Discolouration IVAS 2013 .pdf


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Charcoal treatment on Pinot noir
sparkling base wine: effect on colour,
protein content and foaming properties
Parmentier M. 1, Salmon T. 1, Kemp B. 2, Foss C. 2, Deroy A. 1, Marchal R. 1,2
1,2

Laboratoire d’Oenologie et Chimie Appliquée, URVVC EA 4707, Université de Reims Champagne Ardenne, France.
2 Plumpton College – Wine Centre, East Sussex, BN7 3AE, United Kingdom.
*richard.marchal@univ-reims.fr

Foaming properties are altered by carbon treatment (fig. 5), as previously observed [1,2]. The
contribution of proteins in wine-foaming properties has been known for a long time [3] but the
adsorption of proteins during a charcoal treatment has never been precisely investigated. In
previous studies, proteins were quantified using a direct colorimetric method in which
interferences due to phenolic compounds play a major role.
In this experiment, a modified Bradford method was used for the fisrt time to estimate the
impact of carbon treatment on wine protein content (fig. 6). Nevertheless, we observe a
surprising logarithmic relationship between the wine protein content and the pink colour
decrease, exhibiting a R2=0.987.

Introduction : why this study?
In the Champagne area, 70% of the sparkling wine is produced with Pinot noir and Pinot meunier
varieties (fig. 1). The release of anthocyanins from the skin during the pressing cycle (especially
from the 3rd cycle, fig. 2) gives a pink colour to the juice/wine that is sometimes too marked to
produce a wine as expected by the consumers (fig. 3).

1,6

1st "tailles"

1,4

Série A
Série B
Série C

1,2

100

1
0,8
0,6
0,4
0,2
0
0

50

100

150

200

Figure 1 : Pinot noir
grape berries.

Time (min)

Figure 2 : Press cycle used in region
producing sparkling base wine

Figure 3 : Pinot noir
grape juice released
during the 3rd press cycle.
This juice presents a
marked pink colour.

13
12
11

80

10
9

60

y=
R² = 0,889

40

8

y = 3,1219 ln(x) + 18,939
R² = 0,987

104,73e-0,017x

7
6

20

5

Carbon g/hL
0,2

0
0

10

20

30

0,15

0,1

0,05

0

A520

40

Figure 5 : Relationship between carbon
treatment and the decrease in total proteins.

To reduce or remove this colour, a charcoal treatment is often practiced. Oenological charcoals are
activated carbons, i.e nonspecific adsorptive agents made from wood. The sponge-like carbon binds
with weakly polar molecules, especially those containing benzene rings.
In this study, we try to evaluate in a first part the impact of a carbon treatment on two different Pinot
noir base wine characteristics : the colour and the foamability. In the second part, we try to see if
these changes could be explain by the wine protein content.

[Proteins] mg/L

Wine
before
treatment

120

Cuvée

Foam (mm)

Pressure (bars)

2nd "tailles"

Figure 6 : Relationship between wine discolouration
and the decrease in total proteins.

To better understand the deleterious impact of a carbon treatment on the wine protein content,
a SDS-PAGE + silver nitrate staining was achieved (fig. 7). Five main proteins were quantified
for each of the charcoal doses experienced. For a 60 g/hL treatment, the protein contents only
decreased by 10 to 37%, whereas the wine foamability decreased by 54%. Furthermore, a 40
g/hL charcoal treatment induced a protein content decrease lower than 20% (fig. 8) whilst the
foamability decreased by 50% (fig. 5) and the anthocyanins content by 86% (fig. 4). These
results contradict some of the studies previously published and clearly indicate that proteins
are only a part of the compounds implicated the foaming properties of Pinot noir sparkling wine.

Materials and Methods
Bande 64 kDa
3,0E+06

Carbon dose ( g/hL)
MW

0

10

20

30

40

Aire du pic

Charcoal treatment : A “pink” base wine produced with Pinot noir grape juice was treated with 0 to
60 g/hL charcoal (Carbion – IOC) in triplicate. Trials were conducted at controlled temperature
(18°C) in 100mL bottles. 24hrs after treatment, the carbon treated wines were centrifuged (5 min,
5000g) and filtered using 0.45µm pore size acetate cellulose membrane.

2,5E+06

60

MW
(KDa)

1,5E+06

y = -8948,1x + 2E+06
R² = 0,9264

1,0E+06
5,0E+05

charbon g/hL
0,0E+00

75

Colour : The pink colour was estimated with the A520.

2,0E+06

0

20

40

60

50
37
3,5E+06

Foamability : wine foamability was estimated using a sparging procedure (Krüss) with CO2.

2,5E+06

25

2,0E+06
Aire de pic

Protein concentration : the total protein content was estimated using a modified Bradford method.
The wines are ultrafiltered at 3 kDa. The interference with the CBB reagent observed for the
“ultrafiltrate + CBB” was substracted from the value obtained for the sample “wine + CBB”.

20

1,5E+06

y = -12909x + 3E+06
R² = 0,867

1,0E+06
5,0E+05
0,0E+00
0

15

Protein characteristics : Wine proteins are separated using a classical SDS-PAGE technique
(C=13.5%) + silver nitrate staining + Quantity one (Bio-Rad) analysis.

Bande 19kDa

3,0E+06

Figure 7 : Change in protein content according to an
increasing carbon treatment. SDS-PAGE and silver nitrate
staining. MW : molecular weight markers (kDa).

10

20

30

40

50
60
charbon g/hL

Fig. 8 : Changes in the concentration of specific
bands (from fig. 7.) Densitometric integration
after silver nitrate staining.

Colour and Foam
Conclusions and further investigations
As previously observed [1], carbon effectively removes wine phenolic compounds. The pink colour
decrease follows a negative exponential law with a R2>0.98 characteristic of a physisorption
phenomenon. For this wine and this carbon, a 40g/hL treatment was necessary to reduce the pink
colour and produce a wine as expected by the consumers.

Dose of carbon added to the Pinot noir wine (g/hL)
0

5

10

15

20

30

40

60

Carbon is the only oenological product capable to remove phenolic compounds from a
wine in the aim to reduce its pink colour. Nevertheless, stripping of wine is often a
problem with carbon because of the low selectivity.
Further investigations will focus on the possibility to use new products able to remove
anthocyanins with a high selectivity thus avoiding the deleterious effect on foaming
properties. This is of prime importance for Pinot noir (and Pinot meunier) base wine
produced for the elaboration of sparkling wines.

Water

References
A 520

[1] Maujean A., Poinsaut P., Dantan H., Brissonnet F. and Cossiez E., 1990. Etude de la teneur et de la qualité de
mousse des vins effervescents. II. Mise au point d'une technique de mesure de la moussabilité, de la tenue et de la
stabilité de la mousse des vins effervescents. Bull. O.I.V., 711-712, 405-427.

0,20
0,16

[2] Marchal R., Chaboche D., Douillard R. and Jeandet P. 2002. Influence of lysozyme on Champagne base wine
foaming properties. Journal of Agricultural and Food Chemistry, 50, 1420-1428.

y = 0,1536e-0,048x
R² = 0,9837

0,12

[3] Brissonnet F. and Maujean A. 1993. Characterization of foaming proteins in a Champagne base wine. Am. J.
Enol. Vitic., 44, 297-301.

0,08
0,04
0,00
0

5

10

15

20

25

30

35

40

[4] Marchal R., Seguin V. et Maujean A. Quantification of interferences in the direct measurement of proteins in
wines from the Champagne region using the Bradford method. Am. J. Enol. Vitic., 1997, 48, 303-309
.

Carbon (g/hL)
For their support, the authors thank the
Figure 4 : Relationship between the dose of charcoal and the wine pink
colour (A520)after a discolouration treatment

Champagne Palmer
Association Recherche Oenologie Champagne et Université

3-5 July 2013
Reims - France


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