Food Chemistry 211 (2016) 1–7

Contents lists available at ScienceDirect

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

Study of quinones reactions with wine nucleophiles by cyclic

voltammetry
Carla M. Oliveira a,b, António S. Barros a, António C.S. Ferreira b,c, Artur M.S. Silva a,⇑
a
Departamento de Química & QOPNA, Universidade de Aveiro, 3810-193 Aveiro, Portugal
b
Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Rua Arquiteto Lobão Vital, Apartado 2511, 4202-401 Porto, Portugal
c
Stellenbosch University, Private Bag X1, Matieland, 7602 Stellenbosch, South Africa

a r t i c l e i n f o a b s t r a c t

Article history: Quinones are electrophilic species which can react with various nucleophiles, like wine antioxidants,
Received 12 December 2015 such as sulfur dioxide or ascorbic acid, thiols, amino acids, and numerous polyphenols. These reactions
Received in revised form 3 May 2016 are very important in wine aging because they mediate oxygen reactions during both production and bot-
Accepted 4 May 2016
tle aging phases. In this work, the major challenge was to determine the interaction between ortho-
Available online 4 May 2016
quinones and wine nucleophiles (amino acids, thiols, and the antioxidants SO2 and ascorbic acid), by cyc-
lic voltammetry. Wine-model solutions with gallic acid, caffeic acid, or (+)-catechin and nucleophilic
Chemical compounds studied in this article:
compounds were used. To understand the effect of nucleophilic addition in wine, a white wine with
Gallic acid (PubChem CID: 370)
Caffeic acid (PubChem CID: 689043)
the same added nucleophiles was also analysed. Cyclic voltammograms were taken with glassy carbon
(+)-Catechin (PubChem CID: 9064) electrode or screen-printed carbon electrodes, respectively, for wine-model and white wines solutions,
3-Sulfanylhexan-1-ol (PubChem CID: in the absence and in the presence of nucleophiles. A nucleophilic order profile related to the cathodic
521348) current intensity decrease was observed.
4-Methyl-4-sulfanylpentan-2-one Ó 2016 Elsevier Ltd. All rights reserved.
(PubChem CID: 88290)
Furan-2-ylmethanethiol (PubChem CID:
7363)
Ascorbic acid (PubChem CID: 54670067)
Sodium metabisulfite (PubChem CID:
656671)
L-Phenylalanine (PubChem CID: 6140)
L-Methionine (PubChem CID: 6137)

Keywords:
Oxidation
Quinones
Thiols
Amino acids
Antioxidants
Wine
Cyclic voltammetry
Phenolics

1. Introduction

The oxidative processes of wine begin by the oxidation of

Abbreviations: 3SH, 3-sulfanylhexan-1-ol; 4MSP, 4-methyl-4-sulfanylpentan-2-
one; 2FMT, furan-2-ylmethanethiol; AA, ascorbic acid; Ep,a, anodic peak potential;
Ep,c, cathodic peak potential; Ip,a, anodic peak current; Ip,c, cathodic peak current;
SO2, sulfur dioxide.
⇑ Corresponding author at: Department of Chemistry, University of Aveiro, 3810-
193 Aveiro, Portugal.
E-mail address: artur.silva@ua.pt (A.M.S. Silva).
polyphenols containing a catechol or a galloyl group such as (+)-c
atechin/( )-epicatechin, gallocatechin, gallic acid and its esters,
and caffeic acid, which are the most readily oxidized wine
constituents. These substrates are sequentially oxidized to semi-
quinone radicals and quinones while oxygen is reduced to hydro-
gen peroxide (Oliveira, Silva Ferreira, De Freitas, & Silva, 2011).

http://dx.doi.org/10.1016/j.foodchem.2016.05.020
0308-8146/Ó 2016 Elsevier Ltd. All rights reserved.
2 C.M. Oliveira et al. / Food Chemistry 211 (2016) 1–7
Quinones are therefore key reactive electrophilic oxidation inter-
mediates in wine and can undertake many reactions with wine
nucleophiles like sulfur dioxide, ascorbic acid, thiols, amino acids
and others, including the disappearance of undesirable aromas
(Nikolantonaki & Waterhouse, 2012), or the loss of varietal aromas
(Makhotkina, 2011; Waterhouse & Nikolantonaki, 2015) and the
formation of off-flavors (Bittner, 2006). These reactions are very
important in wine aging because they mediate oxygen consump-
tion during both production and bottle aging phases.
Cyclic voltammetry allows the study of both the type and amount
of substances possessing antioxidant activity, given by the poten-
tials at which oxidation occurs, and the current intensity, respec-
tively. This technique has been used for the characterization of
antioxidant properties of wine and wine phenolics (Kilmartin, Zou,
& Waterhouse, 2001; Makhotkina & Kilmartin, 2009, 2012;
Rodrigues, Silva Ferreira, Guedes de Pinho, Bento, & Geraldo, 2007;
Roginsky et al., 2006). Likewise, cyclic voltammetry was also used
to evaluate the ‘‘degradation status” of wines based on the effect
of oxygen regimes and oxidizable species (Martins et al., 2008).
The influence of sulfur dioxide, glutathione and ascorbic acid on
the cyclic voltammograms of wine polyphenols and wines has
already been investigated using a glassy carbon electrode
(Makhotkina & Kilmartin, 2009). The results showed that sulfur
dioxide increased the anodic current and decreased the cathodic
current, pointing to a rapid interaction of SO2 with the oxidized
polyphenol quinones. However, in this study, ascorbic acid pro-
duced no additional effect on the cyclic voltammograms of wine
polyphenols and wines (Makhotkina & Kilmartin, 2009).
Oxidation of wine polyphenols is thought to play a major role in
the degradation of wine thiols. Volatile thiols such as 4-methyl-4-
sulfanylpentan-2-one (4MSP) and 3-sulfanylhexan-1-ol (3SH) are
aromatic molecules having an important organoleptic impact on
white wines with respectively boxwood/broom or grapefruit/pas-
sion fruit notes. These molecules decrease dramatically with wine
aging (Blanchard, Darriet, & Dubourdieu, 2004; Brajkovich et al.,
2005; Lopes et al., 2009; Makhotkina, 2011). On the another hand,
other thiols with undesirable aromas, like 3-(methylsulfanyl)
propan-1-ol (methionol) or 2-sulfanylethanol (2-
mercaptoethanol) also decrease with wine oxidation. Port wine
studies have concluded that old Port wine (barrel aged) never
develops ‘‘off-flavors” associated with the presence of methionol
(cauliflower) or 2-mercaptoethanol (rubber/burnt). In fact, temper-
ature and oxygen are the major two factors in the consumption of
these molecules (Silva Ferreira, Hogg, & Guedes de Pinho, 2003;
Silva Ferreira, Rodrigues, Hogg, & Guedes de Pinho, 2003).
Sulfur dioxide can convert ortho-quinones back to ortho-
dihydroxyphenols (catechols) and react directly with ortho-
quinones to form sulfonic acids. Interestingly, the potential of
ascorbic acid to recycle ortho-quinones back to catechols has also
been suggested by various authors (Danilewicz, 2003; Isaacs &
van Eldik, 1997). Furthermore, wine preservatives like sulfur diox-
ide, ascorbic acid, glutathione or phloroglucinol are all highly reac-
tive with the quinone, 4-methyl-1,2-benzoquinone (Q4MeC), in
wine model-solutions, but showed only additive antioxidant
effects (Nikolantonaki, Magiatis, & Waterhouse, 2014). A synergic
antioxidant effect between these wine preservatives, in wine like
acidic conditions, was not observed (Nikolantonaki et al., 2014).
ortho-Quinones formed during the oxidation process of wine
phenolic compounds can react with amino acids (Rizzi, 2006,
2008). In this reaction (Rizzi, 2006, 2008) an initial Michael addi-
tion of a a-amino acid to an ortho-quinone to form a a-
aminobenzoquinone is proposed. Nevertheless, the formation of
the a-aminobenzoquinone intermediate was not an essential step
in the aldehyde formation. The ortho-quinone can react with a
molecule of a a-amino acid to form an amine/carbonyl condensa-
tion product that can decarboxylate to form an intermediate that
will hydrolyze in a Strecker aldehyde and a transamination
product. Furthermore, under low pH model wine conditions,
the reaction products of a-amino acids with the 4-methyl-1,
2-benzoquinone (Q4MeC) was studied (Nikolantonaki &
Waterhouse, 2012). In this work, the 1,4-conjugate addition
(Michael addition) of one molecule of a-amino acid to the ortho-
quinone giving a 4-amino catechol intermediate was not observed.
Thus, the authors stated that under low pH model wine conditions
the 1,4-conjugate addition of one molecule of a-amino acid to the
ortho-quinone could not be observed, and suggested a Fenton-type
oxidation of the related alcohols as the source of the aldehydes,
affecting wine aroma (Loscos, Hernandez-Orte, Cacho, & Ferreira,
2010; Silva Ferreira, Guedes de Pinho, Rodrigues, & Hogg, 2002;
Wildenradt & Singleton, 1974). Nevertheless, under wine pH
conditions, this reaction has to be confirmed.
In this work, the challenge was to determine the interaction
between ortho-quinones and wine nucleophiles, like the amino
acids phenylalanine and methionine, the thiols 3-sulfanylhexan-
1-ol (3SH), 4-methyl-4-sulfanylpentan-2-one (4MSP), and furan-
2-ylmethanethiol (2FMT), and the antioxidants sulfur dioxide and
ascorbic acid, by cyclic voltammetry. The use of phenylalanine
and methionine is due to their important wine formation of potent
Strecker aldehydes. It was observed that wines stored at high
temperatures and supplemented with high levels of dissolved oxy-
gen suffered a rapid and pronounced oxidative spoilage aroma,
which were related with the presence of 3-(methylthio)
propionaldehyde (methional), responsible for ‘‘boiled-potato”
odour notes, and phenylacetaldehyde, with ‘‘honey-like” odour
notes (Silva Ferreira et al., 2002; Silva Ferreira, Hogg, et al., 2003;
Silva Ferreira, Rodrigues, et al., 2003). The use of 3-
sulfanylhexan-1-ol (3SH) and 4-methyl-4-sulfanylpentan-2-one
(4MSP) was due to their important contribution to wine varietal
aroma. Conversely, the use of furan-2-ylmethanethiol (2FMT) is
due to its negative wine aroma impact with an odoriferous smel-
ling of cooked meat. Sulfur dioxide and ascorbic acid were used
due to their well-known wine antioxidant activity. Experiments
were conducted in wine-model solutions supplemented with gallic
acid, caffeic acid, or (+)-catechin, and in a white wine.
2. Materials and methods
2.1. Chemicals
The following molecules were purchased to Sigma-Aldrich, (Por-
tugal): gallic acid (G7384) (100%); caffeic acid (C0625) (98%); (+)-cat
echin hydrate (C1251) (98%); 3-sulfanylhexan-1-ol (CDS001520)
(100%); 4-methyl-4-sulfanylpentan-2-one (CDS009189) (100%);
furan-2-ylmethanethiol (W249300) (97%); ascorbic acid (A0278)
(100%); and sodium metabisulfite (08982) (98%).
2.2. Model-wine solutions
Wine-model systems were composed by ethanol 12% (v/v) and
0.033 M of tartaric acid, with a pH of 3.6. Gallic acid, caffeic acid, or
(+)-catechin were added at a concentration of 0.6 mM. The nucle-
ophilic compounds [phenylalanine (Phe), methionine (Met), 3-
sulfanylhexan-1-ol (3SH), 4-methyl-4-sulfanylpentan-2-one
(4MSP), furan-2-ylmethanethiol (2FMT), sulfur dioxide (SO2), and
ascorbic acid (AA)] were added at a concentration of 2.4 mM.
2.3. Wine
A commercial white wine (‘‘Casal da Eira”) from Estremadura
region of Portugal (11.5% Vol.) was used for the evaluation of
the interaction between wine ortho-quinones and the added
 C.M. Oliveira et al. / Food Chemistry 211 (2016) 1–7
nucleophiles. The nucleophilic compounds (Phe, Met, 3SH, 4MSP,
2FMT, SO2, and AA) were added to the white wine in a concentra-
tion of 2.4 mM.
2.4. Cyclic voltammetry
2.4.1. Model-wine solutions
Experiments were performed using a potentiostat (microAuto-
lab Type III with an Autolab Faraday Cage) and voltammograms
were obtained with a scan rate of 100 mV/s with an increment
potential of 2.4 mV, between 0.5 V and 1.2 V. The working elec-
trode was a 3 mm glassy carbon disk in combination with a
Metrohm tipholder, cleaned by polishing with 3 lm alumina pow-
der during 30 s follow by fixing the potential to 1.5 V in a 0.1 mol/L
NaOH solution during 5 s, between acquisitions. A saturated calo-
mel electrode was used as a reference electrode in conjunction
with a platinum counter electrode. Each acquisition required
15 mL of sample. Oxygen was removed with a N2 current flow dur-
ing 5 min prior to analysis. Cyclic voltammetry experiments were
controlled by the GPES 4.9 software provided by Ecochemie. Cyclic
voltammograms were taken in the absence and in the presence of
nucleophilic compounds, and the study was made in triplicate.
2.4.2. Wine solutions
Experiments were performed using a potentiostat (microAuto-
lab Type III with an Autolab Faraday Cage) and voltammograms
were obtained with a scan rate of 100 mV/s with an increment
potential of 2.4 mV, between 0.5 V and 1.2 V. Screen-printed car-
bon electrodes were used from DropSens (110-CNT). The working
electrode was a 4 mm carbon with a silver reference electrode in
conjunction with a carbon counter electrode (ceramic substrate:
L33  W10  H0.5 mm). These electrodes exhibit a high electro-
chemical activity and are useful for complex matrices. Each acqui-
sition required 20 lL of sample. Cyclic voltammetry experiments
were controlled by the GPES 4.9 software provided by Ecochemie.
Cyclic voltammograms were taken in the absence as well as in the
presence of nucleophilic compounds, and the study was made in
triplicate.
2.5. Statistical analysis
Paired t-test using Excel software from Windows 10 was
applied to the experimental data; the results were considered to
be significant if the associated p-value was < 0.05.
3. Results and discussion
Cyclic voltammetry is a method to produce polyphenol qui-
nones in a controlled manner (oxidation curves; anodic curves)
(Fig. 1). If a rapid interaction of nucleophiles with polyphenol qui-
nones occurs, there will be less available for its reduction at the
carbon electrode on the reverse potential sweep (reduction curves;
cathodic curves) (Fig. 1). Conversely, if no interaction of nucle-
ophiles with polyphenol quinones occurs, the cathodic peaks will
be retained on the reverse potential sweep.
During the cyclic voltammetry experiments of wine-model
solutions, the anodic and cathodic peak potentials at pH 3.6, of
the studied polyphenols [Ep,a – anodic peak potential (V); Ep,c –
cathodic peak potential (V)], as well the anodic and cathodic peak
currents at pH 3.6 [Ip,a – anodic peak current (A); Ip,c – cathodic
peak current (A)] were evaluated (Table 1). Also, during the cyclic
voltammetry experiments of wine solutions, the anodic and catho-
dic peak potentials as well the anodic and cathodic peak currents
were also evaluated (Table 2).
3
Figs. 2–4 depicts the cyclic voltammograms of the amino acids
phenylalanine and methionine, the thiols 3SH, 4MSP, and 2FMT,
and the antioxidants sulfur dioxide and ascorbic acid, in wine-
model solutions of three representative phenolic compounds
(gallic acid, caffeic acid, and (+)-catechin). Fig. 5 shows cyclic
voltammograms of the former nucleophiles added to a white wine,
where the voltammograms are a combination effect of several
compounds with slightly different formal potentials.
From Tables 1 and 2 one can see that the decreases on the
cathodic peaks intensities were common to all the tested nucle-
ophiles, suggesting that an interaction of polyphenol quinones
with the studied nucleophiles did occur.
3.1. Amino acids
Concerning amino acids, phenylalanine and methionine had no
significant influence on the anodic and cathodic peak current
intensities of gallic and caffeic acids (p-value > 0.05),
Fig. 2a and b; and Table 1. On another hand, phenylalanine and
methionine decreased the current intensities of the cathodic peak
of (+)-catechin by 19 and 29%, respectively (p-values < 0.005);
Fig. 2c and Table 1, as well the two anodic peaks by 21–30%,
respectively (p-values < 0.01).
Regarding amino acids additions to the white wine, phenylala-
nine and methionine decreased the anodic current intensities by
around 20 and 10% for Ep1,a at 0.274 (V) and the Ep2,a at 0.701
(V), respectively (p-values < 0.001), Fig. 5 and Table 2. On another
hand, phenylalanine seemed to have no significant influence
on the cathodic peak current intensity of the wine solution
(p-value > 0.05), but a significant decrease (13%) was observed
for methionine (p-value < 0.001), Fig. 5 and Table 2.
These results point to an interaction of the above amino acids
with the (+)-catechin quinone and to wine quinones, and moreso-
ever with methionine.
3.2. Thiols
Concerning thiols, 3-sulfanylhexan-1-ol (3SH) and 4-methyl-4-
sulfanylpentan-2-one (4MSP) decreased the cathodic current
intensities of all the analysed polyphenols. Furthermore, the
decrease rate follows, respectively for 3SH and 4SMP, the order:
(+)-catechin (in 33–44%; p-values < 0.001) > caffeic acid (in 18–
25%; p-values < 0.05) > gallic acid (in 5–7%; p-values < 0.05),
Fig. 3 and Table 1. These results indicate that 4MSP is more reactive
with quinones than 3SH. Considering furan-2-ylmethanethiol
(2FMT), the decrease of the cathodic current intensities of all the
analysed polyphenols was greater than with the other thiols (14%
for gallic acid, 52% for caffeic acid, and 48% for (+)-catechin; p-
values < 0.001). Moreover, 2FMT decreased the current intensities
of the two anodic peaks of gallic acid, in 25% for Ep1,a at 0.354
(V) (p-value < 0.01), and in 5% for Ep2,a at 0.735 (V), although these
were not significant (Fig. 3a and Table 1). For caffeic acid, 2FMT
decreased the current intensity of the anodic peak by 43% (p-
value < 0.001), Fig. 3b and Table 1, and likewise this thiol has
decreased the current intensities of the two anodic peaks of (+)-
catechin, by 41% for Ep1,a at 0.354 (V) (p-value < 0.005); and in
25% for Ep2,a at 0.747 (V) (p-value < 0.01), Fig. 3c and Table 1.
These results suggest a fast interaction of 2FMT with the caffeic
acid and (+)-catechin quinones.
Regarding thiols additions to the white wine, 3-sulfanylhexan-
1-ol (3SH), 4-methyl-4-sulfanylpentan-2-one (4MSP), and furan-
2-ylmethanethiol (2FMT) decreased the cathodic current intensi-
ties of the wine solutions, although not significant in the case of
3SH (Fig. 5 and Table 2). As for 4SMP and 2FMT the decrease on
the cathodic current intensities of the wine solutions was around
25% (p-values < 0.001). Moreover, 3SH and 2FMT have decreased
4 C.M. Oliveira et al. / Food Chemistry 211 (2016) 1–7

3.5E-05
3.0E-05
Oxidation curve -
2.5E-05
anodic current intensity
2.0E-05

Intensity
Current

I/ A
1.5E-05
1.0E-05
5.0E-06 Reduction curve -
0.0E+00 cathodic current intensity

-5.0E-06
0 0.4 0.8 1.2
Applied Potential Difference
E/ V

Fig. 1. Cyclic voltammogram.

Table 1
Evaluation of the anodic and cathodic peak potentials at pH 3.6, of the studied
polyphenols [Ep,a – anodic peak potential (V); Ep,c – cathodic peak potential (V)], as
well the anodic and cathodic peak currents at pH 3.6 [Ip,a – anodic peak current (A);
Ip,c – cathodic peak current (A)] during the cyclic voltammetry experiment.
Table 2
Evaluation of the anodic and cathodic peak potentials of the studied wine solutions
[Ep,a – anodic peak potential (V); Ep,c – cathodic peak potential (V)], as well the
anodic and cathodic peak currents [Ip,a – anodic peak current (A); Ip,c – cathodic peak
current (A)] during the cyclic voltammetry experiment.

Phenolic compound pH = 3.6 Wine solutions White wine

Ep1,a Ep2,a Ep1,c Ep1,a Ep2,a Ep1,c
Gallic acid 0.354 0.735 0.273 0.274 0.701 0.225
Caffeic acid 0.374 0.327 6 6 6
Ip1,a (10 ) Ip2,a (10 ) Ip1,c (10 )
Catechin 0.354 0.747 0.283
6 6 6 Wine 34.4 ± 0.30 41.0 ± 0.05 14.9 ± 0.30
Ip1,a (10 ) Ip2,a (10 ) Ip1,c (10 )
Wine + Phe 28.0 ± 0.48 36.0 ± 0.27 13.7 ± 0.69
Gallic acid 8.08 ± 0.50 6.14 ± 0.07 1.00 ± 0.04 Wine + Met 26.6 ± 0.83 36.2 ± 0.43 13.0 ± 0.42
Gallic + Phe 8.17 ± 0.60 6.10 ± 0.09 1.00 ± 0.02 Wine + 3SH 25.2 ± 1.57 36.9 ± 0.73 13.7 ± 0.95
Gallic + Met 8.12 ± 0.22 6.02 ± 0.03 0.95 ± 0.03 Wine + 4MSP 34.6 ± 0.15 41.3 ± 0.53 11.2 ± 0.35
Gallic + 3SH 7.01 ± 0.10 6.25 ± 0.15 0.95 ± 0.01 Wine + 2FMT 25.3 ± 1.47 37.0 ± 0.78 11.4 ± 0.32
Gallic + 4MSP 7.59 ± 0.37 6.65 ± 0.12 0.93 ± 0.01 Wine + SO2 35.7 ± 1.23 44.1 ± 0.13 10.8 ± 0.87
Gallic + 2FMT 6.03 ± 0.29 5.85 ± 0.23 0.86 ± 0.02 Wine + AA 39.3 ± 2.34 43.3 ± 1.82 8.20 ± 0.23
Gallic + SO2 9.70 ± 0.43 8.02 ± 0.15 0.82 ± 0.01
Ep1,a – first anodic peak potential; Ep2,a – second anodic peak potential; Ep1,c – first
Gallic + AA 12.95 ± 0.03 9.66 ± 0.03 0.58 ± 0.03
cathodic peak potential; Ip1,a – first anodic peak current; Ip2,a – second anodic peak
Caffeic acid 9.20 ± 0.67 5.38 ± 0.59
current; Ip1,c – first cathodic peak current; (±SD) and n = 3.
Caffeic + Phe 8.79 ± 1.02 5.23 ± 0.68
Caffeic + Met 9.26 ± 0.38 5.44 ± 0.35
Caffeic + 3SH 8.38 ± 0.10 4.40 ± 0.04
Caffeic + 4MSP 10.61 ± 0.51 4.02 ± 0.33 polyphenols, pointing to a rapid interaction of this antioxidant
Caffeic + 2FMT 5.24 ± 0.33 2.57 ± 0.22 with the oxidized polyphenol quinones. The increases of the anodic
Caffeic + SO2 7.73 ± 0.15 1.15 ± 0.02 peaks intensity were respectively of 60% for Ep1,a at 0.354 (V) and
Caffeic + AA 13.64 ± 0.34 2.82 ± 0.19
57% for Ep2,a at 0.735 (V) (p-values < 0.001) for gallic acid; 48% (p-
Catechin 5.08 ± 0.46 6.39 ± 0.46 3.36 ± 0.24
Catechin + Phe 4.03 ± 0.18 4.97 ± 0.14 2.72 ± 0.11 value < 0.001) for caffeic acid; and 51% for Ep1,a at 0.354 (V) (p-
Catechin + Met 3.55 ± 0.13 4.84 ± 0.14 2.39 ± 0.09 value < 0.001) and 16% for Ep2,a at 0.747 (V) (p-value < 0.01) for
Catechin + 3SH 3.80 ± 0.29 5.51 ± 0.18 2.26 ± 0.16 (+)-catechin, Fig. 4 and Table 1. In contrast, this antioxidant
Catechin + 4MSP 4.29 ± 0.28 5.98 ± 0.01 1.89 ± 0.11
decreased the current intensities of the cathodic peaks of gallic
Catechin + 2FMT 2.98 ± 0.41 4.82 ± 0.55 1.74 ± 0.31
Catechin + SO2 3.97 ± 0.41 6.83 ± 0.38 1.71 ± 0.18
acid, caffeic acid, and (+)-catechin, by respectively, 42, 48, and
Catechin + AA 7.64 ± 0.23 7.44 ± 0.21 1.59 ± 0.03 53% (p-values < 0.001), Fig. 4 and Table 1, pointing to a rapid inter-
Ep1,a – first anodic peak potential; Ep2,a – second anodic peak potential; Ep1,c – first
action of ascorbic acid with the oxidized polyphenol quinones. This
cathodic peak potential; Ip1,a – first anodic peak current; Ip2,a – second anodic peak antioxidant begins to oxidize at the carbon electrode prior to the
current; Ip1,c – first cathodic peak current; (±SD) and n = 3. tested phenolics. However, ascorbic acid continues to be accessible
into the electrode meaning that the reduced form should still be
available to react with phenolic quinones. These results do not con-
the current intensities of the two anodic peaks of the wine solu- firm the latest findings by Makhotkina & Kilmartin, 2009 where
tions in respectively, 27% for Ep1,a at 0.274 (V) (p-value < 0.001), ascorbic acid was not see to rapidly interact with caffeic acid and
and in 10% for Ep2,a at 0.701 (V) (p-value < 0.001), Fig. 5 and rutin quinones by cyclic voltammetry. Moreover, the increases in
Table 2. Analogous to the tested model-wine solutions, these the anodic peak current intensities for the tested phenolics and
results also indicate a rapid interaction of 2FMT and 4MSP with ascorbic acid may be related with same interference from the oxi-
wine quinones. dation of ascorbic acid, in dehydroascorbic acid, at lower poten-
tials. Otherwise, it will be expected that both the anodic current
response and the subsequent cathodic response will be lower.
3.3. Antioxidants
Sulfur dioxide increased the anodic current peaks intensities of
gallic acid by 20% for Ep1,a at 0.354 (V) (p-value < 0.01), and 31% for
Concerning antioxidants, ascorbic acid increased the anodic
Ep2,a at 0.735 (V) (p-value < 0.001), and decreased its cathodic
current and decreased the cathodic current for all analysed
 C.M. Oliveira et al. / Food Chemistry 211 (2016) 1–7 5

Gallic (a) Caffeic (b) Catechin (c)

Gallic+Phe Caffeic+Phe Catechin+Phe
30 30 30
Gallic+Met Caffeic+Met Catechin+Met
25 25 25

Current intensity x10-6 (A)

Current intensity x10-6 (A)

Current intensity x10-6 (A)

20 20 20
15 15 15
10 10 10
5 5 5
0 0 0
-5 -5 -5
-10 -10 -10
-15 -15 -15
-0.5 0 0.5 1 1.5 -0.5 0 0.5 1 1.5 -0.5 0 0.5 1 1.5
E (VS SCE) / V E (VS SCE) / V E (VS SCE) / V

Fig. 2. Cyclic voltammograms taken with a 3 mm glassy carbon electrode at 100 mV/s of a): 0.6 mM gallic acid in the absence (blue curve) and in the presence of 2.4 mM Phe
(black curve) and 2.4 mM Met (red curve); b): 0.6 mM caffeic acid in the absence (blue curve) and in the presence of 2.4 mM Phe (black curve) and 2.4 mM Met (red curve);
and c): 0.6 mM (+)-catechin in the absence (blue curve) and in the presence of 2.4 mM Phe (black curve) and 2.4 mM Met (red curve). (For interpretation of the references to
colour in this figure legend, the reader is referred to the web version of this article.)

Gallic (a) Caffeic

(b) Catechin
(c)
40 Gallic+2FMT 40 Caffeic+2FMT 40 Catechin+2FMT
Gallic+3SH Caffeic+3SH Catechin+3SH

Current intensity x10-6 (A)

Current intensity x10-6 (A)

30
Current intensity x10-6 (A)

30 30
Gallic+4MSP Caffeic+4MSP Catechin+4MSP

20 20 20

10 10 10

0 0 0

-10 -10 -10

-20-0.5 0 0.5 1 1.5 -20-0.5 0 0.5 1 1.5 -20-0.5 0 0.5 1 1.5

E (VS SCE) / V E (VS SCE) / V E (VS SCE) / V

Fig. 3. Cyclic voltammograms taken with a 3 mm glassy carbon electrode at 100 mV/s of a): 0.6 mM gallic acid in the absence (blue curve) and in the presence of 2.4 mM
2FMT (black curve), 2.4 mM 3SH (red curve), and 2.4 mM 4MSP (green curve); b): 0.6 mM caffeic acid in the absence (blue curve) and in the presence of 2.4 mM 2FMT (black
curve), 2.4 mM 3SH (red curve), and 2.4 mM 4MSP (green curve); and c): 0.6 mM (+)-catechin in the absence (blue curve) and in the presence of 2.4 mM 2FMT (black curve),
2.4 mM 3SH (red curve), and 2.4 mM 4MSP (green curve). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this
article.)

Gallic (a) Caffeic (b) Catechin

(c)
Gallic+AA Caffeic+AA
25 25 25 Catechin+AA
Gallic+SO2 Caffeic+SO2
Catechin+SO2
20 20 20
Current intensity x10-6 (A)
Current intensity x10-6 (A)
Current intensity x10-6 (A)

15 15 15

10 10 10

5 5 5

0 0 0

-5 -5 -5

-10 -10 -10

-15 -15 -15

-0.5 0 0.5 1 1.5 -0.5 0 0.5 1 1.5 -0.5 0 0.5 1 1.5
E (VS SCE) / V E (VS SCE) / V E (VS SCE) / V

Fig. 4. Cyclic voltammograms taken with a 3 mm glassy carbon electrode at 100 mV/s of a): 0.6 mM gallic acid in the absence (blue curve) and in the presence of 2.4 mM AA
(black curve) and 2.4 mM SO2 (red curve); b): 0.6 mM caffeic acid in the absence (blue curve) and in the presence of 2.4 mM AA (black curve) and 2.4 mM SO2 (red curve); and
c): 0.6 mM (+)-catechin in the absence (blue curve) and in the presence of 2.4 mM AA (black curve) and 2.4 mM SO2 (red curve). (For interpretation of the references to colour
in this figure legend, the reader is referred to the web version of this article.)

current peak intensity by 18% (p-value < 0.001), Fig. 4a and Table 1. (p-value < 0.01), as well the cathodic current intensity by 49% (p-
For caffeic acid, sulfur dioxide decreased the current intensity of value < 0.001), Fig. 4c and Table 1. These results, showed, once
the anodic peak by 16% (p-value < 0.001), as well the cathodic more, a rapid interaction of sulfur dioxide with the gallic acid, caf-
current intensity by 79% (p-value < 0.001), Fig. 4b and Table 1. feic acid and (+)-catechin quinones, and corroborate the latest find-
Likewise, this antioxidant decreased the current intensity of the ings by Makhotkina & Kilmartin, 2009, where sulfur dioxide had a
first anodic peak of (+)-catechin (Ep1,a at 0.354 (V) by 22% rapid interaction with ortho-quinones by cyclic voltammetry.
6 C.M. Oliveira et al. / Food Chemistry 211 (2016) 1–7

140 Wine (a) 140 Wine

(b) 140 Wine
(c)
Wine+AA Wine+Phe Wine+2FMT
120 120 120
Wine+SO2 Wine+Met Wine+3SH
100 100 100
Current intensity x10-6 (A)

Current intensity x10-6 (A)

Current intensity x10-6 (A)
Wine+4MSP
80 80 80
60 60 60
40 40 40
20 20 20
0 0 0
-20 -20 -20
-40 -40 -40
-0.5 0 0.5 1 1.5 -0.5 0 0.5 1 1.5 -0.5 0 0.5 1 1.5
E (VS Ag) / V E (VS Ag) / V E (VS Ag) / V

Fig. 5. Cyclic voltammograms taken with screen-printed carbon electrodes (4 mm) at 100 mV/s of a): white wine in the absence (blue curve) and in the presence of 2.4 mM
AA (black curve) and 2.4 mM SO2 (red curve); b): white wine in the absence (blue curve) and in the presence of 2.4 mM Phe (black curve) and 2.4 mM Met (red curve); and c):
white wine in the absence (blue curve) and in the presence of 2.4 mM 2FMT (black curve), 2.4 mM 3SH (red curve), and 2.4 mM 4MSP (green curve). (For interpretation of the
references to colour in this figure legend, the reader is referred to the web version of this article.)

Regarding antioxidants additions to the white wine, results aldehydes, while hydrogen peroxide will undergo further oxidation
have shown that ascorbic acid and sulfur dioxide have increased reactions.
the current intensities of the two anodic peaks of the wine A nucleophilic gauge related to the cathodic current intensity
solutions in respectively, 4 and 14% for Ep1,a at 0.274 (V) decrease is suggested: i) for the gallic acid quinone: phenylalanine <
(p-values < 0.05), and by 8 and 6% for Ep2,a at 0.701 (V) methionine  3-sulfanylhexan-1-ol < 4-methyl-4-sulfanylpentan-
(p-values < 0.05), Fig. 5 and Table 2. In contrast, these antioxidants 2-one < furan-2-ylmethanethiol < sulfur dioxide < ascorbic acid; ii)
decreased the current intensities of the wine solutions by 27 and considering the caffeic acid quinone: methionine < phenylalanine <
53% for, respectively, AA and SO2 (p-values < 0.001), Fig. 5 and 3-sulfanylhexan-1-ol < 4-methyl-4-sulfanylpentan-2-one < ascorbic
Table 2. Analogous to the tested model-wine solutions, these acid  furan-2-ylmethanethiol < sulfur dioxide; iii) concerning the
results point to a rapid interaction of AA and SO2 with wine catechin quinone: phenylalanine < methionine < 3-sulfanylhexan-
quinones. 1-ol < 4-methyl-4-sulfanylpentan-2-one < furan-2-ylmethanethiol
 sulfur dioxide < ascorbic acid; and iv) for wine: phenylalanine 
3-sulfanylhexan-1-ol < methionine < 4-methyl-4-sulfanylpentan-
4. Conclusion 2-one  furan-2-ylmethanethiol < sulfur dioxide < ascorbic acid.
These results corroborate recent studies (Nikolantonaki &
This work shows that all the tested wine nucleophiles react Waterhouse, 2012) that have compared the first-order rate con-
with quinones and thus are protective antioxidants by capturing stants, measured by UV–vis absorbance at 400 nm, of a quinone
the quinones, avoiding oxidative degradation reactions. Neverthe- of a non-wine like phenol 4-methylcatechol (4MeC) with several
less, this observation should be addressed in the context of the nucleophiles.
reactivity of nucleophiles, as well as in the framework of chemical Moreover, considering the phenolic compounds used in this
transformations promoted in wines. work, (+)-catechin was the phenolic that had the highest quinone
In the current study, results have shown that the lowest reactiv- reactivity, follow by caffeic acid. Conversely, gallic acid was the
ity of quinones with wine nucleophiles was attributed to amino phenolic that had promoted the lowest quinone reactivity with
acids. Nevertheless, this finding is very important because of their wine nucleophiles.
formation of potent Strecker aldehydes with low oxidative spoilage
odour detection thresholds, meaning that amino acids have a false Funding sources
protection on phenolic degradation reactions.
Browning reactions due to polyphenol oxidation will be inhib- The authors wish to thank FCT/MEC for the financial support of
ited in the presence of thiols, as these compounds are expected the QOPNA research Unit (FCT UID/QUI/00062/2013) through
to disrupt phenolic polymerization reactions. As such, all the tested national founds and, where applicable, co-financed by the FEDER,
thiols (3SH, 4MSP, 2FMT) showed a protective effect by intercept- within the PT2020 Partnership Agreement.
ing the quinones, avoiding oxidative degradation reactions. How-
ever, in the case of 3SH and 4MSP, this oxidative protection
Acknowledgements
promotes a decrease of these varietal aromas. Concerning 2FMT,
which was the tested thiol with the highest reactivity on capturing
The authors wish to thank FCT – Portugal (Fundação para a
quinones, the wine could be protected from both a negative aroma
Ciência e Tecnologia) and POPH/FSE for the doctoral grant to C.M.
and phenolic oxidation.
Oliveira (SFRH/BD/64097/2009).
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