Food Chemistry 136 (2013) 224236

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Pre-bottling use of dehydrated waste grape skins to improve colour, phenolic

and aroma composition of red wines
Miguel Angel Pedroza a, Manuel Carmona b, Gonzalo Luis Alonso a, Maria Rosario Salinas a,
Amaya Zalacain a,
a
b

Escuela Tcnica Superior de Ingenieros Agrnomos, Universidad de Castilla-La Mancha, Avda. de Espaa, s/n, Albacete E-02071, Spain
Albacete Science & Technology Park Foundation, Universidad de Castilla La Mancha, Albacete E-02071, Spain

a r t i c l e

i n f o

Article history:
Received 16 April 2012
Received in revised form 21 June 2012
Accepted 25 July 2012
Available online 4 August 2012
Keywords:
Waste grape skins
Wine
Colour
Phenolic compounds
Volatiles
Pre-bottling

a b s t r a c t
Different dehydrated waste grape skins from the juice industry were added into aged and young red
wines as an innovative way of compensating for colour loss before bottling. After addition of grape skins,
colour intensity of wines increased a mean 11% and a maximum of 31% with predominance of the red
component. Total polyphenols mean increase was 10% with a maximum value of 20%. Analysis of low
molecular weight phenolic compounds by HPLCDAD showed a signicant (p < 0.05) content increase
of the bioactive compounds gallic acid, (+)-catechin, ( )-epicatechin, and (E)-resveratrol. Anthocyanins
content also increased at an average of 50 mg/l. The volatile prole of wines analysed by SBSEGCMS
was only moderately inuenced by the treatments. Mixtures of dehydrated waste grape skins were useful
to improve the colour and polyphenol prole of red wines, considering them a useful tool for correcting
colour loss before bottling.
2012 Elsevier Ltd. All rights reserved.

1. Introduction
Dynamic consumer preferences and market trends require wine
producers to be constantly innovative in terms of sensory and
quality parameters. However, many wine cellars deal with limited
budgets, where little is left for approaching constantly changing
consumer demands. Sometimes this translates into accumulation
of out-of-trend wines before bottling, generating a low revenue
perspective. Therefore, considering strategies to overcome this
problem should be taken into account by regulating authorities.
Many sensory and quality parameters of wines are related to
the composition and concentration of avonoids, phenolic acids,
and volatile compounds extracted from grapes during winemaking
(Ribreau-Gayon, Dubourdieu, Donche, & Lonvaud, 2004). Harvesting and oenological techniques used for producing wine also
play a fundamental role in dening the polyphenol and volatile
proles of wines. Moreover, biochemical and chemical reactions
taking place during winemaking and ageing, such as oxidation,
polymerization and complexation reactions, have a signicant effect on the wine prole (Bayonove, Baumes, Crouzet, & Gnata,
2003; Cheynier, Moutounet, & Sarni-Manchado, 2003).
Young red wines have a characteristic bright red colour associated to a higher content of anthocyanins in an equilibrium state
Corresponding author. Tel.: +34 967599310; fax: +34 96 599338.
E-mail address: Amaya.Zalacain@uclm.es (A. Zalacain).
0308-8146/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodchem.2012.07.110

called copigmentation (Boulton, 2001), as well as to a lower exposure to oxygen and derived colour degradation reactions than aged
wines (Oliveira, Ferreira, De Freitas, & Silva, 2011). The optimal sensorial attributes, where colour has a major role, is known to last for
shorter time in standard wines than correct, high quality, and premium wines (Zamora, 2003). The decline of their overall attributes
is partly related to a low concentration of polyphenols, as well as to
oxidative degradation and polymerization-condensation phenomena (Cheynier et al., 2003; Li, Guo, & Wang, 2008). Moreover, since
most standard and low quality wines are made from grapes with
moderate or decient contents of polyphenols, they are doomed
to the use of widely recognized additives like SO2 and ascorbic acid
to reduce oxidative reactions (Oliveira et al., 2011).
The volatile prole of wines is also considered one of the main
quality attributes, where varietal compounds like monoterpenes
and norisoprenoids play a fundamental role (Ebeler & Thorngate,
2009; Francis & Newton, 2005; Zalacain, Marn, Alonso, & Salinas,
2007). These compounds have been associated with the fruity, oral and citrus aroma of wine and some authors have related them
with the expression of terroir or typicity from particular viticultural
regions (Bayonove et al., 2003). Given its importance and considering that most regular quality wines have a limited concentration of
such compounds, strategies for increasing their concentration in
regular quality wines are important.
The use of additives is becoming a common practice in the wine
industry for improving the sensory prole of wines in order to

M.A. Pedroza et al. / Food Chemistry 136 (2013) 224236

make them more competitive and to reduce defects. Wood chips,

enzymes, enological tannin, are some examples of internationally
approved enological practices (OIV, 2012) which modify the composition and sensory characteristics of wines. On the other hand,
other products obtained from grape skins like additive E-163, also
known as Anthocyanins or Enocyanin, are currently commercialized
globally for use in the food industry. Recently, consumer demand
for labeling wine ingredients and additives has brought to debate
whether wine cellars should indicate their use. In this direction,
the use of natural and vegetal based additives, such as enological
tannin from grape seeds and grape skins, is commonly better perceived by consumers (Cheng, Bekhit, Sedcole, & Hamid, 2010).
Process sustainability is progressively becoming a mandatory
standard in developed countries making relevant the proposals
for reusing or exploiting current industrial wastes. In the case of
the wine industry, one of the major wastes is grape marc (mixture
of grape skins, seeds and stalks). This waste and its components
have been recently studied for their potential use as raw materials,
ingredients and antioxidants (Arvanitoyannis, Ladas, & Mavromatis, 2006; Bekhit et al., 2011; Casazza, Aliakbarian, De Faveri, Fiori,
& Perego, 2011; Fiori, 2010; Pinelo, Arnous, & Meyer, 2006; Ping,
Brosse, Chrusciel, Navarrete, & Pizzi, 2011; Spigno & De Faveri,
2007). Waste grape skins from the juice industry are commonly
obtained after shorter processing time (4 days of maceration) than
those from red winemaking. This fact implies that juice industry
grape marc is not as exhausted as that from winemaking and thus
a richer source of several compounds.
Recently Pedroza, Carmona, Salinas, and Zalacain (2011), evaluated the production of ros wines by macerating dehydrated waste
grape skins into white wines, resulting in a stable product with tailor made characteristics according to those obtained in model wine
solutions. Furthermore, this approach made it possible to reuse an
agroindustrial byproduct offering an alternative for commercialization of both; the white wine (converted into ros) and the waste
grape skins. The aim of this work was to use dehydrated waste
grape skins as a new oenological tool for compensating colour,
phenolic and aroma degradation in red wines before bottling. Mixtures of waste grape skins were assayed for producing different
compositional proles, evaluating their immediate impact and
storage behaviour.
2. Materials and methods

225

to table wines commercialized as bulk product with a shelf life no

longer than three years after bottling. These wines were result
from traditional winemaking (510 days of maceration; fermentation temperature 25 C in stainless steel tanks) of several red grape
varieties cultivated regionally (Tempranillo, Cabernet Sauvignon,
Merlot, etc.). Aged wines were produced similarly as young wines,
but with longer maceration times (1020 days) and subjected to at
least 6 months of storage in American oak barrels (250 L). All wines
had pH range = 3.53.9 and alcohol content = 13.514% vol. In order to obtain representative samples of wines before bottling, sampling was performed at the bottling line in 750 ml amber glass
bottles (four bottles of each wine with synthetic cork closures).
Wines were then taken to the laboratory and prepared for its
treatment.
2.3. Maceration conditions
DWGS were macerated according to conditions established by
Pedroza et al. (2011). Briey, a dosage of 5 g DWGS/L was macerated into each wine during 3 days at 18 C. Amber crystal asks
of 125 ml with screwcap plastic closures and 5 0.5 ml headspace
were used during the experiment. After maceration, DWGS were
removed with a strainer and the wines stored in the same type
of crystal bottles at 18 C. Wine evolution analysis started immediately after maceration, and then after 3 and 6 months (T0, T3, and
T6 respectively).
2.4. Chemicals and standards
FolinCiocalteu reagent from Merck (Darmstadt, Germany). Sodium Carbonate from Panreac (Barcelona, Spain). Caffeic acid, (+)catechin, p-coumaric acid, ( )-epicatechin, gallic acid, and (E)-resveratrol, from SigmaAldrich (Steinheim, Germany) were used as
standards for low molecular weight phenolic compound analysis.
Malvidin-3-glucoside (Mv-3-G) standard from Extrasynthse (Geneay, France) was used for anthocyanins quantication. HPLC-grade
acetonitrile was from Panreac (Barcelona, Spain). Eugenol, farnesol,
1-hexanol, b-ionone, isoamyl acetate, D-limonene, nerolidol, (E)whiskylactone, (Z)-whiskylactone, supplied by SigmaAldrich
(Steinheim, Germany), and b-damascenone supplied by Firmenich
(Geneva, Switzerland) were used as calibration standards in wine
model solution (12% v/v ethanol, pH = 3.6, 5 g/l tartaric acid) for
volatile analysis.

2.1. Dehydrated waste grape skins (DWGS)

2.5. Sample characterization
Waste grape marcs of Vitis vinifera Bobal (red variety) and a mixture of 70% Airn (white variety) with an unknown red variety,
namely AMIX, were obtained from a juice concentrate factory in
Castilla-La Mancha (Julian Soler, Cuenca, Spain). Samples obtained
immediately after pressing the macerated must were collected in
plastic bags (60 kg) and frozen at 20 C. In the laboratory, waste
grape marcs were thawed at 25 C and oven dried at 60 C, according
to Pedroza, Carmona, Pardo, Salinas, and Zalacain (2012) until a constant weight was achieved (35% moisture content). Dried samples
were sieved through 3 mm mesh to remove seed and stalks. Dehydrated waste grape skins (DWGS) were then ground in a cutting mill
MS 100 (Retsch, GmbH & Co. KG, Denmark) and sieved to 1.0 mm
particles. Four types of DWGS were then prepared: 100% Bobal (Bobal), 100% AMIX (AMIX), 75% AMIX + 25% Bobal (GM75), and 50%
AMIX + 50% Bobal (GM50). The later mixtures were selected to obtain intermediate values between plain AMIX and Bobal DWGS.
2.2. Wines
Two aged (A05 and A07) and two young (Y08 and Y09) red
wines were provided by a local winery. Young wines corresponded

2.5.1. UVvis spectrophotometry

Standard colour parameters and total phenolic compounds were
measured in a Lambda 25 UVVis spectrophotometer (Perkin Elmer, Norwalk, CT) with quartz cells. All samples were rst ltered
through a PVDF Durapore lter of 0.45 lm (Millipore, Bedford,
MA). Colour was determined following Glories method (Glories,
1984), measuring absorbance at 420, 520, and 620 nm. Total polyphenols (TP) were determined at 750 nm according to Singleton
and Rossi (Singleton & Rossi, 1965) and expressed as mg/l of gallic
acid equivalents (GAE) according to a calibration curve with the
equation TP = (0.2735 Absorbancy 0.036) 1000), R2 = 0.994
and with a mean relative standard deviation below 5%.
2.5.2. Phenolic compounds determination by HPLCDAD
Phenolic compound analysis was carried out according to Cozzolino et al. (2004). The samples were ltered through a PVDF
Durapore lter of 0.45 lm (Millipore, Bedford, MA) and injected
into an Agilent 1100 HPLC chromatograph (Palo Alto, CA) equipped
with a Phenomenex (Torrance, CA) Synergi 4 l Hydro-RP column
(4 lm particle size, 80 pore size, 150  2.0 mm) at 25 C.

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M.A. Pedroza et al. / Food Chemistry 136 (2013) 224236

Solvents were: (A) 1% v/v acetonitrile, 1.5% v/v phosphoric acid in

water and (B) 20% v/v solvent A, 80% v/v acetonitrile. Gradient elution at a constant ow rate of 0.4 ml/min was: 0 min (14.5% solvent
B), 18 min (27.5% solvent B), 20 min (27.5% solvent B), 21 min
(50.5% solvent B), 22 min (50.5% solvent B), 26 min (100% solvent
B), and 28 min (100% solvent B). The injection volume used was
20 ll. Compound detection was carried out with a diode array
detector by comparison with the corresponding UVvis spectra
and retention time of pure standards in the chromatogram. Gallic
acid, (+)-catechin, vanillic acid, syringic acid and ( )-epicatechin
were identied at 280 nm; ferulic acid and caffeic acid were identied at 324 nm, (trans)-resveratrol and p-coumaric acid were
identied at 308 nm; malvidin-3-G, was identied at 520 nm while
delphinidin-3-G, cyanidin-3-G, petunidin-3-G, peonidin-3-G were
identied according to the literature (Alcalde-Eon, Escribano-Bailon, Santos-Buelga, & Rivas-Gonzalo, 2006) and quantied as malvidin-3-G equivalents. Quantication was based on 5-point
calibration curves of respective standards (R2 > 0.95, tted to origin) in wine model solution previously described. Relative standard deviation of all calibration points was below 1%. Data
reported represent the mean of two replicates.

Tukeys test. ANOVA was performed using two sided p 6 0.05 and
Levenes test for assessing equality of variance.

2.5.3. Volatile compound determination by SBSEGCMS

Ten millilitres of wines were used in duplicate to determine the
free volatile fraction (Pedroza, Zalacain, Lara, & Salinas, 2010) by
immersion of a polydimethylsiloxane coated stir bar [Twister,
0.5 mm lm thickness, 10 mm length from Gerstel, (Mlheim an
der Ruhr, Germany)] and stirring at 500 rpm during 1 h at 25 C.
After this time, the stir bar was removed from samples, rinsed with
distilled water, dried with cellulose tissue and nally transferred
into thermal desorption tubes for the GC/MS analysis.
Volatile compounds were desorbed from the stir bar in an ATD
400 (Perkin Elmer, Norwalk, CT) under the following conditions:
oven temperature at 290 C; desorption time, 4 min; cold trap temperature, 30 C; helium inlet ow, 45 ml min 1. After this, the
compounds were transferred into the HewlettPackard 6890 (Palo
Alto, CA) gas chromatograph coupled to an HewlettPackard 3D
mass detector (Palo Alto, CA) with a fused silica capillary column
SGE BP21 (stationary phase 30 m length, 0.25 mm i.d., and
0.25 lm lm thickness) (Ringwood, Australia). The chromatographic program was set at 40 C (held for 2 min), raised to
230 C at 10 C min 1 and held for 15 min. Electron impact mode
at 70 eV was used for mass spectrometry analysis. The mass range
varied from 35 to 500 u (Scan Monitoring) and the detector temperature was 150 C. Identication was carried out using the NIST
library and standard spectra. Quantication was based on 5-point
calibration curves of respective standards (R2 > 0.95, tted to origin) in synthetic wine solution previously described. Mean relative
standard deviation of calibration curves was in all cases below 6%
except for 1-hexanol (45%) and farnesol (15%). Data reported represent the mean of two replicates. To avoid matrix interferences
between the volatile compounds, the MS quantication was carried out in the single ion monitoring (SIM) mode using their characteristic m/z values reported in the NIST library and those from
Zalacain et al. (2007).

3.1. Colour

2.6. Statistical analysis

An experimental design considering wine (A05, A07, Y08, Y09),
DWGS type (control, AMIX, Bobal, GM75, GM50), and storage time
(0, 3, 6 months) as categorical factors. Main effect of each factor
and second order interactions between factors on the dependent
variables (composition of samples) were evaluated by means of
multifactor analysis of Variance (ANOVA) with Statgraphics Centurion 16.1. SPSS Statistics 19.0 Software (Chicago, IL) was used for
identifying homogeneous subsets according to ANOVA post hoc

3. Results and discussions

Colour loss is a natural process experienced by all types of
wines due to chemical and biochemical transformations related
with the polyphenols content and the presence of oxygen. Such
changes are particularly important for red wines because colour
is an important quality parameter that may inuence consumer
preferences (Parpinello, Versari, Chinnici, & Galassi, 2009). Previous work on dehydration and characterization of waste grape skins
from the juice industry (Pedroza et al., 2012) and extraction conditions in wine model solution and white wines (Pedroza et al., 2011)
pointed out favourable parameters for releasing colour, polyphenols and aroma compounds. These works also suggested that the
use of DWGS mixtures could be studied in order to t the needs
of certain types of wines like those having colour loss before bottling. The following ndings evaluated the effect of DWGS in different types of red wine that experienced colour loss before bottling.

The addition of DWGS into wines caused a signicant impact in

the Colour Intensity (CI) and Shade of all wines (Table 1). Bobal
DWGS produced the highest CI increase with respect to control,
having an overall average increase of 15% followed by GM50
(13%), GM75 (10%), and nally AMIX (5%). The maximum CI increase was observed in Bobal-Y09 (31%). The improvement of CI
was mainly ascribed to the increase of absorbance at 520 nm (data
not shown), which is commonly related with the amount of anthocyanins of wines and in this case with those released by each type
of DWGS. Grape skins had the least inuence on CI of A07 wine, as
no signicant increase with respect to control was observed. On
the other hand, all types of DWGS produced a signicant increase
of CI in Y09 and A05 wines. According to these results, it seems that
the type of wine regulates the impact of DWGS, where the initial CI
is not a restrictive variable for colour release. Moreover, this remarks that the treatment may improve the colour of both aged
and young wines.
In red wines, Shade represents a measure of colour degradation
since it is the balance between the yellow and red colour (Zamora,
2003) where the higher the value, the higher the inuence of yellow component and thus lower quality. All DWGS produced a diminution of Shade, where Bobal had smaller values than the rest of
treatments, although slight differences were found with the other
grape skins (Table 1). Shade changes were slightly more positive in
A07, Y08 and Y09 than in A05, where GM50 and GM75 had the
closest values compared to those of control. It is important to note
that A05 control wine had the highest shade value, with a predominant yellow component and thus with higher colour degradation.
Evaluation of CI and Shade at T3 and T6 revealed that DWGS
had different evolution trends in each type of wine. CI predominantly increased with time in all samples, where Bobal had the
highest average increase (16% at T3 and 20% at T6). The most inuenced wine was Y09 at T6, where GM75 reached a maximum increase of 45% with respect to control wine (Table 1). DWGS also
caused a signicant increase in A05 wines after 6 months (21
25%). In contrast, the use of DWGS had a low increase of CI in
A07, where evolution was mostly stable. All second order interactions for CI and shade were statistically signicant, however, interaction plots comparing wine and grape skins revealed that the
highest impact of grape skins was achieved in Y09 wine, where Bobal and GM75 were the DWGS producing the highest values during
the whole experiment.

Table 1
UVvis determinations of Colour and Total polyphenols from young (Y08, Y09) and aged (A05, A07) red wines exhibiting colour loss and added with Dehydrated Waste Grape Skins (GM75: mixture of 75% AMIX + 25% Bobal; GM50:
mixture of 50% AMIX and 50% Bobal).

M.A. Pedroza et al. / Food Chemistry 136 (2013) 224236

227

228

Table 2
Monomeric Antocyanin glycosides composition (mg/l) from young red wines (Y08, Y09) added with Dehydrated Waste Grape Skins (GM75: mixture of 75% AMIX + 25% Bobal; GM50: mixture of 50% AMIX and 50% Bobal).

M.A. Pedroza et al. / Food Chemistry 136 (2013) 224236

Table 3
Monomeric Antocyanin glycosides composition (mg/l) from aged red wines (A05, A07) added with Dehydrated Waste Grape Skins (GM75: mixture of 75% AMIX + 25% Bobal; GM50: mixture of 50% AMIX and 50% Bobal).

M.A. Pedroza et al. / Food Chemistry 136 (2013) 224236

229

230

Table 4
Low molecular weight phenolic compounds composition (mg/l) of young red wines (Y08, Y09) added with Dehydrated Waste Grape Skins (GM75: mixture of 75% AMIX + 25% Bobal; GM50: mixture of 50% AMIX and 50% Bobal).

M.A. Pedroza et al. / Food Chemistry 136 (2013) 224236

Table 5
Low molecular weight phenolic compounds composition (mg/l) of aged red wines (A05, A07) added with Dehydrated Waste Grapeskins (GM75: mixture of 75% AMIX + 25% Bobal; GM50: mixture of 50% AMIX and 50% Bobal) (See
above-mentioned references for further information).

M.A. Pedroza et al. / Food Chemistry 136 (2013) 224236

231

232

Table 6
Volatile compounds (lg/l) of young red wines (Y08, Y09) added with Dehydrated Waste Grape Skins (GM75: mixture of 75% AMIX + 25% Bobal; GM50: mixture of 50% AMIX and 50% Bobal).

M.A. Pedroza et al. / Food Chemistry 136 (2013) 224236

Table 7
Volatile compounds (lg/l) of aged red wines (A05, A07) added with Dehydrated Waste Grape Skins (GM75: mixture of 75% AMIX + 25% Bobal; GM50: mixture of 50% AMIX and 50% Bobal). (See above-mentioned references for further
information.).

M.A. Pedroza et al. / Food Chemistry 136 (2013) 224236

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M.A. Pedroza et al. / Food Chemistry 136 (2013) 224236

On the other hand, Shade values of wines with DWGS had a tendency to increase with time while control wines were more stable.
This result was mainly attributed to the increase of the yellow
component which may be ascribed to oxidative browning of polyphenols (Li et al., 2008). Such browning may be directly related
with the decrease of total polyphenols subject of the following
discussion.
3.2. Total polyphenols
After maceration (T0), Total polyphenols (TP) were signicantly
increased (520%) by all treatments, where Bobal had the higher
impact, closely followed by the mixtures GM75 and GM50 (Table 1). It was appreciated that DWGS released the highest amount
of TP in A05 wines (98 6 mg GAE/L) in contrast with Y09 wines
(31 3 mg GAE/L). A previous work macerating Bobal and AMIX
in wine model solution and using the same extraction parameters
(Pedroza et al., 2011), showed that both DWGS were able to release
up to 180 mg GAE/L. This fact suggests that the maximum release
of TP may be primarily controlled by the matrix, leaving a residual
role to the grape skin type. Moreover, when evaluating white
wines, it was observed that DWGS released up to 391 mg GAE/L,
indicating that white wine matrix (with lower content of polyphenols) have more favourable equilibrium conditions thus accepting
a higher amount of polyphenols in solution. It appears that after
DWGS addition, TP reached an equilibrium concentration similar
within all types of wines regardless of treatment, where an average
value of 618 11 mg GAE/L could be characteristic of our samples.
However, these results remark the potential of DWGS for improving the phenolic content of wines to the extent of matching the
concentration of aged wines to that of treated young wines.
Regarding evolution of TP, DWGS-wines had a similar behaviour
as control ones in all treatments (Table 1). This was in agreement
with previous observations on the evolution of TP in ros wines
elaborated with DWGS (Pedroza et al., 2011). A higher decrease
of TP was observed in Y09 after 6 months where AMIX treatment
had the higher loss. Such a decrease could be related to the increase of the shade values previously discussed. It can also be
noted from the evolution data (Table 1) that TP may increase at
T3 with a following decrease after T6, remarking the continuously
changing dynamic equilibrium occurring during wine storage. Results suggest that using DWGS do not cause particular alteration in
the evolution of total polyphenols, and that their positive effect is
stable for up to 6 months. All second order interactions were statistically signicant, showing that all wines treated with grape skins
had superior mean TP values than control wines during the whole
experiment. In addition, the concentration of TP was similar between all wines-grape skin combinations, where bobal had not signicant superior values.
3.3. Anthocyanins
After maceration, DWGS released into all wines an average of
50 mg/l of total monoglucoside anthocyanins. Bobal was the DWGS
producing the highest release (63 mg/l) in A05 wine, while AMIX
had the lowest (37%) in A07 wine (Table 2). However, when evaluating the average release of each DWGS in all wines, it was found
that Bobal and GM50 had similar yields (52 mg/l) followed by
AMIX (47 mg/l) and GM75 (46 mg/l). It was remarkable that these
values were similar in all wines since the release of anthocyanins
depends on the equilibrium concentration of each wine, and it
would be expected that young wines, with higher endogenous concentration of anthocyanins before treatment, had a less efcient
extraction than aged wines (with signicantly lower anthocyanins
and therefore a less saturated solution). This average anthocyanins
yield was similar to that obtained in model wine solutions with

Bobal (50 mg/l) and lower to that obtained when macerating AMIX
with white wines (68 mg/l) (Pedroza et al., 2011) Such facts suggest that the white wine matrix has other variables participating
in the solubility of anthocyanins.
Anthocyanins experienced a signicant decrease during storage,
accounting for a 5070% loss after 3 months and a complementary
640% loss after 6 months (Tables 2 and 3). Evolution patterns of
the different anthocyanins were similar within all wines. AMIX
and GM75 had the lowest decrease of anthocyanins (77 2%) in
A05, A07, and Y08. Apparently, DWGS with higher amount of white
grape skins were able to keep a higher concentration of free anthocyanins in solution for a longer time. However, this behaviour not
observed in Y09 wine, as it had the highest decrease (8589%) of
anthocyanins regardless of DWGS type. Observed changes may
be ascribed either to polymerization-stabilization reactions (Boulton, 2001; Cheynier et al., 2003) and/or degradation due to chemical oxidation phenomena (Li et al., 2008). Since the colour of
samples does not change as abruptly as the anthocyanins concentration, we considered that the rst hypothesis may predominantly
occur, supporting the concept of the partial role of monoglucoside
anthocyanins in colour (Zamora, 2003). In spite of the degradation,
samples with DWGS had signicantly higher concentration of total
anthocyanins at the end of the experiment. All factor interactions
were statistically signicant, however, the most important differences were noted in the concentration of anthocyanins between
treated and control wines, regardless of the grape skin type.

3.4. Low molecular weight phenolic compounds

After maceration treatment (T0), DWGS were able to signicantly increase the concentration of total low molecular weight
phenolic compounds (LMWPC) in young and aged wines (Tables
4 and 5). This was particularly important for A05 wine, where
GM50 produced the highest increase, followed by AMIX, GM75
and nally Bobal (Table 4). It was noted that AMIX, GM50, and
GM75 produced always higher yields than Bobal, regardless of
the type of wine. This was in agreement with previous ndings
on model wine solutions where AMIX released a higher amount
of LMWPC, (Pedroza et al., 2011; Pedroza et al., 2012) therefore
mixtures of DWGS are a good strategy for balancing the deciencies of composition. Since LMWPC relate to sensory attributes like
the acid taste of wines and bitterness, the use of DWGS may allow
winemakers to design particular taste proles by adjusting the
proportion of different grape skins. Regarding the copigmentation
phenomena, Boulton (2001) suggests that an increase in the concentration of co-factor molecules, such as gallic acid, catechin, caffeic acid, epicatechin, etc., during maceration of wines may play a
central role for increasing the solubility of anthocyanins in the
wine solution and therefore achieving higher extraction yield of
anthocyanins and improved colour. Moreover, the increase of cofactor concentration has been considered to prevent colour degradation in grape juice (Brenes, Del Pozo-Insfran, & Talcott, 2005).
Main LMWPC released by DWGS in wines were gallic acid, (+)catechin, ( )-epicatechin, and (E)-resveratrol (Tables 4 and 5). Gallic acid had the highest increase in Y09 wine (7074%). Catechin
and ( )-epicatechin were better released in A05 (74353% and
168263% respectively). The former three compounds have been
reported to have antioxidant activity with suitable use as dietary
supplements (Yilmaz & Toledo, 2004). Caffeic and coumaric acids
had signicantly higher yields in both young wines. (E)-resveratrol
had a signicant increase in Y08 (115136%). The later molecule is
of signicant relevance since it is being attributed with health promoting properties such as prevention of cardiovascular diseases
and antioxidant activity (Fernndez-Mar, Mateos, Garca-Parrilla,
Puertas, & Cantos-Villar, 2012; Frombaum, Le Clanche,

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235

Bonnefont-Rousselot, & Borderie, 2012). Coumaric acid concentration decreased after DWGS addition in A07 (Table 4).
In general, after 6 months of storage all wines with DWGS had
superior content of total LMWPC than control wines. DWGS with
higher proportion of white grape skins continued having the higher
amount of LMWPC. This was also conrmed by interaction plots
comparing average values of total LMWPC during the whole experiment. It was observed that the composition of aged wines was
more unstable than that of young wines; the evolution of Y08
wines showed that gallic acid, (+)-catechin and caffeic acid remained stable during the trial, while ( )-epicatechin, and (E)-resveratrol reached a maximum concentration at T6. On the other
hand, Y09 wines with DWGS had higher concentration of caffeic
acid (T6), ( )-epicatechin (T3), and (E)-resveratrol (T6). Increasing
with time of the later compound is of particular interest since they
might be released by hydrolysis from their glycosilated precursors
(Gmez-Gallego, Garca-Carpintero, Snchez-Palomo, HermosnGutirrez, & Gonzlez Vias, 2012). Interaction plots revealed that
A05 was the most affected wine during storage, although it was
also the wine most favoured by the DWGS treatment.

Wood derived compounds (Table 7) in control wines were

mostly unaffected by DWGS treatment and remain stable during
storage, corroborating previous ndings on the limited inuence
of treatment on these compounds (Pedroza et al., 2011). This is
an important feature given that these compounds are responsible
for highly appreciated notes of coconut, vanilla, oak and spiced
descriptors as well as indicators of wood origin (Daz-Plaza, Reyero, Pardo, Alonso, & Salinas, 2002; Garde-Cerdn & Ancn-Azpilicueta, 2006).
The dehydration pretreatment of DWGS was a cause of the low
volatile yield in the wines. It is also important that the source grape
varieties are considered of a non aromatic nature, because of their
low concentration of monoterpenes (Mateo & Jimnez, 2000). The
use of more aromatic cultivars such as Moscatel may be an interesting alternative for releasing more odourants and producing
wines with novel characteristics. Although the volatile prole of
wines with DWGS was mostly similar to that of control samples,
the best results were observed during the rst 3 months when volatiles released are still over their corresponding OT.

3.5. Volatile composition

4. Conclusions

The volatile composition of wines was modied after addition

of DWGS by the increase of b-ionone (oral descriptor). This compound is an important impact odourant because of its low olfactory
threshold and appreciated descriptor. The increase of b-ionone was
of the most importance in the case of aged wines (Table 7), where
this compound was absent or below its olfactory threshold (OT)
(0.09 lg/l (Francis & Newton, 2005)) in control wine and after
addition of DWGS, it increased to a concentration 26 times over
the OT. Bobal was the DWGS releasing the highest amount of b-ionone (0.54 lg/l in A05 wine). Wine-time was the only statistically
signicant interaction of this compound, describing a decrease
with time, particularly important in the case of young wines.
On the other hand, nerolidol and b-damascenone concentration
decreased after treatment regardless of the DWGS type. This phenomena was also observed when adding DWGS to white wines
(Pedroza et al., 2011) However, in the case of b-damascenone,
the concentration (0.772.33 lg/l) remained over the OT
(0.05 lg/l (Francis & Newton, 2005)), thus its characteristic oralfruity note (Bayonove et al., 2003) may persist in the wine.
Although statistically signicant interactions (wine-DWGS and
wine-time) were observed for this compound, the most important
differences were always associated to control wines having higher
mean concentration than those with DWGS.
The herbaceous compounds represented by 1-hexanol remained mostly without signicant differences or in some cases
in a lower concentration than control wines. Also this compound
was not over its OT. Nerolidol and Farnesol were not detected in
Y09 after addition of DWGS, indicating that in this particular wine
these compounds could be transformed during treatments as a result of hydrolysis and cyclation reactions taking place at wine conditions (Marco, 2006). This was not observed in the rest of wines,
thus this behaviour was not clear. When evaluating the evolution
of wines in terms of chemical families (Tables 6 and 7), an increase
with time of isoamyl acetate (banana note) was observed in all
treated wines, but no particular DWGS seems to induce such
behaviour. Since this compound is produced by yeast (Styger, Prior,
& Bauer, 2011), this could be related with refermentation of wines,
although no visual sign of this was observed. Terpenes in A05, A07
and Y08 increased after 6 months of storage. Increasing of these
compounds with storage may be due to their existence in glycosilated forms and subsequent chemical hydrolysis. It may as well be
related to their susceptibility to different reactions and transformations (isomerization, cyclation, oxidation, etc.) (Gnata, 2003).

Reaching a conclusion, the use of dehydrated waste grape skins

from the juice industry proved being a useful tool for ameliorating
quality parameters of colour and phenolic compounds before bottling. Firstly, the red colour of wines increased by all DWGS regardless of the wine type, together with Total polyphenols content. The
complementary mixing of red and white grape skins was useful to
release signicant amounts of anthocyanins and low molecular
weight phenolic compounds, characteristic each grape skin variety.
Important bioactive compounds such as (E)-resveratrol and catechins were signicantly released by all types of DWGS. The volatile
prole of wines was moderately inuenced by DWGS although
quality impact odourants like b-ionone were released. Moreover
compounds related to wood ageing such as whiskylactones and
eugenol were not particularly affected by treatment. The type of
wine inuenced the release of compounds quantitatively, indicating that chemical equilibrium of each wine may help or hinder
the extraction. Addition of DWGS allowed preventing signicant
loss of phenolic compounds occurring during storage. Although
the use of DWGS is currently not authorized as enological practice
because of its novelty, probably, the most important challenges for
its aproval as wine additive will come from political and cultural
concerns related with the usage conditions.

Acknowledgements
M.A.P. has received a CONACYT grant from the Mexican Government. This Study has been funded by Junta de Comunidades de
Castilla-La Mancha (Project PAI08-0148-9842). Thanks to Ana Soler
from Julian Soler S.A. Juice Concentrate Factory for supplying waste
grape skins. Thanks to Kathy Walsh for proofreading the
manuscript.

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