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OENO One 2021, 1, 337-347 337
© 2021 International Viticulture and Enology Society - IVES
Received: 12 June 2020 y Accepted: 28 January 2021 y Published: 22 March 2021
DOI:10.20870/oeno-one.2021.55.1.3695
Changes in the avan-3-ol and polysaccharide content during the
fermentation of Vitis vinifera Cabernet-Sauvignon and cold-hardy
Vitis varieties Frontenac and Frontenac blanc
Paméla Nicolle1,2,4, Kyle A. Williams3, Paul Angers2, Karine Pedneault1,2,4*
1Centre de Développement Bioalimentaire du Québec, La Pocatière, QC, Canada, G0R 1Z0
2Département Sciences des aliments, Université Laval, Québec, QC, Canada, G1V 0A6
3Malvern Panalytical Inc., Westborough, MA 01581, United States
4Département des Sciences, Université Sainte-Anne, NS, Canada, B0W 1M0First
*corresponding author: karine.pedneault@usainteanne.ca
a b s t r a c t
Grape variety has a signicant impact on wine avan-3-ol and polysaccharide prole. The main objective of this
work was to study differences in avan-3-ol and polysaccharide diffusion from grape to wine during the fermentative
alcoholic maceration of three Vitis sp. varieties: the cold-hardy hybrid varieties Frontenac and Frontenac blanc,
and the V. vinifera variety Cabernet-Sauvignon. Polysaccharides from must and wine were precipitated by ethanol
and quantied using the phenol-sulfuric method of Dubois. Flavan-3-ol concentration and prole were analysed by
HPLC-FLD. Results showed that wines from Frontenac and Frontenac blanc had less oligomeric and polymeric
avan-3-ols than those from V. vinifera Cabernet-Sauvignon. Wines made from Frontenac also had a higher
concentration in total polysaccharides. Preliminary results from GPC/SEC analyses suggested that Frontenac wine
had a higher content in mannoproteins and rhamnogalacturonan-2 polysaccharides compared to the other studied
varieties. Overall, wines of Frontenac showed the highest content in total polysaccharides, and the lowest content
in condensed tannins. As polysaccharides are known to negatively impact wine perceived astringency, these results
suggest that signicant attention should be given to the polysaccharide composition of cold-hardy cultivars in the
context of cold climate wine production. Such knowledge may help winemakers from cold climate areas to improve
the winemaking processes and nal wine composition when working with cold-hardy Vitis sp. varieties.
Knowledge on interspecic hybrid polysaccharide and avan-3-ol kinetic during the alcoholic fermentative maceration
may help the winemakers from cold climate areas to improve winemaking processes and nal wine composition.
k e y w o r d s
cold-hardy grape, Vitis vinifera, cold climate, winemaking, avan-3-ol, polysaccharide
Supplementary data can be downloaded through: https://oeno-one.eu/article/view/3695
© 2021 International Viticulture and Enology Society - IVES
338 OENO One 2021, 1, 337-347
Paméla Nicolle et al.
INTRODUCTION
Cold-hardy Vitis cultivars issues from crosses
between V. vinifera and native American
species have been largely implemented for wine
production in northern areas such as Eastern
Canada, Eastern and Northern Europe and
Midwestern United States (Ehrhardt et al., 2014;
Liu et al., 2015; Ma et al., 2017;
Manns et al., 2013; Pedneault et al., 2013;
Zhang et al., 2015). Cold-hardy grapevines
present certain specications making them well
suited for northern climates, including resistance
to winter temperatures reaching below –30 °C and
a generally short growing season (Fennell, 2004;
Londo and Kovaleski, 2017). Along with cold
resistance, most of them also show a high degree
of resistance to fungal diseases (Pedneault and
Provost, 2016). In most cases, the specic genetic
of cold-hardy cultivars translates into certain
berry characteristics such as thick skins and high
rmness in berries, even at full ripeness (Pedneault
and Provost, 2016). These characteristics highly
contrast with traditional V. vinifera berries that
usually soften signicantly along the ripening
process (Maury et al., 2009; Robin et al., 1997).
Changes in berry rmness are mainly attributable
to modications in the mechanical properties
of cell walls occurring during ripening. Those
involve berry cell wall components such as
hemicellulose, pectin and cellulose that undergo
solubilisation and depolymerisation processes,
but also rearrangements of their associations
(Goulao and Oliveira, 2008). Both the nature and
the extent of these changes are inuenced by the
grapevine’s genotype as well as its interactions
with the environment (Rihan et al., 2017). Berries
from interspecic hybrid cultivars that present
cold-hardy and fungus-resistance properties have
long been known for the particularly high pectin
content of their skin cell walls when compared
to V. vinifera (Apolinar-Valiente et al., 2017;
Lee et al., 1975; Springer and Sacks, 2014).
The winemaking process partly aims at extracting
grape berry components such as tannins and aroma.
However, along the process, macromolecules
such as polysaccharides and proteins are
also extracted from berries as well as from
fermenting microorganisms (yeast, bacteria). Pectic
polysaccharides mostly originate from the grape
berry cell wall, whereas microorganisms provide
wine with glycoproteins such as mannoproteins
(Dols-Lafargue et al., 2007; Guadalupe and
Ayestarán, 2007; Vidal et al., 2003). The structure,
concentration and interactions between proteins,
tannins and polysaccharides play a crucial role
in the sensory properties of wine, especially
regarding the mouthfeel and taste of red wine.
The role of tannins in the sensory properties of
wine has been largely studied over the years and
extensively reviewed (Bajec and Pickering, 2008;
Ma et al., 2014; McRae and Kennedy, 2011;
Scollary et al., 2012; Soares et al., 2017). In
contrast, knowledge of wine polysaccharides and
proteins remains scarce. Recently, polysaccharides
have been shown to inhibit polyphenol-
protein aggregation (including tannin-protein
aggregation) and hence authors suggested that
polysaccharides can modulate wine astringency
(Brandão et al., 2017; Lankhorst et al., 2017;
Watrelot et al., 2017).
Poor astringency is the main issue in red
wine production from cold-hardy and
fungus-resistant cultivars in cold-climate regions
(Nicolle et al., 2018, 2019; Springer et al., 2016a).
Tannin extractability from interspecic hybrids
is known to be lower than that of Vitis vinifera
(< 6 % vs. 8 – 22 %) and usually results in wines
with fewer tannins (< 100 mg/L catechin equivalent)
with a low mean degree of polymerisation
(mDP ≤ 4) (Manns et al., 2013; Springer and
Sacks, 2014). From a sensory perspective, this
translates into bitterness rather than astringency.
Recent progress has highlighted the impact of
proteins on tannin retention in hybrid red wine
(Nicolle et al., 2019; Springer et al., 2016b),
but, thus far, little attention has been given to
polysaccharides in this context. Differences
between the respective cell wall composition
of cold-hardy and V. vinifera cultivars suggest
that polysaccharide content and composition of
cold-hardy berries might contribute to the poor
astringency of the resulting wines.
In this preliminary study, we followed the
changes in polysaccharide and tannin content
and prole during the alcoholic fermentation of
two red cultivars typically very different from
each other: V. vinifera Cabernet-Sauvignon
(high tannin extractability, 22 %; high tannin
content, up to 1900 mg/L catechin equivalent) and
cold-hardy cultivar Vitis sp. Frontenac (low tannin
extractability; low tannin content, < 160 mg/L
epicatechin equivalent) (Harbertson et al., 2008;
Nicolle et al., 2019; Springer and Sacks, 2014).
Whites fermented like red wines can show
lower viscosity and different mouthfeel sensory
attributes than red wines, differences that have
been both attributed to the absence of anthocyanins
during fermentation (Oberholster et al., 2009).
OENO One 2021, 2, 337-347 339
© 2021 International Viticulture and Enology Society - IVES
Yet, Frontenac blanc, a white cultivar derived
from white-fruited mutations of the varieties
Frontenac and Frontenac gris, typically produces
high viscosity wines. For this reason, we chose to
include this variety in this study.
The main objective of this study was to determine
the effect of grape variety on skin and seed
avan-3-ol and polysaccharide diffusion from
grapes to wine during the fermentative alcoholic
maceration of Vitis vinifera Cabernet-Sauvignon
and cold-hardy Vitis cultivars Frontenac and
Frontenac blanc and to determine the qualitative
and quantitative composition of the nal wines.
Knowledge of interspecic hybrid kinetics
during the winemaking may help the winemakers
from cool climate areas to improve winemaking
processes et wine composition.
MATERIALS AND METHODS
1. Grape material
The cold-hardy hybrid grape varieties, Frontenac
(FR, red variety) and Frontenac blanc (FB,
white variety) (both issued from Landot
(L. 4511) x Vitis riparia 89) were harvested in a
commercial vineyard located in Saint-Rémi (QC,
Canada) (45° 16′ 0″ N, 73° 37′ 0″ W). Berries
from Vitis vinifera Cabernet-Sauvignon (CS, red
variety) were imported from California (CA, USA)
through a local dealer. All berries were harvested
in 2015 and stored at –30 °C under controlled
atmosphere until the experiment, as carried out by
Springer et al. (2016b).
2. Winemaking trials
The grapes were thawed at 4 °C and then manually
destemmed and pressed. The must and pomace
were placed in a 10 L fermenter bucket, treated
with SO2 (30 mg/L, sulphur dioxide as potassium
metabisulte) and cold-soaked (4 °C, overnight).
The must and pomace were transferred in a 10
L fermentation unit equipped with a removable
head plate tted with two ports, one for sampling
and the other one for carbon dioxide discharge.
Temperature regulation in the fermentation unit
was carried out by circulating water through
two hoses connected to a temperature-controlled
water bath. Fermentations were performed as
follows: Alcoholic fermentative maceration
(AFM) was induced by a commercial dry yeast
Saccharomyces cerevisiae (Lalvin BM 4X4®;
Lallemand Inc., Montreal, Canada) at 250 mg/L
and carried out at 24 °C until dryness. The cap
was punched twice a day for the rst two days and
then once a day. Alcoholic fermentation level was
checked daily by measuring the concentration in
total soluble solid (°Brix). Fermenting must was
sampled daily for 11 days and stored at –30 °C
for future analyses. At the end of the process,
wines were pressed manually using cotton
cheesecloth, packed in hermetically sealed bags
under argon and stored at 4 °C. Fermentations
were performed in triplicate for each variety.
The composition of musts and nal wines (alcohol
concentration, % v/v; titratable acidity, g tartaric
ac. eq./L; pH; primary amino nitrogen, mg/L; and
ammonia, mg/L) is provided in Table 1.
3. Sugars and ethanol analysis
Ethanol, glucose and fructose contents were
quantied as described by Nicolle et al. (2019).
Briey, analyses were performed on an HPLC
system (WatersTM, Millipore Corp., Milford, Mass.
USA) equipped with a refractive index detector
(Hitachi model L-7490, Foster City California,
USA), using a WatersTM Sugar Pack-I column
(6.5 mm x 300 mm) from WatersTM (Millipore
Corp., Milford, Mass. USA). Analyses were
performed in duplicate.
4. Flavan-3-ol analysis
Flavan-3-ol content and composition were
measured as described by Nicolle et al. (2018).
Briey, analyses were carried out on an
Agilent 1260 Innity HPLC system (Agilent
Technologies, Santa Clara, CA, USA) equipped
with a uorescence detector (G1321C, Agilent,
Santa Clara, CA, USA). The separation was
performed on a Develosil® Diol column (250 mm
× 4.6 mm; 5 μm particle size) tted with a Cyano
Security Guard column (Phenomenex®, Torrance,
CA, USA). Analyses were performed in duplicate.
5. Polysaccharide analysis
Total polysaccharides were precipitated
as described by Segarra et al. (1995)
and quantied by UV-Vis spectroscopy
(UV-Vis spectrophotometer UV-2700; Shimadzu,
Quebec, Canada) using the phenol-sulfuric method
of Dubois et al. (1956). Galactose was used as a
standard for quantication. Total polysaccharide
precipitation and quantication were carried out
in triplicate.
Some preliminary and complementary gel
permeation/size exclusion chromatography
(GPC/SEC) assays were conducted using the
Malvern Panalytical OMNISEC® system (Malvern
Panalytical Ltd, Malvern, UK). The OMNISEC®
GPC/SEC system combines multiple detectors
© 2021 International Viticulture and Enology Society - IVES
340 OENO One 2021, 1, 337-347
Paméla Nicolle et al.
(differential refractive index, diode-array-based
UV/Vis spectrophotometer, right angle and low
angle light scattering and four-capillary differential
viscometer) to quantify polysaccharides and
measure their intrinsic viscosity (representative of
molecular structure, density and branching) and
absolute molecular weight. Given the interest of
this method and results and their relative novelty
to the characterisation of polysaccharide in Vitis
sp., the details of the method and the results are
presented as supplementary material.
6. Statistical analysis
ANOVA analyses of the must and wine basic
parameters (primary fermentable sugars,
alcohol concentration, titratable acidity, pH,
primary amino nitrogen and ammonia) were
analysed using the MIXED procedure of the
SAS® software (version 3.5 Basic Edition;
SAS Institute Inc., Cary, NC, USA). The DIFF option
in an LSMEANS (least-squares means) statement
was used and means were compared using the
Tukey HSD (“Honestly Signicant Difference”)
post-hoc test.
Flavan-3-ol and polysaccharide concentrations
were analysed with the SAS® software
(version 3.5 Basic Edition; SAS Institute Inc.,
Cary, NC, USA) using ANOVA methods with
PROC MIXED statement, analysing the main and
interaction effects of the two following factors:
cultivar and day of fermentation. Since each wine
was sampled during AFM, a repeated-measures
model was used, along with the DIFF option in
an LSMEANS (least-squares means) statement.
Multiple comparisons were made using the
Tukey HSD (“Honestly Signicant Difference”)
post-hoc test.
TABLE 1. Composition of musts and wines made from the cold-hardy Vitis sp. Frontenac blanc, Frontenac
and V. vinifera Cabernet-Sauvignon (Primary fermentable sugars, g/L; alcohol concentration, % v/v;
titratable acidity, g/L tartaric acid eq.; pH; primary amino nitrogen, mg/L; ammonia, mg/L).
1 For a given matrix (must, wine) and parameter, values in the same column followed by different letters are signicantly different
according to Tukey’s honest signicance test at the 0.05 probability level. n = 3 samples per variety X matrix (must, wine).
2 Not available.
Parameter Variety Must Wine
Primary fermentable sugars
(g/L)
Frontenac blanc 237.44 ± 1.85 A11.28 ± 0.38 A
Frontenac 227.85 ± 16.62 A 0.99 ± 0.01 A
Cabernet-Sauvignon 256.35 ± 39.95 A 1.68 ± 0.38 A
Alcohol (%, v/v)
Frontenac blanc 0.00 A 15.11 ± 0.49 A
Frontenac 0.00 A 13.99 ± 0.00 B
Cabernet-Sauvignon 0.00 A 15.56 ± 0.04 A
Titrable acidity
(g/L tartaric acid eq.)
Frontenac blanc 13.24 ± 0.19 A 11.41 ± 0.04 B
Frontenac 14.14 ± 1.49 A 14.97 ± 0.95 A
Cabernet-Sauvignon 4.25 ± 0.24 B 8.66 ± 0.88 C
pH
Frontenac blanc 3.09 ± 0.06 B 3.18 ± 0.09 B
Frontenac 3.08 ± 0.12 B 3.25 ± 0.05 B
Cabernet-Sauvignon 3.65 ± 0.08 A 3.86 ± 0.05 A
Primary amino nitrogen
(mg/L)
Frontenac blanc 272.33 ± 27.65 A n.a.2
Frontenac 206.67 ± 5.86 B n.a.
Cabernet-Sauvignon 118.00 ± 14.73 C n.a.
Ammonia (mg/L)
Frontenac blanc 11.67 ± 3.21 C n.a.
Frontenac 31.12 ± 6.87 B n.a.
Cabernet-Sauvignon 49.00 ± 2.83 A n.a.
OENO One 2021, 2, 337-347 341
© 2021 International Viticulture and Enology Society - IVES
TABLE 2. Monomeric, oligomeric (2 - 5 avan-3-ol units) and polymeric (≥ 6 avan-3-ol units)
avan-3-ol content (mean ± SD, mg/L epicatechin equivalent) during the alcoholic fermentative maceration
(AFM) of V. vinifera Cabernet-Sauvignon and cold-hardy Vitis sp. cultivars Frontenac and Frontenac blanc.
1Values on the same row (lower-case letters) or the same column (capital letters) followed by different letters are signicantly
different according to Tuckey’s honest signicance test at the 0.05 probability level.
Cultivar
Parameter Day of AFM Cabernet-Sauvignon Frontenac Frontenac blanc
Mean ± SD Mean ± SD Mean SD
Monomeric
avan-3-ol
(mg/L EC eq.)
0 15.29 ± 0.76 G1b 17.93 ± 2.16 FG ab 25.88 ± 4.97 H a
1 16.99 ± 0.26 G b 14.04 ± 1.00 G b 27.92 ± 3.42 GH a
2 28.98 ± 0.32 F a 14.79 ± 0.42 G b 32.67 ± 4.87 G a
3 30.17 ± 0.02 F b 22.14 ± 3.68 F b 46.46 ± 2.52 F a
4 36.39 ± 0.79 Eb 34.97 ± 4.52 Eb 59.20 ± 4.54 Ea
5 41.69 ± 1.23 D b 49.99 ± 4.76 D b 78.30 ± 8.20 D a
6 47.37 ± 2.65 C b 55.77 ± 6.20 BC b 87.05 ± 16.33 BC a
7 48.65 ± 4.50 BC b 56.67 ± 3.22 ABC b 89.34 ± 9.23 AB a
8 51.23 ± 2.22 ABC b 58.45 ± 3.45 ABC b 82.45 ± 6.45 CD a
9 48.40 ± 5.30 C b 53.45 ± 4.64 CD b 89.45 ± 8.37 AB a
10 53.64 ± 2.45 AB b 59.45 ± 2.45 AB b 91.23 ± 9.34 AB a
11 55.83 ± 7.99 A b 60.98 ± 12.34 A b 94.34 ± 4.56 A a
Oligomeric
avan-3-ol
(mg/L EC eq.)
0 27.99 ± 5.81 I a 17.41 ± 4.47 F a 15.26 ± 9.87 G a
1 30.85 ± 2.93 I a 11.28 ± 1.14 F b 11.18 ± 2.29 G b
2 49.22 ± 2.72 H a 13.49 ± 0.73 F b 28.85 ± 6.99 F b
3 61.31 ± 0.75 G a 20.05 ± 11.16 F b 45.03 ± 1.32 Ea
4 77.58 ± 1.10 F a 39.22 ± 6.85 Eb 52.02 ± 3.56 Eb
5 96.16 ± 3.34 Ea 45.05 ± 6.12 Ec 75.50 ± 7.22 D b
6 123.38 ± 2.55 D a 63.35 ± 6.18 D c 82.65 ± 13.44 D b
7 128.42 ± 15.60 D a 77.31 ± 4.23 C c 96.45 ± 11.20 C b
8 139.99 ± 13.67 C a 87.34 ± 2.34 BC b 98.39 ± 4.34 C b
9 144.27 ± 22.24 C a 93.45 ± 5.34 B b 109.34 ± 9.39 B b
10 165.77 ± 22.76 B a 109.34 ± 2.34 A b 119.98 ± 2.39 AB b
11 189.50 ± 27.35 A a 112.88 ± 5.39 A b 125.90 ± 19.30 A b
Polymeric
avan-3-ol
(mg/L EC eq.)
0 54.25 ± 13.04 Eb 68.74 ± 6.47 B b 101.74 ± 7.27 AB a
1 37.37 ± 9.52 F c 61.92 ± 10.86 BC b 84.48 ± 24.21 DE a
2 39.72 ± 5.98 F ab 28.06 ± 4.12 F b 53.41 ± 13.40 G a
3 42.69 ± 6.66 F ab 29.62 ± 5.78 EF b 54.35 ± 9.32 G a
4 60.39 ± 9.32 Eab 36.74 ± 0.82 Eb 58.89 ± 8.72 G a
5 55.99 ± 0.00 Ea 49.67 ± 9.90 D a 61.34 ± 12.90 G a
6 95.66 ± 0.00 C a 54.83 ± 5.58 CD b 60.50 ± 19.67 G b
7 86.64 ± 11.33 D a 65.60 ± 0.10 B b 71.12 ± 3.24 F ab
8 103.76 ± 12.22 BC a 69.09 ± 2.46 B b 79.34 ± 4.34 EF b
9 79.78 ± 14.65 D a 78.90 ± 6.78 A a 88.34 ± 8.45 CD a
10 113.94 ± 0.00 A a 82.34 ± 9.23 A b 95.43 ± 10.58 BC b
11 105.85 ± 17.56 AB a 84.34 ± 5.90 A b 109.12 ± 7.77 A a
© 2021 International Viticulture and Enology Society - IVES
342 OENO One 2021, 1, 337-347
Paméla Nicolle et al.
RESULTS AND DISCUSSION
1. Flavan-3-ols
In this study, avan-3-ol analyses were achieved
by HPLC-FLD using correction factors to
adjust the respective responses of small to large
proanthocyanidins in uorescence as outlined by
Nicolle et al. (2018). HPLC-FLD is much less used
than the traditional protein precipitation method
of Adams-Harberston for tannin quantication
in oenology. However, we recently showed that
results from both methods are highly correlated
(r2 = 0.8579) (Nicolle et al., 2019). Monomeric,
oligomeric (2-5 avan-3-ol units) and polymeric
(≥ 6 avan-3-ol units) avan-3-ol content during
AFM of V. vinifera Cabernet-Sauvignon (CB)and
cold-hardy Vitis sp. cultivars, Frontenac (FR) and
Frontenac blanc (FB) are presented in Table 2.
The concentration in polymeric avan-3-ols was
signicantly higher in FB musts when compared to
CS and FR musts. After 11 days of AFM, wines from
all three varieties showed a similar concentration
in polymeric avan-3-ols but the concentration in
monomeric avan-3-ols was signicantly higher
in FB wines compared to CS and FR wines.
CS wines showed a signicantly higher
concentration in oligomeric avan-3-ols compared
to FR and FB wines and overall, a higher
concentration in condensed tannins (oligomeric and
polymeric avan-3-ols) (295 mg/L EC equivalent
vs. 197 and 235 mg/L EC equivalent, respectively).
The tannin content and composition of the grape
skin and seed of all three varieties, as well as their
cell wall structure (involved in tannin diffusion and
retention in wine) could explain those differences.
The winemaking process (e.g., maceration time
and temperature), which was similar for all
cultivars in this study and the alcohol content
(e.g., involved in disorganization of seed protection
outer lipidic layer), which differed along the AFM
between varieties, also play an important role on
cell wall disruption and, therefore, the percentage
of extractable tannins from grape seeds and skins
(Rousserie et al., 2019).
The kinetics of avan-3-ol extraction during
AFM varied between cultivars but some
similarities were also observed. For instance,
the concentration in monomeric avan-3-ols
tripled in wines from all three varieties when
compared to musts and the concentration in
oligomeric avan-3-ols increased by six to eight
times during AFM. Previous studies on V. vinifera
cultivars showed that avan-3-ol monomers and
small avan-3-ol oligomers (2 - 3 avan-3-ol
units) are primarily extracted at the end of the
cold prefermentative maceration whereas larger
avan-3-ol oligomers (4 - 5 avan-3-ols units) are
mostly extracted during further winemaking stages
(González-Manzano et al., 2006). In contrast
with mono- and oligomers, the concentration in
polymeric avan-3-ols doubled in CS wines during
AFM, whereas both FB and FR wines showed
little to no signicant difference in this aspect.
FIGURE 1. Kinetic of fermentable sugar consumption during the alcoholic fermentative maceration of the
cold-hardy Vitis sp. Frontenac blanc, Frontenac and V. vinifera Cabernet-Sauvignon.
OENO One 2021, 2, 337-347 343
© 2021 International Viticulture and Enology Society - IVES
Our results show a dramatic fall in polymeric
avan-3-ol concentration in both FB and FR
wines (up 2.5 and 1.9 times less, for FR and FB
wines, respectively) 2 days after the beginning
of AFM, suggesting that a physicochemical
phenomenon occurred. Previous studies showed
that the rise in ethanol concentration during
AF weakens the hydrophobic interactions
between cell wall components and polymeric
avan-3-ols, thereby facilitating their extraction
(Casassa and Harbertson, 2014). However, based
on the kinetics of fermentable sugar consumption
by yeast, both FR and FB ended their fermentation
faster than CS (5 days versus 9 days) but yet
showed a dramatic decrease in their polymeric
avan-3-ol content (Figure 1 and Table 2).
Cell wall components from berries of cold-hardy
cultivars have been shown to bind tannins at
a higher rate than those from V. vinifera berries
(Springer and Sacks, 2014; Springer et al., 2016a).
Results on the negative impact of pomace on
tannin retention in Frontenac wines recently
suggested that skin cell wall components,
including polysaccharides, could have a larger
role than initially anticipated on tannin retention
in cold-hardy wines (Nicolle et al., 2019).
2. Total polysaccharides
In this study, direct precipitation of the total
must/wine colloids with ethanol acid, followed by
the traditional colorimetric phenol-sulfuric
assay were used for the determination of total
polysaccharides. This direct quantication
method reacts with both neutral and acidic
polysaccharides, although the sensitivity for
neutral polysaccharides is 2.5 times higher than
for acidic polysaccharides (Segarra et al., 1995).
Total polysaccharide content during AFM of
V. vinifera Cabernet-Sauvignon and cold-hardy
Vitis sp. cultivars Frontenac and Frontenac blanc
are presented in Table 3.
Musts from FB, FR and CS showed no signicant
difference in total polysaccharide concentration,
but signicant differences between wines.
On day 10, FR wines had a signicantly higher
concentration in polysaccharides than CS wines
(1321.4 mg/L compared to 921.7 mg/L galactose
equivalent; Table 3) whereas FB showed similar
content to both CS and FR.
The kinetics of polysaccharide extraction
during the winemaking process showed that
the concentration of polysaccharides increased
TABLE 3. Polysaccharide concentration (mean ± SD, mg/L galactose equivalent) during the alcoholic
fermentative maceration (AFM) of V. vinifera Cabernet-Sauvignon and cold-hardy Vitis sp. cultivars
Frontenac and Frontenac blanc.
1Values on the same row (lower-case letters) and the same column (capital letters) followed by different letters are signicantly
different according to Tuckey’s honest signicance test at the 0.05 probability level.
Cultivar
Parameter Day of
AFM Cabernet-Sauvignon Frontenac Frontenac blanc
Mean ± SD Mean SD Mean SD
Polysaccharide
(mg/L
galactose eq.)
0 439.81 ± 193.58 FG1a 476.99 ± 59.75 G a 279.95 ± 85.20 F a
1 364.41 ± 90.58 G b 582.51 ± 79.74 G ab 698.31 ± 130.93 Ea
2 567.43 ± 98.98 Eb 956.24 ± 227.77 F a 974.35 ± 190.97 DE a
3 831.20 ± 97.53 BC b 1200.11 ± 204.38 Ea 1095.49 ± 215.73 CDE ab
4 603.54 ± 17.94 Eb 1201.87 ± 171.89 Ea 1056.72 ± 198.98 BCD a
5 555.09 ± 121.23 EF c 1432.90 ± 123.34 A a 1160.70 ± 123.37 AB b
6 626.73 ± 17.47 DE b 1672.93 ± 249.02 AB a 1077.73 ± 137.21 ABCD a
7 1009.34 ± 107.47 A b 1411.50 ± 232.90 A a 1110.13 ± 156.40 ABC b
8 745.67 ± 76.54 CD c 1567.90 ± 301.87 AB a 1190.34 ± 130.44 A b
9 751.49 ± 111.09 C b 1459.98 ± 274.98 BC a 990.45 ± 120.40 CDE b
10 921.65 ± 116.29 AB b 1321.37 ± 201.85 DE a 989.45 ± 198.40 CDE ab
© 2021 International Viticulture and Enology Society - IVES
344 OENO One 2021, 1, 337-347
Paméla Nicolle et al.
progressively up to 4.2 times for FB and
3.5 for FR between 0 and 7 days of AF, reaching
more than 1000 mg/L galactose equivalent,
whereas it only increased by 2.3 times in CS during
the same period (Table 3). On-skin fermentation
performed in red winemaking is indeed known to
strongly favour the extraction of polysaccharides
in wine (Garrido-Bañuelos et al., 2019; Guadalupe
and Ayestarán, 2008). In the second half of
the winemaking process, the concentration in
polysaccharides slightly decreased or stabilized.
Similarly, Guadalupe and Ayestarán (2007) found
that Tempranillo red wine total polysaccharide
concentration increased progressively by 90 %
in the rst two-thirds of the AFM and reached
more than 800 mg/L. However, the same authors
observed a substantial decrease at the end of
the AFM, during post maceration (4 days) and
malolactic fermentation (20 days), reaching
around 400 mg/L. This value is much lower than
the ones obtained in our experiment. A possible
explanation for this difference could be that
frozen grapes were used for the current study.
Freezing and unfreezing disrupts cell structure
and is known to increase the extraction of cell
wall components such as condensed tannins
and anthocyanins (Sacchi et al., 2005). In our
experiment, this phenomenon likely impacted
the dynamic of compound extraction during the
AFM (e.g., softer cell walls earlier than usual
in the fermentation process) and increased the
polysaccharide solubilisation, thus limiting the
rate of precipitation of polysaccharides later on.
Spectrophotometric methods are providing a global
reading of the polysaccharide content of must
and wine but, in certain conditions, interferences
may occur from other macromolecules.
For instance, the Dubois method has been
shown to overestimate polysaccharide content
when wine protein content exceeds 100 mg/L
(Segarra et al., 1995). In previous studies, wine
protein concentration ranging from 4 to 49 mg/L
(n = 33) have been reported in nished CS wine
(Segarra et al.,1995) and from 29 to 49 mg/L
in unned experimental and commercial wines
(Fukui and Yokotsuka, 2003), suggesting that our
CS wine polysaccharide measurements are likely
accurate, at least during the last fermentation days,
in the nished wine. On the other side, higher
polysaccharide concentrations (1.8 to 3.1 g/L,
n = 22) have been reported in commercial CS
wines from Chile and France, using the Dubois
method (Matsuhiro et al., 2009).
Data about the protein content of interspecic
hybrid are scarcer than those for traditional
V. vinifera varieties. Yet, data from
Springer et al. (2016b) showed protein content
ranging from 37 to 133 mg/L in unned
experimental hybrid wines made from different
varieties. The berries used for the current study
were also analysed for their protein content in
another experiment (same vintage, same
vineyard), using a similar fermentation process
(Nicolle et al., 2019) and concentrations of
129, 102 and 43 mg/L corresponding to day 4
(completed fermentation), day 8 and day 15
of the winemaking process, respectively, were
found. These data suggest that overestimation
of polysaccharide content could have occurred
from day 1 to circa day 6 of the current
experiment, but the signicantly higher
polysaccharide concentrations found from day
7 to 10 should be quite accurate.
To our knowledge, the polysaccharide
concentration of FR and FB wine is reported for
the rst time in this short communication.
Preliminary GPC/SEC assays conducted on
must, mid-AF and wine samples from all three
varieties suggest that FR wines could contain a
signicant proportion of low-molecular-weight
polysaccharides and could have higher content
in larger polysaccharides when compared to CS
wines (supplemental material). Polysaccharides
from both FB and FR wine also appeared to
be more branched than those from CS wines.
Although the GPC/SEC approach seems a
powerful tool to understand grape and wine
polysaccharide structure, more work is needed to
conrm those ndings and validate this approach.
CONCLUSIONS
Wines made from the cold-hardy cultivars
Frontenac and Frontenac blanc showed lower
content in oligomeric and polymeric avan-3-ols
than those from Vitis vinifera Cabernet-Sauvignon.
The total polysaccharide concentration of these
wines increased during the alcoholic fermentative
maceration before decreasing or stabilising by
the end of the process. The wines made from the
cold-hardy hybrid Frontenac showed a higher
concentration in total polysaccharide compared
to Frontenac blanc and Cabernet-Sauvignon
wines. This suggests that specic attention should
be brought to the impact of the polysaccharide
composition of cold-hardy cultivars, such as
Frontenac, as these polysaccharides could
strongly contribute to lower the astringency of
hybrid wines.
OENO One 2021, 2, 337-347 345
© 2021 International Viticulture and Enology Society - IVES
Acknowledgements: The authors thank the
Ministère de l’Agriculture, des Pêcheries et
de l’Alimentation du Québec (MAPAQ) and
Agriculture and Agri-Food Canada (AAFC) for
nancing this study. We also thank the Centre de
Recherche Bioalimentaire du Québec (CDBQ),
the Conseil de Recherche en Sciences Naturelles
et en Génie du Canada (CRSNG) and the Fonds
de Recherche Nature et Technologies du Québec
(FQRNT) for supporting the scholarship of
Dr. Pamela Nicolle, PhD student at the time of
this experiment. We also thank (1) Véronique
Richard from Institut sur la Nutrition et les
Aliments Fonctionnels (INAF) for her help on
proanthocyanidin analyses and (2) Charlène
Marcotte for her help during the experiment.
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