Eur Food Res Technol (2005) 220:607–614
DOI 10.1007/s00217-004-1108-x
ORIGINAL PAPER
Mar
a Monagas · Carmen Gmez-Cordovs ·
Begoa Bartolom
Evolution of polyphenols in red wines from Vitis vinifera L.
during aging in the bottle
I. Anthocyanins and pyranoanthocyanins
Received: 14 September 2004 / Published online: 21 January 2005

Springer-Verlag 2005
Abstract The evolution of anthocyanins and pyranoan-
thocyanins (as measured by high-performance liquid
chromatography) in young red wines from Vitis vinifera
L. cv Tempranillo, Graciano and Cabernet Sauvignon
(vintage 2000) from Navarra (Spain) was studied during
26 months of aging in the bottle. For the anthocyanin
pigments of grape origin, a progressive decrease in their
concentration, corresponding to first-order kinetics, was
observed during this period. Independently of the antho-
cyanin structure studied, grape anthocyanins in Tem-
pranillo wine presented twofold slower kinetics than those
in Graciano and Cabernet Sauvignon wines, which ex-
hibited a very similar disappearance rate. Acylated an-
thocyanins presented a slightly higher disappearance rate
than the nonacylated ones, indicating the possible hy-
drolysis of the former into the latter forms. However, no
distinction was observed in the kinetics of the different
anthocyanidin forms (delphinidins, petunidins, peonidins
and malvidins). These results indicate that during aging
under nonoxidative conditions (bottle), the chemical re-
activity of grape anthocyanins in wine is influenced by
the grape variety, a factor that imposes over the stability
associated with the chemical structure of each anthocya-
nin form. In relation to the evolution of pyranoantho-
cyanins, anthocyanin–pyruvic acid adducts presented a
similar or lower disappearance rate than their corre-
sponding anthocyanin precursors during the first months
of aging in the bottle, while anthocyanin–vinylphenol and
anthocyanin–vinylflavanol adducts did not exhibit sig-
nificant variations during the whole period studied.
Keywords Red wine · Aging in the bottle ·
Anthocyanins · Pyranoanthocyanins · Disappearance
kinetics
M. Monagas · C. Gmez-Cordovs ()) · B. Bartolom
Instituto de Fermentaciones Industriales,
CSIC,
Juan de la Cierva 3, 28006 Madrid, Spain
e-mail: cgcordoves@ifi.csic.es
Tel.: +34-91-5622900
Fax: +34-91-5644853
Introduction
Grape anthocyanin pigments are mainly located in the
skin, being largely responsible for the color of red wine.
The main anthocyanins identified in Vitis vinifera spp.
grapes and wines are the 3-O-glucosides, 3-O-acetyl glu-
cosides and 3-O-p-coumaroyl glucosides of delphinidin,
cyanidin, petunidin, peonidin and malvidin, as well as the
3-O-caffeoyl glucosides of malvidin and peonidin [5, 8,
13, 15, 18]. Recently, the cis isomer of malvidin-3-O-p-
coumaroyl glucoside has been identified in grapes and
wines from V. vinifera [15, 18]. The anthocyanin profile
and concentration is largely dependent on grape variety,
but also on seasonal or climatic conditions and soil
composition. Finally, the winemaking technology (mac-
eration and fermentation) employed also influences the
anthocyanin content in red wine. However, during wine
maturation and aging there is a progressive loss of an-
thocyanin pigments, mainly due to their participation in
numerous chemical reactions. Some of these reactions
result in the formation of stabler oligomeric and/or
polymeric pigments which give rise to important wine
color changes (from bright red to brick red hues). Among
these pigments are the products resulting from the direct
and acetaldehyde-mediated anthocyanin–tannin conden-
sation reactions [29, 34], as well as the products origi-
nated from the C-4/C-5 cycloaddition reaction of antho-
cyanins with other molecules bearing a polarizable double
bond (pyruvic acid, 4-vinylphenols, hydroxycinnamic
acids, vinylflavanols, acetaldehyde, acetone, among oth-
ers), giving rise to so-called pyranoanthocyanins [4, 6, 9,
10, 12, 15, 20, 27]. The concentration of anthocyanins,
copigments, acetaldehyde and other yeast metabolites, as
well as the pH, temperature, and presence of oxygen and
sulfur dioxide, among others, are factors that affect the
progress of anthocyanin condensation reactions in wine,
and therefore their rate of disappearance during matura-
tion and aging [2, 7, 22–25, 30, 32, 33].
Previous studies related to the disappearance kinetics
of anthocyanins during wine conservation had mainly
concentrated on the kinetics of total anthocyanins or on
608
that of major anthocyanins, malvidin-3-glucoside and its
acylated forms with acetic and p-coumaric acids [3, 7, 11,
24]. However, there is no information in the literature in
relation to the kinetics of the remaining anthocyanidin
forms (cyanidins, delphinidins, petunidins and peonidins)
found in wines from V. vinifera. Several discrepancies
also exist among authors concerning whether the acylated
forms are more susceptible to having a higher disap-
pearance rate than the nonacylated ones [7, 11, 14, 17,
24]. In relation to the evolution of pyranoanthocyanins
during wine aging, anthocyanin–pyruvic acid adducts
have presented different evolution trends depending on
the fermentation (temperature, yeast strain, SO2 concen-
tration) and aging conditions employed [1, 2, 11, 16, 19,
21, 24, 26], whereas very scarce or no literature data exist
with respect to the evolution of anthocyanin–vinylphenol
[28] and anthocyanin–vinylflavanol adducts. The aim of
the present work was to study the influence of the an-
thocyanin chemical structure, wine anthocyanin profile
and grape variety on the evolution of anthocyanins and
pyranoanthocyanins during wine aging in the bottle. For
that, grapes from varieties exhibiting different anthocya-
nin profiles (Tempranillo, with a relatively high propor-
tion of cinnamoyl derivatives; Graciano, with a high
proportion of peonidin derivatives and a balanced pro-
portion of acetyl and cinnamoyl derivatives; and Cabernet
Sauvignon, with a high proportion of acetyl derivatives
[18]), were vinified in the same manner and the antho-
cyanin evolution of their resulting wines was studied
during 26 months of aging in the bottle.
Materials and methods
Winemaking
Monovarietal young red wines made from grapes of V. vinifera cv.
Tempranillo, Graciano and Cabernet Sauvignon grown in the same
geographical area and elaborated at the Viticulture and Enology
Station of Navarra (EVENA), Spain (vintage 2000), were used for
this study. A lot of 220 kg of grapes of each variety was des-
temmed, crushed and collected in 200-l stainless steel wine vats.
Semi-industrial-scale fermentations were performed with a yeast
inoculum of 25 g/hl (80% EVENA Saccharomyces cerevisiae Na33
yeast strain; 20% Lallemand S. bayanus EC118 yeast strain) at a
temperature up to 27 C. The cap was punched down twice a day
until it remained submerged during a 14-day maceration period. At
the end of the alcoholic fermentation, the wines were racked and
stabilized for a period of 1 month at –2 C. The wines were then
filtrated through SEITZ K250 filters (2.5–3.0 mm) (Sert Schenk
Filter System, Bad Krevznach, Germany) and finally bottled after
correcting the free SO2 level to 30 mg/l. Two wine samples from
each variety were analyzed after 1.5, 3, 7, 9, 12, 19.5 and 26 months
of bottling and storage at 13 C and 80–85% relative humidity. The
optimum storage temperature for wines ranges between 13 and
17 C.
High-performance liquid chromatography–diode-array
detection analysis
A Waters (Milford, MA) liquid chromatography system equipped
with a 600-MS controller, a 717Plus autosampler, and a 996 pho-
todiode-array detector was used. Separation was performed on a
reverse-phase Waters Nova-Pak C18 (150 mm3.9 mm, 4 mm) at
room temperature. A gradient consisting of solvent A (water–for-
mic acid, 90/10, v/v) and solvent B (water–methanol–formic acid,
45/45/10, v/v/v) was applied at a flow rate of 0.8 ml/min as follows:
15–80% solvent B linear from 0 to 30 min, 80% solvent B isocratic
from 30 to 43 min, followed by washing (methanol) and re-equil-
ibration of the column from 43 to 75 min. One-hundred microliters
of wine, previously filtered through a 0.45-mm membrane, was
injected onto the column. Diode-array detection was performed
from 260 to 600 nm. Quantification was carried out by area mea-
surements at 530 nm and the anthocyanin content was expressed as
malvidin-3-glucoside (Estrasynthse, France) by a standard cali-
bration curve.
Statistical analysis
Analysis of variance (ANOVA) and linear regression analysis of the
data were performed using the PC software package Statgraphics
Plus 2.1 (Graphics Software Systems, Rockwille, MD, USA).
Results and discussion
Anthocyanidin-3-glucosides, anthocyanidin-3-(6-acetyl)-
glucosides, anthocyanidin-3-(6-p-coumaroyl)-glucosides
and anthocyanidin-3-(6-caffeoyl)-glucosides, as well as
several groups of more complex anthocyanin-derived
pigments, including pyranoanthocyanins resulting from
the C-4/C-5 cycloaddition of anthocyanins with pyruvic
acid, 4-vinylphenols and vinylflavanols, as well as the
dimers originated from the direct and acetaldehyde-me-
diated anthocyanin–flavanol condensation reactions, were
identified in the different wines according to the method
of Monagas et al. [15].
Evolution of anthocyanins of grape origin
The evolution of delphidin, petunidin, peonidin and mal-
vidin-3-glucosides, delphinidin, petunidin and malvidin-3-
(6-acetyl)-glucosides, petunidin, peonidin and malvidin-3-
(6-p-coumaroyl)-glucosides, and malvidin-3-(6-caffeoyl)-
glucoside in the wines from Tempranillo, Graciano and
Caberent Sauvignon, after 1.5, 3, 7, 9, 19.5 and 26 months
of aging in the bottle, is shown in Fig. 1. Cyanidin
derivatives, peonidin-3-(6-caffeoyl)-glucoside and mal-
vidin-3-(6-p-coumaroyl)-glucoside-cis isomer, were not
considered in the study owing to their low content. Del-
phinidin-3-(6-p-coumaroyl)-glucoside and peonidin-3-
(acetyl)-glucoside were not included since their corre-
sponding peak integration was very complicated as the
aging time progressed. Figure 2 shows the evolution of the
anthocyanins grouped according to their acylation [simple
glucosides (Fig. 2a), acetyl glucosides (Fig. 2b) and cin-
namoyl glucosides (Fig. 2c)] and anthocyanidin [del-
phinidins (Fig. 2e), petunidins (Fig. 2f), peonidins
(Fig. 2g) and malvidins (Fig. 2h)] patterns, as well as that
of total anthocyanins (Fig. 2d) quantified in the different
wines. Cinnamoyl glucosides included both p-coumaroyl
and caffeoyl anthocyanins. Simple glucosides and mal-
vidins were the most abundant groups of the anthocyanins
in the different wines studied (Fig. 2), with the total an-
thocyanin concentration being higher in Tempranillo wine
than in Cabernet Sauvignon and Graciano wines (Fig. 2d).
 609

Fig. 1 Evolution of individual anthocyanins during aging in the bottle. a Simple glucosides; b acetyl glucosides; c cinnamoyl glucosides.
TEM Tempranillo; GRA Graciano; CS Cabernet Sauvignon
610

Fig. 2 Evolution of the different groups of anthocyanins and of total anthocyanins during aging in the bottle. a Simple glucosides;
b acetyl glucosides; c cinnamoyl glucosides; d total anthocyanins; e delphinidins; f petunidins; g peonidins; h malvidins
 611
Table 1 Dissapearance rate of anthocyanins in Tempranillo, Graciano and Cabernet Sauvignon wines.
Tempranillo Graciano Cabernet S.
3 2 3 2
t1/4 t1/2 k10 R t1/4 t1/2 k10 R t1/4 t1/2 k103 R2
Individual anthocyanins
Delphinidin-3-glucoside 7.9 18.9 36.6a 0.946 5.5 13.3 52.3 a
0.937 6.1 14.7 47.2 a
0.970
Petunidin-3-glucoside 7.9 18.9 36.6 a 0.936 4.3 10.3 67.2 b
0.973 3.7 8.9 77.8 b
0.989
Peonidin-3-glucoside 8.5 20.5 33.8 a 0.902 4.6 11.1 62.2 b
0.971 4.1 9.9 69.8 ab
0.914
Malvidin-3-glucoside 8.4 20.3 34.1 a 0.957 4.7 11.3 61.2 b
0.967 5.2 12.6 55.0 b
0.963
Delphinidin-3-(6-acetyl)- 9.3 22.5 30.9 a 0.958 4.1 9.8 70.9 ab
0.997 5.4 12.9 53.7 b
0.983
glucoside
a b b
Petunidin-3-(6-acetyl)- 9.8 23.7 29.2 0.977 5.2 12.5 55.5 0.985 4.2 10.0 69.3 0.972
glucoside
a b b
Malvidin-3-(6-acetyl)- 8.2 19.7 35.2 0.968 4.0 9.6 72.1 0.991 4.0 9.7 71.1 0.992
glucoside
a b b
Malvidin-3-(6-caffeoyl)- 7.9 19.1 36.4 0.963 3.7 8. 8 78.7 0.996 4.1 9.8 70.5 0.993
glucoside
a b ab
Petunidin-3-(6-p-coumaroyl)- 8.4 20.3 34.1 0.948 5.8 14.1 49.3 0.992 4.8 11.5 60.3 0.943
glucoside
a b b
Peonidin-3-(6-p-coumaroyl)- 8.7 21.1 32.9 0.957 3.9 9.5 73.2 0.980 3.1 7.4 93.9 0.991
glucoside
a b b
Malvidin-3-(6-p-coumaroyl)- 9.2 22.1 31.3 0.964 3.8 9.2 75.3 0.986 4.0 9.7 71.4 0.986
glucoside
Groups according to the acylation pattern
a b b
Simple glucosides 8.4 20.2 34.3 0.955 4.7 11.3 61.2 0.967 5.1 12.4 56.0 0.963
a b b
Acetyl glucosides 8.9 21.3 32.5 0.935 4.1 9.8 70.5 0.991 4.1 9.8 70.7 0.993
a b b
Cinnamoyl glucosides 8.8 21.2 32.7 0.966 3.8 9.3 74.8 0.986 4.1 9.9 70.2 0.989
Groups according to the anthocyanidin pattern
Delphinidins 8.3 20.1 34.5 a 0.921 5.1 12.3 56.4 a
0.921 5.9 14.1 49.0 a
0.945
Petunidins 8.0 19.3 35.9a 0.947 4.4 10.7 64.7 b
0.980 3.9 9.4 73.9 b
0.988
Peonidins 8.2 19.8 35.0a 0.956 4.4 10.7 64.7 b
0.974 4.4 10.5 66.1 ab
0.912
Malvidins 8.5 20.5 33.8a 0.965 4.5 10.9 63.6 b
0.973 4.7 11.3 61.2 b
0.978
Total anthocyanins 8.6 20.6 33.6a 0.963 4.5 10.9 63.6 b
0.973 4.7 11.3 61.2 b
0.978
t1/4 is the time required for a 25% reduction of the initial anthocyanin concentration (months).
t1/2 is the time required for a 50% reduction of the initial anthocyanin concentration (months).
k is the rate constant (per month).
Different letters in the same row indicate no overlapping of the confidence intervals for k at 95% resulting from the lineal regression
analysis.

A progressive decrease in the content of the different
anthocyanins in Tempranillo, Graciano and Cabernet
Sauvignon wines was observed during aging in the bottle,
with this decrease being more pronounced between 3 and
9 months. As observed by other authors [3, 7, 11, 24],
the anthocyanin decline followed first-order kinetics:
ln½A
¼ kt þ ln½A
0 , where [A] is the pigment concen-
tration (milligrams per liter) and t is the period (months) of
aging in the bottle. The reaction rate constant (k) for each
pigment was determined by calculating the slope of the
curve ln[A] versus t by linear regression analysis. The
reaction quarter-life (t1/4) and half-life (t1/2), corresponding
to the times required for a 25 and a 50% reduction of the
initial anthocyanin concentration, respectively, were also
calculated by the equation t1=x ¼ ½ln x  ln ðx  1Þ
=k,
where [A]0/x is the reduced concentration. Table 1 sum-
marizes the disappearance kinetics data for the different
anthocyanins, either individual, grouped or as a total.
By the application of linear regression analysis, the
reaction rate constant (k) was statistically significant
(p<0.05) for all the anthocyanins studied (results not
shown). As presented in Table 1, k values were much
lower for Tempranillo wine than for Graciano and
Cabernet Sauvignon wines, which exhibited similar ki-
netics (Table 1). This fact was observed for each of the
anthocyanins evaluated, independently of their chemical
structure or acylation pattern, although in some cases (i.e.,
delfinidin-3-glucoside) the confidence levels for k at 95%
resulting from the linear regression analysis overlapped
(Table 1). In general, the disappearance kinetics for an-
thocyanins in Graciano and Cabernet Sauvignon wines
was approximately twofold faster than that for antho-
cyanins in Tempranillo wine, as can be observed from the
t1/4 and t1/2 values (Table 1). The time required for a 25%
(t1/4) reduction of the initial concentration in Tempranillo
wine was approximately of the order required for a 50%
(t1/2) reduction of the initial anthocyanin content in wines
from Graciano and Cabernet Sauvignon (Table 1). Mc-
Closkey and Yengoyan [14] also found differences in the
anthocyanin kinetics of wines manufactured from two
different grape varieties (Zinfandel and Cabernet Sauvi-
gnon). Similarly, Mateus and De Freitas [11] also re-
ported differences in the anthocyanin kinetics in port
wines from Touriga Nacional and Touriga Francesa grape
612
Fig. 3 Evolution of pyranoanthocyanins during aging in the bottle. a Anthocyanin–pyruvic acid adducts; b anthocyanin–vinylphenol and
anthocyanin–vinylflavanol adducts
varieties. Both authors attributed the difference found to
the particular chemical composition of each wine.
According to McCloskey and Yengoyan [14], the order
of the disappearance rate was malvidin-3-(6-p-coumar-
oyl)-glucoside>malvidin-3-(6-acetyl)-glucoside>mal-
vidin-3-glucoside, in wines from Zinfandel stored for 3
years at 4 C. Later on, Bakker [3], and more recently
Mateus and De Freitas [11], also found that malvidin-3-
glucoside acylated with either acetic or p-coumaric acids
presented a disappearance rate slightly higher than the
nonacylated form during the aging of port wine. Dallas et
al. [7] also arrived at the same conclusions during the
aging of Tinta Roriz (Tempranillo) wine for 3 months at
12 C. However, when the kinetics study was performed
at higher temperatures (22–42 C), no significant differ-
ences were found among the different anthocyanin forms,
also coinciding with the results first reported by Nagel
and Wulf [17] in accelerated aging studies performed at
22 C. The results found in this study (26 months of aging
in the bottle at 13 C) are in accordance with these ob-
servations in the case of Graciano and Cabernet Sauvi-
gnon wines, in that nonacylated anthocyanins (simple
glucosides) presented a lower k value than the acylated
ones (acetyl glucosides and cinnamoyl glucosides) (Ta-
ble 1), although no statistically significant differences
were found between the two forms (results not shown). In
Tempranillo wine, both forms showed very similar k
values. Differences between acylated and nonacylated
anthocyanins would only be found in wines from those
varieties that show a higher disappearance rate during
aging. The nonacylated forms would be effectively stabler
than the acylated ones, although it could also be postu-
lated that the acylated forms are hydrolyzed and con-
verted into simple glucosides, finally explaining their
higher relative disappearance rate. In fact, the concen-
tration of free trans-p-coumaric acid substantially in-
creased during the 26 months of aging in the bottle (ac-
companying paper), indicating that p-coumaroyl gluco-
side anthocyanins could be an extra source of this com-
pound besides that corresponding to the hydrolysis of p-
coumaroyltartaric acid, which is known to occur during
wine aging [28, 31].
In terms of the anthocyanidin profile, no statistically
significant differences (results not shown) were found
between the k values of delphinidins, petunidins, peoni-
dins and malvidins (Table 1), despite the fact that an in-
crease in the number of methoxy groups in the B ring of
the molecule suggests a higher structural stability [13].
Finally, the kinetics parameters of total anthocyanins in
the wines from each variety were very similar to those of
malvidins, since this group represented the higher pro-
portion of total pigments (74–89%). In accordance with
the kinetics data, losses in total anthocyanins registered
after the 26 months of bottle-aging were lower in Tem-
pranillo wine (58%) than in Graciano (79%) and Cabernet
Sauvignon (79%) wines.
Evolution of anthocyanin-derived pigments:
pyranoanthocyanins
Anthocyanin–pyruvic acid adducts
Anthocyanin–pyruvic acid adducts represented the most
abundant group of pyranoanthocyanin pigments studied
(Fig. 3). The main anthocyanin–pyruvic acid adducts
present in the different wines were malvidin-3-glucoside
pyruvate for the wines from the three grape varieties;
malvidin-3-(6-acetyl)-glucoside pyruvate for Cabernet
Sauvignon wine; malvidin-3-(6-p-coumaroyl)-glucoside
pyruvate for Tempranillo wine, and peonidin-3-glucoside
pyruvate for Graciano wine, which were the derivatives
corresponding to the main anthocyanin precursors in each
variety [15] (Fig. 1). As a group, anthocyanin–pyruvic
acid adducts decreased during the first months of bottle-
aging, although a slight stabilization was observed after 9

months of aging in the case of Graciano and Cabernet
Sauvignon wines (Fig. 3a). Therefore, the evolution of
these adducts could not be modeled by first-order kinet-
ics. The two-way ANOVA analysis indicated statistically
significant differences (p<0.05) according to both time (p
=0.0184) and variety (p = 0.0000) factors.
By calculating the time required for a 25% reduction
of the initial concentration of each anthocyanin–pyruvic
acid adduct, it was found that the losses registered by
these pigments during the first months of aging were
similar or inferior to those of their anthocyanin precur-
sors, and were superior for Graciano and Cabernet
Sauvignon wines than for Tempranillo wine. The time
required for a 25% reduction of malvidin-3-glucoside was
8.4, 4.7 and 5.2 months in Tempranillo, Graciano and
Cabernet Sauvignon wines, respectively (Table 1),
whereas for malvidin-3-glucoside pyruvate it was 9.6, 7.2
and 5.1 months, respectively. For peonidin-3-glucoside
versus peonidin-3-glucoside pyruvate in Graciano wine, it
was 4.6 versus 8.1 months, respectively; for malvidin-3-
(6-acetyl)-glucoside versus malvidin-3-(6-acetyl)-gluco-
side pyruvate in Cabernet Sauvignon wine, it was 4.0
versus 6.6 months; and finally for malvidin-3-(6-p-cou-
maroyl)-glucoside versus malvidin-3-(6-p-coumaroyl)-
glucoside pyruvate in Tempranillo wine, it was 9.2 versus
8.7 months. Other authors [11, 26] have also described a
lower loss of malvidin-derived pyruvic acid adducts in
comparison with their anthocyanin precursors. Consider-
ing the 26 months of aging in the bottle, the losses reg-
istered in the total concentration of anthocyanin–pyruvic
acid adducts were also lower (49% in Tempranillo, 32%
in Graciano and 48% in Cabernet Sauvignon wines) than
those reported here earlier for total anthocyanins of grape
origin.
According to the literature data, the evolution of an-
thocyanin–pyruvic acid adducts during wine aging seems
to follow different patterns as a function of the fermen-
tation and aging conditions employed during the wine-
making process. For instance, in wines treated with pec-
tolitic enzymes, Revilla et al. [21] found an increase in the
concentration of malvidin-3-glucoside pyruvate during
the first 6 months of aging in the bottle, followed by a
slight decrease. Similarly, Atanasova et al. [2] found an
increase in these pigments both in nonoxygenated and
oxygenated wines during a 7-month period, concluding
that the anthocyanin–pyruvic acid condensation reaction
was not influenced by oxygen. On the other hand, Mateus
and De Freitas [11] reported losses (9–18%) of malvidin-
derived pyruvic acid adducts in port wines during 38
months in bottles, although losses were higher (70%) in
wines stored in oak barrels (oxidative conditions) during
the same period of aging. However, Prez-Maragio and
Gonz lez-San Jos [19] recently reported an increment of
these derivatives in red wines during the first 8 months of
storage in oak barrels.
613
Anthocyanin–vinylphenol
and anthocyanin-vinylflavanol adducts
Both vinylphenol and vinylflavanol derivatives were
identified in Graciano and Cabernet Sauvignon wines,
whereas only anthocyanin–vinylphenol adducts were de-
tected in Tempranillo wine [15], findings which are in
accordance with the concentration of the non-anthocyanin
precursors (hydroxycinnamic acids and flavanols) of
these pigments in the different wines (accompanying
paper). Figure 3b shows the evolution of the group of
anthocyanin–vinylphenol adducts in Tempranillo (sum of
malvidin-3-glucoside–vinylphenol, malvidin-3-gluco-
side–vinylcatechol, and malvidin-3-glucoside–vinylgua-
iacol), and of anthocyanin–vinylphenol and anthocyanin–
vinylflavanol adducts in Graciano (sum of malvidin-3-
glucoside–vinylphenol and malvidin-3-glucoside–
vinylepicatechin) and Cabernet Sauvignon [sum of mal-
vidin-3-glucoside–vinylphenol, malvidin-3-glucoside–
vinylepicatechin and malvidin-3-(6-acetyl)-glucoside–
vinylepicatechin] wines. For Tempranillo and Cabernet
Sauvignon wines a slight increase in concentration was
registered between 7 and 19.5 months of aging, whereas
Graciano wine presented a slight decrease between 9 and
23 months of aging in the bottle (Fig. 3b). However, the
two-way ANOVA revealed no significant differences
(p>0.05) in relation to the time factor (p =0.3756), being
significant only in the function of the variety factor (p
=0,000).To date, there is not much literature data con-
cerning the evolution of anthocyanin–vinylphenol and
anthocyanin–vinylflavanol adducts in wine, under oxida-
tive or nonoxidative conditions, although Schwarz et al.
[27] did not find any influence of oxygen on the rate of
formation of vinylphenol derivatives in model solutions.
Recently, the same group found that in wines from
Pinotage (vintages 1998-2001), a variety very abundant in
caffeic acid, the major synthesis of the pigment malvidin-
3-glucoside–vinylcatechol occurred after 2.5–4 years of
aging in the bottle, mainly depending on the concentration
of caffeic acid but not on that of malvidin-3-glucoside
[28]. These observations agree with the results found in
this work, since no significant changes were registered in
the total concentration of vinylphenol derivatives after 26
months (2.2 years) of aging in the bottle, even in the
presence of a sufficient amount of both precursors, mal-
vidin-3-glucoside and hydroxycinnamic acids (accompa-
nying paper).
Conclusions
The results found in this study demonstrate that the dis-
appearance kinetics of grape anthocyanins during wine
aging under nonoxidative conditions (bottle) is influenced
by the grape variety, a factor that imposes over the
chemical structure of each anthocyanin. In other words,
for a particular grape variety, all the anthocyanins showed
approximately the same level of reactivity in wine inde-
pendent of the stability associated with the chemical
614
structure of their different forms. It is also concluded that
the wine anthocyanin profile is not related to the slower
anthocyanin kinetics in Tempranillo wine in comparison
with that in Graciano and Cabernet Sauvignon wines,
since these latter wines presented very similar kinetics
data but a very distinct anthocyanin profile (Figs. 1, 3).
On the other hand, the anthocyanin content is not the
factor conditioning these results, considering that Tem-
pranillo wine presented the highest total anthocyanin
concentration but the lowest disappearance rate of these
compounds during aging. The existence of other compo-
nents and/or factors characteristic of the wine from each
variety that may influence the relative development of
certain chemical reactions seems to be responsible for the
differences found in the anthocyanin kinetics as a function
of the grape variety employed, as is described in the ac-
companying paper.
New data is provided concerning the concentration and
evolution of different types of pyranoanthocyanin pig-
ments in wine. Whereas the initial concentration and
distribution of anthocyanin–pyruvic acid adducts in the
wines studied is mainly influenced by the anthocyanin
profile of the grape variety employed, that of anthocya-
nin–vinylphenol and anthocyanin–vinylflavanol adducts
also depends on the concentration of their non-anthocy-
anin precursors (see accompanying paper). Different
evolution patterns were also found between the different
type of pyranoanthocyanins during wine aging in bottle:
anthocyanin–pyruvic acid adducts presented a similar or
lower disappearance rate than their corresponding an-
thocyanin precursors during the first months of aging in
bottle, while anthocyanin-vinylphenol and anthocyanin-
vinylflavanol adducts did not exhibited significant varia-
tions during the whole period studied.
Acknowledgements The authors are grateful to Juli n Suberviola
(EVENA, Navarra, Spain) for providing the wine samples and to
Pedro J. Mart
n (CSIC, Madrid, Spain) for his kind assistance with Chem 51:7951–7961
the graphs. The authors also thank the Agencia Espaola de Co-
operacin International for a MUTIS predoctoral scholarship to
M.M. and the Spanish Comisin Interministerial de Ciencia y
Tecnolog
a (projects AGL2000-1427-C02-02 and AGL2003-
07394-C02-02) for funding.
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