The AWRI methodology

applied by ADVID and

comparison with the Glories
Method

Dr Paul Smith
Research Manager - Chemistry
 How does the MCP tannin method
work?

Treatment
Allow Allow
to Addition to Phenolics
Stand of salt Stand without Tannin
Methyl Cellulose for 2 and for 10
polymer solution mins water mins
Tannin /
Centrifuge methylcellulose
Wine or grape pellet
extract
Measure A280

Control Phenolics
incl. Tannin
Addition
of salt
and A280 (Control) - A280 (Treatment) = A280 (Tannin)
Wine or grape water
extract only (expressed as
Measure
g/L epicatechin
A280
equivalents)
 How does the Glories method work?

pH1 Addition Phenolics

of HCl incl. Tannin A520 (pH1 – pH3.2)
and
water A280 and A520 = extractable
Grape pH1
homogenate
anthocyanins
Measure
4 hours
(known mass) A280
A520
A280 (pH1)
Addition
pH3.2 of Phenolics =IPT (total phenolics)
tartaric incl. Tannin
acid and
water
A280 and A520
pH3.2 A280:A520 (pH3.2)
Grape
homogenate
Measure = seed maturity
4 hours
(known mass) A280
A520
 How do the methods differ?

http://www.awri.com.au/wp-content/uploads/2014/01/measuring-grape-tannins.pdf
How do the results compare?
How do the results compare?
 How do the results compare?

Fragoso, S., M. Mestres, O. Busto, and J. Guasch (2010) Comparison of Three Extraction Methods Used To Evaluate Phenolic
Ripening in Red Grapes. Journal of Agricultural and Food Chemistry.

NOTE: Ultra Turrax homogenisation of 300 berries, extracted 25g of homogenate. Did not use acid for AWRI method.
Why do the results differ?
 New metrics:
‘Wine extractable’ tannin

• Whole grape extracts, crushed berries in acidified 15% ethanol

• Harsh grape extractions (50% ethanol) following homogenisation

• 1 kg fermentations

HYPOTHESIS:

Extract dilute ethanol

>
Extract with high solvent:water
 A new way of assessing
Grape tannin extractability

Bindon, K., S. Kassara, W. Cynkar, E. Robinson, N. Scrimgeor, and P.A. Smith (2014) Comparison of
Extraction Protocols To Determine Differences in Wine-Extractable Tannin and Anthocyanin in Vitis vinifera L.
cv. Shiraz and Cabernet Sauvignon Grapes. Journal of Agricultural and Food Chemistry 62, 4558−4570. doi:
dx.doi.org/10.1021/jf5002777.
 Wine-extractable grape tannin

• A strong correlation was found with wine-like extraction and

wine tannin

• Small differences were found between cultivars

 Regionality affects
wine-extractable grape tannin

• Large differences were found between regions

• Riverland = lowest tannin; McLaren Vale = highest tannin

• Highest tannin grapes/wines were also the ‘ripest’ samples

3

2.5 y = 1.0484x - 0.6157

R² = 0.9273
Wine tannin (g/L)

1.5
Shiraz
Cabernet S.
1 Langhorne Creek
Riverland
McLaren Vale
0.5 Clare
Padthaway
Linear (Shiraz)
0
0.5 1 1.5 2 2.5 3 3.5
Wine-like tannin extract (mg/g)
 Homogenate extracts still work OK...

• Correlation grape – wine for homogenate extracts was not

as clear-cut as for wine-like extracts

• BUT when the data was separated for cultivar, homogenate

tannin strongly correlated with wine tannin
A sneak peek…
 A280 Tannin

What does this number mean?

In the Vineyard
 Tannin (epicat. eq. mg/g)

2
4
8

6
10
Barossa

Limestone Coast

Lower Murray
CAS

Murray Darling

SW Australia

Tannin (epicat. eq. mg/g)

2
4
6
8
10
concentration

Barossa

Limestone Coast
SHZ

Lower Murray
Regional differences in Grape Tannin

SW Australia
 Vintage differences in Grape Tannin
concentration

CAS SHZ
10 10
Tannin (epicat. eq. mg/g)

Tannin (epicat. eq. mg/g)

8 8

6 6

4 4

2 2
2006 2007 2006 2007

Vintage N Mean Vintage N Mean

2006 81 5.5 2006 97 4.1
2007 32 7.2 2007 64 5.5
 Varietal differences in Grape Tannin
concentration
10
Tannin (epicat. eq. mg/g)
8

CAS MER SHZ

Variety N Low Med High Mean

CAS 116 2.6 – 5.2 mg/g 5.2 - 6.8 mg/g 6.8 - 9.8 mg/g 5.98 mg/g
MER 81 1.9 – 4.8 mg/g 4.8 – 6.7 mg/g 6.7 – 8.7 mg/g 5.68 mg/g
SHZ 167 2.2 - 3.6 mg/g 3.4 - 5.4 mg/g 5.4 - 10.7 mg/g 4.63 mg/g
 90
Tannins 2009 (mg/g
dm)
80 Tannins 2010 (mg/g
dm)
70 Tannins 2011 (mg/g
dm)
Tannin concentration (mg/g dm)

Tannins 2012 (mg/g

60 dm)

50

40

30

20

10

0
70 60 50 40 30 20 10 0

Days before harvest

How does vintage variation

affect tannin and colour?
Grapes 2009 2010 2011 2012
Colour (fw) 2.36b 2.57a 2.22b 1.92c
Tannins (fw) 6.33b 6.54b 4.09c 7.23a
Colour (dm) 7.13b 8.20a 8.49a 6.26c
Tannins (dm) 21.19b 20.91b 15.66c 23.28a
Wine
Colour (mg/L) 686b 1035a 479d 592c
Tannins (mg/L) 1.47b 1.89a 0.74c 1.35b
Wine score 15 - 17 14 – 17.5 11 - 15 ???

These values are averages of 3 clones : CL10, SA125 and Reynella grown on TR

How does vintage variation

affect tannin and colour?
 Colour Density vs. Tannin
Cool 2006

Cabernet Shiraz
5
Tannin (epicat. eq. g/L)

Tannin (epicat. eq. g/L)

R= 0.47 R= 0.42
4

3
2

1 1
8 9 10 11 12 13 14 15 9 10 11 12 13 14 15

Colour density (AU) Colour density (AU)

* Colour and data analysis performed by Dr. R. Dambergs

 Colour Density vs. Tannin
Warm 2004

Cabernet Shiraz
R = 0.66 2 R = 0.78
Tannin (epicat. eq. g/L)

2
1

5 10 15 20 5 10 15

Colour density (AU) Colour density (AU)

* Colour and data analysis performed by Dr. R. Dambergs

In the Winery
 Tannin (epicat. eq. g/L)

4
5

1
2
3
Barossa
Fleurieu
Greater Perth
Limestone Coast
Lower Murray
CAS

Mt Lofty Ranges
NE Victoria
NW Victoria
Sth WA
Sth NSW
4

2
3

1
5

Barossa
Fleurieu
concentration

Greater Perth
Limestone Coast
Lower Murray
SHZ

Mt Lofty Ranges
NE Victoria
Regional differences in Wine Tannin

NW Victoria
Sth WA
Sth NSW
 Vintage differences in Wine Tannin
concentration

Tannin (epicat. eq. g/L)

4
SHZ CAS
3

1 2004

2005

2006

2007
2004 2005 2006 2007
Variety N Mean N Mean N Mean N Mean
CAS 89 1.8 57 1.7 189 2.2 209 2.8
SHZ 69 1.3 62 1.6 290 2.0 603 2.5

Concentrations in g/L
 Varietal differences in Wine Tannin
concentration

Tannin (epicat. eq. g/L)

4

0
CAS MER PIN SHZ

Variety N Low Med High Mean

CAS 629 0.1 – 1.7 g/L 1.7 – 2.8 g/L 2.8 – 5.3 g/L 2.9 g/L
MER 188 0.3 - 1.4 g/L 1.4 – 2.2 g/L 2.2 – 3.3 g/L 1.8 g/L
PIN 112 0.4 – 0.8 g/L 0.8 – 1.4 g/L 1.4 – 3.5 g/L 1.2 g/L
SHZ 1107 0.4 – 1.7 g/L 1.7 – 2.7 g/L 2.7 – 4.9 g/L 2.2 g/L
 Sensory
and
Consumer Preferences
 MCPT assay and Sensory

6.0

5.5

5.0
Drying

4.5

4.0

3.5
r2=0.85

3.0
1.0 1.5 2.0 2.5

Wine Tannin by MCPT assay (epicat. eq. g/L)

Sensory data: Consumer Study - Kate Lattey, Leigh Francis
 Acknowledgements

• Keren Bindon
• Jacqui McRae
• Alex Schulkin
• Bob Dambergs
• Sheridan Barter
• Mark Solomon
• Ruchi Shah
• Leigh Francis

• Stella Kassara
• Wies Cynkar
• Ella Robinson
• Neil Scrimgeour
• Peter Godden The Australian Wine Research Institute, a member of the
• Eric Wilkes Wine Innovation Cluster in Adelaide, is supported by
Australian grapegrowers and winemakers through their
investment body, the Australian Grape and Wine Authority
with matching funds from the
Australian Government.
 Understanding grape and wine tannins

Grape tannins
•proanthocyanidins or condensed tannins
•polymeric flavan-3-ols (catechins)
•colourless polyphenols
•precipitate proteins  astringent

Red wine tannins

•grape tannins (30%) & modified grape tannins (70%)
•plus oak-derived hydrolysable tannins & oenotannins
•include also pigmented tannins (pigmented polymers)
 Colour properties of wines made with
S. cerevisiae and S. bayanus

S. cerevisiae S. bayanus S. cerevisiae S. bayanus

Day 8 Day 8 Day 387 Day 387

WCD 11.93±0.25 11.97±0.13 8.80±0.23 10.34±0.25

M3G 357.7±4.9 300.8±3.0 125.6±2.6 88.5±3.3

PP 11.9±0.2 12.6±0.2 20.5±0.5 31.5±1.1

• WCD not correlated with anthocyanins on either day

• Pigmented polymers account for much of the day

387 WCD
WCD: wine colour density (sum of absorbance 420nm + absorbance 520 nm)
M3G: malvidin-3-glucoside (mg/L)
PP: pigmented polymers by HPLC as M3G equivalents (mg/L)
 Tannin addition trial

• No effect on colour parameters

• Small but significant effect on astringency

Panel score for 'overall astringency'
3.0
Average wine colour density (A420 +A 520 )
Panel Score (Mean)

6
2.5
WCD (a.u.)

5
2.0 4
1.5 3

1.0 2
1
0.5
0
0.0
control - - - fermentation
pre post - fermentation
control pre -
posttreatment
200 mg/L 500treatment
mg/L 1000 mg/L
fermentation fermentation
treatment treatment tannin added at sensory
 Wine tannins, colour and age

25000 14.0
anthocyanins
12.0
20000

wine colour density 10.0

15000
8.0

6.0
10000

4.0
5000
2.0

0 0.0
2000 1990 1980 1970 1960 1950

Coonawarra Cabernet Sauvignon vintage 1954 – 2004, S. Hodder

analysed 12/2004 by M. Parker & B. Dambergs
 Wine tannins, colour and age

25000 14.0
anthocyanins
anthocyanins
12.0
20000

wine colour
wine colourdensity
density 10.0

15000
8.0

Pigmented tannins
pigmented tannins
6.0
10000

4.0
5000
2.0

0 0.0
2000 1990 1980 1970 1960 1950

Coonawarra Cabernet Sauvignon vintage 1954 – 2004, S. Hodder

analysed 12/2004 by M. Parker & B. Dambergs
 Current Research

Are there Relationships between

Tannin Concentration and
Variety,
Quality and
Consumer Preference?
 Quality vs. Wine Tannin

CAS SHZ

Tannin (epicat. eq. g/L)

Tannin (epicat. eq. g/L)

Higher Lower Higher Lower

Quality Grading Quality Grading

Analysis of Variance Analysis of Variance

F Ratio = 12.1 F Ratio = 42.2
p Value = <.0001 p Value = <.0001
N = 349 N = 634
One-Way ANOVA - plot shows mean and mean error bars and Std dev – (2004 –2007)
 Consumer Preference

7.5
Tannin ‘sweet spot’
(1.3 – 1.9 g/L)
Mean liking score

1.43
1.58
1.18
1.33 1.32 1.92
6.5
1.98 2.30
1.18 2.38
1.18 1.84

5.5
2.5 3.0 3.5 4.0 4.5 5.0

Astringency (drying) intensity

 Wine tannins, colour and age

30000

25000

20000

15000

10000

5000
 wine tannin concentration is not related to wine age
0

2000 1990 1980 1970 1960 1950

Coonawarra Cabernet Sauvignon vintage 1954 – 2004, S. Hodder

analysed 12/2004 by M. Parker & B. Dambergs
 Quality vs. Wine Tannin
Tannin (epicat. eq. g/L)

Quality Grading Quality Grading Quality Grading

Analysis of Variance Analysis of Variance Analysis of Variance
F Ratio = 11.8 F Ratio = 10.9 F Ratio = 8.3
p Value = <.0001 p Value = <.0001 p Value = <.0001
N = 273 N = 125 N = 190

One-Way ANOVA - plot shows mean and mean error bars and Std dev – (SHZ 2007 only)
 • Correlate very highly for 50% grape homogenate extracts
• Correlate highly for red wines

Red wine Grape homogenate extract

800 300

BSA (cat eq. mg/L)

y = 319.13x - 256.07 y = 0.3603x - 43.043
BSA (cat eq. mg/L)

600 R2 = 0.80 R2 = 0.96

200
400

100
200

0.50 1.00 1.50 2.00 2.50 3.00 100 300 500 700 900

MCPT (epicat eq. (g/L) MCPT (epicat eq. mg/L)

• The two assays show good precision

• Showed strong correlations with sensory attributes
• The MCP tannin assay is significantly faster (at least 5x), simpler
and cheaper to perform