Accepted Manuscript Influence of bottle storage time on colour, phenolic composition and sensory properties of sweet red wines Ana Marquez, Maria P. Serratosa, Julieta Merida PII: DOI: Reference:
S0308-8146(13)01368-X http://dx.doi.org/10.1016/j.foodchem.2013.09.103 FOCH 14731
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Food Chemistry
Received Date: Revised Date: Accepted Date:
15 July 2013 10 September 2013 16 September 2013
Please cite this article as: Marquez, A., Serratosa, M.P., Merida, J., Influence of bottle storage time on colour, phenolic composition and sensory properties of sweet red wines, Food Chemistry (2013), doi: http://dx.doi.org/ 10.1016/j.foodchem.2013.09.103
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1
Influence of bottle storage time on colour, phenolic
2
composition and sensory properties of sweet red wines
3
4
Ana Marquez, Maria P. Serratosa, Julieta Merida*
5
Department of Agricultural Chemistry. Faculty of Sciences. University of Cordoba.
6
Edificio Marie Curie. Campus of Rabanales. E-14014. Cordoba, Spain.
7 8 9
* Corresponding Author. E-mail: [email protected]. Tel: +34 957 218 612; Fax: +34 957 212 146
1
10
ABSTRACT
11
Changes in colour and phenolic composition in sweet red wines made from Merlot,
12
Syrah and Tempranillo grapes were studied in order to assess the influence of bottle
13
storage over a period of 12 months. For this purpose, wine colour parameters, sensory
14
analysis and concentrations of monomeric anthocyanins, pyranoanthocyanins,
15
methylmethine-mediated condensation adducts, flavan3-ol derivatives and flavonols
16
were measured. Hue increased and red colours decreased with the storage time,
17
particularly over the first 3 months. The concentrations of low molecular weight flavan-
18
3-ol derivatives decreased with time due to the effect of their conversion into tannins of
19
high molecular weight. In addition, the glycosylated flavonols decreased through
20
hydrolysis to give the corresponding aglycones. Overall, the concentration of phenolic
21
compounds decreased markedly with storage time, whereas the antioxidant activity in
22
the wines remained constant throughout. A panel of expert tasters judged the colour,
23
aroma and flavour of all initial and final wines to be acceptable.
24 25
Keywords: Sweet red wines, Bottle storage, Phenolic compounds, Colour
2
26
1.
27
The colour of red wines is one of the most important characteristics and it is the first
28
attribute assessed by consumers during the tasting. The colour of wine reflects some of
29
its virtues and defects, its age and whether it has been subjected to a specific ageing
30
process.
Introduction
31
Phenolic compounds are the main agents responsible of the colour of wine,
32
whether white or red. Also, they influence other sensory properties such as astringency
33
and bitterness. Anthocyanins constitute the most important polyphenol family in red
34
wines and are responsible for their colour. In fact, anthocyanins are gradually extracted
35
from grape skins during the early stages of winemaking, and give musts and young
36
wines their typical bluish red colour. During the subsequent vinification and ageing
37
steps, the wine colour changes by the effect of these compounds undergoing mainly
38
copigmentation, cycloaddition, polymerization and oxidation reactions (Cheynier &
39
Ricardo da Silva, 1991; Monagas, Bartolome & Gomez-Cordoves, 2005; Alcalde-Eon,
40
Escribano-Bailon, Santos-Buelga & Rivas Gonzalo, 2006).
41
During the bottle storage time of the red wine, spontaneous clearing, colour
42
stabilisation and reactions that give the formation of more complex compounds have
43
been found (Del Alamo-Sanza & Nevares Dominguez, 2006). As the storage time in
44
bottle increases, copigmentation and polymerization anthocyanins reactions take place
45
(Eiro & Heinonen, 2002) to form more stable substances that change the initially bluish
46
red hues of young wines to orangish red hues in aged wines (Atasanova, Fulcrand,
47
Cheynier & Moutounet, 2002). Among others, anthocyanins can react with flavan-3-ol
48
derivatives, whether directly or via acetaldehyde (Vivar-Quintana, Santos-Buelga &
49
Rivas-Gonzalo, 2002). Also, anthocyanins can condense with other compounds of lower
50
molecular weight such as pyruvic acid, vinylphenol or glyoxylic acid (Marquez, 3
51
Serratosa & Merida, 2013). The resulting pigments, known as pyranoanthocyanins, are
52
more stable and keep the wine colour intensity along the time (Revilla & Gonzalez-
53
Sanjose, 2001), but these pigments have more orangish hues than the starting
54
anthocyanins. On the other hand, during these wine evolution steps the flavan-3-ol
55
derivatives undergo oxidative browning reactions and interact with proteins to form
56
substances that cause wine clouding (Cheynier & Ricardo da Silva, 1991). These
57
reactions alter the colour but also reduce astringency and bitterness, thereby leading to
58
softer tasting wines.
59
In recent years, the demand and consumption of red wines has risen at the
60
expense of white wines. Also, there has been an increasing interest in sweet wines.
61
Traditionally, sweet wines have been made from white grapes, known as ice wines,
62
noble rot wines or raisin wines (Cliff, Yuksel, Girard & King, 2002; Guarrera, Campisi
63
& Asmundo, 2005; Thibon, Dubourdieu, Darriet & Tominaga, 2009). At present,
64
however, some Spanish viticultural areas are increasingly producing sweet red wines
65
obtained by using special procedures involving on-vine or off-vine drying of the grapes,
66
fermentation and/or fortification, maceration, etc. Most of these wines are not aged, but
67
seemingly require storage for some time. Although some authors have examined
68
changes during the red wine bottle stabilisation (Perez-Magariño & Gonzalez San-Jose,
69
2004; Del Alamo Sanza & Nevares Dominguez, 2006), none studied this evolution in
70
sweet red wines.
71
In this work, changes in phenolic compounds and colour in sweet wines made
72
from red grapes grown in the Montilla–Moriles region (southern Spain) have been
73
examined during the stabilisation time in bottle. The aim was to assess the development
74
of the different reactions involving these compounds and the sensory impact of the
75
storage time.
4
76 77
2.
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2.1. Samples
Material and methods
79
Young sweet red wines from Merlot, Syrah and Tempranillo varieties, with
80
approximately 300 g/L of reducing sugar and 15% (v/v) alcohol were used. For the
81
elaboration of the wines, the grapes were dried at controlled temperature. The raisins
82
were pressed, the musts were fortified and a maceration step with the skins was carried
83
out. The final wines were stored in black bottles and the headspace was replaced by
84
nitrogen before closing. The bottles were labeled and stored in the dark at 20 °C for 12
85
months and the wines were analyzed at 0, 3, 6, 9 and 12 months during their
86
stabilisation. All the samples were centrifuged at 3000 rpm, filtered and analysed in
87
triplicate.
88
2.2. Reagents
89
Phenolic compounds standards were obtained from Extrasynthese (Genay, France).
90
Methanol, formic acid, hydrochloric acid and acetonitrile were purchased from Merck
91
(Madrid, Spain).
92
2.3. Extraction of phenolic compounds
93
For the anthocyanin extraction, a volume of 2 mL of wine was passed through a
94
Sep-Pak C18 cartridge packed with 900 mg of material (Long Body Sep-Pak Plus,
95
Waters Corporation, Milford, MA) previously activated with methanol and HCl 0.01%.
96
The cartridge was washed with HCl 0.01% and ethyl acetate, and the anthocyanins were
97
recovered with methanol acidified to pH 2.
5
98
For the extraction of flavan-3-ol derivatives and flavonols, 15 mL of wine adjusted
99
to pH 7 were passed through Sep-Pak C18 cartridge activated with methanol and water
100
at pH 7. The cartridge was washed with water at pH 7 and the flavan-3-ol derivatives
101
were eluted with 16% acetonitrile at pH 2. Subsequently, the flavonols were recovered
102
with ethyl acetate, evaporated to dryness and dissolved in methanol.
103
The collected fractions were concentrated on a vacuum centrifuge thermostated at
104
35 ºC and passed through a filter of 0.45 µm pore size for injection into a HPLC
105
instrument.
106
2.4. HPLC analysis
107
The identification of the phenolic compounds was achieved by UV spectra obtained
108
by diode array HPLC (Spectra-Physics UV6000LP) and it was confirmed by
109
HPLC/ESI-MS analysis (TermoQuest Finnigan AQA quadrupole mass spectrometer),
110
according to the methods proposed by Marquez, Serratosa, Lopez-Toledano and Merida
111
(2012). The column used in the analyses was a 250 mm x 4.6 mm i.d., 5 µm,
112
LiChrospher 100 RP-18, using 10% aqueous formic acid (A) and acetonitrile/formic
113
acid/H2O (45:45:10) (B) as mobile phases at a flow rate of 1 mL/min. The mobile phase
114
gradient for the anthocyanin identification was as following: gradient elution from 15%
115
to 30% B in 17 min, gradient elution up to 73% B in 28 min, gradient elution up to
116
100% B in 3 min and isocratic elution for 3 min, using absorbance at 520 nm to
117
quantify. For the flavan-3-ol derivatives identification, the absorbance at 280 nm was
118
used with a gradient elution from 5% to 10% B in 25 min, gradient elution up to 20% B
119
in 10 min, gradient elution up to 30% B in 10 min, gradient elution up to 100% B in 15
120
min, and isocratic elution for 10 min. The flavonols were identified at 360 nm and this
121
fraction was obtained by gradient elution from 5% to 30% B in 5 min, gradient elution
6
122
up to 40% B in 14 min, gradient elution up to 80% B in 1 min and isocratic elution for 5
123
min.
124
2.5. Spectrophotometric measurements
125
A PerkinElmer (Waltham, MA) Lambda 25 spectrophotometer, with quartz cells of
126
1 mm light path was used. All measurements were corrected for a path length of 1 cm.
127
Samples were previously passed through Millipore (Billerica, MA) HA filters of 0.45
128
µm pore size. Absorbances at 420, 520 and 620 nm were measured and colour intensity
129
(A420+A520+A620), hue (A420/A520) and percentages of yellow, red and blue colours were
130
calculated. Polymeric pigments colour (PPC) was obtained as the absorbance at 520 nm
131
after 45 min of 5 mL of wine previously supplied with 15 mg of Na2S2O5. Wine
132
coloured anthocyanins (WCA) was determined by diluting 1 mL of sample ten times
133
with 1 M HCl and measuring the absorbance at 520 nm after 45 min at 25 ºC. The total
134
tannin index was determined by measurement of the absorbance at 550 nm in a cell of 1
135
cm light path after acid hydrolysis of the samples.
136
2.6. Antioxidant activity
137
The DPPH assay was realized for the antioxidant activity determination. The wines
138
were diluted with a solution containing 12% ethanol, and an aliquot of 200 µL was
139
placed in a cell with 3 mL of a 45 mg/L solution of DPPH in methanol. A blank (200 µL
140
dilution sample and 3 mL methanol), a control sample (200 µL of 12% ethanol in water
141
and 3 mL of DPPH solution) and a Trolox standard (200 µL of 80 mg/L Trolox solution
142
and 3 mL of DPPH solution) were also prepared in parallel. The absorbances at 517 nm
143
of the control sample and blank were measured and the absorbances of the standard and
144
samples were measured after 10 min of incubation. The antioxidant activity, expressed
7
145
in millimoles of Trolox (mmol TE) per liter, was calculated according to Serratosa,
146
Marquez, Lopez-Toledano, Medina and Merida (2011).
147
2.7. Sensory analysis
148
The initial and final sweet red wines were assessed for colour, aroma and flavour
149
acceptability by 15 tasters in a panel in accordance with ISO 8586-1:1993. The tasting
150
room was kept at 20ºC and wines served in tasting glasses certified and coded.
151
Evaluation of the quality of the wines was made using the method according to ISO
152
4121:2003, with options of desirable (5-6), acceptable (3-4) and undesirable (1-2). The
153
final punctuations were calculated as the mean, taking into account the evaluation of
154
each taster.
155
2.8. Statistical Procedures
156
The results for all samples were subjected to multifactor analysis of variance
157
(ANOVA) and discriminant analysis in triplicate, using the Statgraphics Computer
158
Package v. 5.0 from Statistical Graphics Corp (Warrenton, Virginia).
159 160
3.
Results and discussion
161
Wines undergo a large number of chemical reactions that alter their phenolic
162
composition and colour during ageing. In this work, changes in anthocyanins during the
163
storage of sweet red wines from Merlot, Syrah and Tempranillo grapes in bottle were
164
examined. Fig. 1 shows the chromatograms obtained after 0, 3, 6, 9 and 12 months of
165
storage in Merlot wine. As can be seen, the anthocyanin profile changed markedly with
166
time and the peak areas for monomeric anthocyanins decreased considerably with time.
8
167
Table 1 shows the concentration of each individual anthocyanin and those
168
homogeneous groups obtained with an analysis of variance at the 99.9% confidence
169
level. As can be seen, the concentrations of monomeric anthocyanins decreased
170
markedly with time, which is consistent with the results of many previous studies on the
171
storage of red wines in bottle (Perez-Magariño & Gonzalez San-Jose, 2004; Del Alamo
172
Sanza & Nevares Dominguez, 2006). The greatest decrease was observed during the
173
first 3 months in the wines from Merlot, Syrah and Tempranillo grapes (64, 77 and 83%
174
respectively). Most anthocyanin monomers disappeared after 3-9 months of storage. As
175
a result, the obtained wines contained malvidin-3-glucoside and malvidin-3-
176
coumaroylglucoside,
177
acetylglucoside in Merlot and Syrah wines, and petunidin-3-glucoside and peonidin-3-
178
acetylglucoside in Merlot wine. The final concentration of monomeric anthocyanins
179
was 13.6 mg/L in Merlot wine, 8.68 mg/L in Syrah wine and 3.52 mg/L in Tempranillo
180
wine. Therefore, the reduction in the anthocyanin contents was 93, 96 and 98%
181
respectively, and consistent with previous results of Garcia-Falcon, Perez-Lamela,
182
Martinez-Carballo and Simal-Gandara (2007), who reported a decrease by 72 and 85%
183
in Mencia and Brancellao wines respectively, after 12 months of storage in bottle.
in
addition
to
peonidin-3-glucoside
and
malvidin-3-
184
Glucosylated derivatives were the major monomeric anthocyanins present, with
185
63, 58 and 60% in the initial Merlot, Syrah and Tempranillo wines respectively, and 20,
186
17 and 14% respectively, after 12 months of storage (Fig. 2). The other three
187
monomeric anthocyanin families (acetylated, coumaroylated and caffeoylated
188
derivatives) also decreased or even disappeared during storage.
189
The decreases in the anthocyanin contents during storage must have resulted
190
from the gradual conversion of monomeric compounds into more stable oligomers or
191
polymers (Monagas, Gomez-Cordoves & Bartolome, 2006). As can be seen in Table 2,
9
192
polymeric pigment colour (PPC) increased gradually during storage, especially during
193
the first 3 months, when the absorbance of the Merlot, Syrah and Tempranillo wines
194
rose by 1.21, 1.10 and 1.08 a.u. respectively. This trend continued throughout the
195
storage period in the Merlot and Syrah wines, but ceased after 3 months in the
196
Tempranillo wines. The three types of wine showed no statistically significant
197
differences in this respect after 6 months.
198
A linear correlation between the variation of the content in monomeric
199
anthocyanins and PPC provided a Pearson linear correlation coefficient of -0.9504 (p
3) by the tasters. The initial wines were judged slightly different in colour and
343
flavour, but very similar in aroma. Syrah and Merlot wine were the best scored for
344
colour (4.4) and flavour (4.4) respectively, before storage. By contrast, the aged wines
345
differed markedly in colour, with the Merlot wine as the best scored (4.5) and
346
Tempranillo wine as the worst (2.8). These sensory results are consistent with the
347
analytical results. Thus, Merlot wine was that with the reddest, most stable colour, and
348
Tempranillo wine that with the brownest colour. On the other hand, Tempranillo wine
349
was judged the best in terms of aroma (4.2), well above Merlot wine (2.9) and Syrah
350
wine (3.2).
351
In summary, the storage of sweet red wines in bottle for a year caused the
352
degradation of monomeric anthocyanins and the completely disappearance of most of
353
them between 6 and 9 months. Based on the results, the decrease in the contents of these
354
compounds was a result of various gradual reactions leading to more stable oligomers or
355
polymers, and others yielding anthocyanin derivatives such as pyranoanthocyanins and
356
methylmethine-mediated condensation adducts. These changes in anthocyanin
357
compounds caused the red colour of the young wines to evolve to browner hues as
358
storage time increased. This was especially so in Tempranillo wine and, to a lesser
359
extent, in Merlot wine. By the effect of their polymerization to compounds of higher
360
molecular weights and their participation in reactions with anthocyanins to form
361
adducts, flavan-3-ol derivatives decreased during storage in bottle. Simultaneously,
362
glycosylated flavonols were hydrolysed to their corresponding aglycones. Despite their
363
changes in phenolic concentrations, the wines retained their antioxidant activity and
364
hence their beneficial effects on health. Finally, both young and aged wines were judged
365
acceptable
by
expert
tasters
on
the
16
basis
of
their
sensory
properties.
366 367
Acknowledgments
368
The authors gratefully acknowledge financial support from the Spanish Government,
369
Minister of Education (FPU scholarship of A. Marquez) for the realization of this work.
370 371
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3000
UV6000-520nm
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-200
468
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Minutes
469 470
Fig. 1. Chromatographic anthocyanin profile of Merlot wine after 0, 3, 6, 9 and 12
471
months of bottle storage.
472 473 474
22
100% 80% 60% 40% 20% 0% 0
475
3
6
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9
12 months
% Methylmethine-mediated condensation products % Pyranoanthocyanins %Caffeoylglucosides %Coumaroylglucosides %Acetylglucosides %Glucosides
476 477
Fig. 2. Relative contents of the anthocyanin compounds in wines during the bottle
478
storage.
479 480 481
23
Antioxidant Activity (mmol TE/L)
9 8 7 6 5 4 3 Merlot Syrah Tempranillo
2 1 0
482 483
0
3
6
9
12
Fig. 3. Antioxidant activity in wines during the bottle storage.
484 485 486
24
months
497
(a)
Function 2 (5.5%)
Merlot wines
Syrah wines
12 months
0 months 9 months
Tempranillo wines
6 months
498 499
3 months
(b) Function 2 (17.5%)
487 488 489 490 491 492 493 494 495 496
Function 1 (92.2%)
Function 1 (82.5%)
500 501
Fig. 4. Multivariate discriminant analysis. Byplot using as dependent variable the grape variety (a) and the bottle storage time (b).
502 503
26
Table 1. Anthocyanins (mg/L), flavan-3-ol derivatives (mg/L), total tannins (g/L) and flavonols (mg/L) concentrations in wines during the bottle storage (means, standard deviations and homogenous groups for an analysis of variance with a confidence level of 99.9%). Merlot Months
0
3 c
6 b
12
0
3 b
12
0
3 c
9 a
12
n.d.
n.d.
5.53±0.110
n.d.
n.d.
7.17±0.210
0.981±0.101
n.d.
n.d.
3.97±0.272b 5.56±0.221c 11.6±0.839b 29.3±0.436c n.d. n.d. n.d. 5.10±0.140c 7.00±0.185c n.d. n.d. n.d. 2.19±0.057b n.d 67.1±2.50c
2.99±0.066a 3.30±0.186b 5.43±0.095a 11.5±0.321b n.d. n.d. n.d. 4.01±0.217bc 5.56±0.235b n.d. n.d. n.d. 2.35±0.020b n.d. 37.7±1.13b
n.d. 1.69±0.136a 2.76±0.336a 5.40±0.416a n.d. n.d. n.d. 2.85±0.168ab 2.86±0.533a n.d. n.d. n.d. 1.14±0.036a n.d. 16.7±0.919a
n.d 1.20±0.068a 2.09±0.167a 4.15±0.174a n.d. n.d. n.d. 2.64±0.066a 2.23±0.271a n.d. n.d. n.d. 1.32±0.021a n.d. 13.6±0.514a
6.54±0.105b 9.42±0.146c 39.0±1.060c 77.9±2.63c 5.53±0.085 4.97±0.165 6.14±0.173b 13.3±0.379c 28.3±0.624d 5.20±0.114 4.98±0.100 7.22±0.085c 10.9±0.379 7.17±0.170 224.9±5.60d
2.56±0.061a 3.02±0.026b 7.67±0.271b 15.5±0.265b n.d. n.d. 3.34±0.090a 5.45±0.165b 6.90±0.104c n.d. n.d. 2.93±0.032b 3.30±0.332 n.d. 52.2±2.00c
2.35±0.090a 2.78±0.093b 4.15±0.212a 6.43±0.329a n.d. n.d. n.d. 3.28±0.150a 4.23±0.339b n.d. n.d. 2.39±0.145a 2.50±0.046 n.d. 30.2±0.817b
n.d. 1.38±0.147a 2.57±0.301a 3.41±0.030a n.d. n.d. n.d. n.d. 2.71±0.032a n.d. n.d. n.d. 1.13±0.020 n.d. 11.2±0.285a
n.d. n.d. 1.89±0.176a 2.83±0.146a n.d. n.d. n.d. n.d. 2.66±0.375a n.d. n.d. n.d 1.30±0.056 n.d. 8.68±0.727a
5.50±0.053c 11.7±0.153c 11.9±0.265b 75.7±2.32c 6.37±0.340 4.78±0.183 5.47±0.234 6.15±0.111 9.62±0.095c 4.27±0.021 5.92±0.178b 4.93±0.13 12.2±0.153c 8.31±0.151b 171.7±3.34d
2.21±0.017b 3.07±0.038b 2.75±0.153a 11.1±1.01b n.d. n.d. n.d. n.d. 3.57±0.279b n.d. n.d. n.d. 3.20±0.031b 2.74±0.146a 28.5±1.32c
1.18±0.066a 2.01±0.300a 2.14±0.146a 5.50±0.363a n.d. n.d. n.d n.d. 2.66±0.350ab n.d. n.d. n.d. 1.39±0.142a n.d. 15.9±1.13b
n.d. n.d. n.d. 3.16±0.251a n.d. n.d. n.d. n.d. 1.85±0.235a n.d. n.d. n.d. 1.07±0.050a n.d. 6.08±0.501a
n.d. n.d. n.d. 2.44±0.129a n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. 1.08±0.106a n.d. 3.52±0.124a
A vitisin Malvidin-3-acetylglucoside A vitisin Peonidin-3-acetylglucoside B vitisin Malvidin-3-glucoside B vitisin Peonidin-3-glucoside B vitisin Peonidin-3-acetylglucoside Pyranoanthocyanins
1.20±0.097a n.d. n.d. n.d. n.d. 1.20±0.097a
7.70±0.181c 3.94±0.155d 3.23±0.346c 2.30±0.140a 4.71±0.355c 22.6±0.827c
5.03±0.118b 2.31±0.225c 3.34±0.181b 2.25±0.032a 3.70±0.185b 16.6±0.489d
4.76±0.477b 1.95±0.046b 1.78±0.122a n.d. 3.01±0.437a 11.5±1.01b
4.46±0.123b 1.52±0.055a 1.79±0.305a n.d. 2.75±0.240a 10.5±0.667b
n.d. n.d. 0.296±0.034a n.d. 1.22±0.114a 1.52±0.146a
1.39±0.010a 0.744±0.023a 0.959±0.064b n.d. 1.74±0.093ab 4.83±0.131b
2.38±0.204b 2.23±0.078c 1.51±0.067c n.d. 2.26±0.316b 8.38±0.654c
3.05±0.144b 1.99±0.121c 1.98±0.213c 1.57±0.147a 1.69±0.160ab 10.3±0.270d
2.97±0.255b 1.60±0.095b 1.85±0.183c 1.43±0.075a 1.88±0.322ab 9.73±0.719cd
2.82±0.386a n.d. 0.846±0.072a n.d. n.d. 3.66±0.414a
3.35±0.093a n.d. 1.04±0.098a n.d. 1.33±0.121a 5.71±0.295ab
5.46±0.399b 1.57±0.125a 2.54±0.230c n.d. 3.02±0.430b 12.6±0.849d
4.02±0.478a 1.14±0.127a 1.81±0.266b n.d. 0.966±0.025a 7.94±0.782c
3.25±0.166a 1.34±0.079a 1.11±0.100a n.d. 1.23±0.136a 6.93±0.367bc
Malvidin-3-glucosidemethylmethine-(epi)catechin Malvidin-3-glucosidemethylmethine -(epi)catechin Malvidin-3-glucosidemethylmethine-(epi)catechin Peonidin-3-glucosidemethylmethine-(epi)catechin Peonidin-3-glucosidemethylmethine-(epi)catechin Malvidin-3-acetylglucosidemethylmethine-(epi)catechin Malvidin-3-acetylglucosidemethylmethine-(epi)catechin Methylmethine adducts
0.935±0.077a 4.29±0.061bc
5.08±0.242c
3.88±0.408b
4.03±0.079b
0.792±0.032a 1.78±0.151b
4.11±0.228c
3.36±0.310c
2.41±0.258b
0.663±0.034a 1.46±0.097ab 4.48±0.430d
2.62±0.185c
2.08±0.095bc
0.730±0.105a 5.34±0.106c
2.66±0.089b
3.03±0.412b
2.63±0.151b
0.788±0.042a 1.76±0.095b
1.97±0.204bc 2.27±0.105cd 2.59±0.130d
0.681±0.016a 2.21±0.163b
2.23±0.255b
2.42±0.215b
1.92±0.321b
0.433±0.040a 1.37±0.114bc
1.24±0.057b
1.76±0.188cd
1.89±0.117d
0.527±0.019a 0.672±0.032a 1.34±0.157b
1.67±0.142b 1.61±0.180b
0.653±0.077a 1.93±0.095b
1.99±0.101b
1.72±0.194b
1.59±0.140b
2.21±0.241b
1.49±0.108a
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
0.903±0.032
n.d. 2.01±0.087
n.d.
n.d. a
18.8±0.198
c
n.d. a
n.d.
n.d.
n.d.
1.89±0.135
a
0.378±0.016
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
2.00±0.116a
6.93±0.141b
11.7±0.958c
7.73±0.410b
6.82±0.093b
0.901±0.025
b
1.51±0.074
b
0.458±0.005
0.641±0.070
n.d.
1.50±0.074
n.d.
0.395±0.032a 0.669±0.044b
n.d.
n.d.
n.d.
n.d.
12.3±1.41b
3.33±0.126a
8.29±0.192c
12.3±0.702b
10.9±0.372b 10.0±0.400b
14.6±0.506
0.928±0.080
12.2±0.259b
1
b
n.d.
a
c
b
b
0.841±0.014a 1.33±0.181a
2.50±0.078
6 b
5.23±0.010c 8.13±0.172d 23.9±0.200c 80.7±1.57d n.d. 4.61±0.113 5.60±0.110 7.95±0.444d 24.2±0.265d 4.38±0.100 4.47±0.10 5.16±0.280b 7.16±0.185c 6.23±0.030b 186±1.55d
c
2.13±0.023
9 a
4.95±0.046
n.d.
2.37±0.040
6 a
Cyanidin-3-glucoside Petunidin-3-glucoside Peonidin-3-glucoside Malvidin-3-glucoside Delphinidin-3-acetylglucoside Cyanidin-3-acetylglucoside Petunidin-3-acetylglucoside Peonidin-3-acetylglucoside Malvidin-3-acetylglucoside Cyanidin-3-coumaroylglucoside Petunidin-3-coumaroylglucoside Peonidin-3-coumaroylglucoside Malvidin-3-coumaroylglucoside Malvidin-3-caffeoylglucoside Anthocyanin monomers
n.d.
2.63±0.050
9 a
Tempranillo
Delphinidin-3-glucoside
n.d.
3.52±0.155
Syrah
(+)-Catechin (-)-Epicatechin Procyanidins B1+B3 Procyanidins B2+B4 B2-O-gallate dimer B2-O’-gallate dimer B1-O-gallate dimer Flavan-3-ol derivatives
79.4±1.34b 25.7±5.03a 14.3±1.37b 16.3±1.18c 12.2±1.05a n.d. 6.10±0.464a 154±7.03b
138.2±10.6c 35.7±0.551b 12.4±0.245b 21.6±1.25d 24.9±1.85b 10.6±1.10b 18.5±1.41d 262±12.3b
73.1±0.405ab 12.7±0.513b 9.07±0.245a 5.16±0.291b 15.5±0.924a n.d. 8.35±0.950ab 124±0.401a
75.1±5.17ab n.d. 9.26±0.644a 4.57±0.339ab 32.9±0.569c n.d. 12.1±0.341c 134±4.40ab
58.6±2.22a 4.68±0.719a 7.06±0.229a 1.84±0.305a 36.3±0.503c 3.05±0.497a 10.4±1.00bc 122±1.94a
75.0±3.69c 42.0±1.66b 7.66±0.261c 15.5±1.20b 5.22±0.882a 1.42±0.172a n.d. 150±4.07c
62.2±2.92bc 12.7±0.153a 5.02±0.600ab 6.88±0.287a 4.18±0.225a 7.14±0.374c 6.78±0.482a 105±3.73b
44.6±3.97a n.d. 5.39±0.026ab 7.57±1.20a 13.4±1.31b 3.52±0.511b n.d. 74.5±1.27a
48.0±7.05ab n.d. 5.91±0.026b 7.88±0.155a 23.2±0.945c 3.21±0.280ab n.d. 88.2±7.50ab
35.3±1.85a n.d. 4.02±0.522a 7.72±0.287a 20.2±1.67c 4.91±0.905b 18.1±0.611b 90.2±4.10ab
86.6±5.97d 43.3±0.723c 9.20±0.613b 10.0±0.455b 11.1±0.819a 4.89±0.391a 4.58±0.359a 170±4.13d
69.7±0.615cd 20.8±0.557b 6.21±0.223a 8.71±0.761ab 12.9±0.889ab 7.72±0.748a 6.51±0.756a 133±2.73c
46.7±7.54b 4.85±0.758a 4.73±0.835a 7.43±0.519a 17.0±1.38b n.d. 15.2±1.10b 95.9±6.30b
51.0±8.28bc n.d. 4.24±0.354a 7.70±0.171a 24.6±1.66c n.d. 14.8±0.252b 102±9.81b
5.74±0.424a n.d. 5.38±0.745a 8.70±0.625ab 25.3±0.777c 4.95±0.646b 17.1±1.10b 67.2±2.49a
Total tannins (g/L)
8.81±0.208ª
13.2±0.196c
12.9±0.302c
11.5±0.184b
8.70±0.200a
6.35±0.093a
8.90±0.040b
13.3±0.519d
12.0±0.237c
11.2±0.375c
9.93±0.028b
9.18±0.077a
13.3±0.169c
10.1±0.018b
10.0±0.117b
Myricetin-3-glucoside Quercetin-3-glucuronide + quercetin-3-galactoside Quercetin-3-glucoside Laricitrin-3-glucoside Kaempferol-3-glucoside Isorhamnetin-3-glucoside + kaempferol-3-glucuronide Syringetin-3-glucoside Myricetin Quercetin Laricitrin Kaempferol Isorhamnetin Syringetin Flavonols
0.295±0.010b 0.511±0.041c 0.125±0.007a 0.599±0.012c n.d.
n.d.
0.315±0.038b n.d.
4.77±0.198
n.d.
n.d.
n.d.
5.98±0.212
b
5.24±0.352
ab
4.33±0.191
2.45±0.123 0.455±0.073 1.14±0.036 1.03±0.099b 0.273±0.003a n.d. 0.798±0.046a n.d. n.d.
4.66±0.186b
0.492±0.041a 0.360±0.021a 0.256±0.038a n.d.
11.8±0.850c
0.756±0.023a
1.39±0.055c
0.660±0.006b 0.211±0.015a n.d.
n.d.
n.d. 0.376±0.012a 2.87±0.050a 1.04±0.085a 0.471±0.016a 2.51±0.026a 0.502±0.072a 8.53±0.139a
c
2.34±0.044c 0.906±0.049b 2.75±0.192cd 1.20±0.106b 1.27±0.081c 0.971±0.069b n.d. 17.9±0.827d
n.d. 0.151±0.008a 2.15±0.038b 0.838±0.046a 1.01±0.001b 0.484±0.020a 0.243±0.009a 4.88±0.054a
3.91±0.153 0.249±0.019b 1.92±0.089a n.d. 0.520±0.029ab 0.514±0.018a n.d. 24.0 ±0.797c
3.83±0.284 0.604±0.027d 5.55±0.337c 1.88±0.068b 1.11±0.101d 3.87±0.192d n.d. 29.4±1.96d
2.46±0.061 0.261±0.024b 3.55±0.107b 1.02±0.023a 0.799±0.073bc 1.86±0.052b 0.429±0.023a 13.5±0.336b
2.61±0.035 0.496±0.027c 4.87±0.117c 1.29±0.050a 1.06±0.117cd 2.54±0.338c 0.543±0.095a 14.9±0.605b
n.d. 0.108±0.017a 2.43±0.146a 0.944±0.152a 0.446±0.013a 0.908±0.017a 0.379±0.031a 5.22±0.314a
5.41±0.257 0.427±0.024ab 2.67±0.131a n.d. 0.481±0.023a 1.49±0.085a n.d. 37.7±2.01c
3.24±0.047b b
4.81±0.263 0.896±0.008c 6.74±0.326d 1.97±0.202b 0.835±0.058c 9.67±0.380c n.d. 37.5±2.41c
n.d., not detected
2
3.49±0.286b a
0.596±0.015 0.526±0.039b 3.77±0.097b 1.22±0.079a 0.623±0.025b 4.55±0.113b 1.08±0.053b 22.7±0.686b
n.d. 0.799±0.042c 5.74±0.323c 1.46±0.053a 0.945±0.040c 5.31±0.569b 0.581±0.036a 24.8±0.686b
2.45±0.115 n.d. 0.874±0.063a n.d. 0.760±0.075a n.d. n.d. 22.3±1.13e
a
1.16±0.107a 1.26±0.131c 2.44±0.180bc 0.934±0.091ab 1.13±0.040bc 0.621±0.054a 0.528±0.020c 9.99±0.312b
a
n.d.
7.98±0.470 1.95±0.129c 2.22±0.202b
4.80±1.03b
b
n.d.
0.915±0.066a 4.06±0.042bc
n.d. n.d. n.d.
b
c
3.51±0.227
0.061±0.006a n.d. b
7.57±0.373 n.d. 1.08±0.021 n.d. 1.52±0.060c 0.840±0.040b 0.532±0.039a 0.415±0.049a 0.657±0.034c 0.525±0.013b 0.262±0.010a 0.333±0.027a
a
a
n.d.
c
10.4±0.987b 1.84±0.065 n.d. n.d. 0.657±0.110b 0.317±0.033a 0.320±0.029a n.d. 0.523±0.031b 0.528±0.085b 0.298±0.002a n.d. a
b
4.79±0.170
a
5.05±0.173 1.11±0.057c 1.02±0.021c
b
a
5.24±0.313
ab
b
b
a
n.d.
0.410±0.046a 0.468±0.006a 0.440±0.013a 0.519±0.086a
1.78±0.083b 1.34±0.026c 3.12±0.031d 1.25±0.055b 1.21±0.021bc 0.631±0.004a 0.366±0.014b 14.9±0.106c
n.d. n.d. n.d.
Table 2. Colour parameters in wines during the bottle storage (means, standard deviations and homogenous groups for an analysis of variance with a confidence level of 99.9%). Merlot Months A420 (a.u.) A520 (a.u.)
0
3
3.03±0.027
b
4.14±0.037
d
Syrah
6
2.84±0.014ª 2.97±0.005ª
9
3.70±0.015
c
3.96±0.017
c c
A620 (a.u.)
0.801±0.009ª
0.799±0.003ª
1.12±0.008
% yellow
38.0±0.031ª
43.0±0.091c
42.1±0.032b
c
44.9±0.062
b
45.1±0.024
b
12.1±0.037
b
12.7±0.029
d
5.82±0.018
a
7.65±0.032
d
% red % blue IC (a.u.) Hue
51.9±0.039
10.0±0.021ª 7.18±0.064
b
0.732±0.001 b
WCA (a.u.)
11.1±0.919
PPC (a.u.)
1.06±0.023ª
b
0.957±0.003 5.00±0.081
a
2.27±0.063b
c
0.934±0.001 4.97±0.047
a
2.64±0.092c
12
4.02±0.007
e
3.99±0.008
c
1.21±0.004
d
3.82±0.036
d
3.71±0.025
b
1.05±0.015
b
3
2.79±0.004
a
4.55±0.012
d a
2.90±0.003
b
3.10±0.001
b
0.743±0.003
44.5±0.033e
34.7±0.076a
43.3±0.042ª
43.2±0.088ª
56.6±0.063
d
13.1±0.033
e
12.3±0.062
c
8.76±0.027
a
8.00±0.014
e
7.52±0.061
c
7.33±0.012
d
1.01±0.001
d
1.03±0.003
e
0.613±0.002
4.03±0.073ª
3.97±0.238ª
12.0±0.124
d
2.76±0.013cd
2.84±0.030d
0.953±0.042a
a
Tempranillo
6
0.704±0.003
43.6±0.009d
b
0
b
9
3.47±0.011
c
3.23±0.053
c d
12
3.55±0.041
c
3.01±0.013
b
0.861±0.001
0.814±0.005
43.0±0.009b
45.9±0.394c
48.1±0.333d
46.0±0.035
c
42.7±0.465
b
40.8±0.267
a
11.0±0.033
b
11.4±0.070
c
11.0±0.070
b
5.99±0.004
a
6.71±0.043
c
6.55±0.039
b
1.07±0.021
c
1.18±0.016
d
4.05±0.104
b
3.35±0.074
a
0.935±0.001 7.05±0.336
c
2.05±0.043b
b
2.20±0.054c
1
c
2.32±0.015cd
0
3.54±0.021
c
2.91±0.018
a
0.801±0.013
c
3
2.87±0.014
a
3.91±0.013
c a
6
3.52±0.008
b
3.31±0.006
b
51.9±0.347d
53.1±0.015e
43.1±0.045
39.2±0.020
c
37.4±0.289
b
36.6±0.003a
10.9±0.081
d
11.0±0.039
d
10.8±0.059
c
10.3±0.011b
6.83±0.014
b
7.64±0.023
d
8.02±0.052
e
7.57±0.032c
1.07±0.001
b
1.27±0.001
c
1.39±0.020
d
1.45±0.001e
3.37±0.271
ab
3.75±0.058
b
3.09±0.043
ab
2.78±0.104a
52.0±0.046
11.1±0.108
b
9.98±0.016
a
6.44±0.039
b
6.78±0.027
a
1.22±0.001
d
3.45±0.120
ab
2.41±0.023d
1.41±0.075a
3.09±0.013a
49.8±0.030c
40.1±0.037
9.16±0.340
4.48±0.019d
d
e
c
3.36±0.036
b
45.9±0.038b
38.1±0.046a
a
2.49±0.223b
d
4.66±0.040
0.872±0.003c
48.8±0.075d
0.733±0.002
3.37±0.010
b
0.967±0.009
0.839±0.009
a
4.27±0.013
12 e
0.944±0.001
0.751±0.004
b
9 c
2.59±0.015b
d
2.68±0.033b
2.65±0.025b
504
Bottle storage influences the evolution of sweet red wines
505
The anthocyanin monomers are converted into anthocyanin adducts and polymeric
506
pigments
507
The red colour of the young sweet wines evolves to browner hues during the storage
508
Flavanols decrease due to the effect of the polymerization on tannins and adducts
509
The sweet red wines retained their antioxidant activity after one year of bottling
510
27
S0308-8146(13)01368-X http://dx.doi.org/10.1016/j.foodchem.2013.09.103 FOCH 14731
To appear in:
Food Chemistry
Received Date: Revised Date: Accepted Date:
15 July 2013 10 September 2013 16 September 2013
Please cite this article as: Marquez, A., Serratosa, M.P., Merida, J., Influence of bottle storage time on colour, phenolic composition and sensory properties of sweet red wines, Food Chemistry (2013), doi: http://dx.doi.org/ 10.1016/j.foodchem.2013.09.103
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1
Influence of bottle storage time on colour, phenolic
2
composition and sensory properties of sweet red wines
3
4
Ana Marquez, Maria P. Serratosa, Julieta Merida*
5
Department of Agricultural Chemistry. Faculty of Sciences. University of Cordoba.
6
Edificio Marie Curie. Campus of Rabanales. E-14014. Cordoba, Spain.
7 8 9
* Corresponding Author. E-mail: [email protected]. Tel: +34 957 218 612; Fax: +34 957 212 146
1
10
ABSTRACT
11
Changes in colour and phenolic composition in sweet red wines made from Merlot,
12
Syrah and Tempranillo grapes were studied in order to assess the influence of bottle
13
storage over a period of 12 months. For this purpose, wine colour parameters, sensory
14
analysis and concentrations of monomeric anthocyanins, pyranoanthocyanins,
15
methylmethine-mediated condensation adducts, flavan3-ol derivatives and flavonols
16
were measured. Hue increased and red colours decreased with the storage time,
17
particularly over the first 3 months. The concentrations of low molecular weight flavan-
18
3-ol derivatives decreased with time due to the effect of their conversion into tannins of
19
high molecular weight. In addition, the glycosylated flavonols decreased through
20
hydrolysis to give the corresponding aglycones. Overall, the concentration of phenolic
21
compounds decreased markedly with storage time, whereas the antioxidant activity in
22
the wines remained constant throughout. A panel of expert tasters judged the colour,
23
aroma and flavour of all initial and final wines to be acceptable.
24 25
Keywords: Sweet red wines, Bottle storage, Phenolic compounds, Colour
2
26
1.
27
The colour of red wines is one of the most important characteristics and it is the first
28
attribute assessed by consumers during the tasting. The colour of wine reflects some of
29
its virtues and defects, its age and whether it has been subjected to a specific ageing
30
process.
Introduction
31
Phenolic compounds are the main agents responsible of the colour of wine,
32
whether white or red. Also, they influence other sensory properties such as astringency
33
and bitterness. Anthocyanins constitute the most important polyphenol family in red
34
wines and are responsible for their colour. In fact, anthocyanins are gradually extracted
35
from grape skins during the early stages of winemaking, and give musts and young
36
wines their typical bluish red colour. During the subsequent vinification and ageing
37
steps, the wine colour changes by the effect of these compounds undergoing mainly
38
copigmentation, cycloaddition, polymerization and oxidation reactions (Cheynier &
39
Ricardo da Silva, 1991; Monagas, Bartolome & Gomez-Cordoves, 2005; Alcalde-Eon,
40
Escribano-Bailon, Santos-Buelga & Rivas Gonzalo, 2006).
41
During the bottle storage time of the red wine, spontaneous clearing, colour
42
stabilisation and reactions that give the formation of more complex compounds have
43
been found (Del Alamo-Sanza & Nevares Dominguez, 2006). As the storage time in
44
bottle increases, copigmentation and polymerization anthocyanins reactions take place
45
(Eiro & Heinonen, 2002) to form more stable substances that change the initially bluish
46
red hues of young wines to orangish red hues in aged wines (Atasanova, Fulcrand,
47
Cheynier & Moutounet, 2002). Among others, anthocyanins can react with flavan-3-ol
48
derivatives, whether directly or via acetaldehyde (Vivar-Quintana, Santos-Buelga &
49
Rivas-Gonzalo, 2002). Also, anthocyanins can condense with other compounds of lower
50
molecular weight such as pyruvic acid, vinylphenol or glyoxylic acid (Marquez, 3
51
Serratosa & Merida, 2013). The resulting pigments, known as pyranoanthocyanins, are
52
more stable and keep the wine colour intensity along the time (Revilla & Gonzalez-
53
Sanjose, 2001), but these pigments have more orangish hues than the starting
54
anthocyanins. On the other hand, during these wine evolution steps the flavan-3-ol
55
derivatives undergo oxidative browning reactions and interact with proteins to form
56
substances that cause wine clouding (Cheynier & Ricardo da Silva, 1991). These
57
reactions alter the colour but also reduce astringency and bitterness, thereby leading to
58
softer tasting wines.
59
In recent years, the demand and consumption of red wines has risen at the
60
expense of white wines. Also, there has been an increasing interest in sweet wines.
61
Traditionally, sweet wines have been made from white grapes, known as ice wines,
62
noble rot wines or raisin wines (Cliff, Yuksel, Girard & King, 2002; Guarrera, Campisi
63
& Asmundo, 2005; Thibon, Dubourdieu, Darriet & Tominaga, 2009). At present,
64
however, some Spanish viticultural areas are increasingly producing sweet red wines
65
obtained by using special procedures involving on-vine or off-vine drying of the grapes,
66
fermentation and/or fortification, maceration, etc. Most of these wines are not aged, but
67
seemingly require storage for some time. Although some authors have examined
68
changes during the red wine bottle stabilisation (Perez-Magariño & Gonzalez San-Jose,
69
2004; Del Alamo Sanza & Nevares Dominguez, 2006), none studied this evolution in
70
sweet red wines.
71
In this work, changes in phenolic compounds and colour in sweet wines made
72
from red grapes grown in the Montilla–Moriles region (southern Spain) have been
73
examined during the stabilisation time in bottle. The aim was to assess the development
74
of the different reactions involving these compounds and the sensory impact of the
75
storage time.
4
76 77
2.
78
2.1. Samples
Material and methods
79
Young sweet red wines from Merlot, Syrah and Tempranillo varieties, with
80
approximately 300 g/L of reducing sugar and 15% (v/v) alcohol were used. For the
81
elaboration of the wines, the grapes were dried at controlled temperature. The raisins
82
were pressed, the musts were fortified and a maceration step with the skins was carried
83
out. The final wines were stored in black bottles and the headspace was replaced by
84
nitrogen before closing. The bottles were labeled and stored in the dark at 20 °C for 12
85
months and the wines were analyzed at 0, 3, 6, 9 and 12 months during their
86
stabilisation. All the samples were centrifuged at 3000 rpm, filtered and analysed in
87
triplicate.
88
2.2. Reagents
89
Phenolic compounds standards were obtained from Extrasynthese (Genay, France).
90
Methanol, formic acid, hydrochloric acid and acetonitrile were purchased from Merck
91
(Madrid, Spain).
92
2.3. Extraction of phenolic compounds
93
For the anthocyanin extraction, a volume of 2 mL of wine was passed through a
94
Sep-Pak C18 cartridge packed with 900 mg of material (Long Body Sep-Pak Plus,
95
Waters Corporation, Milford, MA) previously activated with methanol and HCl 0.01%.
96
The cartridge was washed with HCl 0.01% and ethyl acetate, and the anthocyanins were
97
recovered with methanol acidified to pH 2.
5
98
For the extraction of flavan-3-ol derivatives and flavonols, 15 mL of wine adjusted
99
to pH 7 were passed through Sep-Pak C18 cartridge activated with methanol and water
100
at pH 7. The cartridge was washed with water at pH 7 and the flavan-3-ol derivatives
101
were eluted with 16% acetonitrile at pH 2. Subsequently, the flavonols were recovered
102
with ethyl acetate, evaporated to dryness and dissolved in methanol.
103
The collected fractions were concentrated on a vacuum centrifuge thermostated at
104
35 ºC and passed through a filter of 0.45 µm pore size for injection into a HPLC
105
instrument.
106
2.4. HPLC analysis
107
The identification of the phenolic compounds was achieved by UV spectra obtained
108
by diode array HPLC (Spectra-Physics UV6000LP) and it was confirmed by
109
HPLC/ESI-MS analysis (TermoQuest Finnigan AQA quadrupole mass spectrometer),
110
according to the methods proposed by Marquez, Serratosa, Lopez-Toledano and Merida
111
(2012). The column used in the analyses was a 250 mm x 4.6 mm i.d., 5 µm,
112
LiChrospher 100 RP-18, using 10% aqueous formic acid (A) and acetonitrile/formic
113
acid/H2O (45:45:10) (B) as mobile phases at a flow rate of 1 mL/min. The mobile phase
114
gradient for the anthocyanin identification was as following: gradient elution from 15%
115
to 30% B in 17 min, gradient elution up to 73% B in 28 min, gradient elution up to
116
100% B in 3 min and isocratic elution for 3 min, using absorbance at 520 nm to
117
quantify. For the flavan-3-ol derivatives identification, the absorbance at 280 nm was
118
used with a gradient elution from 5% to 10% B in 25 min, gradient elution up to 20% B
119
in 10 min, gradient elution up to 30% B in 10 min, gradient elution up to 100% B in 15
120
min, and isocratic elution for 10 min. The flavonols were identified at 360 nm and this
121
fraction was obtained by gradient elution from 5% to 30% B in 5 min, gradient elution
6
122
up to 40% B in 14 min, gradient elution up to 80% B in 1 min and isocratic elution for 5
123
min.
124
2.5. Spectrophotometric measurements
125
A PerkinElmer (Waltham, MA) Lambda 25 spectrophotometer, with quartz cells of
126
1 mm light path was used. All measurements were corrected for a path length of 1 cm.
127
Samples were previously passed through Millipore (Billerica, MA) HA filters of 0.45
128
µm pore size. Absorbances at 420, 520 and 620 nm were measured and colour intensity
129
(A420+A520+A620), hue (A420/A520) and percentages of yellow, red and blue colours were
130
calculated. Polymeric pigments colour (PPC) was obtained as the absorbance at 520 nm
131
after 45 min of 5 mL of wine previously supplied with 15 mg of Na2S2O5. Wine
132
coloured anthocyanins (WCA) was determined by diluting 1 mL of sample ten times
133
with 1 M HCl and measuring the absorbance at 520 nm after 45 min at 25 ºC. The total
134
tannin index was determined by measurement of the absorbance at 550 nm in a cell of 1
135
cm light path after acid hydrolysis of the samples.
136
2.6. Antioxidant activity
137
The DPPH assay was realized for the antioxidant activity determination. The wines
138
were diluted with a solution containing 12% ethanol, and an aliquot of 200 µL was
139
placed in a cell with 3 mL of a 45 mg/L solution of DPPH in methanol. A blank (200 µL
140
dilution sample and 3 mL methanol), a control sample (200 µL of 12% ethanol in water
141
and 3 mL of DPPH solution) and a Trolox standard (200 µL of 80 mg/L Trolox solution
142
and 3 mL of DPPH solution) were also prepared in parallel. The absorbances at 517 nm
143
of the control sample and blank were measured and the absorbances of the standard and
144
samples were measured after 10 min of incubation. The antioxidant activity, expressed
7
145
in millimoles of Trolox (mmol TE) per liter, was calculated according to Serratosa,
146
Marquez, Lopez-Toledano, Medina and Merida (2011).
147
2.7. Sensory analysis
148
The initial and final sweet red wines were assessed for colour, aroma and flavour
149
acceptability by 15 tasters in a panel in accordance with ISO 8586-1:1993. The tasting
150
room was kept at 20ºC and wines served in tasting glasses certified and coded.
151
Evaluation of the quality of the wines was made using the method according to ISO
152
4121:2003, with options of desirable (5-6), acceptable (3-4) and undesirable (1-2). The
153
final punctuations were calculated as the mean, taking into account the evaluation of
154
each taster.
155
2.8. Statistical Procedures
156
The results for all samples were subjected to multifactor analysis of variance
157
(ANOVA) and discriminant analysis in triplicate, using the Statgraphics Computer
158
Package v. 5.0 from Statistical Graphics Corp (Warrenton, Virginia).
159 160
3.
Results and discussion
161
Wines undergo a large number of chemical reactions that alter their phenolic
162
composition and colour during ageing. In this work, changes in anthocyanins during the
163
storage of sweet red wines from Merlot, Syrah and Tempranillo grapes in bottle were
164
examined. Fig. 1 shows the chromatograms obtained after 0, 3, 6, 9 and 12 months of
165
storage in Merlot wine. As can be seen, the anthocyanin profile changed markedly with
166
time and the peak areas for monomeric anthocyanins decreased considerably with time.
8
167
Table 1 shows the concentration of each individual anthocyanin and those
168
homogeneous groups obtained with an analysis of variance at the 99.9% confidence
169
level. As can be seen, the concentrations of monomeric anthocyanins decreased
170
markedly with time, which is consistent with the results of many previous studies on the
171
storage of red wines in bottle (Perez-Magariño & Gonzalez San-Jose, 2004; Del Alamo
172
Sanza & Nevares Dominguez, 2006). The greatest decrease was observed during the
173
first 3 months in the wines from Merlot, Syrah and Tempranillo grapes (64, 77 and 83%
174
respectively). Most anthocyanin monomers disappeared after 3-9 months of storage. As
175
a result, the obtained wines contained malvidin-3-glucoside and malvidin-3-
176
coumaroylglucoside,
177
acetylglucoside in Merlot and Syrah wines, and petunidin-3-glucoside and peonidin-3-
178
acetylglucoside in Merlot wine. The final concentration of monomeric anthocyanins
179
was 13.6 mg/L in Merlot wine, 8.68 mg/L in Syrah wine and 3.52 mg/L in Tempranillo
180
wine. Therefore, the reduction in the anthocyanin contents was 93, 96 and 98%
181
respectively, and consistent with previous results of Garcia-Falcon, Perez-Lamela,
182
Martinez-Carballo and Simal-Gandara (2007), who reported a decrease by 72 and 85%
183
in Mencia and Brancellao wines respectively, after 12 months of storage in bottle.
in
addition
to
peonidin-3-glucoside
and
malvidin-3-
184
Glucosylated derivatives were the major monomeric anthocyanins present, with
185
63, 58 and 60% in the initial Merlot, Syrah and Tempranillo wines respectively, and 20,
186
17 and 14% respectively, after 12 months of storage (Fig. 2). The other three
187
monomeric anthocyanin families (acetylated, coumaroylated and caffeoylated
188
derivatives) also decreased or even disappeared during storage.
189
The decreases in the anthocyanin contents during storage must have resulted
190
from the gradual conversion of monomeric compounds into more stable oligomers or
191
polymers (Monagas, Gomez-Cordoves & Bartolome, 2006). As can be seen in Table 2,
9
192
polymeric pigment colour (PPC) increased gradually during storage, especially during
193
the first 3 months, when the absorbance of the Merlot, Syrah and Tempranillo wines
194
rose by 1.21, 1.10 and 1.08 a.u. respectively. This trend continued throughout the
195
storage period in the Merlot and Syrah wines, but ceased after 3 months in the
196
Tempranillo wines. The three types of wine showed no statistically significant
197
differences in this respect after 6 months.
198
A linear correlation between the variation of the content in monomeric
199
anthocyanins and PPC provided a Pearson linear correlation coefficient of -0.9504 (p
3) by the tasters. The initial wines were judged slightly different in colour and
343
flavour, but very similar in aroma. Syrah and Merlot wine were the best scored for
344
colour (4.4) and flavour (4.4) respectively, before storage. By contrast, the aged wines
345
differed markedly in colour, with the Merlot wine as the best scored (4.5) and
346
Tempranillo wine as the worst (2.8). These sensory results are consistent with the
347
analytical results. Thus, Merlot wine was that with the reddest, most stable colour, and
348
Tempranillo wine that with the brownest colour. On the other hand, Tempranillo wine
349
was judged the best in terms of aroma (4.2), well above Merlot wine (2.9) and Syrah
350
wine (3.2).
351
In summary, the storage of sweet red wines in bottle for a year caused the
352
degradation of monomeric anthocyanins and the completely disappearance of most of
353
them between 6 and 9 months. Based on the results, the decrease in the contents of these
354
compounds was a result of various gradual reactions leading to more stable oligomers or
355
polymers, and others yielding anthocyanin derivatives such as pyranoanthocyanins and
356
methylmethine-mediated condensation adducts. These changes in anthocyanin
357
compounds caused the red colour of the young wines to evolve to browner hues as
358
storage time increased. This was especially so in Tempranillo wine and, to a lesser
359
extent, in Merlot wine. By the effect of their polymerization to compounds of higher
360
molecular weights and their participation in reactions with anthocyanins to form
361
adducts, flavan-3-ol derivatives decreased during storage in bottle. Simultaneously,
362
glycosylated flavonols were hydrolysed to their corresponding aglycones. Despite their
363
changes in phenolic concentrations, the wines retained their antioxidant activity and
364
hence their beneficial effects on health. Finally, both young and aged wines were judged
365
acceptable
by
expert
tasters
on
the
16
basis
of
their
sensory
properties.
366 367
Acknowledgments
368
The authors gratefully acknowledge financial support from the Spanish Government,
369
Minister of Education (FPU scholarship of A. Marquez) for the realization of this work.
370 371
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Thibon, C., Dubourdieu, D., Darriet, P., & Tominaga, T. (2009). Impact of noble rot on
457
the aroma precursor of 3-sulfanylhexanol content in Vitis vinifera L. cv Sauvignon
458
blanc and Semillon grape juice. Food Chemisty, 114, 1359-1364.
459
Vivar-Quintana, A. M., Santos-Buelga, C., & Rivas-Gonzalo, J. C. (2002).
460
Anthocyanin-derived pigments and colour of red wines. Analytica Chimica Acta,
461
458, 147-155.
20
462
Zafrilla, P., Morillas, J., Mulero, J., Cayuela, J. M., Martinez-Cacha, A., Pardo, F., &
463
Lopez Nicolas, J. M. (2003). Changes during storage in conventional and
464
ecological wine: Phenolic content and antioxidant activity. Journal of Agricultural
465
and Food Chemisty, 51, 4694-4700.
466 467
21
3000
UV6000-520nm
UV6000-520nm
UV6000-520nm
PDA-520nm
PDA-520nm 1400
5
2500 1200
1000 2000
20
4
0 months
1
2
800
3 6 4
3 months
1000
2
1
7 8
3
12 13
9 9
11 10 12
13
14
16 19 17
21
22
23
26
mAU
mAU
1500
5
27
600
19 20
16 15
17
15
17 20 16 19
24
27
5
6 months
1
2
3
4
500
3
9 months
3
12 months
400
7
5
4
5
4
7
7
11 9 10 12 8
9 10 12
9 10 12
15
200
19 17 20
24
16
19 17 20
24
15
27
24
16
25
27 0
25
27
0
-200
468
5
10
15
20
25
30
35
40
45
Minutes
469 470
Fig. 1. Chromatographic anthocyanin profile of Merlot wine after 0, 3, 6, 9 and 12
471
months of bottle storage.
472 473 474
22
100% 80% 60% 40% 20% 0% 0
475
3
6
9
12
0
3
6
9
12
0
3
6
9
12 months
% Methylmethine-mediated condensation products % Pyranoanthocyanins %Caffeoylglucosides %Coumaroylglucosides %Acetylglucosides %Glucosides
476 477
Fig. 2. Relative contents of the anthocyanin compounds in wines during the bottle
478
storage.
479 480 481
23
Antioxidant Activity (mmol TE/L)
9 8 7 6 5 4 3 Merlot Syrah Tempranillo
2 1 0
482 483
0
3
6
9
12
Fig. 3. Antioxidant activity in wines during the bottle storage.
484 485 486
24
months
497
(a)
Function 2 (5.5%)
Merlot wines
Syrah wines
12 months
0 months 9 months
Tempranillo wines
6 months
498 499
3 months
(b) Function 2 (17.5%)
487 488 489 490 491 492 493 494 495 496
Function 1 (92.2%)
Function 1 (82.5%)
500 501
Fig. 4. Multivariate discriminant analysis. Byplot using as dependent variable the grape variety (a) and the bottle storage time (b).
502 503
26
Table 1. Anthocyanins (mg/L), flavan-3-ol derivatives (mg/L), total tannins (g/L) and flavonols (mg/L) concentrations in wines during the bottle storage (means, standard deviations and homogenous groups for an analysis of variance with a confidence level of 99.9%). Merlot Months
0
3 c
6 b
12
0
3 b
12
0
3 c
9 a
12
n.d.
n.d.
5.53±0.110
n.d.
n.d.
7.17±0.210
0.981±0.101
n.d.
n.d.
3.97±0.272b 5.56±0.221c 11.6±0.839b 29.3±0.436c n.d. n.d. n.d. 5.10±0.140c 7.00±0.185c n.d. n.d. n.d. 2.19±0.057b n.d 67.1±2.50c
2.99±0.066a 3.30±0.186b 5.43±0.095a 11.5±0.321b n.d. n.d. n.d. 4.01±0.217bc 5.56±0.235b n.d. n.d. n.d. 2.35±0.020b n.d. 37.7±1.13b
n.d. 1.69±0.136a 2.76±0.336a 5.40±0.416a n.d. n.d. n.d. 2.85±0.168ab 2.86±0.533a n.d. n.d. n.d. 1.14±0.036a n.d. 16.7±0.919a
n.d 1.20±0.068a 2.09±0.167a 4.15±0.174a n.d. n.d. n.d. 2.64±0.066a 2.23±0.271a n.d. n.d. n.d. 1.32±0.021a n.d. 13.6±0.514a
6.54±0.105b 9.42±0.146c 39.0±1.060c 77.9±2.63c 5.53±0.085 4.97±0.165 6.14±0.173b 13.3±0.379c 28.3±0.624d 5.20±0.114 4.98±0.100 7.22±0.085c 10.9±0.379 7.17±0.170 224.9±5.60d
2.56±0.061a 3.02±0.026b 7.67±0.271b 15.5±0.265b n.d. n.d. 3.34±0.090a 5.45±0.165b 6.90±0.104c n.d. n.d. 2.93±0.032b 3.30±0.332 n.d. 52.2±2.00c
2.35±0.090a 2.78±0.093b 4.15±0.212a 6.43±0.329a n.d. n.d. n.d. 3.28±0.150a 4.23±0.339b n.d. n.d. 2.39±0.145a 2.50±0.046 n.d. 30.2±0.817b
n.d. 1.38±0.147a 2.57±0.301a 3.41±0.030a n.d. n.d. n.d. n.d. 2.71±0.032a n.d. n.d. n.d. 1.13±0.020 n.d. 11.2±0.285a
n.d. n.d. 1.89±0.176a 2.83±0.146a n.d. n.d. n.d. n.d. 2.66±0.375a n.d. n.d. n.d 1.30±0.056 n.d. 8.68±0.727a
5.50±0.053c 11.7±0.153c 11.9±0.265b 75.7±2.32c 6.37±0.340 4.78±0.183 5.47±0.234 6.15±0.111 9.62±0.095c 4.27±0.021 5.92±0.178b 4.93±0.13 12.2±0.153c 8.31±0.151b 171.7±3.34d
2.21±0.017b 3.07±0.038b 2.75±0.153a 11.1±1.01b n.d. n.d. n.d. n.d. 3.57±0.279b n.d. n.d. n.d. 3.20±0.031b 2.74±0.146a 28.5±1.32c
1.18±0.066a 2.01±0.300a 2.14±0.146a 5.50±0.363a n.d. n.d. n.d n.d. 2.66±0.350ab n.d. n.d. n.d. 1.39±0.142a n.d. 15.9±1.13b
n.d. n.d. n.d. 3.16±0.251a n.d. n.d. n.d. n.d. 1.85±0.235a n.d. n.d. n.d. 1.07±0.050a n.d. 6.08±0.501a
n.d. n.d. n.d. 2.44±0.129a n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. 1.08±0.106a n.d. 3.52±0.124a
A vitisin Malvidin-3-acetylglucoside A vitisin Peonidin-3-acetylglucoside B vitisin Malvidin-3-glucoside B vitisin Peonidin-3-glucoside B vitisin Peonidin-3-acetylglucoside Pyranoanthocyanins
1.20±0.097a n.d. n.d. n.d. n.d. 1.20±0.097a
7.70±0.181c 3.94±0.155d 3.23±0.346c 2.30±0.140a 4.71±0.355c 22.6±0.827c
5.03±0.118b 2.31±0.225c 3.34±0.181b 2.25±0.032a 3.70±0.185b 16.6±0.489d
4.76±0.477b 1.95±0.046b 1.78±0.122a n.d. 3.01±0.437a 11.5±1.01b
4.46±0.123b 1.52±0.055a 1.79±0.305a n.d. 2.75±0.240a 10.5±0.667b
n.d. n.d. 0.296±0.034a n.d. 1.22±0.114a 1.52±0.146a
1.39±0.010a 0.744±0.023a 0.959±0.064b n.d. 1.74±0.093ab 4.83±0.131b
2.38±0.204b 2.23±0.078c 1.51±0.067c n.d. 2.26±0.316b 8.38±0.654c
3.05±0.144b 1.99±0.121c 1.98±0.213c 1.57±0.147a 1.69±0.160ab 10.3±0.270d
2.97±0.255b 1.60±0.095b 1.85±0.183c 1.43±0.075a 1.88±0.322ab 9.73±0.719cd
2.82±0.386a n.d. 0.846±0.072a n.d. n.d. 3.66±0.414a
3.35±0.093a n.d. 1.04±0.098a n.d. 1.33±0.121a 5.71±0.295ab
5.46±0.399b 1.57±0.125a 2.54±0.230c n.d. 3.02±0.430b 12.6±0.849d
4.02±0.478a 1.14±0.127a 1.81±0.266b n.d. 0.966±0.025a 7.94±0.782c
3.25±0.166a 1.34±0.079a 1.11±0.100a n.d. 1.23±0.136a 6.93±0.367bc
Malvidin-3-glucosidemethylmethine-(epi)catechin Malvidin-3-glucosidemethylmethine -(epi)catechin Malvidin-3-glucosidemethylmethine-(epi)catechin Peonidin-3-glucosidemethylmethine-(epi)catechin Peonidin-3-glucosidemethylmethine-(epi)catechin Malvidin-3-acetylglucosidemethylmethine-(epi)catechin Malvidin-3-acetylglucosidemethylmethine-(epi)catechin Methylmethine adducts
0.935±0.077a 4.29±0.061bc
5.08±0.242c
3.88±0.408b
4.03±0.079b
0.792±0.032a 1.78±0.151b
4.11±0.228c
3.36±0.310c
2.41±0.258b
0.663±0.034a 1.46±0.097ab 4.48±0.430d
2.62±0.185c
2.08±0.095bc
0.730±0.105a 5.34±0.106c
2.66±0.089b
3.03±0.412b
2.63±0.151b
0.788±0.042a 1.76±0.095b
1.97±0.204bc 2.27±0.105cd 2.59±0.130d
0.681±0.016a 2.21±0.163b
2.23±0.255b
2.42±0.215b
1.92±0.321b
0.433±0.040a 1.37±0.114bc
1.24±0.057b
1.76±0.188cd
1.89±0.117d
0.527±0.019a 0.672±0.032a 1.34±0.157b
1.67±0.142b 1.61±0.180b
0.653±0.077a 1.93±0.095b
1.99±0.101b
1.72±0.194b
1.59±0.140b
2.21±0.241b
1.49±0.108a
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
0.903±0.032
n.d. 2.01±0.087
n.d.
n.d. a
18.8±0.198
c
n.d. a
n.d.
n.d.
n.d.
1.89±0.135
a
0.378±0.016
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
n.d.
2.00±0.116a
6.93±0.141b
11.7±0.958c
7.73±0.410b
6.82±0.093b
0.901±0.025
b
1.51±0.074
b
0.458±0.005
0.641±0.070
n.d.
1.50±0.074
n.d.
0.395±0.032a 0.669±0.044b
n.d.
n.d.
n.d.
n.d.
12.3±1.41b
3.33±0.126a
8.29±0.192c
12.3±0.702b
10.9±0.372b 10.0±0.400b
14.6±0.506
0.928±0.080
12.2±0.259b
1
b
n.d.
a
c
b
b
0.841±0.014a 1.33±0.181a
2.50±0.078
6 b
5.23±0.010c 8.13±0.172d 23.9±0.200c 80.7±1.57d n.d. 4.61±0.113 5.60±0.110 7.95±0.444d 24.2±0.265d 4.38±0.100 4.47±0.10 5.16±0.280b 7.16±0.185c 6.23±0.030b 186±1.55d
c
2.13±0.023
9 a
4.95±0.046
n.d.
2.37±0.040
6 a
Cyanidin-3-glucoside Petunidin-3-glucoside Peonidin-3-glucoside Malvidin-3-glucoside Delphinidin-3-acetylglucoside Cyanidin-3-acetylglucoside Petunidin-3-acetylglucoside Peonidin-3-acetylglucoside Malvidin-3-acetylglucoside Cyanidin-3-coumaroylglucoside Petunidin-3-coumaroylglucoside Peonidin-3-coumaroylglucoside Malvidin-3-coumaroylglucoside Malvidin-3-caffeoylglucoside Anthocyanin monomers
n.d.
2.63±0.050
9 a
Tempranillo
Delphinidin-3-glucoside
n.d.
3.52±0.155
Syrah
(+)-Catechin (-)-Epicatechin Procyanidins B1+B3 Procyanidins B2+B4 B2-O-gallate dimer B2-O’-gallate dimer B1-O-gallate dimer Flavan-3-ol derivatives
79.4±1.34b 25.7±5.03a 14.3±1.37b 16.3±1.18c 12.2±1.05a n.d. 6.10±0.464a 154±7.03b
138.2±10.6c 35.7±0.551b 12.4±0.245b 21.6±1.25d 24.9±1.85b 10.6±1.10b 18.5±1.41d 262±12.3b
73.1±0.405ab 12.7±0.513b 9.07±0.245a 5.16±0.291b 15.5±0.924a n.d. 8.35±0.950ab 124±0.401a
75.1±5.17ab n.d. 9.26±0.644a 4.57±0.339ab 32.9±0.569c n.d. 12.1±0.341c 134±4.40ab
58.6±2.22a 4.68±0.719a 7.06±0.229a 1.84±0.305a 36.3±0.503c 3.05±0.497a 10.4±1.00bc 122±1.94a
75.0±3.69c 42.0±1.66b 7.66±0.261c 15.5±1.20b 5.22±0.882a 1.42±0.172a n.d. 150±4.07c
62.2±2.92bc 12.7±0.153a 5.02±0.600ab 6.88±0.287a 4.18±0.225a 7.14±0.374c 6.78±0.482a 105±3.73b
44.6±3.97a n.d. 5.39±0.026ab 7.57±1.20a 13.4±1.31b 3.52±0.511b n.d. 74.5±1.27a
48.0±7.05ab n.d. 5.91±0.026b 7.88±0.155a 23.2±0.945c 3.21±0.280ab n.d. 88.2±7.50ab
35.3±1.85a n.d. 4.02±0.522a 7.72±0.287a 20.2±1.67c 4.91±0.905b 18.1±0.611b 90.2±4.10ab
86.6±5.97d 43.3±0.723c 9.20±0.613b 10.0±0.455b 11.1±0.819a 4.89±0.391a 4.58±0.359a 170±4.13d
69.7±0.615cd 20.8±0.557b 6.21±0.223a 8.71±0.761ab 12.9±0.889ab 7.72±0.748a 6.51±0.756a 133±2.73c
46.7±7.54b 4.85±0.758a 4.73±0.835a 7.43±0.519a 17.0±1.38b n.d. 15.2±1.10b 95.9±6.30b
51.0±8.28bc n.d. 4.24±0.354a 7.70±0.171a 24.6±1.66c n.d. 14.8±0.252b 102±9.81b
5.74±0.424a n.d. 5.38±0.745a 8.70±0.625ab 25.3±0.777c 4.95±0.646b 17.1±1.10b 67.2±2.49a
Total tannins (g/L)
8.81±0.208ª
13.2±0.196c
12.9±0.302c
11.5±0.184b
8.70±0.200a
6.35±0.093a
8.90±0.040b
13.3±0.519d
12.0±0.237c
11.2±0.375c
9.93±0.028b
9.18±0.077a
13.3±0.169c
10.1±0.018b
10.0±0.117b
Myricetin-3-glucoside Quercetin-3-glucuronide + quercetin-3-galactoside Quercetin-3-glucoside Laricitrin-3-glucoside Kaempferol-3-glucoside Isorhamnetin-3-glucoside + kaempferol-3-glucuronide Syringetin-3-glucoside Myricetin Quercetin Laricitrin Kaempferol Isorhamnetin Syringetin Flavonols
0.295±0.010b 0.511±0.041c 0.125±0.007a 0.599±0.012c n.d.
n.d.
0.315±0.038b n.d.
4.77±0.198
n.d.
n.d.
n.d.
5.98±0.212
b
5.24±0.352
ab
4.33±0.191
2.45±0.123 0.455±0.073 1.14±0.036 1.03±0.099b 0.273±0.003a n.d. 0.798±0.046a n.d. n.d.
4.66±0.186b
0.492±0.041a 0.360±0.021a 0.256±0.038a n.d.
11.8±0.850c
0.756±0.023a
1.39±0.055c
0.660±0.006b 0.211±0.015a n.d.
n.d.
n.d. 0.376±0.012a 2.87±0.050a 1.04±0.085a 0.471±0.016a 2.51±0.026a 0.502±0.072a 8.53±0.139a
c
2.34±0.044c 0.906±0.049b 2.75±0.192cd 1.20±0.106b 1.27±0.081c 0.971±0.069b n.d. 17.9±0.827d
n.d. 0.151±0.008a 2.15±0.038b 0.838±0.046a 1.01±0.001b 0.484±0.020a 0.243±0.009a 4.88±0.054a
3.91±0.153 0.249±0.019b 1.92±0.089a n.d. 0.520±0.029ab 0.514±0.018a n.d. 24.0 ±0.797c
3.83±0.284 0.604±0.027d 5.55±0.337c 1.88±0.068b 1.11±0.101d 3.87±0.192d n.d. 29.4±1.96d
2.46±0.061 0.261±0.024b 3.55±0.107b 1.02±0.023a 0.799±0.073bc 1.86±0.052b 0.429±0.023a 13.5±0.336b
2.61±0.035 0.496±0.027c 4.87±0.117c 1.29±0.050a 1.06±0.117cd 2.54±0.338c 0.543±0.095a 14.9±0.605b
n.d. 0.108±0.017a 2.43±0.146a 0.944±0.152a 0.446±0.013a 0.908±0.017a 0.379±0.031a 5.22±0.314a
5.41±0.257 0.427±0.024ab 2.67±0.131a n.d. 0.481±0.023a 1.49±0.085a n.d. 37.7±2.01c
3.24±0.047b b
4.81±0.263 0.896±0.008c 6.74±0.326d 1.97±0.202b 0.835±0.058c 9.67±0.380c n.d. 37.5±2.41c
n.d., not detected
2
3.49±0.286b a
0.596±0.015 0.526±0.039b 3.77±0.097b 1.22±0.079a 0.623±0.025b 4.55±0.113b 1.08±0.053b 22.7±0.686b
n.d. 0.799±0.042c 5.74±0.323c 1.46±0.053a 0.945±0.040c 5.31±0.569b 0.581±0.036a 24.8±0.686b
2.45±0.115 n.d. 0.874±0.063a n.d. 0.760±0.075a n.d. n.d. 22.3±1.13e
a
1.16±0.107a 1.26±0.131c 2.44±0.180bc 0.934±0.091ab 1.13±0.040bc 0.621±0.054a 0.528±0.020c 9.99±0.312b
a
n.d.
7.98±0.470 1.95±0.129c 2.22±0.202b
4.80±1.03b
b
n.d.
0.915±0.066a 4.06±0.042bc
n.d. n.d. n.d.
b
c
3.51±0.227
0.061±0.006a n.d. b
7.57±0.373 n.d. 1.08±0.021 n.d. 1.52±0.060c 0.840±0.040b 0.532±0.039a 0.415±0.049a 0.657±0.034c 0.525±0.013b 0.262±0.010a 0.333±0.027a
a
a
n.d.
c
10.4±0.987b 1.84±0.065 n.d. n.d. 0.657±0.110b 0.317±0.033a 0.320±0.029a n.d. 0.523±0.031b 0.528±0.085b 0.298±0.002a n.d. a
b
4.79±0.170
a
5.05±0.173 1.11±0.057c 1.02±0.021c
b
a
5.24±0.313
ab
b
b
a
n.d.
0.410±0.046a 0.468±0.006a 0.440±0.013a 0.519±0.086a
1.78±0.083b 1.34±0.026c 3.12±0.031d 1.25±0.055b 1.21±0.021bc 0.631±0.004a 0.366±0.014b 14.9±0.106c
n.d. n.d. n.d.
Table 2. Colour parameters in wines during the bottle storage (means, standard deviations and homogenous groups for an analysis of variance with a confidence level of 99.9%). Merlot Months A420 (a.u.) A520 (a.u.)
0
3
3.03±0.027
b
4.14±0.037
d
Syrah
6
2.84±0.014ª 2.97±0.005ª
9
3.70±0.015
c
3.96±0.017
c c
A620 (a.u.)
0.801±0.009ª
0.799±0.003ª
1.12±0.008
% yellow
38.0±0.031ª
43.0±0.091c
42.1±0.032b
c
44.9±0.062
b
45.1±0.024
b
12.1±0.037
b
12.7±0.029
d
5.82±0.018
a
7.65±0.032
d
% red % blue IC (a.u.) Hue
51.9±0.039
10.0±0.021ª 7.18±0.064
b
0.732±0.001 b
WCA (a.u.)
11.1±0.919
PPC (a.u.)
1.06±0.023ª
b
0.957±0.003 5.00±0.081
a
2.27±0.063b
c
0.934±0.001 4.97±0.047
a
2.64±0.092c
12
4.02±0.007
e
3.99±0.008
c
1.21±0.004
d
3.82±0.036
d
3.71±0.025
b
1.05±0.015
b
3
2.79±0.004
a
4.55±0.012
d a
2.90±0.003
b
3.10±0.001
b
0.743±0.003
44.5±0.033e
34.7±0.076a
43.3±0.042ª
43.2±0.088ª
56.6±0.063
d
13.1±0.033
e
12.3±0.062
c
8.76±0.027
a
8.00±0.014
e
7.52±0.061
c
7.33±0.012
d
1.01±0.001
d
1.03±0.003
e
0.613±0.002
4.03±0.073ª
3.97±0.238ª
12.0±0.124
d
2.76±0.013cd
2.84±0.030d
0.953±0.042a
a
Tempranillo
6
0.704±0.003
43.6±0.009d
b
0
b
9
3.47±0.011
c
3.23±0.053
c d
12
3.55±0.041
c
3.01±0.013
b
0.861±0.001
0.814±0.005
43.0±0.009b
45.9±0.394c
48.1±0.333d
46.0±0.035
c
42.7±0.465
b
40.8±0.267
a
11.0±0.033
b
11.4±0.070
c
11.0±0.070
b
5.99±0.004
a
6.71±0.043
c
6.55±0.039
b
1.07±0.021
c
1.18±0.016
d
4.05±0.104
b
3.35±0.074
a
0.935±0.001 7.05±0.336
c
2.05±0.043b
b
2.20±0.054c
1
c
2.32±0.015cd
0
3.54±0.021
c
2.91±0.018
a
0.801±0.013
c
3
2.87±0.014
a
3.91±0.013
c a
6
3.52±0.008
b
3.31±0.006
b
51.9±0.347d
53.1±0.015e
43.1±0.045
39.2±0.020
c
37.4±0.289
b
36.6±0.003a
10.9±0.081
d
11.0±0.039
d
10.8±0.059
c
10.3±0.011b
6.83±0.014
b
7.64±0.023
d
8.02±0.052
e
7.57±0.032c
1.07±0.001
b
1.27±0.001
c
1.39±0.020
d
1.45±0.001e
3.37±0.271
ab
3.75±0.058
b
3.09±0.043
ab
2.78±0.104a
52.0±0.046
11.1±0.108
b
9.98±0.016
a
6.44±0.039
b
6.78±0.027
a
1.22±0.001
d
3.45±0.120
ab
2.41±0.023d
1.41±0.075a
3.09±0.013a
49.8±0.030c
40.1±0.037
9.16±0.340
4.48±0.019d
d
e
c
3.36±0.036
b
45.9±0.038b
38.1±0.046a
a
2.49±0.223b
d
4.66±0.040
0.872±0.003c
48.8±0.075d
0.733±0.002
3.37±0.010
b
0.967±0.009
0.839±0.009
a
4.27±0.013
12 e
0.944±0.001
0.751±0.004
b
9 c
2.59±0.015b
d
2.68±0.033b
2.65±0.025b
504
Bottle storage influences the evolution of sweet red wines
505
The anthocyanin monomers are converted into anthocyanin adducts and polymeric
506
pigments
507
The red colour of the young sweet wines evolves to browner hues during the storage
508
Flavanols decrease due to the effect of the polymerization on tannins and adducts
509
The sweet red wines retained their antioxidant activity after one year of bottling
510
27
