential alcohol.

All of these factors can contribute to the differences observed between juice samples and the final wine composition.

C ALIFO R NIA GLO B A L DATA

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Ye a r

B rix

Glu/Fru

pH

TA

M ali c

Tar t ar i c

Pot as s i um

A m m oni a

N OPA

YAN

2019

24.0

2 49.5

3.62

5.0

2.14

4.5

1656

61

122

175

2018

24.2

2 52.5

3.53

5.7

2.88

5.3

1563

56

110

156

2017

24.5

2 54.0

3.62

5.3

2.19

5.1

1859

69

120

174

2016

24.6

2 58.2

3.59

5.4

2.38

4.5

1690

69

147

203

2015

24.8

2 62.9

3.56

5.5

2.31

5.0

1748

75

120

180

2014

24.7

2 62.5

3.60

5.1

2.14

4.9

1665

66

112

166

C A LIFOR NIA R ED G LO B A L DATA Ye a r

B rix

Glu/Fru

pH

TA

M ali c

Tar t ar i c

Pot as s i um

A m m oni a

N O PA

YAN

2019

25.1

261.0

3.73

4.5

1.92

4.16

1778

53

1 13

156

2018

25.4

265.5

3.64

4.9

2.34

4.20

1654

45

95

131

2017

25.4

264.0

3.73

4.7

1.93

4.73

2011

62

1 08

153

2016

25.6

269.2

3.69

4.9

2.22

4.15

1845

66

1 35

189

2015

26.0

276.3

3.69

4.9

2.23

4.54

1943

65

1 08

160

2014

25.9

275.4

3.74

4.4

1.97

4.39

1805

58

99

145

C A LIFOR NIA W H I T E G LO B A L DATA Ye a r

B rix

Glu/Fru

pH

TA

M ali c

Tar t ar i c

Pot as s i um

A m m oni a

N O PA

YAN

2019

23.0

238.1

3.51

5.5

2.36

4.8

1535

69

1 31

188

2018

23.1

239.5

3.42

6.4

3.42

6.4

1472

67

1 25

180

2017

23.6

244.0

3.52

5.9

2.46

5.47

1707

76

1 32

194

2016

23.6

247.3

3.48

5.9

2.53

4.87

1534

72

1 58

217

2015

23.7

249.8

3.43

6.1

2.39

5.39

1553

84

1 31

200

2014

23.6

249.5

3.46

5.8

2.31

5.39

1525

75

1 25

187 7

A N A LY Z I N G WINE DURING F E R M E N TAT I O N Because the must components are in a state of flux from cold soak to post malolactic fermentation, many winemakers prefer to make incremental adjustments rather than rely on one initial or massive adjustment.

Winemakers who are targeting a certain TA and pH or ethanol level, for instance, often check their wine chemistry again at the fermentation midpoint.

These mid-point numbers are used to make ongoing and final fermentation adjustments.

It's important to note that in ripe fruit, glucose + fructose numbers often appear higher than the corresponding °Brix results. This is because °Brix is measured as a percentage by weight, meaning brix values are greatly influenced by the density of juice. Glucose + Fructose is measured as weight by volume and is independent of juice density. A must with 23.3 °Brix will not have 23.3% by volume fermentable sugar.

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S U G A R CO N T E N T °Brix is not a true measure of fermentable sugar. Two juices with identical °Brix may have ver y different fin al alco hol concentrations due to var ying amounts of fermentable sugars.

Sugar concentration increases rapidly in grapes as they mature.

This increase is usually due to sugar movement from the leaves to the fruit. During the final stages of berry development, berry dehydration may also contribute significantly to the final sugar concentration. °Brix is a measure of soluble solids in juice and must. The soluble solids in grape juice are primarily sugars. Organic acids, however, have a significant impact on brix, especially with unripe grapes. °Brix is used as an estimate of sugar

concentration and often as a predictor of potential alcohol, but is not a true measure of fermentable sugar. Two juices with identical °Brix may have very different final alcohol concentrations due to varying amounts of fermentable sugars. Observations over the last five vintages indicate that the amount of glucose + fructose per degree brix varies slightly from vintage to vintage. This can have a significant impact on potential ethanol predictions based solely on Brix.

The sum of glucose + fructose measures the two main sugars present in juice that can be fermented by yeast. This analysis provides a sound basis for estimates of potential ethanol in the wine. This additional analysis is an important supplement to °Brix testing when final ethanol predictions are critical.

Glu + F r u ( g /L): D egr e e B r i x R at i o, Wh i t e Va r i et a l s

Glu + F r u ( g /L): D e gr e e B r i x R at i o, R e d Va r i et a l s

*Due to the wildfires, 2020 Juice Panel data is not included

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NITROGEN CO M P O U N D S

Yeast assimilable nitrogen includes both alpha amino nitrogen (NOPA) and ammonia.

Analysis of only alpha amino nitrogen or only ammonia nitrogen does not provide an accurate indication of total nitrogen status for a given must.

Ammonia and NOPA can change independently of each other resulting in different ratios of these two YAN components.

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mg/ L

Knowledge of nitrogen status is critical for effective fermentation management.

Nitrogen compounds are essential macronutrients for yeast, and are required for cell growth, multiplication, and yeast activity.

As with other juice chemistry components, YAN values fluctuate vintage to vintage due to changes in ammonia and/or NOPA.

YA N Differenc es in Red Varietals by Vintage

mg/L

Sluggish and stuck fermentations, coupled with sulfide formation, have become increasingly common and are often associated with deficiencies of yeast assimilable nitrogen in the must. However, excessive concentrations of certain nitrogen compounds have been associated with microbial spoilage and other fermentation problems.

YA N Differenc es in White Varietals by Vintage

AMMONIA

m g/L

Ammonia Differenc es in Red Varietals by Vintage

mg/L

Ammonia Differenc es in White Varietals

by Vintage

Ammonia is the form of nitrogen nutrition most easily assimilated by yeast.

Wineries routinely supplement nitrogen deficient musts with diammonium phosphate at the start of or during fermentation to provide adequate nitrogen levels. Additional ammonia analysis and adjustments during fermentation may also be beneficial in minimizing the risk of stuck fermentations and sulfide formation. Ammonia results are expressed as mg NH3 per liter. These values may be expressed as nitrogen equivalents by multiplying NH3 results by 0.82.

*Due to the wildfires, 2020 Juice Panel data is not included

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N O PA Alpha amino nitrogen, otherwise referred to as “Nitrogen by OPA”, or NOPA, is determined using a method specific for alpha amino groups. It is a measurement of primary amino acids usable by yeast. NOPA does not include proline, which is not utilized by yeast, or ammonia. NOPA results are expressed as mg nitrogen per liter.

N OPA Differenc2014-2019 es in White NOPA Varietals by Vintage 250

mg/L

200

CA 2014

150

CA 2015 CA 2016

100

CA 2017 CA 2018

50 0

12

CA 2019 Pinot Gris

Chardonnay

Sauvignon Blanc

Riesling

Viognier

mg/L

N OPA Differenc es in Red Varietals by Vintage

*Due to the wildfires, 2020 Juice Panel data is not included 13

ACID BALANCE The acid composition of must is a complex balance of free hydrogen ions, acids, acid salts, and cations.

Concentrations of these various components and their interactions influence many winemaking parameters. Vintage variations in any of the components that impact acid balance can result in unexpected changes in the final pH and TA of a wine.

The principal objective of acid management is to achieve and maintain a pH favorable to optimum wine balance and stability.

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TA R TA R I C AC I D Tartaric acid is one of the two major organic acids found in grapes. It accumulates in grape tissue early during development and declines during ripening due to berry growth and dilution.

Tartaric acid is not usually metabolized in grapes. It is present in grapes, must, and wine as a free acid and weak acid-salt complex.

Tartaric acid-salts may precipitate, primarily as potassium bitartrate and calcium tartrate.

An increase in the ratio of the free tartaric acid to the tartaric acid salts will cause a decrease in pH. This will affect the flavor, balance, and stability of the final product.

Tartaric acid is commonly used to adjust the acid balance of juices and wines. Understanding tartrate interactions is important in designing appropriate acidification strategies.

Both the formation and solubility of salts are affected by a balance of components that are in flux throughout the early life of a wine.

Tar taric Differenc es in White Varietals by Vintage

g /L

g /L

Tar taric Differenc es in Red Varietals by Vintage

PH pH is a measure of free hydrogen ions in solution (which corresponds to the chemical definition of acidity) and is used as a gauge of wine acidity.

Wine color, potassium bitartrate stability (cold stability), calcium stability, and molecular SO2 level are directly related to wine

pH Differenc es in White Varietals by Vintage

pH. pH is also critical in relationship to microbial stability, interactions of phenolic compounds, and color expression.

Overall, the 2018 vintage had lower pH as compared to the 2017 vintage.

pH Differenc es in Red Varietals by Vintage

*Due to the wildfires, 2020 Juice Panel data is not included 15

M A L I C AC I D

Malic acid is converted to lactic acid during malolactic fermentation, causing the loss of an acid group. The effect of this

acid reduction on pH depends upon the initial amount of malic acid and buffer capacity of the wine. Malolactic fermentation in wines containing low levels of malic acid and high buffer capacity will have little impact on wine pH. Malolactic conversion in wines with high malic acid and low buffer capacity can result in a substantial pH increase. Malic acid tends to be higher in cooler vintages. That said, 2018 had the highest malic acid levels for all varietals except Syrah as compared to the previous four vintages.

M alic Differenc es in White Varietals by Vintage

M alic Differenc es in Red Varietals by Vintage

g /L

g/L

Malic acid accumulates early in berry development and declines during ripening due to dilution and respiration. Viticultural practices and grape cluster environments may directly affect respiration rates of malic acid. Malic acid levels affect pH and titratable acidity.

T I T R ATA B L E AC I D I T Y Titratable acidity (TA) measures total available hydrogen ions in solution. This measurement includes both the free hydrogen ions and the undissociated hydrogen ions from acids that can be neutralized by sodium hydroxide.

TA is the most widely used measurement of acidity in wine.

Although generally considered a simple parameter, titratable

TA Differenc es in White Varietals by Vintage

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acidity is actually a reflection of complex interactions between the hydrogen ions, organic acids, organic acid-salts, and cations in solution. Often there is no direct correlation between TA and pH. Two musts with similar titratable acidity may have very different pH values.

TA Differenc es in Red Varietals by Vintage

P O TA S S I U M

Potassium moves into cells in exchange for hydrogen ions from organic acids. Potassium concentration is highest near the

grape skin. Crushing, skin contact, and pressing all influence potassium levels. Potassium is a critical factor in pH, acid salt formation, tartrate precipitation, and buffer capacity. Decreases in juice potassium are associated with decreased juice pH. For example, 2018 vintage California Cabernet Sauvignon harvested in November, had approximately 22% lower potassium values as compared to 2017. The pH for the 2018 vintage averaged 3.78 versus 3.95 for the 2017 vintage.

Potassium Differenc es in White Varietals by Vintage

Potassium Differenc es in Red Varietals by Vintage

mg/L

m g/ L

Potassium is the primary cation present in grape tissue.

Potassium concentration in the berry is a function of root uptake and translocation. Both are strongly affected by viticultural factors including choice of rootstock, potassium fertilization, and canopy management.

*Due to the wildfires, 2020 Juice Panel data is not included

CO N C L U S I O N S Juice chemistry is the foundation for the resulting wine.

Producing wines with specific targets for ethanol and acid balance, while avoiding fermentation-related problems, requires a thorough understanding of the juice chemistry.

Individual components of the juice can and do change from vintage to vintage. Winemakers aware of these changes will be able to adjust their winemaking process, where necessary, to achieve their targeted wine styles.

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Measuring Proteins in Juice and Fermentation Samples ETS has received an increasing number of requests to develop a method for determining the efficacy of bentonite fining in juice or during fermentation. Traditional Heat Stability tests can be difficult to perform on juice and fermenting samples. ETS has developed methods to directly quantify two proteins produced

by Vitis vinifera in response to biotic and abiotic stress. These proteins, chitinase and thaumatin-like protein (TLP), are very resilient and persist through the fermentation process. Both proteins have been implicated in haze formation although chitinase may play a larger role in heat stability issues. ETS has developed Enzyme-Linked Immunosorbent Assays (ELISA) to detect the individual proteins. Extensive testing last year indicates a linear response between addition of bentonite and removal of both chitinase and TLP. The ELISA based analyses are applicable to monitoring bentonite fining of juice, as well as bentonite fining during fermentation.

Chit inase and N TU R e sp o n se t o B e n t o n i t e A d d i t i o n i n J u i c e 70.0 60.0 50.0 40.0 30.0

Chit inas e (ug/L )

N TU

20.0 10.0 0.0

0

1

4

2 3 Be n t o n i t e ( LB S / K G A L)

5

Response of C hit inas e a n d T LP t o B e n t o n i t e A d d i t i o n i n J u i c e 14.0 12.0 10.0 8.0 6.0

Chit inas e

TL P

N TU

4.0 2.0 0.0

0

1

2 3 4 Bent o n i t e ( LB S / K G A L)

5

Juice Protein Panel • • 18

Chitinase Thaumatin-Like Protein

JUICE SCORPIONS Indigenous microb es com i ng i nt o t h e wi ner y on fruit is one of the m os t i m por t ant ent r y rout es for spoila ge orga nisms t h at c an c aus e s t uc k and s luggi s h fermenta tio ns a nd VA problem s .

Identifying and quantifying yeast and bacteria that can cause spoilage during the winemaking process is the first step in preventing these spoilage problems.

Using Scorpions to see the full picture of spoilage microbes in the juice from incoming fruit gives the winemaker better situational awareness for potential problems that can occur during the cold soak process, in stuck or sluggish fermentations, or later during wine aging.

Volatile acidity in juice Volatile acidity, measured as acetic acid, can be formed throughout the winemaking process. Both acetic acid bacteria and strains of wild yeast – particularly Hanseniaspora and Pichia – are commonly linked to volatile acidity production prior to and in the early stages of fermentation.

Elevated VA levels often occur during the cold soak process, or between cold soak and fermentation during red wine production. The VA-producing spoilage microorganisms grow quickly during this time, producing increasing levels of acetic acid until fermentation conditions inhibit their growth.

Production of high levels of volatile acidity prior to fermentation can also cause

problems later in the production process, including possible impacts on the fermentation performance and wine sensory attributes.

Large numbers of Acetic Acid bacteria on incoming fruit can carry through the fermentation and cause problems with VA production when exposed to air during barrel aging.

Effects on fermentation In addition to causing sensory impacts, large populations of wild yeast can deplete the YAN in the must, resulting in a YAN deficiency for the Saccharomyces cerevisiae driving the fermentation. Winemakers who detect high levels of Hanseniaspora or Pichia in a must usually recheck YAN before yeast inoculation, and supplement YAN if necessary.

Likewise, if the Scorpions assay detects heterofermentative lactic acid bacteria, such as Lactobacillus brevis, L. kunkeei, L. hilgardii, L. fermentum, and Oenococcus oeni, in a juice, winemakers usually increase their monitoring of malic acid and microbe levels if the fermentation becomes sluggish or stuck. Early identification of the presence of these bacteria and recognizing the risk they pose to difficult fermentations is key to preventing VA formation in stuck fermentations.

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W H AT W E ' R E LOOKING FOR...

HANSENIASPORA

Hanseniaspora (Kloeckera) is a wild apiculate yeast that is often present at high levels on incoming fruit. Hanseniaspora can initiate fermentation in the must and produce high levels of volatile acids, including acetic acid ACETIC ACID and ethyl acetate. It has been associated PICHIA BACTERIA with acid rot in grapes infected by Acetic acid bacteria are Botrytis cinerea. Population levels Pichia is a wild yeast that is often commonly associated with grapes usually decline as alcohol present at high levels on incoming and the winery environment. The concentration increases. fruit. Pichia can initiate fermentation, three groups of commonly detected resulting in production of high levels of acetic acid bacteria are Gluconobacter, volatile acids, including acetic acid and ethyl Gluconacetobacter and Acetobacter. Both acetate. These yeast have been associated Gluconacetobacter and Acetobacter can with fflms formed in barrels and tanks generate acetic acid from ethanol in the during storage. presence of oxygen. The presence cause can of these organisms elevated volatile acidity in wines exposed to air.

Z YG O S A C C H A R O M YC E S MANY WINERIES AROUND THE WORLD USE GRAPE C O N C E N T R AT E I N T H E P R O D U C T I O N O F T H E I R W I N E S . A LT H O U G H G R A P E C O N C E N T R AT E I S N O T T H E O N LY S O U R C E O F Z YG O S A C C H A R O M Y C E S Y E A S T, I T I S C E R TA I N LY A C O M M O N S O U R C E F O R Z YG O S A C C H A R O M YC E S T O B E I N T R O D U C E D I N T O T H E W I N E R Y. The primary species of Zygosaccharomyces typically associated with grape concentrate are Zygosaccharomyces bailii and Zygosaccharomyces bisporus.

Recently, a different species, Zygosaccharomyces rouxii, was isolated from a fermenting concentrate sample provided by a client. In response to this finding, we modified the design of the Zygosaccharomyces Scorpions primer/probe combination to detect more species. The new Zygosaccharomyces Scorpions diagnostic detects an additional 7 species of Zygosaccharomyces for a total of 9 species.

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W H AT W E ' R E L O O K I N G FOR...

Zygosaccharomyces bailii

Zygosaccharomyces kombuchaensis

Zygosaccharomyces bisporus

Zygosaccharomyces parabailii

Zygosaccharomyces rouxii

Zygosaccharomyces pseudobailii

Zygosaccharomyces lentus

Zygosaccharomyces pseudorouxii

Zygosaccharomyces mellis

N O N - S A CC H A R O M YC E S

YEASTS DIAGNOSTICS Several providers of commercial yeast to the wine industry are offering strains of non-Saccharomyces yeast for use in the winemaking process. These yeasts can be used as bioprotective agents to reduce the impact of indigenous non-Saccharomyces strains as well as a way to reduce SO2 use prior to fermentation. In addition, these yeast strains may provide improvements via increased aroma complexity, wine structure and mouthfeel.

In response to the increased use of these yeast strains by our clients, ETS has developed a series of PCR-based diagnostics to detect the presence of the individual strains; Metschnikowia pulcherrima, Lachancea (Kluyveromyces) thermotolerans, Torulapsora delbrueckii, and Pichia kluyveri.

The PCR-based diagnostics can be used to determine implant success with the individual strains. They can also be used with the ETS Juice Yeast Scorpion Panel to look at the efficacy of these strains at reducing levels of Hanseniaspora uvarum and Pichia membranifaciens, commonly observed in must samples. The diagnostics can be requested for individual yeast species, or as a complete panel.

W H AT W E ' R E L O O K I N G FOR...

Non-Saccharomyces Yeast Panel •

Metschnikowia pulcherrima

•

Lachancea (Kluyveromyces) thermotolerans

•

Torulapsora delbrueckii

•

Pichia kluyveri

Individual yeast analyses •

Metschnikowia pulcherrima

•

Lachancea (Kluyveromyces) thermotolerans

•

Torulapsora delbrueckii

•

Pichia kluyveri

21

VOLATILE ACIDITY 22

The two components commonly associated with "VA taints " , Acetic Acid and Ethyl Acetate, can be formed by both yeast and bacteria - and in the case of bacteria, can be formed with or without oxygen present.

VOLATILE ACIDITY (VA)

ETHYL ACETATE

is strictly speaking a measure of the volatile acids in wine, although in the real world the contribution of volatile acids other than acetic acid is negligible. VA is a normal component of wines at moderate levels (normal concentrations range from 0.3-0.9 g/L of acetic acid), but very quickly becomes undesirable as levels rise. The sensory threshold is around 0.91.0 g/L, depending on the style of wine, and the U.S. government sets legal limits of 1.2 g/L in white wine and 1.4 g/L in red wine.

Although not an acid, ethyl acetate is considered by some to be a component of VA. As an ester formed by ethanol and acetic acid, it is often linked to increased production of VA. From a sensory point of view, ethyl acetate is often classified as a “VA taint”, and its "nail polish remover” odor is often a telltale sign of high VA. Similar to acetic acid, there is a fine line between complexity and spoilage: small amounts of ethyl acetate can contribute “fruitiness/sweetness” or other positive characteristics to a wine at low levels.

Normal concentrations are usually less than 100 mg/L, while the sensory threshold is generally 130150 mg/L, depending on the wine style.

COLD SOAK / EARLY STAGE FERMENTATION Acetic Acid can be produced prior to fermentation by Acetic Acid Bacteria and wild yeast in compromised fruit.

It’s unusual to see alcohol present in juice before fermentation, but if clusters experience fungal rot or other types of damage (such as bird or insect damage), wild yeast in the vineyard can begin fermenting the juice that is leaked out. Acetic Acid bacteria can then convert the alcohol to acetic acid, causing “sour rot” and leading to high VA levels before fermentation has even begun.

Ethyl Acetate is often produced in the early stages of fermentation, and can be a particular problem in native fermentations with a slow start. Native yeast, especially Hanseniaspora, are the main Ethyl Acetate producers at this stage. Note that the Hanseniaspora can consume most or all of the YAN in the must very early in the fermentation, although they will only produce alcohol up to around 6%. High levels of Hanseniaspora and low YAN concentrations can contribute to stuck fermentations.

PRIMARY FERMENTATION Acetic acid production in primary fermentation is generally caused by yeast, including Saccharomyces and other species, but can also be formed by bacteria.

The native yeasts Hanseniaspora and Pichia can drive fermentations up until around 6-7% alcohol, at which point they become stressed by the alcohol.

Saccharomyces is more competitive as it is tolerant to and produces higher alcohol levels. In certain situations, the native yeasts respond to the changing fermentation conditions by producing elevated levels of acetic acid.

Ethyl Acetate production during fermentation is significantly impacted by the yeast strain and fermentation temperature. Although Saccharomyces cerevisiae will produce ethyl acetate, research has indicated that some of the Saccharomyces bayanus strains are more likely to form ethyl acetate in cold fermentations.

Bacteria, usually Lactobacillus, can also generate acetic acid from sugar and can often produce high levels of VA in stuck and sluggish fermentations.

Oenococcus oeni, the bacteria used for inoculating most malolactic fermentations can also produce acetic acid from fructose. This frequently occurs at the end of malolactic fermentation if there is still fermentable sugar remaining in the wine.

23

24

POTENTIAL ALCOHOL

The amount of fermentable sugar (glucose and fructose) in juice and the average conversion rate of sugar into alcohol can be used to predict the potential alcohol level in wines.

Q: HOW ACCURATE ARE POTENTIAL ALCOHOL ESTIMATES? A: Our clients have reported that glucose + can be a challenge and can affect potential fructose values improve the quality of their predictions, but it is important to remember that yeast populations and fermentation conditions vary, and any prediction of potential alcohol is only an approximation.

Alcohol conversion ratios can be variable, so it is possible your actual alcohol may be lower or higher than the estimate.

Many of our clients have found that the conversion rates observed for their own yeasts and fermentation conditions remain relatively constant, and they use their internally observed conversion rates to calculate potential alcohol content based on their glucose + fructose values.

With white wines, predictions are usually fairly accurate. With red wine, however, getting a truly representative juice sample

alcohol predictions. A juice sample taken soon after a tank is filled may not take into account un-popped berries, unripe berries (less sugar and more acids), and raisins (sometimes an overlooked source of large amounts of sugar, acid, and potassium).

We suggest sampling after an initial 10°Brix drop, and analyzing the fermenting sample for glucose + fructose and alcohol simultaneously for a more accurate potential alcohol estimate. Proper sample preparation is key to accurate results. At ETS, juice samples are centrifuged before analysis, and then mixed by inversion to avoid stratification, ensuring the most accurate results. Particulates have a minimal impact on refractometry, but can have a large impact on densitometry results.

Q: WHAT'S THE BEST WAY TO PREDICT POTENTIAL ALCOHOL LEVELS? A: Predicting

potential alcohol levels in finished wines sounds simple, but there is more than one way to measure “sugar”, and formulas to convert this sugar into potential alcohol often miss the mark.

either hydrometers or digital instruments), and other secondary measurements. The differences among the various measurement techniques are quite unpredictable depending on sample composition.

The “old school” method was to multiply °Brix by 0.6. One degree Brix is defined as 1 gram of sucrose in 100 grams of aqueous solution.

However, grape juice does not naturally contain sucrose, but rather glucose, fructose, and a variety of organic acids and other dissolved solids. So when used for grape juice, °Brix is actually just an approximation of dissolved sugar, not an accurate representation of the fermentable sugars, and using ºBrix for estimating potential alcohol adds an additional layer of uncertainty to alcohol predictions. Differences between ºBrix and actual fermentable sugar content are even more pronounced in high ºBrix fruit and in fruit affected by fungal growth.

A more modern calculation that has proven to be more accurate uses glucose+fructose analysis, which provides a more accurate measurement of the levels of fermentable sugar compared to using ºBrix.

How ºBrix is measured also has an influence. Differences exist between ºBrix by refractometry, densitometry (using

Note that in ripe fruit, glucose + fructose numbers often appear higher than the corresponding ºBrix results, because ºBrix is measured as a percentage by weight, meaning ºBrix values are greatly influenced by the density of the juice, while glucose + fructose is measured as weight by volume and is independent of juice density.

An official conversion rate formula used in Europe is: Potential Alcohol (% vol) = glucose + fructose (g/L) / 16.83. In practice, rounding the 16.83 conversion factor to 17 is common.

In the wine industry, a term like " sugar " can mean different things. Clients often request testing for "RS" , but this term can be very ambiguous.

REDUCING SUGAR

°Brix is a measurement of the apparent concentration of sugar. It is commonly used for grape juice and must and is expressed as a percentage by weight (% w/w). One degree Brix is defined as 1 gram of sucrose in 100 grams of aqueous solution. When the solution contains dissolved solids other than pure sucrose, as is the case for grape juice and must, the °Brix is only an approximation of dissolved sugar.

Historically, “Residual Sugar” was measured by the Reducing Sugar method. This test derives its name from the ability of most sugars in juice or wine to ‘reduce’ other compounds. The most common reducing sugars are glucose and fructose. However, the method does not distinguish between fermentable and non-fermentable sugars, or other ‘reducing’ compounds for that matter, and these other compounds may contribute to reported results.

GLUCOSE + FRUCTOSE In grape juice, Glucose + Fructose analysis measures the combined concentrations of the two main sugars present that can be consumed by yeast, also known as "fermentable sugars." Compared to °Brix, Glucose + Fructose can provide a better estimate of potential alcohol concentration after fermentation. In wine, “residual sugar” usually refers to the sum of Glucose + Fructose, an indication the amount of fermentable sugars remaining post fermentation, which is also an indication of ‘dryness’.

Sucrose is not captured by this test. If it has been used in the winemaking process (such as for chaptalization of must, secondary fermentation of sparkling wine or added as a sweetener) measurement of Glucose + Fructose alone is usually not adequate – instead see Glucose + Fructose (Inverted)

Because of these limitations, the Reducing Sugar method is no longer the preferred choice to monitor completion of primary fermentation.

GLUCOSE AND FRUCTOSE PANEL The Glucose and Fructose Panel provides the individual levels of glucose and fructose, in addition to their combined concentration.

This test is often requested to investigate or remedy stuck or sluggish fermentations.

GLUCOSE + FRUCTOSE (INVERTED) Inverted Glucose + Fructose provides the sum of the concentrations of glucose and fructose after “inversion” of the sample.

Inversion is a process by which sucrose is broken apart into glucose and fructose, so that it can be measured and included in the reported results. Hence, this test is useful when “Residual Sugar” is required after chaptalization of must, secondary fermentation of sparklings or whenever sucrose has been used as a sweetener in wine, other alcohol beverages or spirits.

SUGAR ANALYSIS

°BRIX

25

BUILDING A PHENOLICS PROGRAM 26

ETS offers a full suite of advanced HPLC-based analytical tools to evaluate phenolic compounds in grapes, juice, fermenting must and wine.

The range of phenolic analyses allows flexible use and implementation to suit individual needs.

VI N E YA RD DECISIONS

GRAPE PHENOLIC PANEL Phenolic compounds in red wine grapes are directly linked to eventual wine flavor, color and aging characteristics.

The grape phenolic panel can characterize site to site variation as well as within site differences. It works well as a prediction tool for describing vintage effects on potential phenolics and is a great tool for vineyard research projects. It is very sensitive to changes in phenolic compounds occurring during grape maturation. The

changes in grape tannin are particularly important for red wine picking decisions. The grape phenolic panel can track changes in seed ripening, skin tannin extractability and tannin modification.

Dilution and concentration effects on tannin and other phenolic components can be monitored with the Grape Phenolic Panel particularly when used in conjunction with Grape Water Content.

UN D E RS TAN DIN G R AW MATERIA LS GRAPE PHENOLIC PANEL

Successful winemaking strategies require accurate information on grape composition. Winemakers use this panel during fermentation to reach target levels of tannin for specific wine styles, to monitor seed extraction, for adjusting tannin modification through oxygenation and for decisions on extended maceration and pressing.

M A NAG IN G FE R M ENTATIO N

RAPID PHENOLIC PANEL FOR WINE Phenolic compounds are extracted from grapes during fermentation and maceration. Monitoring the phenolic composition of the must during fermentation can greatly enhance a winemaker’s control of the process.

and press timings can all be fine-tuned with feedback on changes in phenolic composition. With this information, winemakers can adapt winemaking practices to fit the vintage and successfully create wines of a target style.

Juice bleeds, fermentation temperatures, pump-over or punch down regimes, the use of rack-and-return, oxygen or air additions

S E T T I N G TARGE T S

RAPID PHENOLIC PANEL Phenolic composition is one of the main components of red wine style. The amount of tannin and its composition is the foundation of a wine’s structure. There are no “correct” values for these parameters. A winery must define their own style as a brand and for individual products within that brand.

Integrating phenolic information into stylistic choices requires an understanding of the impact of tannin on the sensory profile of wines. For a winery new to this information a good strategy

is to analyze a selection of recent products. Recent production lots, finished wines and competitor’s products are good choices.

Tasting products with analytical information allows winery staff to build the connections between taste and analytical information.

Taste is the final arbiter of style but a clear understanding of the relationship of taste and analysis is needed to turn analytical information into action.

W I N E LOT CHAR ACTERIZATIO N

RAPID PHENOLIC PANEL After the completion of fermentation/maceration, a wine lot typically represents a specific vineyard and fermentation tank.

This is an excellent point for collecting quality control data. A comprehensive review of production lots is a powerful tool for monitoring block to block variation and the effects of winemaking practices. Analysis of finished production lots early in the vintage is very useful for changing fermentation practices and targets later in the vintage.

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BL EN DI N G

RAPID PHENOLIC PANEL FOR WINE OR RED WINE PHENOLI C PROFILE Winemakers interested in consistent tannin and color levels benefit by comparing the phenolic profiles of bulk wines prior to blending. Potential blends can be compared to target phenolic levels and benchmarks prior to final blend.

BOTT LED WINE C HARAC TERI ZATI O N RED WINE PHENOLI C PROFILE

Many wineries establish QC benchmarks for phenolic content immediately after bottling. This is especially useful for determining product consistency and for monitoring wine development during aging.

F INIS HED WINE EVALUATI O N S RED WINE PHENOLIC PROFILE

A historical review of products from within a winery and evaluation of similar products from other producers is an excellent way to establish phenolic benchmarks.

This is often the first step in building an integrated program of phenolic analyses. A careful review of finished wines combined with sensory evaluation and market feedback can identify program strengths and weaknesses. The identification of desirable levels for key phenolic components creates targets that can be incorporated into process control points in the vineyard and winery.

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E T S R AP I D P HEN OLI C PAN ELS GRAPE PHENO LIC PA N EL The rapid phenolic panel for grape berries includes a 'wine-like' extraction of grape berries along with analysis of total anthocyanins, polymeric anthocyanins, quercetin, tannins and catechin in the resulting extract. It is an excellent tool for monitoring phenolics development in grapes, evaluating vineyard and vintage variation, viticultural experiments, and can help guide tannin management decisions during winemaking. Submitting samples as berries is recommended.

CO MPO NENTS NAME

UNIT

TECHNIQUE

Cate c h i n

mg /L

H PL C

Cate c h i n/ Tanni n Rat io Po ly m e r ic Ant h o c yani n s

Ca lcul at ion mg /L

Po ly m e r ic Ant h o c yani n s/Ta n n in rat io

H PL C Ca lcul at ion

Q ue rc e t i n glyc o s id e s

mg /L

H PL C

Tanni ns

mg /L

H PL C

Tot al Ant h o c yani ns

mg /L

H PL C

RA PID PHENO LIC PA N EL - W I N E The rapid phenolic panel for wine provides a snapshot of important phenolic compounds with next day results. This information can help guide decisions on adjusting fermentation and maceration processes and blending decisions and works well as a complement to the rapid phenolic panel for grape berries to monitor vineyard effects on wine phenolic composition.

CO MPO NENTS NAME

UNIT

TECHNIQUE

Cate c h i n

mg /L

H PL C

Cate c h i n/ Tanni n Rat io Po ly m e r ic Ant h o c yani n s

Ca lcul at ion mg /L

Po ly m e r ic Ant h o c yani n s/Ta n n in rat io

H PL C Ca lcul at ion

Tanni ns

mg /L

H PL C

Tot al Ant h o c yani ns

mg /L

H PL C

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EUCALYPTOL Since the discovery of eucalyptol (1,8-cineole) in red wines by ETS Laboratories in 2003, we’ve detected and measured eucalyptol in a large variety of wines exhibiting “eucalyptus-like” aromas. Flavors perceived during tasting were usually strongly related to concentrations of eucalyptol. Regardless of the grape variety, trace levels close to 1 ppb are associated with “fresh”, slightly “minty” notes. In the low ppb range, “minty” or “fresh bayleaf” aromas become stronger, and more easily identifiable as “eucalyptus” as concentrations increase. Wines with strong “eucalyptus” odors may contain more than 20ppb of eucalyptol.

Eucalyptol’s sensory impact in wine is considered more or less desirable depending on the grape variety. Wines from southern Rhône and Mediterranean varieties seem better able to accommodate the characteristic well. Moderate levels can be appreciated in wines from Bordeaux varieties. Whereas with Pinot Noir, even trace levels can detract from varietal expression.

Application: As eucalyptol may be mostly contributed by eucalyptus-derived MOG (leaves, bark debris…) hiding in grapes, analyzing grape samples before harvest is mostly pointless, and routine testing is not offered by ETS Laboratories.

ROMAS

On the other hand, wine analysis assists winemaking teams in objectively documenting their sensory impressions and managing a strong flavor component. Unlike IBMP and smoke-derived compounds (see p. 38), eucalyptol is relatively slow to extract during red winemaking, making shorter macerations a valid strategy to minimize impact. Winemakers who wish to minimize or maintain consistent levels of “eucalyptus” character will also benefit by determining eucalyptol concentrations in distinct wine lots prior to blending.

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Eucalyptol- Eucalyptus traits are considered typical in some “cult” wines; on the other hand, in excess this character can be overwhelming.

Even a slight “eucalyptus” note can interfere with delicate varietal aromas, and can have a detrimental influence on certain grape varieties.

MONITORING GLUTATHIONE Glutathione is not an aroma compound itself, but is a powerful antioxidant that protects white wines and rosés from oxidation and loss of aroma or flavor.

A low level of glutathione in grapes leads to lower levels in the juice, and early losses of aroma compounds.

Glutathione levels fluctuate during production, as the compound can be absorbed by yeast and then released after fermentation.

If final glutathione levels are low in young wines, the wines will experience faster loss of fresh varietal and fruity aromas, and poor aging potential.

Monitoring glutathione levels can be beneficial through out the winemaking process to maximize white wine aroma and flavor, and prevent premature aging.

1

The glutathione content in grapes indicates their antioxidant potential, and can be influenced by a number of factors including soil nitrogen, vineyard practices, and grape maturity levels.

2

Analyzing changes in glutathione levels during production helps to pinpoint where in the process glutathione is being lost – often from contact with air or exposure to copper residues.

3

A testing program can also identify winemaking processes that boost glutathione release after fermentation, and increase levels in wines.

IBMP Isobutylmethoxypyrazine (IBMP) is the most important methoxypyrazine, a group of molecules responsible for very distinctive vegetal aromas in Sauvignon Blanc and a variety of red wines, mainly from the Cabernet family. In Sauvignon Blanc, these compounds add an often desired “grassy” character.

In red wines however, the “green bell pepper” flavor is largely unpopular. Excessive IBMP levels lead to disappointing ratings and mixed success in the marketplace.

The “green bell pepper” flavor in wine depends primarily on IBMP levels in harvested grapes. Once grapes have been picked, IBMP levels are not easily altered by standard winemaking processes.

Application: The intensity of “green bell pepper/grassy” characters in wines can be predicted by measuring IBMP in grapes right before harvest. Grape screening of IBMP helps identify “problem” vineyards or blocks.

Since IBMP decreases during grape maturation, monitoring IBMP levels throughout ripening is a unique tool for assessing “aromatic maturity”. It allows targeting harvest dates based on desired aroma characteristics.

Monitoring IBMP from the early stages of the ripening process can greatly improve fruit quality from underperforming vineyards. Levels in grapes are well known to be linked to vine vigor, canopy and water availability, with severe heat occasionally causing IBMP’s natural degradation to stop. Once the kinetics of IBMP accumulation and degradation in specific sites are understood, viticultural practices can be modified accordingly.

Once grapes have been picked, IBMP levels are not easily altered by standard winemaking processes.

Glutathione, a natural grape antioxidant, can protect the aroma and flavor of white and rosé wines and prevents premature aging.

IBMP (2-Isobutyl-3methoxypyrazine) is the main compound responsible for the “green bell pepper” aroma in wine.

The IBMP potential of grapes can be grossly underestimated from juice samples, making whole berries the preferred sample in most cases.

Analyzing juice samples may be relevant in white winemaking, however.

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THE I M PA C T O F WILDFIRES WHAT WE'VE LEARNED, OUR NEW FINDINGS, AND HOW WE SHOULD PROCEED FOR HARVEST 2021.

32

WILD FIRE IMPAC T PANELS Volatile Markers - Basic Panel

Volatile Markers Extended Panel

Glycosylated Markers Panel

ETS has been offering the Basic Panel of volatile (“free”) markers, comprised of guaiacol and 4-methylguaiacol since 2008.

This extended panel of volatile markers complements our basic panel. In addition to guaiacol and 4-methylguaiacol (guaiacols), this extended panel measures secondary markers of impact including o-cresol, m-cresol, p-cresol, phenol, syringol and 4-methylsyringol.

In 2020, ETS further perfected a unique panel of glycosylated (“bound”) smoke markers using a state-ofthe-art combination of solid phase extraction, liquid chromatography, and triple quadrupole mass spectrometry (SPE/HPLC/MS/MS – QQQ.

Despite its apparent simplicity, this panel has an excellent track record for assessing wildfire impact with preharvest grape samples, small-scale fermentation samples (micro-ferments), and production wines before any contact with oak.

The value and efficacy of this panel was confirmed by the 2018 Mendocino Complex Fire Lake County Winegrape Commission Study, with the participation of the University of California Cooperative Extension (UCCE) and the Australian Wine Research Institute (AWRI) (1). Also, volatile guaiacol results on grape samples are typically requested by crop insurance providers.

This extended panel is available for grapes, small-scale fermentation samples (micro-ferments), and wines. Compared to the Basic Panel, additional markers allow more complete assessments of wildfire impact, and provide useful information with moderately oaked wines (e.g. from so-called “neutral” barrels, which often contribute low amounts of guaiacols making interpretation of results very difficult).

This Basic Panel was our “fall back” panel during the 2020 Harvest due to the unprecedented number of samples received during the catastrophic wildfire events. For the 2021 harvest, ETS also is offering an Expanded Volatile Markers Panel and a Glycosylated Markers Panel.

Our new Wildfire Glycosylated Markers panel includes, for each of the volatile markers listed in our Expanded Panel, its main glycosylated (sugar-bound) form – See figure 1. Note that glycosylated compounds are not contributed by toasted oak, making them particularly useful to assess wildfire impact in oaked wines.

They are not directly odor-active, but may contribute lingering aftertastes often experienced with impacted wines. It is also possible (but not yet substantiated by data) that they hydrolyze in wine, slowly releasing volatile "free" forms and causing wildfire flavors to become more noticeable with time.

ETS has worked behind the scenes with the Wine Institute Technical Committee, the AWRI and several major wineries to reach an agreement on a common list of glycosylated markers. We also joined forces to have pure reference compounds (and their isotopic analogues) synthesized for each of the markers in that list.

This goal was achieved in December 2020. These analytical standards allow extremely reproducible quantitative results comparable between laboratories. ETS has offered an updated panel employing these standards since January 2021 for wines, and will offer it for grapes and micro-ferments during the 2021 harvest season.

Volatile (“Free”) Markers

Glycosylated (“Bound") Forms

Guaiacol

Guaiacol Rutinoside

4-Methylguaiacol

4-Methylguaiacol Rutinoside

ortho-, meta- and para- Cresol

Cresol Rutinoside

Phenol

Phenol Rutinoside

Syringol

Syringol Gentiobioside

4-Methylsyringol

4-Methylsyringol Gentiobioside

Figure 1: Compounds included in the ETS Wildfire Expanded Volatile Markers (left column) and their main glycosylated (sugar-bound) forms included in the Glycosylated Markers panel (right column).

(1) 2018 Mendocino Complex Fire Lake County Winegrape Commission Study - with the participation of the University of California Cooperative Extension (UCCE) and the Australian Wine Research Institute (AWRI) - see “From Blaze to Bottle: Smoke Gets in Your Wine”by Glenn McGourty in the January 2020 Issue of Wine Business Monthly.

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WILDFIRE IMPACT: WHAT SAMPLES TO TEST?

Wildfire impacts in grapes and wines are caused by a wide range of volatile phenols found in wildfire smoke. These compounds are absorbed and accumulate in berries. They eventually end up in wine where they can cause unwanted flavors. These off-flavors, described as “smoky”, “bacon”, “campfire” and “ashtray”, are usually long lasting and linger on the palate even after the wine is swallowed.

Wildfire impact in wine was identified as a serious problem after the 2003 wildfires in Australia and British Columbia. The California wine industry was also affected following the wildfires of summer 2008, and wildfire impact has been recognized as a concern for growers and wineries ever since. Here is a review of the various types of samples that may be submitted for testing:

1. Berries During the 2008 California wildfires, ETS developed an analytical tool to screen grapes for the risk of wildfire impact. The analysis measures trace levels of free guaiacol and 4-methylguaiacol in whole berries.

Knowing the levels of these indicators in berries enables winemakers to assess the risk of wildfire smoke impact and choose an appropriate course of action to mitigate the effects in their wines. In 2021, an extended panel of volatile (“free”) wildfire markers and a glycosylated markers panel is also available for berry samples.

Exposure of vines to wildfire smoke can widely vary within a small geographic area, depending mainly on proximity with the fires and wind conditions. In effect, getting representative samples can be challenging.

Mixing grape varieties in composite samples should be avoided, as grape cultivars often react differently to a similar exposure to smoke. Syrah grapes contain naturally occurring guaiacol and should never be mixed with grapes from other varieties.

34

Submit 200 to 300 loose berries, keeping them cold and undamaged as much as possible (do not crush them).

When shipping samples, use hard plastic containers with icepacks in an insulated package. Avoid submitting cluster samples, which trigger additional fees and may delay getting results. A typical strategy for berry sampling is to collect berries from at least 50 clusters, taking four berries from each cluster from the top, bottom, front and back of the cluster.

Berry samples are often requested by crop insurance providers, although micro-ferment samples have been more widely accepted during the 2020 fire events. It is advisable to keep backup samples in a freezer.

2. Juice It is possible to measure wildfire impact markers in juice samples, but since compounds are mostly located in skins, whole berry testing is the preferred method for pre-harvest screening. Please avoid submitting fermenting samples which may constitute a safety hazard.

3. Small-Scale Fermentations (Micro-Ferments) In order to complement pre-harvest grape tests, wines from small-scale fermentations (“micro-ferments” or “bucket ferments”) may be tested for volatile wildfire markers (basic or extended panels), and for glycosylated markers. The pros and cons of both tests are outlined in Figure 2.

4.

Production Wines Analyzing immediately after completion of primary fermentation allows a first assessment of wildfire impact in production wines. It is preferable to sample and analyze wines that have not come in contact with oak or oak-derived products, which can contribute volatile smoke markers.

With barreled wines, it is still possible to get useful information from the volatile markers by choosing the extended panel of volatile (“free”) smoke markers rather than the basic volatile marker panel (guaiacols only), and taking samples from the most “neutral” barrels available. Keep in mind that there is no issue analyzing oaked wines for glycosylated markers, as oak or oak products do not contain these compounds. Analyzing for glycosylated markers in wine is always relevant regardless of contact with oak.

G r a p e S a mp l e s

M i cr o - fe r me nts

I m m e d i at e

>1 We ek

S e ns o r y Ev a l u at i o n

N ot v e r y u s e fu l

Use ful , but d ifficul t (ne ed fo r mul tipl e t r aine d taste r s i nc l ud ing se nsitiv e ind iv idual s)

Ana l y s i s Tu r na r o u nd Ti m e

1 - 2 d ay s

1- 2 d ays

Pr e di c t i o n o f S m o k e C ha r a c t e r s i n Pr o du c t i o n Wi ne s

I ndi r e c t ( v a r i ab l e “ mu l t i p l i e r s ” b et w e e n g r a p e a nd w i ne r e s u l t s )

R e ds: mo r e d ir e ct ( but d e l aye d ) White s: unc e r tain (f e r me nt with s k ins fo r “wo r st c a se sc e nar i o ” ?)

S a mp l e Pr e p a r at i o n Ti m e ( b e fo r e s e ndi ng t o t he l ab o r at o r y )

Figure 2: Grape Samples vs. Small Scale Ferments (Micro-ferments) as preharvest testing options for wildfire smoke impact.

35

&

EXT E N D E D VOLAT ILE MA R K E RS GLY C OS Y L AT E D MA R K E R S WHAT LEVELS AND PATTERNS TO EXPECT IN WINES? Following the 2018 Mendocino Complex Fire Study (1), the selection of volatile (“free”) and glycosylated (“bound”) markers currently offered by ETS and the AWRI has shown to be relevant in the context of a major California wildfire event (with the exception of volatile syringol and 4-methylsyringol, which did not appear closely related to the level of exposure to wildfire smoke).

The strength of correlations observed between markers, however, left the following question open: “Is testing for other markers than “free” guaiacol and 4-methylguaiacol really useful from a practical standpoint?”

After having analyzed to date more than a thousand wines for both volatile extended markers and glycosylated markers, we can answer this question. So, let’s present examples of results, typical and less typical, that we have observed with 2020 wines from California and Oregon.

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Ty p i c a l … a n d Le s s Ty p i c a l Pa tter ns To put it simply… as a general rule when volatile markers are high, glycosylated markers are also high. This is illustrated figure 3, which shows a range of levels observed in Cabernet Sauvignon wines, from normal low baseline levels observed without exposure to wildfires, to extreme levels resulting from severe exposure. According to our observations so far, this range can be expected with most red wines, while levels in white and rosé wines are generally lower due to reduced skin contact.

A notable exception is of course Syrah, now well known to contain naturally significant baseline levels of volatile guaiacol.

With this variety however, baseline levels of other volatile markers appear to be normal, making them useful to assess wildfire smoke impact.

Another oddity is Petit Verdot, which doesn’t appear to contain naturally unusual baseline levels of wildfire markers, but seems

year after year to be much more sensitive to wildfire smoke.

With Petit Verdot wines, the observed range is much larger than with any other variety. We have measured volatile guaiacol exceeding 200 ug/L, and combined levels of glycosylated markers approaching 1,000 ug/L. Examples of “typical” patterns, meaning with volatile marker levels accompanied by relatively expectable levels of glycosylated

markers, can be observed in impacted white, rosé or red wines from various varieties. Figure 4 shows a few examples of wines all containing volatile guaiacol in the 4-6 ug/L range, usually associated with noticeable characters from the sensory standpoint, for which glycosylated markers match this most commonly observed pattern.

N orm a l

Mod e rate

Hig h

Ver y Hig h

E xtreme

guaiacol

1. 4

2. 9

7.0

1 4.2

2 1.4

4-Methylguaiacol

<1. 0

<1. 0

1.8

3 .1

4.9

Cresols (sum)

1. 0

3. 4

10.6

2 2 .4

2 6.3

P h e n ol

2. 5

7. 8

2 0.6

3 4.9

2 8.7

Syringol

<5 . 0

<5 . 0

1 3 .4

10.5

3 0.1

4-Methylsyringol

<5 . 0

<5 . 0

<5.0

<5.0

8.0

Guaiacol Rutinoside

0.0

1. 4

3 .0

3 .6

8.5

4-Methylguaiacol Rutinoside

1. 0

1. 3

3 .9

5.5

8.1

Cresol Rutinoside

2. 0

6.5

1 5.3

2 3 .8

4 8.2

Phenol Rutinoside

1. 0

8.3

1 5.5

2 1.9

63 .2

Syringol Gentiobioside

2. 0

2. 4

11.8

1 8.7

2 2 .0

4-Methylsyringol Gentiobioside

<1. 0

<1. 0

1.1

3 .4

4.8

Figure 3: Range of volatile and glycosylated markers observed in 2020 Cabernet Sauvignon wines, from normal baseline to extreme levels resulting from severe exposure to wildfire smoke.

Please note, these are examples of data collected from 2020 wine samples. Levels and ranges observed for the various markers measured may or may not apply to current or future events.

37

PG

CH

PN

ME

PV

guaiacol

4. 7

5.7

5.8

4.1

4.8

4-Methylguaiacol

1. 0

1.6

1.5

1.0

1.4

Cresols (sum)

5.1

5.8

8.4

5.3

3 .1

Phenol

10 . 9

11.4

11.4

4.1

6.5

Syringol

<5 . 0

< 5.0

7.6

5.3

8.3

4-Methylsyringol

<5 . 0

< 5.0

<5.0

<5.0

<5.0

Guaiacol Rutinoside

4. 0

2 .9

2 .8

3 .1

2 .5

4-Methylguaiacol Rutinoside

8.2

3 .2

3 .4

5.1

5.6

Cresol Rutinoside

21. 2

1 4.4

7.5

1 5.2

6.0

Phenol Rutinoside

6.5

7.6

9.8

11.0

6.0

Syringol Gentiobioside

1. 2

1.7

6.1

8.8

10.5

4-Methylsyringol Gentiobioside

1. 1

1.1

1.0

1.9

1.2

Less typical are patterns where volatile markers are relatively low, sometimes barely suggesting wildfire impact, or even no impact at all, co-occurring with relatively high levels of glycosylated markers. These patterns are quite uncommon, but particularly worrisome to winemakers, due to the fact that glycosylated (“bound”) compounds, which are odorless, 38

Figure 4: 2020 white and red wines from various grape varieties (Whites: PG = Pinot Grigio, CH = Chardonnay, Reds: PN = Pinot Noir, ME = Merlot, PV = Petit Verdot) displaying a typical pattern between volatile and glycosylated markers. Note that all these wines contain volatile guaiacol in the 4-6 ug/L range, usually associated with noticeable sensory wildfire characters.

Please note, these are examples of data collected from 2020 wine samples. Levels and ranges observed for the various markers measured may or may not apply to current or future events.

are suspected to contribute unpleasant aftertastes and degrade with time, releasing volatile (“free”) odor-active compounds. Examples of such patterns are presented in figure 5 - three examples on the left.

On the other hand, opposite patterns, showing high volatile markers but relatively low levels of glycosylated

markers, are observed with some regularity, especially with 2020 Pinot Noir wines – see figure 5 - two examples on the right. We do not know at this point if this is a recurrent occurrence with Pinot Noir, which might be observed year after year. Low levels of glycosylated markers may be caused by a weakened defense mechanism from the plant

against smoke volatiles. This could be related to the severe heat wave we experienced all across the Western United States in mid-August 2020, causing a slowed or even “shutdown” metabolism with Pinot Noir grapes during most of their exposure to wildfire smoke.

CH

CB

GR Ro se

OR PN

CA PN

guaiacol

1. 2

1. 5 0

1.0

5.3

6.0

4-Methylguaiacol

<1. 0

<1. 0

<1.0

1.7

1.9

Cresols (sum)

2. 0

<3. 0

<3 .0

6.9

6.5

Phenol

<2. 0

<2. 0

<2 .0

9.2

6.2

Syringol

<5 . 0

<5 . 0

<5.0

<5.0

6.0

4-Methylsyringol

<5 . 0

<5 . 0

<5.0

<5.0

<5.0

Guaiacol Rutinoside

4.5

10 . 3

5.8

1.4

1.0

4-Methylguaiacol Rutinoside

2. 5

8.0

5.7

1.8

1.1

Cresol Rutinoside

1 2. 0

33. 6

2 6.9

3 .1

2 .0

Phenol Rutinoside

15.5

19.5

2 8.9

2 .8

2 .5

Syringol Gentiobioside

1. 0

20. 9

5.0

2 .0

1.5

4-Methylsyringol Gentiobioside

1. 0

2. 1

<1.0

<1.0

<1.0

Figure 5: 2020 white and red wines from various grape varieties (Whites: CH = Chardonnay, CB = Chenin Blanc, Rosé: GR = Grenache, Reds: PN = Pinot Noir) displaying atypical patterns between volatile and glycosylated markers. The three wines on the left have very low levels of volatile markers with high glycosylated markers, while the two wines on the right show opposite patterns.

Please note, these are examples of data collected from 2020 wine samples. Levels and ranges observed for the various markers measured may or may not apply to current or future events.

To conclude, additional volatile markers and glycosylated markers clearly allow more complete assessments of wildfire impact in production wines. They have the potential to provide useful and actionable information when measured in grapes and micro-ferments before harvest.

39

40

QU E STI O NS ANSWERS

&

Q: I WANT TO TEST GRAPES FOR WILDFIRE IMPACT, WHAT KIND OF SAMPLE SHOULD I SUBMIT? A: The preferred sample for red or white grapes is a representative 200-300 loose berry sample, with

berries as intact as possible. Transport in small rigid “sandwich boxes” works well – this is necessary if you are shipping samples. Avoid submitting cluster samples: the additional sample preparation time in the lab will delay results. We do not recommend submitting juice samples.

Q: SHOULD I CRUSH BERRIES AND LET THEM SOAK IN THEIR JUICE BEFORE BRINGING THEM? A: We do not recommend doing so. When we have control of the sample preparation, our interpretation guidelines are more applicable.

Q: WITH WHITE GRAPES, SHOULDN’T I BRING JUICE SAMPLES? A: Since wildfire impact compounds are mostly located in skins, we still prefer whole berry samples

for white grapes. Our risk interpretation guidelines are similar for white and red grapes. Of course with white grapes, the risk of off characters materializing in wine can be mitigated to some degree by minimizing skin contact (with red grapes, making rosé by the direct pressing method can be a successful approach too), but in case of maceration with skins (e.g. following machine harvesting, destemming, or intentional skin contact) the risk level may be equivalent.

Q: CAN I MIX BERRIES FROM DIFFERENT VARIETIES AND BRING A COMPOSITE SAMPLE? A: This is not advisable. Over the years we’ve seen drastic differences in pick-up of wildfire impact

compounds between grape varieties. For example, Petit Verdot is often much more impacted than other Bordeaux cultivars. In 2020 we saw very substantial differences in behavior between Chardonnay and Pinot Noir.

Q: CAN YOU TEST SYRAH GRAPES? A: Syrah naturally contains variable amounts of guaiacol, the main marker. This makes it impossible

to assess wildfire impact based upon guaiacol only, whether with grapes or micro-ferments. The only but very imperfect strategy available so far has been to use other varieties grown next to Syrah blocks as “proxies”. The availability of the extended volatile markers panel and the glycosylated markers panel should be a great help assessing directly Syrah samples.

Q: CAN YOU TEST FOR MORE THAN JUST “FREE” GUAIACOLS (GUAIACOL AND 4-METHYLGUAIACOL)?

A: Yes. We have offered our enhanced and extended volatile (free) markers panel for wines in November

2020, and in December 2020 we began offering an updated version of our glycosylated (bound) glycosylated markers panel. For the 2021 harvest we will offer both tests for grapes, micro-ferments, and production wines.

41

Q: DOES ETS MEASURE “TOTAL” MARKERS, OR ALL FORMS OF “BOUND” MARKERS? A: Some laboratories offer

testing after various acid and heat treatments, in an attempt to measure “Total” or “Bound” forms in their entirety. We have serious reservations about such tests and do not offer them. This concern is shared by our international colleagues such as Australian Wine Research Institute (AWRI) and they do not offer them either. Instead, for each of the volatile markers listed in our Expanded Panel, we can measure directly by HLPC/MS/MS its main glycosylated (sugar-bound) form (see figure 2). This is the strategy also adopted by the AWRI.

Figure 6: Glycosylated Wildfire Markers and their isotopic analogues detected by LC/ MS/MS (ETS Laboratories December 2020). These standards make quantitative, accurate and reproducible results (comparable over a long period of time and between different laboratories) possible.

Q: WHAT IS THIS UPDATED ANALYSIS FOR GLYCOSYLATED (“BOUND”) MARKERS AT ETS? A: In 2019-2020 ETS enhanced its own method for the determination of glycosylated (sugar-bound) smoke markers, but

quantitative results had to be expressed as “equivalent” ug/L units - a strategy commonly used when exact reference compounds are not available (for example just as tannins are commonly expressed as catechin or epicatechin equivalents). Following the 2020 firestorms, ETS joined resources with the AWRI and others from around the world to have pure reference compounds synthesized, so that results can now be reported as “true” concentrations (see Fig. 6). Accurate, quantitative and reproducible results (comparable over a long period of time and between different laboratories) are now achievable and a reality at ETS.

Q: I’D LIKE TO HAVE MICROFERMENTS TESTED. WILL ETS PREPARE THEM FOR ME? A: Small-scale fermentations or “micro ferments” can be a good test for

smoke exposure, from which wildfire characters can be detected by sensory evaluation and confirmed by quantitative laboratory analysis.

Unfortunately, due to practical constraints, we cannot offer to perform micro-ferments. For guidance, we recommend watching the tutorial “Step-by-Step: How to do small-scale fermentations for the evaluation of grape smoke exposure risk” available on the UC Davis website. Another good resource is the small-lot fermentation method protocol as specified by the Australian Wine Research Institute (“AWRI“). At completion of fermentation, transfer the fermented wine into a bottle, let settle in fridge for a few hours, decant and submit sample in a 60 mL plastic tube.

If a crop insurance claim is considered, check with your insurance provider to determine if they accept results from micro-ferments.

A step-by-step procedure for small-scale fermentations (micro-ferments) is available on the UC Davis Website: https://wineserver.ucdavis.edu/multimedia/ step-step-how-do-small-scale-fermentationsevaluation-grape-smoke-exposure-risk The AWRI protocol is found at: https://www.awri.com.au/wp-content/uploads/ small_lot_fermentation_method.pdf

Q: WHEN IS THE BEST TIME TO BRING SAMPLES? A: For grape samples the typical recommendation is about 7-10 days prior to harvest. Keep in mind that the impact of wildfires is

cumulative and that “negative” results early in the season may give a false sense of security, especially if more exposure happens.

Of course with micro-ferments several days need to be accounted for after grape sampling, in order to produce fermented samples that can be analyzed.

Q: WILL YOU HELP ME WITH RESULTS INTERPRETATION? A: Of course. Although each fire event is unique, our interpretation guidelines based on volatile (free) guaiacol derived from our

experience since 2008 have stood the test of time, and have proven to be quite robust (Fig. 7). The presence of additional markers, with the extended volatiles marker panel, and the availability of a glycosylated markers panel, allow refining and confirming diagnostics. Always feel free give us a call for assistance with result interpretation.

42

INTERPRETATION GUIDELINES FOR WHOLE BERRY TESTS (EXCLUDING SYRAH): ANTICIPATED RISK OF WILDFIRE IMPACT IN WINE LOW

MODERATE 0.5

HIGH

VERY HIGH

1.0

2.0

FREE GUAIACOL (ug/kg) INTERPRETATION GUIDELINES FOR MICROFERMENTS AND UNOAKED WINES (AGAIN EXCLUDING SYRAH): PREDICTABLE SENSORY IMPACT UNLIKELY 1.O

2.O

POSSIBLE 3.O

LIKELY 4.O

MOST LIKELY 6.O

FREE GUAIACOL (ug/L) Figure 7: Interpretation guidelines for volatile (free) guaiacol, the main marker of wildfire smoke impact, in grapes and micro-ferments and unoaked wines. Additional markers in the extended volatile panel as well as glycosylated markers allow refining and confirming assessments.

IMPORTANT: THESE GUIDELINES ARE FOR GUAIACOL, NOT SUM OF COMPOUNDS. GUIDELINES ARE SIMPLY OBSERVATIONS BASED UPON PAST EVENTS, AND MAY OR MAY NOT APPLY TO CURRENT OR FUTURE EVENTS. ETS DOES NOT, AND WILL NOT, PROPOSE ACCEPTANCE OR REJECTION CRITERIA.

Q: WHAT ARE NORMAL “BASELINE” LEVELS FOR GLYCOSYLATED MARKERS? A: Grapes and wines may naturally contain low “baseline” levels of glycosylated markers, variable by grape variety and

geographic origin. ETS has been engaged for years in building a database for grapes not exposed to wildfire smoke, as well as wines also from grapes not exposed to wildfire smoke. The good news is that despite the fact that this project suffered a setback last year, with fires raging in the western states for a good part of the growing season, we are confident that our current database is sufficient to help our clients with result interpretation.

Q: MY GRAPES (OR MICRO-FERMENTS) TESTED “POSITIVE”. SHOULD I HARVEST? A: Our interpretation guidelines are related to incremental risk scales, as there is no “magic number” below which no risk

is present and above which wines are guaranteed to be impacted. Choosing to harvest or not will always be complex risk management decisions, loaded with painful consequences for both growers and wineries. Analytical results only help in making these difficult decisions.

Q: DOES ETS RECOMMEND KEEPING PRE-HARVEST BACKUP SAMPLES? A: Grape samples and micro-ferments serve two distinct purposes: helping with harvest decisions and serve as proof of wildfire impact for insurance claims. Especially in the second case, it makes sense to keep backup samples, especially in the context of catastrophic wildfires.

Q: WHAT IS YOUR TURNAROUND TIME? A: First, we want to thank the University, State, and other labs that stepped-in during the 2020 fire season. For the 2021

season, we have expanded yet again our GC/MS, GC/MS/MS and LC/MS/MS analytical capabilities. We also have the options of transferring excess samples for rapid analysis by certain trusted partner laboratories under ISO 17025 guidelines. For the most accurate turnaround time we recommend taking a look at our website (Analyses - Smoke Markers). In the case of extraordinary fire events, we will keep our turnaround time information for grape, micro-ferment and wine tests prominently displayed and updated regularly.

Q: DOES IT MAKE SENSE TO SUBMIT SAMPLES FROM VINEYARDS NOT EXPOSED TO SMOKE THIS YEAR? A: It does. The West Coast Smoke Exposure Task Force experts, among others, recommend to have a sampling plan ready

before wildfires happen, and know what “baseline” numbers to expect with your samples. Whether you choose to rely on direct berry tests or micro-ferments, “rehearsing” that plan during a non-smoke year achieves both goals.

43

WIN E

AUT HE NTI C I T Y AT E TS

There are over 10,000 varieties of grape, some of which produce wines that carry premium prices in the fine wine market with some bottles being sold for over $10K. With the global wine market expected to increase to $420B by 2023, and with wine fraud everpresent, it is of great importance that the wine industry has a means to be able to authenticate its products.

At ETS we can analyze the individual components of wine using modern instrumentation, but full characterization of wine for establishing authenticity is not so straightforward.

For example, measurement of glycosylated smoke markers involves sample clean-up to reduce signal interference before measurement using chromatography & mass spectrometry, all of which takes time and money to perform and represents just one small fraction of the wines’ character. Scientists have applied colorimetric, spectrophotometric, and grape DNA techniques to fully characterize wine, but recently the classical physics technique Nuclear Magnetic Resonance (NMR) has come to the fore to address this challenge. The main advantage of NMR is that it can give a full digital wine signature in a matter of a few minutes using less than one milliliter of wine sample. Yet, far from being a signature that simply tracks the wine (as with RFID technology), this digital signature can then be compared with a database to give information about authenticity about the wine itself, and/or to ensure supply chain authenticity.

Introducing the Proton: Wine is a complex hydro-alcoholic solution containing a variety of organic and inorganic compounds made of chemical elements. Of these, Carbon and Hydrogen are the most abundant in nature and are the focus of organic chemistry.

The Hydrogen nucleus (aka Proton) possesses a magnetic moment and is considered NMR active for this reason. Nuclei with odd numbers of protons and neutrons in the nucleus have the property of spin and induce a magnetic field , whereas nuclei with even numbers of Protons and Neutrons, such as Carbon-12, do not induce a magnetic field, and are not NMR active. For clarity, the remainder of this article will focus on the proton.

S ome NMR Theor y: In this classical physics technique, the sample of wine is placed inside a strong magnetic field. In doing so, every atomic nucleus in the sample with an odd number of either protons or neutrons polarize and align parallel to the magnetic field in two different spin states, low and high energy, while spinning and precessing in a manner similar to a spinning bar magnet.

A B I T O F B AC KG RO U N D NMR was discovered in 1938, and the first NMR unit was produced in 1952. NMR technology is widely and routinely used in society today. For example, it is the basis for magnetic resonance imaging, and it is used by synthetic chemists to help elucidate molecular structures of pure organic compounds as part of drug discovery. NMR offers the extreme reproducibility and transferability required for wine authenticity. Even small changes in concentration can be detected for many different compounds.

44

Those that orientate against the magnetic field have higher energy compared to those aligned with the magnetic field, and the difference between the two energy states depends on the type of nucleus and the strength of the applied magnetic field.

As mentioned, the frequency and intensity of the weak resonant signal is characteristic for each Proton in the sample of wine, because it depends largely on the effect of neighboring nuclei.

Given that there are many thousands of different molecules in wine, we would expect to see many different resonance signals emanating from them. All the resonant signals when combined together give what’s known as a Free Induction Decay (FID).

This raw NMR data is then mathematically transformed into interpretable spectra a.k.a. the digital signature. This is nontargeted analysis and does not provide information about wine composition.

With the sample positioned in the center of a strong magnetic field, the nuclei are excited by means of a radio frequency pulse that induces magnetic resonation in the protons. Briefly, the NMR probe, which is a combined RF transmitter/receiver, passes a current through coils to generate a broad-band pulse of radio frequencies directed at the sample. The protons in the sample resonate with the RF frequencies at the precession frequency (also called the Larmor Frequency), and this induces a secondary oscillating magnetic field. The oscillating magnetic fields perturb the orientation of the nuclei, causing them to flip 90 ° to the transverse plane so that the magnetic moment of the nucleus is oriented at right angles to the direction of the magnetic field. When the pulse is stopped the nuclei gradually relax back to their original orientation, aligned parallel to the magnetic field. As they relax back, a small amount of RF energy is released from the nuclei, inducing a current in the probe receiver coils. This in turn produces a weak resonance signal that can be measured. The intensity and frequency of the resonance signal for each proton is different according to their local atomic environment, and decays over time as they re-establish magnetic equilibrium. At this point the system is ready for another pulse (or scan). Typically 32 scans are accumulated during a proton NMR experiment for optimal signal intensity.

Additionally, NMR is capable of performing quantitative targeted analyses of over 70 compounds in wine, which can also aid identification. While not as sensitive as mass spectrometry (generally parts per billion detection levels), NMR can detect down to parts per million levels. NMR is quantitative because the intensity of the resonance signal is directly proportional to the protons present, but a reference must be used for quantitation.

In combination, the unique non-targeted digital signature together with the amount of targeted analyses furnishes us with a powerful package to ensure authenticity. Clearly, NMR has a lot to offer the wine industry, even though the results of some samples may require expert interpretation, and the ability of the system to authenticate a sample of wine relies heavily on the statistics of previously populated wine databases. As with all systems that rely on statistics, the data fed into the database must be comprehensive and of the highest quality possible so that the database improves with each addition. It follows then that the larger the database, the greater the power of the system’s power to derive authenticity.

WfYUh]b[ U XUhU V U g Y The only currently available wine NMR databases are populated with European wines, lessening the relevance of the database for high caliber US wines. As such, there is a current need to establish and populate a database for US varietal wines in a similar manner to our European partners at Bruker. This would increase authenticity testing in the US market with the aim of cutting down on future wine fraud. If you’re interested in joining us in the effort to reduce wine fraud, please contact us.

45

HARVEST G R AP E M ATU RI TY M ON I TORI N G PAN ELS The ETS Grape Maturity Monitoring Panels provide a set of analyses that growers and winemakers request to monitor fruit maturity, including Brix, Glucose + Fructose, TA, pH and malic acid, as well as berry size parameters and average sugar per berry. Monitoring sugar per berry allows growers to determine the duration and rate of active sugar loading, during which vines synthesize and actively transport sugar into berries. The time at which the sugar loading phase ends, and what level of Brix was achieved at that time, are both important indicators of wine growing conditions (excessive or insufficient vigor, water availability and resistance to heat stress).

For more details see pg. 4

46

TOOLKIT E T S S CORP I ON S

TM

Wild yeast and bacteria from the vineyard may be introduced into the winery on the harvested fruit, causing spontaneous fermentation and/or spoilage. ScorpionsTM DNA analysis offers winemakers an early detection tool to identify these spoilage organisms.

Despite the best practice of modern winemaking methods, microbial contamination often occurs during wine production. Spoilage microbes are capable of survival and growth in the wine, potentially producing off-flavors, off aromas, and turbidity. Microbiological contamination is often undetected until related problems in the wine become noticeable by sensory evaluation.

Scorpions™ assays, based on specific genetic targets, detect the full range of wine and juice spoilage organisms. This genetic analysis method detects microbial populations directly in wine or juice. Results are routinely reported within two business days, giving winemakers the ability to address problems before wine defects occur.

Targeted genetic probes give the winemaker the ability to monitor only those specific spoilage organisms that have the potential to adversely impact wine quality, and to accurately measure populations down to extremely low levels.

For more details see pg. 19

47

H A R V E S T TOOLKIT

GRA PE P H E N O LI CS Follow changes in grape phenolics during ripening, using the catechin and tannin "ripeness" index to monitor seed ripening, and quercetin analysis to monitor canopy effects on grape phenolics.

For more details see pg. 26

GRAP E WATER CON TEN T Changes in grape water content influence finished wine composition and can be as important as standard sugar and acid measurements when making picking decisions. Grape water content is also very useful for understanding changes in TA, pH, ºBrix, and other harvest indicators.

MO N I TO R I N G I B M P The compound responsible for a "green bell pepper" aroma in wine, IBMP, decreases quickly during maturation. But once grapes are picked, it is hard to control. Monitoring changes in grape IBMP directly influences final levels in wine, and is crucial in making picking decisions.

For more details see pg. 31

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H A R V E S T TOOLKIT

GLU TAT H I O N E Glutathione, a natural grape antioxidant, can protect the aroma and flavor of white and rosé wines and prevent premature aging. Glutathione levels fluctuate during production, as the compound can be absorbed by yeast and then released after fermentation.

For more details see pg. 31

WI LDFI RE I M PACT The compounds in wildfire smoke are absorbed by vines and can cause unwanted flavors in wine.

Analyzing for these compounds allows winemakers to screen grapes for the risk of smoke impact and work to mitigate its effects.

For more details see pg. 32

HA R VE S T J U I CE PA N E L Our most popular harvest panel offers a full range of grape and must analyses, combining more than 10 tests including fermentable sugar (to help estimate alcohol content) and YAN (yeast-assimilable nitrogen ­­— to help predict sluggish or stuck fermentation and potential sulfide formation.)

For more details see pg. 6

49

H A R V E S T TOOLKIT

B OTRY TI S PAN EL This comprehensive test panel checks grapes for Botrytis (using ScorpionsTM ) and laccase, detecting both the spoilage organism and its byproducts that can harm your wine.

L ACCA S E AC T I VI T Y Laccase is a polyphenol oxidase associated with rot caused by Botrytis. Elevated levels of laccase can result in oxidation of phenolic compounds that may cause color degradation or premature browning in red wines. In addition, laccase mediated oxidation can also affect the aroma profile of the wine.

Y EAS T V I AB I LI TY Our automated method reports yeast viability and total cell count back to the client within hours, and the real-time microscopic flow image analysis examines 1,000 times the volume used in standard microscopic methods, vastly increasing the accuracy of your results.

50

H A R V E S T TOOLKIT

R A P I D PH E N O L I C S By the end of maceration or fermentation, the tannin content of a wine is already fixed. Monitoring phenolics during this critical period allows winemakers to better control tannins by increasing or decreasing phenolic extraction.

For more details see pg. 29

DN A FI N GERP RI N TI N G ETS Laboratories offers DNA fingerprinting to distinguish between closely related strains of Saccharomyces cerevisiae. ETS MLVA technology allows winemakers to monitor yeast population in native fermentations and check the efficiency of inoculations with commercial strains.

E U C A LY P TO L Eucalyptus character is a controversial sensory expression in red wines from California and countries with Mediterranean climates. Even a slight “eucalyptus” note can interfere with delicate varietal aromas, and can have a detrimental influence on certain grape varieties.

For more details see pg. 30

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HARVEST RESOURCES Since we first opened our doors in St. Helena in 1978, we’ve grown alongside the wine industry, partnering with our clients as they’ve gone on to create many of the world’s finest wines. We’re continuing to invest not only in Napa Valley, but also in other rapidly growing wine regions by expanding our local and online services to support winemakers with advanced tools and technical assistance.

Here's what to take advantage of in Harvest 2021: 52

L O C A L J U I C E A N A LY S E S AT S AT E L L I T E L A B O R AT O R I E S A L L S AT E L L I T E S

Samples can be dropped off locally for any analysis ETS offers. These common harvest analyses will be run onsite for quicker turnaround.

A N A LY S I S

TECHNIQUE

TURNAROUND

JUICE PANEL

MINERVA

SAME DAY

BUFFER CAPACITY

MANUAL

SAME DAY

ETHANOL

MINERVA

SAME DAY

FREE SO

FLOW INJECTION

SAME DAY

TOTAL SO

FLOW INJECTION

SAME DAY

TURBIDITY

TURBIDIMETRY

SAME DAY

VOLATILE ACIDITY

SEQUENTIAL ANALYZER

SAME DAY

GRAPE MATURITY MONITORING PANEL

VARIOUS

SAME DAY

RAPID PHENOLIC PANEL

HPLC

1 DAY

GRAPE PHENOLIC PANEL

HPLC

1 DAY

SCORPIONS BACTERIA JUICE PANEL

SCORPIONS™

2 DAYS

SCORPIONS YEAST JUICE PANEL

SCORPIONS™

2 DAYS

SCORPIONS COMBINED JUICE PANEL

SCORPIONS™

2 DAYS

BRIX G LUCOSE + FRUCTOSE PH TA (TITRATABLE ACIDITY) TARTARIC ACID L- M ALIC ACID POTASSIUM NOPA AMMONIA

PA S O R O B L E S , WA L L A WA L L A & NEWBERG

PA S O R O B L E S & NEWBERG

VISIT OUR WEBSITE FOR CURRENT PRICES, AND THE FULL LIST OF O N - S I T E W I N E A N A LY S E S .

W E K N O W Y O U ' R E B U S Y.

WE'LL COME TO YOU. Y l danded W c i rier in b e k berg

We’ve teamed up with our friends at Crush2Cellar to launch our new complimentary sameday courier van. The van will drop off supplies and collect ETS samples along three routes throughout the Willamette Valley, running directly through Carlton and McMinnville every day, and Corvallis on Thursdays.

This new, expanded service will be in addition to our on-demand ETS courier service, which so many of you already utilize, and is available year round.

ETS pickup requests can be placed on our website through your customer account, https:// www.etslabs.com/login, by phone at (503) 537-6245, or by email at infoOR@etslabs.com. If you miss the cutoff for the scheduled routes, just give us a call and we will send out one of our ondemand couriers directly to your facility.

54

ETS

V I N TAG E P O R TA L Winemakers often say they remember the last vintage, and their best and worst vintages. For the last seven years ETS Laboratories has provided Post-Harvest seminars detailing the differences observed in juice chemistry panel components each harvest as compared to previous harvests. Each vintage exhibits differences in juice chemistry with trends that are similar in California, Oregon and Washington. While seminar attendees usually find something interesting in our presentations, many continued to suggest that making real-time available data would be a very useful tool. The ETS Vintage portal provides a new way to identify vintage trends in juice chemistry and grape phenolic development in real time.

Discussions with clients suggest the ability to compare their juice panel and grape phenolic data with that of others would provide a tool to recognize vintage trends. In addition, it would enable them to see if their individual results are similar to others in their region growing a particular varietal. Early identification of trends in a vintage can help with picking and process decisions.

JOIN US We invite our clients to join the Vintage Portal community this harvest. The results from the Juice Panels and Grape Phenolic Panels, with no identifying information other than varietal and AVA, are going to be analyzed and posted daily. The only information we need is the varietal and AVA code for each sample. The ETS harvest labels for Juice and Berry samples have boxes to enter the two letter codes for the AVA and varietal. These codes can be found on the ETS website and at the front counter at each of the ETS locations. We look forward to working with you to provide this new and exciting resource to our clients.

JUICE & BERRY SAMPLES Client #:

Client:

Sample ID:  Juice Panel

Juice Scorpions™

 GLU+FRU  BRIX

 Malic Acid  NOPA

 TA  pH

 Ammonium  Potassium

 Combined  Bacteria

 Tartaric  Other:

N

V

C

 Ethanol

 TSO 2

 VA

 Grape Water Content

VAR

AVA

 Yeast

 FSO2

S

 Grape Maturity Panel  Grape Phenolic Panel

Enter the AVA and Varietal on your juice label.

For more information go to help.etslabs.com and search for Vintage Portal.

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H A NDLE W I T H

CARE

It’s important to collect and handle harvest samples carefully to ensure accurate and representative results.

We’ve collected our recommendations to help you get the most out of your harvest analyses.

JUICE SAMPLING Most harvest samples received at ETS come to the laboratory as juice.

Berries pressed for a juice sample should be selected from at least 20-40 different clusters, and can easily be pressed by hand in their collection bag – pour the juice into a standard ETS 60mL sample tube and label with your ETS client labels.

Samples should be kept cool to prevent fermentation.

GRAPE SAMPLING FOR PHENOLICS For grape phenolic testing, a representative sample is critical to obtain accurate results, especially in varietals with tight clusters. Samples for the Grape Phenolic Panel should include berries from at least 20-40 different clusters. Clusters can be collected either from harvest containers or directly from the vineyard.

To get a representative sample, all the berries must be stripped from the clusters and mixed before bagging a 300-400 berry sample for analysis (about 500g or 16 oz).

Samples should contain only intact and undamaged fruit to ensure accurate results.

BERRY SAMPLING Take 200-400 berries per block. Pick berries from random clusters on both sides of the row. Take berries from the top and bottom of both the front and back of each cluster. *Samples submitted for berry analysis should contain only intact and undamaged fruit.

We encourage clients to submit berry samples rather than whole clusters. If samples are submitted as clusters, ETS will prepare a berry sample for an additional fee.

LABELING Each bag of berries should be clearly labeled with the client name, sample ID, and analyses to be performed.

ETS provides free sample labels that are pre-printed and barcoded with your client ID – visit our website, or give us a call.

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GEAR UP FOR HARVEST

Don’t get caught empty handed – order complimentary tubes, pre-printed labels, and shipping pods to take the headache out of collecting and shipping samples.

1. Login to your ETS account and select the winery you're ordering supplies for.

Outside the range of our sample pickup service?

ETS offers free shipping kits, which include an insulated envelope and an ice pack, to help you easily send in samples no matter where you're located.

S H I PP IN G JU I CE S AM P LES FO R R EGU LAR AN ALYS ES

S HI P P I N G JU I CE SAMPLES FOR S CORP I ON ANALYSIS

To prevent problems from fermentation, juice samples should be frozen or boiled for shipment, and clearly marked as either “FROZEN” or “BOILED” depending on the treatment used.

To ensure accurate results, it's important to avoid damaging DNA or killing yeast and bacteria:

Boiling 2. Use the "Get Supplies" button on the dashboard to place a supply order.

Boil samples with a loosely fitting cap to prevent evaporation and concentration.

Do not over-boil.

° Keep samples cool with ice packs.

Ship by overnight delivery using a °parcel carrier like FedEx, UPS, or GSO. Scorpions samples should not be frozen or boiled.

Freezing Freeze the sample in a plastic ETS sample tube. Do not over-fill the tube – leave a small space for the sample to expand when frozen.

Never freeze samples in glass containers to prevent breakage and injury.

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L O C AT I O N S & SERVICES In addition to our St. Helena headquarters, we operate satellite laboratories across the West coast to bring advanced tools to winemakers' doorsteps and provide local support to other growing wine regions.

These quick guides provide a reference for the 2021 harvest, including weekend hours and convenient services to make it easier than ever to send in harvest samples, including dropbox locations and complimentary courier service.

As always, if you have any questions, just call your local lab and we'll be happy to help.

S T. H E L E N A C A L I F O R N I A

A F T E R H O U R S D RO P B OX S T. H E L E N A

PHONE

(707) 963-4806

Located at our laboratory, next to the main entrance.

COURIER SERVICE Our complimentary courier service is available in Napa, Sonoma, and Mendocino counties every day ETS is open.

Samples left overnight will be processed when we open the following business day.

SHIPPING ADDRESS

REQUEST A PICKUP

899 Adams Street, Suite A St. Helena, CA 94574

You can request a courier pickup by logging in to your ETS account, or by calling our lab.

SEE INSTRUCTIONS FOR SHIPPING JUICE S A M P L E S – P. 4 1

DEADLINE

Please request a pickup by 10 am This allows us to ensure speedy turnaround on time-critical harvest analyses.

HOURS Monday – Friday: 7am – 8pm

DROPBOX IS LOCKED FO R T H E P I N C O D E , C A L L U S , O R LO G I N TO YO U R E T S A C C O U N T A N D V I S I T “ C O N TA C T ”

HARVEST HOURS

WEEKEND SCHEDULE

LO D I D RO P B OX E S TAT E C R U S H

F O L L O W U S O N T W I T T E R F O R U P D AT E S : TWITTER.COM/ETSLABS

S AT U R D AY

S U N D AY

AUG 14 – SEPT 03

On Call*

On Call*

SEPT 04 – SEPT 24

9am - 4pm

On Call*

SEPT 25 – OCT 29

9am - 6pm

9am - 4pm

OCT 30 – NOV 12

9am - 4pm

On Call*

NOV 13 – NOV 21

On Call*

On Call*

* TO SCHEDULE ON-CALL SERVICE, PLEASE C A L L B Y 2 P M O N F R I D AY : ( 7 0 7 ) 9 6 3 - 4 8 0 6

COMING SOON: S O U T H N A PA D R O P B O X RUTHERFORD EQUIPMENT 759 Technology Way Napa, CA

2 W. Lockeford Street Lodi, CA PICKUP TIME

Samples are picked up at 10am each weekday, and Saturdays during harvest.

Harvest is a busy time for everyone, save some time dropping samples closer to where you are.

Updates on our new dropbox can be found on our website: www.etslabs.com/contact.

OPENING HOURS T H E D R O P B O X I S AVA I L A B L E W H E N E S TAT E C R U S H I S O P E N : ( 2 0 9 ) 3 6 8 - 7 5 9 5

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HEALDSBURG CALIFORNIA

A F T E R H O U R S D RO P B OX HEALDSBURG

PHONE

(707) 433-7051

Located at our laboratory, on the north side of the building.

COURIER SERVICE Our complimentary courier service is available in Napa, Sonoma, and Mendocino counties every day ETS is open.

Samples left overnight will be processed when we open the following business day.

ADDRESS

REQUEST A PICKUP

190 Foss Creek Circle, Suite G Healdsburg, CA 95448

You can request a courier pickup by logging in to your ETS account, or by calling our lab.

I F YO U N E E D TO S H I P S A M P L E S , P L E A S E S E N D T H E M D I R E C T LY T O O U R S T . H E L E N A LAB: 899 ADAMS STREET, SUITE A ST. HELENA, CA 94574

DEADLINE

Please request a pickup by 10 am This allows us to ensure speedy turnaround on time-critical harvest analyses.

HOURS Monday – Friday: 7am –7pm

DROPBOX IS LOCKED. FO R T H E P I N C O D E , C A L L U S , O R LO G I N TO YO U R E T S A C C O U N T A N D V I S I T “ C O N TA C T ”

HARVEST HOURS

D R O P B O X L O C AT I O N S

WEEKEND SCHEDULE

SONOMA COUNTY

F O L L O W U S O N T W I T T E R F O R U P D AT E S : TWITTER.COM/ETSLABS

S U N D AY

AUG 14 – SEPT 03

On Call*

On Call*

7795 Bell Rd, Windsor, CA 95492

SEPT 04 – SEPT 24

9am - 4pm

On Call*

• S A M P L E S W I L L B E A C C E P T E D AT E N A R T I S A N D W I L L B E TA K E N TO E T S L A B O R ATO R I E S FOR TESTING.

SEPT 25 – OCT 29

9am - 4pm

9am - 4pm

OCT 30 – NOV 12

9am - 4pm

On Call*

NOV 13 – NOV 21

On Call*

On Call*

* TO SCHEDULE ON-CALL SERVICE, PLEASE C A L L B Y 2 P M O N F R I D AY : ( 7 0 7 ) 9 6 3 - 4 8 0 6

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ENARTIS

S AT U R D AY

• P L E A S E L E AV E S A M P L E S B Y 1 1 A M F O R S A M E - D AY S E R V I C E

PASO ROBLES

CALIFORNIA

A F T E R H O U R S D RO P B OX PA S O R O B L E S

PHONE

(805) 434-9322

Located at our laboratory, next to the front door.

Samples left overnight will be processed when we open the following business day.

ADDRESS 3320 Ramada Drive, Suite B Paso Robles, CA 93446

COURIER SERVICE We're excited to offer complimentary courier service this September for our clients in the Central Coast.

REQUEST A PICKUP

You can request a courier pickup by logging in to your ETS account, or by calling our Paso Robles lab.

DEADLINE

Please request a pickup by 10 am.

I F YO U N E E D TO S H I P S A M P L E S , P L E A S E S E N D T H E M D I R E C T LY T O O U R S T . H E L E N A LAB: 899 ADAMS STREET, SUITE A ST. HELENA, CA 94574

This allows us to ensure speedy turnaround on time-critical harvest analyses.

HOURS

COURIER SERVICE SERVICE AREA

Monday – Friday: 7am – 7pm

DROPBOX IS LOCKED. FO R T H E P I N C O D E , C A L L U S , O R LO G I N TO YO U R E T S A C C O U N T A N D V I S I T “ C O N TA C T ”

HARVEST HOURS

D R O P B O X L O C AT I O N S

WEEKEND SCHEDULE

CENTRAL COAST WINE SERVICES

F O L L O W U S O N T W I T T E R F O R U P D AT E S : TWITTER.COM/ETSLABS

AUG 14 – SEPT 03

We're currently picking up samples in the Paso Robles and SLO areas. We will continue to expand as demand growsplease let us know if you'd like service.

S AT U R D AY

S U N D AY

On Call*

On Call*

2717 Aviation Way #101, Santa Maria, CA 93455

S T O L P M A N V I N E YA R D S

SEPT 04 – SEPT 24

9am - 4pm

On Call*

SEPT 25 – OCT 29

9am - 4pm

9am - 4pm

OCT 30 – NOV 12

9am - 4pm

On Call*

NOV 13 – NOV 21

On Call*

On Call*

1700 Industrial Way, Lompoc, CA 93436

ENARTIS 270 E Hwy 246 #109, Buellton, CA 93427 FOR ALL CENTRAL COAST DROPBOXES, PLEASE DROP SAMPLES BY 11:00 AM FOR S A M E - D AY D E L I V E R Y M O N D AY - F R I D AY

* TO SCHEDULE ON-CALL SERVICE, PLEASE C A L L B Y 2 P M O N F R I D AY : ( 7 0 7 ) 9 6 3 - 4 8 0 6

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NEWBERG OREGON

A F T E R H O U R S D RO P B OX NEWBERG

PHONE

(503) 537-6245

Located at our laboratory, next to the main entrance.

Samples left overnight will be processed when we open the following business day.

ADDRESS

COURIER SERVICE Our complimentary courier service is available in Salem, McMinnville, Newberg, and the surrounding areas every day ETS is scheduled to be open.

REQUEST A PICKUP

214 W. Hancock Street Newberg, OR 97132

You can request a courier pickup by logging in to your ETS account, or by calling our lab.

I F YO U N E E D TO S H I P S A M P L E S , P L E A S E S E N D T H E M D I R E C T LY T O O U R S T . H E L E N A LAB: 899 ADAMS STREET, SUITE A ST. HELENA, CA 94574

DEADLINE

Please request a pickup by 10 am This allows us to ensure speedy turnaround on time-critical harvest analyses.

HOURS Monday – Friday: 7am – 7pm

DROPBOX IS LOCKED. FO R T H E P I N C O D E , C A L L U S , O R LO G I N TO YO U R E T S A C C O U N T A N D V I S I T “ C O N TA C T ”

HARVEST HOURS

WEEKEND SCHEDULE

ROS E B U RG D RO P B OX UMPQUA COMMUNITY COLLEGE

F O L L O W U S O N T W I T T E R F O R U P D AT E S : TWITTER.COM/ETSLABS

S AT U R D AY

S U N D AY

AUG 21 – SEPT 10

On Call*

On Call*

SEPT 11 – OCT 31

9am - 4pm

On Call*

NOV 01 – NOV 21

On Call*

On Call*

* TO SCHEDULE ON-CALL SERVICE,PLEASE C A L L B Y 2 P M O N F R I D AY : ( 5 0 3 ) 5 3 7 - 6 2 4 5

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PICKUP TIME

Samples collected at 11:30 Monday – Thursday.

Anything dropped off after 11:30am will be processed with the next pickup.

912 Umpqua College Rd. Roseburg, OR

WALLA WALLA WASHINGTON

PHONE

HOURS

COURIER SERVICE

(509) 524-5182

Monday – Friday: 7am – 7pm HARVEST HOURS

ADDRESS

A F T E R H O U R S D RO P B OX WA L L A WA L L A

3020 E. Isaacs Ave. Walla Walla, WA 99362 I F YO U N E E D TO S H I P S A M P L E S , P L E A S E S E N D T H E M D I R E C T LY T O O U R S T . H E L E N A LAB: 899 ADAMS STREET, SUITE A ST. HELENA, CA 94574

Located at our laboratory, next to the main entrance.

Samples left overnight will be processed when we open the following business day.

WEEKEND SCHEDULE

Our complimentary courier service is available in Walla Walla and the surrounding areas every day ETS is scheduled to be open.

REQUEST A PICKUP

You can request a courier pickup by logging in to your ETS account, or by calling our lab.

F O L L O W U S O N T W I T T E R F O R U P D AT E S : TWITTER.COM/ETSLABS

S AT U R D AY

S U N D AY

AUG 21 – SEPT 10

On Call*

On Call*

Please request a pickup by 10 am

SEPT 11 – OCT 31

9am - 4pm

On Call*

This allows us to ensure speedy turnaround on time-critical harvest analyses.

NOV 01 – NOV 21

On Call*

On Call*

* TO SCHEDULE ON-CALL SERVICE,PLEASE C A L L B Y 2 P M O N F R I D AY : ( 5 0 9 ) 5 2 4 - 5 1 8 2

DEADLINE

DROPBOX IS LOCKED. FO R T H E P I N C O D E , C A L L U S , O R LO G I N TO YO U R E T S A C C O U N T A N D V I S I T “ C O N TA C T ”

D R O P B O X L O C AT I O N S PROSSER

R E D M O U N TA I N CO O P E R W I N E CO.

AUGUST-JANUARY

RICHLAND CENTRAL INDUSTRIAL SALES

WOODINVILLE W O O D I N V I L L E C U S TO M CRUSH

Dropbox

401 7th St. Prosser, WA

35306 N Sunset Rd. Benton City, WA

2235 Henderson Loop Richland, WA

14030 NE 145th St., Suite B Woodinville, WA

PICKUP TIME: 11AM

PICKUP TIME: 11:30 AM

PICKUP TIME: 12 PM

PICKUP TIME:

PLEASE DROP SAMPLES BY 10:30 A M F O R S A M E - D AY D E L I V E R Y M O N D AY - F R I D AY

PLEASE DROP SAMPLES BY 11 A M F O R S A M E - D AY D E L I V E R Y M O N D AY - F R I D AY

PLEASE DROP SAMPLES BY 11:30 A M F O R S A M E - D AY D E L I V E R Y M O N D AY - F R I D AY

PLEASE DROP SAMPLES BY 2 PM FOR OVERNIGHT SHIPMENT M O N D AY - F R I D AY

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