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solution by standards
at 515 nm. The measurements were carried out using a
spectrophotometer UV-160 (Shimadzu, Japan).
2.5. Statistical analysis
The data are presented as mean SD for triplicate analysis. The
results were subjected to one-way analysis of variation ANOVA
with Fishers Least Signicant Difference test. P < 0.05 was consid-
ered signicant. The Pearson Correlation test for correlation analy-
sis was used. The statistical analysis was performed using Statistica
(Stat Soft, USA).
3. Results and discussion
3.1. The prole of red cabbage anthocyanin
Anthocyanins in red cabbage products were analysed using
HPLC-DAD-MS/MS method. The identication of anthocyanins was
carried out by means of the comparison of their retention time,
UVVis and MS/MS spectra, and the previous data (Charron et al.,
2007; Wiczkowski et al., 2013). As presented in Fig. 1 and Table 2,
anthocyanin proles of red cabbage products obtained in this study
is characterised by twenty derivatives of cyanidin with the main
structureof cyanidin-3-diglucoside-5-glucosides. Twoof themwere
nonacylated while eighteen acylated. Among acylated compounds,
eleven were monoacylated and seven were diacylated. Sinapic,
ferulic, caffeic and p-coumaric acids were found as acyl residues
in the acylated structure. Among twenty anthocyanins identied
and quantied seven of them predominated: one nonacylated
W. Wiczkowski et al. / Food Chemistry 167 (2015) 115123 117
(cyanidin-3-diglucoside-5-glucoside), three monoacylated (cyanidin-3-
(p-coumaroyl)-diglucoside-5-glucoside, cyanidin-3-(feruloyl)-digluco-
side-5-glucoside, cyanidin-3-(sinapoyl)-diglucoside-5-glucoside),
and three diacylated (cyanidin-3-(feruloyl)(feruloyl)-diglucoside-
5-glucoside and cyanidin-3-(feruloyl)(sinapoyl)-diglucoside-5-gluco-
side, and cyanidin-3-(sinapoyl)(sinapoyl)-diglucoside-5-glucoside).
These seven main compounds covered almost 68% of the total
anthocyanins content in fresh red cabbage. The proles of anthocy-
anins containing twenty cyanidin derivatives were observed for all
samples processed, however, it has to be noted that the content of
individual constituents differed depending on treatments applied
(Table 3). Similar prole of anthocyanins in fresh red cabbage was
observedin authors previous study (Wiczkowski et al., 2013). Other
previous studies of red cabbage anthocyanins exhibited differences
when compared with authors study in the following respects: (1)
number of anthocyanin in the prole (from nine to thirty-six
anthocyanins derivatives); (2) type of aglycone appearing in antho-
cyanins structure (pelargonidin, peonidin, andmalvidinexcept from
cyanidin); (3) kind of sugars bonded to anthocyanins aglycone
(xylose besides glucose); (4) type of acyl group (p-hydroxybenzoic
and malonic acids except from cinnamic acid derivatives); and (5)
acylation pattern (triacylation) (Charron et al., 2007; Lin, Li, &
Hwang, 2008; McDougall, Fyffe, Dobson, & Stewart, 2007; Pliszka
et al., 2009; Wu & Prior, 2005; Wu et al., 2004). These disparity
may result from varietal differences and the impact of biotic
(diseases and plant pests) and abiotic factors (temperature,
precipitation, insolation) (Podsedek, 2007).
3.2. Changes in the content of anthocyanins in red cabbage during
processing
Red cabbage constitute a rich source of nonacylated, monoacy-
lated, and diacylated forms of cyanidin and, therefore, provides an
unique ground for investigating the relationship between the con-
tent and prole of anthocyanins versus the type of food processing.
In this study the impact of processing was evaluated by comparing
Fig. 1. HPLC chromatogram of anthocyanins prole of red cabbage detected at 520 nm. Names of anthocyanins identied correspond to a number referred to in Table 1.
Table 2
The UVVis and MS data of anthocyanins from red cabbage products.
Peak Compounds k
vis
(nm) k
acyl
(nm) [M]
+
(m/z) MS/MS (m/z)
1 Cyanidin-3-diglucoside-5-glucoside 513 x 773 611/449/287
2 Cyanidin-3-glucoside-5-glucoside 512 x 611 449/287
3 Cyanidin-3-(sinapoyl)-diglucoside-5-glucoside 527 330 979 817/449/287
4 Cyanidin-3-(sinapoyl)-triglucoside-5-glucoside 525 321 1141 979/449/287
5 Cyanidin-3-(caffeoyl)(p-coumaroyl)-diglucosides-5-glucoside 521 314 1081 919/449/287
6 Cyanidin-3-(feruloyl)-triglucosides-5-glucoside 522 320 1111 949/449/287
7 Cyanidin-3-(sinapoyl)-triglucoside-5-glucoside 525 321 1141 979/449/287
8 Cyanidin-3-(feruloyl)(feruloyl)-triglucoside-5-glucoside 536 321 1287 1125/449/287
9 Cyanidin-3-(feruloyl)-diglucoside-5-glucoside 522 328 949 787/449/287
10 Cyanidin-3-(feruloyl)(sinapoyl)-triglucoside-5-glucoside 536 324 1317 1155/449/287
11 Cyanidin-3-(caffeoyl)-diglucoside-5-glucoside 522 315 935 773/449/287
12 Cyanidin-3-(p-coumaroyl)-diglucoside-5-glucoside 521 312 919 757/449/287
13 Cyanidin-3-(caffeoyl)(p-coumaroyl)-diglucoside-5-glucoside 521 314 1081 919/449/287
14 Cyanidin-3-(feruloyl)-diglucoside-5-glucoside 523 329 949 787/449/287
15 Cyanidin-3-(sinapoyl)-diglucoside-5-glucoside 526 328 979 817/449/287
16 Cyanidin-3-(feruloyl)-glucoside-5-glucoside 522 328 787 449/287
17 Cyanidin-3-(sinapoyl)-glucoside-5-glucoside 527 330 817 449/287
18 Cyanidin-3-(feruloyl)(feruloyl)-diglucoside-5-glucoside 535 328 1125 963/449/287
19 Cyanidin-3-(feruloyl)(sinapoyl)-diglucoside-5-glucoside 535 330 1155 993/449/287
20 Cyanidin-3-(sinapoyl)(sinapoyl)-diglucoside-5-glucoside 535 332 1185 1023/449/287
118 W. Wiczkowski et al. / Food Chemistry 167 (2015) 115123
fresh red cabbage with fermented, stored and stewed red cabbage.
The analysis conducted shown, for the rst time, that processes of
fermentation, storage and stewing affect the concentration and
prole of anthocyanins in red cabbage. Generally, results indicated
that processes applied contributed to decrease in the total content
of anthocyanins in red cabbage products (Table 3). This observa-
tion is in agreement with previous results which showed that both
technological and home treatment of plant materials led to a
decrease in the content of anthocyanins (Ba kowska-Barczak,
2005). In our study, the concentration of anthocyanins in fresh
red cabbage var. Langedijker Polona determined by HPLC-DAD
was 6.30 0.09 mg of cyanidin/g dm. The loss of total anthocya-
nins during fermentation process was at the level of 24%. A similar
reduction in anthocyanins content (25%) was observed during
30 min of the stewing process. When the process of stewing was
set at 60 min the degradation of anthocyanins increased respec-
tively (34%). In case of storage of fermented red cabbage at 4 C,
the intensity of anthocyanins degradation depended on the storage
length. In red cabbage stored for 7 days the intensity of this process
was found at a signicantly low level. On the other hand, after
180 days of storage under the same conditions, a very intense deg-
radation of anthocyanins was observed. The order of anthocyanins
degradation intensity was as follows: 7 days (2%) <30 days (13%)
<60 days (30%) <90 days (36%) <180 days (58%). In other reports
on the impact of the processes such as thawing, mashing, micro-
wave heating, boiling, blanching, juice extraction, pasteurisation,
and storage on the level of anthocyanin, the losses of these com-
pounds varied in a wide range of 395% (Giusti & Wrolstad,
2003; Hartmann, Patz, Andlauer, Dietrich, & Ludwig, 2008;
Sadilova, Carle, & Stintzing, 2007; Scalzo, Genna, Branca, Chedin,
& Chassaigne, 2008). Our ndings can be placed in the lower and
middle ranges of the values cited. On the other hand, according
to Truong et al. (2010) steaming of purple sweet potato slices
had no inuence on the total anthocyanins concentration, but it
was also noted that the previous slicing of fresh materials at room
temperature resulted in a decrease of the total anthocyanins con-
tent. It is generally assumed that red cabbage anthocyanins are
characterised by the best stability when compared to anthocyanins
from other plants (Clifford, 2000; Giusti & Wrolstad, 2003). This
property is ascribed to the chemical structure of red cabbage
anthocyanins, most of which being acylated and having a high
degree of glucosylation. Moreover, diacylated anthocyanins, which
are present inter alia in red cabbage, show a higher stability during
storage than monoacylated compounds (Giusti & Wrolstad, 2003).
Since previous studies showed that during processing leaching
causes losses of polyphenols (Davey et al. 2000), in our study the
produced liquids (fresh juice, fermented juice, stewed juice/water)
were collected and a proportional volume of each was mixed with
tissues of appropriate red cabbage products before using these
materials for further analysis. Consequently, the reduction of
anthocyanins content by leaching was excluded. In this situation,
other factors such as temperature, length of the process, presence
of oxygen, light, plant enzymes and microorganisms activity and
pH value could come into force (Clifford, 2000; Castaneda-
Ovando, Pacheco-Hernandez, Paez-Hernandez, Rodriguez &
Galan-Vidal, 2009; Truong et al., 2010). The strength of the impact
of these factors depends on the kind of the treatment used. In addi-
tion, the appearance of one or several factors at the same time
could have signicant inuence on anthocyanins fate. Therefore,
the effect of synergistic and antagonistic interactions of agents
cited should also be included, taking into consideration the fact
that while one main factor may be impossible to be marked but
the interaction effects may be directly found (Castaneda-Ovando,
Pacheco-Hernandez, Paez-Hernandez, Rodriguez & Galan-Vidal,
2009).
Regarding fermentation, the degradation effect may be attrib-
uted to the temperature throughout the entire process 24 C
during the rst three days and 18 C over the next eleven days.
Table 3
Content and contribution of anthocyanins in fresh, fermented, stored and stewed red cabbage.
Compounds
a
% of contribution in total anthocyanins content
Red cabbage products
Fresh Fermented Stored after fermentation (days) Stewed (min)
7 30 60 90 180 30 60
1 17.3 18.6 21.6 21.3 22.8 23.1 20.4 18.6 19.8
2 3.2 4.0 3.8 3.1 2.9 2.6 3.0 4.2 3.6
3 5.2 4.6 4.5 4.6 4.7 4.9 4.5 4.4 4.6
4 1.1 0.8 0.9 1.2 1.2 1.3 1.5 1.3 1.2
5 3.3 3.3 1.9 1.9 1.9 2.0 2.0 3.8 3.6
6 2.7 2.5 1.9 1.7 1.7 1.6 1.5 3.0 3.4
7 2.4 2.5 1.9 1.9 2.0 2.0 2.0 2.7 2.4
8 1.0 1.0 0.2 0.2 0.2 0.3 0.5 1.1 1.0
9 2.9 2.9 1.9 1.4 1.3 1.3 0.5 2.7 2.4
10 4.1 3.8 1.5 1.7 1.7 2.0 2.0 3.4 3.4
11 1.1 1.0 1.1 1.2 1.2 1.3 1.5 1.1 1.2
12 9.4 10.3 13.5 12.8 12.9 13.0 13.4 11.2 11.6
13 0.8 0.6 1.1 1.0 0.5 0.0 0.0 0.6 0.7
14 7.3 7.9 10.7 10.4 10.4 10.4 10.9 10.5 10.8
15 8.6 7.1 10.7 10.1 10.1 10.1 9.5 5.9 5.3
16 1.7 2.1 1.9 2.2 1.8 0.7 0.0 1.5 2.4
17 2.7 2.9 2.6 1.9 1.7 1.0 0.5 2.3 2.2
18 7.1 7.3 4.3 4.6 5.0 5.2 6.0 6.5 6.5
19 7.8 7.7 6.2 6.3 6.9 7.5 9.5 6.8 6.5
20 10.2 8.8 7.9 8.2 9.1 9.8 10.9 8.4 7.5
Nonacylated 20.6 22.6 25.4 24.9 25.7 25.7 23.4 22.8 23.4
Monoacylated 45.1 44.8 51.5 50.6 49.0 47.6 45.8 46.6 47.5
Diacylated 34.3 32.6 23.1 24.4 25.3 26.7 30.8 30.6 29.2
Total
b
6.30 0.09
A
4.78 0.09
B
4.68 0.10
B
4.05 0.08
C
3.36 0.14
D
3.07 0.13
D
2.01 0.15
E
4.74 0.13
B
4.15 0.09
C
a
Refer to Table 1.
b
All values were expressed as milligrams of cyanidin per gram dry matter of red cabbage. Data are expressed as means SD (n = 3). Means in line related to a total content
of anthocyanins for each products followed by the different letters are signicantly different (P < 0.05).
W. Wiczkowski et al. / Food Chemistry 167 (2015) 115123 119
The negative inuence of room temperature (2022 C) on the con-
centration of phytochemicals during food processing has been pre-
viously reported (Aaby, Wrolstad, Ekeberg & Kkrede, 2007). Also,
the impact of fermentation microbiota may determine anthocya-
nins content and composition of red cabbage. The previous report
of Bisakowski, Atwal, Gardner, and Champagne (2007) on the onion
avonoids indicated that lactic acid fermentation inuences the
composition of these compounds. Moreover, during the preparation
of red cabbage for processes, the bulbs were cut into strips and, con-
sequently, cell structure was disrupted and the anthocyanins
became more prone to enzymatic and nonenzymatic oxidation
(Pourcel, Routaboul, Cheynier, Lepiniec, & Debeaujon, 2007). In this
respect, it can be stated that plant enzymes may affect the content
and composition of anthocyanins until they are deactivated by
environmental factors of the fermentation process.
The impact of all above factors on anthocyanins stability during
fermentation could depend on the pH of the processes environ-
ment (Castaneda-Ovando, Pacheco-Hernandez, Paez-Hernandez,
Rodriguez & Galan-Vidal, 2009; Clifford, 2000). A pH value within
the rst several days of fermentation could constitute another neg-
ative factor for the stability of anthocyanins (Table 1). At the begin-
ning, the pH of fermentation juice was under 6.0, to gradually
decline to a level of 3.8 in the 8 day of the process, and remained
at this level over the next 7 days. This high pH value during the
rst days of fermentation and its rate of change could result in
the degradation of anthocyanins. Previous studies suggest that
pH below 4.0 is favourable for anthocyanins stability, while pH
around neutral is negative (Markakis, 1982). On the other hand,
the reports of Giusti and Wrolstad (2003) and McDougall et al.
(2007) showed that acylated anthocyanins were more stable at
pH near neutral than nonacylated anthocyanins, since acylation
increases anthocyanins stability. Taking this information into
account, it can be stated that a very high percentage of acylated
anthocyanins (approximately 80%) in the studied red cabbage
had limited the rate of degradation of anthocyanins (only 24%
decrease) during 14 days of fermentation (Table 3).
Our ndings revealed that storage time signicantly affected
the level of anthocyanins (Table 3). Also the previous report of
Giusti and Wrolstad (2003) indicated that the length of storage
lead to anthocyanins degradation. In addition, these authors found
that the temperature of storage exerted an effect on the anthocya-
nins degradation kinetics at 25 C it was higher than at 2 C. In
our study, under the storage at 4 C the rate of anthocyanins
decline during 180 day was 0.32% of total anthocyanins per day.
The highest decrease in the anthocyanins content was noted in
the period between 90 and 180 days of storage, while the highest
degradation rate of anthocyanins (0.59% of total anthocyanins/
day) in the period between 7 and 30 days. A similar rate of total
anthocyanins decrease (0.34% of total anthocyanins/day) in a
strawberry juice stored for 77 days at 8 C by Hartmann et al.
(2008) was indicated. Also storage of 112 days at 6 C showed a
considerable loss of total anthocyanins (0.38% of total anthocya-
nins/day) in strawberry puree (Aaby, Wrolstad, Ekeberg &
Kkrede, 2007). Regarding the pH value during storage, it increased
slightly during this process but remained favourable for anthocya-
nins stability (Markakis, 1982), close to 4.0 (Table 1). Therefore,
apart from the storage time and mentioned temperature, other fac-
tors such as uncontrolled growth of microorganisms may inuence
the level of anthocyanins during storage.
A 30 and 60-min process of stewing in low amounts of water
lead to a signicant decrease in the content of red cabbage antho-
cyanins, whereas in case of shorter time of treatment the decline in
anthocyanins concentration was lower (Table 3). In addition, in our
experiment stewing at high temperature (around 100 C) for
60 min was one of the processes applied causing the highest losses
in anthocyanins. This may result from high temperature and dura-
tion of the process. Also in case of different food products heating
lead to a signicant decrease in the total content of anthocyanins.
The heat treatment of strawberry juice at high temperature (85 C)
for a very short time (5 s) indicated a loss of 12% in anthocyanins
content, while prolongation of the process time up to 15 min
resulted in higher losses of anthocyanins (21% diminution)
(Hartmann et al., 2008). In case of other heat treatments, such as
boiling, blanching and steaming, the reduction of monomeric
anthocyanins measured by pH-differential method in red cabbage
was noted in the range of 1222% (Volden et al., 2008). However, in
the report cited a part of recovered anthocyanins (638%) was
found in the processing water. Also Scalzo et al. (2008) indicated
that blanching and cooking had reduced the total cauliower
anthocyanins content. On the other hand, the same authors exhib-
ited that in case of microwave treatment the content of anthocya-
nins had remained almost unchanged. At the stewing process of
red cabbage the plant enzyme system could also operate but to a
very small extent, since upon cutting of cabbage, it was subjected
to high temperatures inactivating enzymes (Jang, Ma, Shin, & Song,
2005).
3.3. Changes in the composition of anthocyanins in red cabbage during
processing
The effect of fermentation, storage and stewing on the
individual cyanidin derivatives of red cabbage was studied in
detail. From the results obtained, shown in Table 3, it was observed
that in fresh red cabbage, cyanidin-3-diglucoside-5-glucoside
was the major compound followed by cyanidin-3-(sinapoyl)
(sinapoyl)-diglucoside-5-glucoside and cyanidin-3-(p-coumaroyl)-
diglucoside-5-glucoside. Similarly, after fermentation and stewing
as well as during storage, the mentioned nonacylated derivative
was the main form, but followed by different acylated derivatives
of cyanidin. Regarding the fermented cabbage, cyanidin-3-
(p-coumaroyl)-diglucoside-5-glucoside and cyanidin-3-(sinapoyl)
(sinapoyl)-diglucoside-5-glucoside were the second and the third
major constituents, respectively. In terms of stored and stewed
products, besides cyanidin-3-diglucoside-5-glucoside and cyani-
din-3-(p-coumaroyl)-diglucoside-5-glucoside, cyanidin-3-(feruloyl)-
diglucoside-5-glucoside was noted as one of the three main
anthocyanin compounds. Both processes, fermentation and stewing,
lead to a decrease in the content of all analysed anthocyanins. How-
ever, the intensity of decrease depended on the chemical structure
of these compounds. Generally, among seven main individual red
compounds found in fermented and stewed red cabbage products,
the lowest rate of decrease was noted in the case of nonacylated
anthocyanins. Among main monoacylated anthocyanins, derivatives
acylated with sinapic acid were characterised by the highest losses
(Table 3). The order of stability of monoacylated anthocyanins for
fermented and stewed red cabbage was similar: cyanidin-3-
(p-coumaroyl)-diglucoside-5-glucoside = cyanidin-3-(feruloyl)-dig-
lucoside-5-glucoside > cyanidin-3-(sinapoyl)-diglucoside-5-glucoside
and cyanidin-3-(feruloyl)-diglucoside-5-glucoside > cyanidin-3-
(p-coumaroyl)-diglucoside-5-glucoside > cyanidin-3-(sinapoyl)-dig-
lucoside-5-glucoside, respectively. These results are in accordance
with study of Sadilova et al. (2007) which showed that anthocya-
nins acylated with sinapic acid have the lowest half-life when
compared to the compounds acylated with ferulic and coumaric
acids. Also, during simulation of neutral pancreatic digestion red
cabbage anthocyanins acylated with sinapic acid were less stable
than those acylated with p-coumaric and ferulic acids
(McDougall et al., 2007). In our study, within the diacylated forms
of cyanidin-3-diglucoside-5-glucoside, anthocyanins conjugated
with two sinapic acids were less stable than forms bond with
two ferulic acids. These ndings stay in agreement with the
observations made for the fate of diacylated anthocyanins during
120 W. Wiczkowski et al. / Food Chemistry 167 (2015) 115123
treatment under small intestine conditions (McDougall et al.,
2007). In contrast to the results obtained for fermentation
and stewing processes, during 7 and 30 days of storage, an
increase in the content of four derivatives (cyanidin-3-digluco-
side-5-glucoside, cyanidin-3-(p-coumaroyl)-diglucoside-5-glucoside,
cyanidin-3-(feruloyl)-diglucoside-5-glucoside, cyanidin-3-(sinapoyl)-
diglucoside-5-glucoside) was found. In the other point of
observation (60, 90, and 180 days of storage) the concentration
of anthocyanins analysed was lower when compared to the
non-stored fermented red cabbage. Moreover, in case of storage,
an entirely different prole of stability of acylated anthocyanins
was found. The derivatives acylated with sinapic acid were
characterised by lower losses when compared to forms conjugated
with ferulic acid.
Since the anthocyanins identied had the same main core
cyanidin glucosides all applied processes could lead to a
conversion of one compound to another. Diacylated derivatives
of cyanidin-3-diglucoside-5-glucoside could degrade to monoacy-
lated forms, and next to nonacylated constituents. Also, a direct
transformation from diacylated to nonacylated derivatives was
possible. In addition, the hydrolysis of glucosidic bond in chemical
structure of anthocyanins present in red cabbage could result in
the formation of anthocyanins with a lower degree of glucosyla-
tion. Previously, the deacylation of vegetable avonoid was found
by DuPont, Mondin, Williamson, and Price (2000). Furthermore, a
cleaving of acylated anthocyanins into the acyl-glycosides and
the aglycone was observed (Sadilova et al., 2007). These phenom-
ena may constitute the basis for explaining the increase of nonacy-
lated anthocyanins content and their contribution to the total
anthocyanins concentration during treatments used when com-
pared to the concentration of acylated forms (Table 3).
3.4. The contribution of nonacylated, monoacylated and diacylated
anthocyanins in the total anthocyanin content of red cabbage products
The percentages of nonacylated, monoacylated and diacylated
cyanidin derivatives in the total anthocyanin content of red cab-
bage products were studied. After the fermentation process, the
contribution of nonacylated compounds to the total content of
anthocyanins increased when compared to the fresh red cabbage
anthocyanins prole. The percentages of both nonacylated antho-
cyanins identied were higher than in fresh red cabbage (Table 3).
In contrast to nonacylated compounds, the presence of acylated
forms in the total anthocyanins concentration decreased during
fermentation. During 180 days of storage the participation of non-
acylated, monoacylated, and diacylated forms in the total anthocy-
anins concentration was variable. In case of nonacylated
compounds, during the rst 7 days of storage an increase in the
presence of these derivatives in the total content of anthocyanins
was observed (Table 3). After 30, 60 and 90 days of storage the
contribution of nonacylated compounds to the total content of
anthocyanins was very similar to this noted after 7 days of storage.
However, after 180 day of storage the value of these parameters
decreased to the level of 23.4%. In terms of monoacylated deriva-
tives of cyanidin, after 7 days of storage, higher contribution of
these compounds to the total content of anthocyanins was
observed when compared to non-stored fermented product. In
the other point of observation (30, 60, 90, and 180 days of storage)
the presence of cyanidin monoacylated derivatives in the total con-
tent of anthocyanins decreased gradually. An entirely opposite
phenomenon was found in case of diacylated anthocyanins. During
the rst 7 days of storage the contribution of these compounds to
the total content of red pigment decreased when compared to non-
stored fermented cabbage. Subsequently, the value of this param-
eter in other points of measurements increased gradually. After
stewing, the contribution of nonacylated derivatives of anthocya-
nins to the total anthocyanins content was higher when compared
to the results obtained for the fresh product. In addition, it was
observed that the presence of monoacylated compounds in the
total concentration of analysed substances increased. An opposite
phenomenon was noted in the case of diacylated derivatives of
cyanidin. During 30 min of stewing the contribution of diacylated
compounds decreased from 34.3% to 30.6%. Subsequent 30 min of
high temperature treatment caused further degradation of diacy-
lated anthocyanins up to level of 29.2% of the total anthocyanins
content. As has been mentioned in previous subpart, anthocyanins
found in red cabbage had identical main core therefore the changes
in the contribution of nonacylated, monoacylated and diacylated
anthocyanins in the total anthocyanin content of red cabbage
during processing, may result from a conversion of one anthocyanin
to another.
3.5. Changes in red cabbage antioxidant capacity during processing
In this study, ve methods (ACW PCL, ACL PCL, ORAC, ABTS and
DPPH) to determine the antioxidant capacity of red cabbage prod-
ucts were used. For a comparison, previous studies (Prior, Wu, &
Schaich, 2005) suggested the use of the three methods (ORAC,
ABTS, and FolinCiocalteu assay) to be applied for standardisation
of antioxidant capacity measurements of food samples. In conse-
quence, the application of these ve methods and the use of nine
different products of red cabbage (fresh, fermented, stored by 7,
30, 60, 90 and 180 days, stewed 30 min and 60 min) for the tests
allowed to obtain a full range of antioxidant capacities of this veg-
etable during food processing. Ultimately, a specic antioxidant
capacity of different red cabbage products was determined
(Table 4). The values of antioxidant capacity of red cabbage
products provided by ABTS assay ranged from 133.1 to 166.5 lmol
Trolox/g dm. In case of ORAC assay, these values ranged from 318.3
to 411.3 lmol Trolox/g dm. The antioxidant capacity estimated by
ACW PCL assay oscillated between 71.6 and 103.5 lmol Trolox/g dm,
while by ACL PCL assay from 413.7 to 632.8 lmol Trolox/g dm.
Table 4
The antioxidant capacity of red cabbage products (fresh, fermented, stored, stewed) determined by ve different assays.
Antioxidant activity assays lmol Trolox/g dm
a
Red cabbage products
Fresh Fermented Stored after fermentation Stewed red cabbage
7 days 30 days 60 days 90 days 180 days 30 min 60 min
TEAC 166.5 2.9
A
137.9 2.5
BC
136.6 2.4
BC
133.1 2.4
C
139.9 1.6
B
136.5 1.9
BC
135.5 1.7
C
135.7 1.9
C
133.8 2.8
C
ORAC
FL
411.3 8.6
A
380.6 5.6
C
397.0 7.1
B
364.2 5.0
D
347.8 7.3
E
357.7 7.1
DE
318.3 2.2
F
397.9 1.5
B
368.4 1.7
D
ACW PCL 103.5 1.2
A
94.0 1.2
B
83.5 1.7
C
80.5 1.7
CD
79.1 1.2
D
80.2 1.7
CD
71.6 0.9
E
100.7 1.3
A
92.0 1.4
B
ACL PCL 632.8 5.4
A
499.6 3.4
D
496.7 1.2
D
478.2 3.4
E
474.5 2.0
E
452.3 1.0
F
413.7 1.1
G
537.4 7.7
B
512.2 5.8
C
DPPH 47.0 1.0
A
45.7 1.3
A
43.6 1.2
B
41.9 1.0
C
38.3 2.8
CD
38.7 0.7
D
38.1 1.2
D
45.7 1.2
A
42.6 0.6
BC
a
Data are expressed as means SD (n = 3). All values were expressed as micromoles of Trolox per gram dry matter of red cabbage. Means in line related to a respective test
followed by the different letters are signicantly different (P < 0.05).
W. Wiczkowski et al. / Food Chemistry 167 (2015) 115123 121
In terms of DPPH method the values of antioxidant capacity ranged
from 38.1 to 47.0 lmol Trolox/g dm. Generally, the highest antiox-
idant capacity of red cabbage varieties was noted for in the case of
evaluation conducted by ACL PCL method, while the lowest by
DPPH. The order of antioxidant capacity of red cabbage was as fol-
lows: ACL PCL > ORAC
FL
> ABTS > ACW PCL < DPPH. The antioxidant
capacity of red cabbage products analysed by ACL PCL was highly
correlated with that provided by ACW PCL (r = 0.865), ORAC
FL
(r = 0.859) and DPPH (r = 0.832). Moreover, high correlation
between ACW PCL: ORAC
FL
(r = 0.846), ACW PCL: DPPH
(r = 0.904) and ORAC
FL
: DPPH (r = 0.900) was found.
All samples of red cabbage products scavenged radicals and
their antiradical potential differed signicantly across the products
studied (P < 0.05). Fresh red cabbage was characterised by a stron-
ger ability to radical scavenge than fermented, stored and stewed
red cabbage. The highest reduction of this parameter occurred for
red cabbage stored for 180 days (average 28.1%). The lowest
decrease in the antioxidant capacity was determined for red cab-
bage stewed for 30 min (average 10.4%). The order of scavenging
capacity was as follows: fresh > stewed 30 min > fermented =
stored 7 days > stewed 60 min > stored 30 days > stored 60 day-
s > stored 90 days > stored 180 days (Table 4). The results obtained
indicate, for the rst time, that differences in the antioxidant
capacity of red cabbage products occurred depending on the type
treatment applied. Other reports also demonstrated that the anti-
oxidant activity of food products can depend on the kind of food
processing used (Aaby et al., 2007). After heating, a decline of TEAC
in black carrot, elderberry and strawberry samples was found by
Sadilova et al. (2007), what the authors assigned among others to
anthocyanins degradation following thermal exposure. Moreover,
authors suggested that the loss of antioxidant capacity supplied
by anthocyanins could not be compensated by the activity of newly
formed phenolics upon heating. Other study indicated that cooking
increased free radical scavenging activity for red cabbage (Wu
et al., 2004).
In our analysis, the antioxidant capacity of red cabbage products
is positively and signicantly correlated (P < 0.05) with the total
anthocyanins concentration, determined by four assays used:
ORAC
FL
(r = 0.949), ACL PCL (r = 0.938), DPPH (r = 0.926) and ACW
PCL (r = 0.861). The results clearly showed that anthocyanins occur-
ring in fermented, stored and stewed red cabbage were responsible
for their antioxidant capacity. In addition, the contribution of seven
main derivatives of cyanidin (cyanidin-3-diglucoside-5-glucoside,
cyanidin-3-(p-coumaroyl)-diglucoside-5-glucoside, cyanidin-3-(feru-
loyl)-diglucoside-5-glucoside, cyanidin-3-(sinapoyl)-diglucoside-
5-glucoside, cyanidin-3-(feruloyl)(feruloyl)-diglucoside-5-glucoside,
cyanidin-3-(feruloyl)(sinapoyl)-diglucoside-5-glucoside, and cyani-
din-3-(sinapoyl)(sinapoyl)-diglucoside-5-glucoside) to the antioxi-
dant capacity of red cabbage products was determined. Upon
Pearson correlation analysis between the concentration of these
major red cabbage anthocyanins and antioxidant capacity of red
cabbage products extracts it was found that the content of cyani-
din-3-diglucoside-5-glucoside indicated positive and signicant
correlation (P < 0.05) with antioxidant capacity provided by the
two methods used (for ORAC
FL
r = 0.934, for ACL PCL r = 0.918).
The level of both cyanidin-3-(p-coumaroyl)-diglucoside-5-gluco-
side and cyanidin-3-(feruloyl)-diglucoside-5-glucoside was corre-
lated (P < 0.05) with the antioxidant activity measured with only
one assay (for ORAC
FL
r = 0.913 and r = 0.884, respectively). The
concentration of cyanidin-3-(feruloyl)(feruloyl)-diglucoside-5-glu-
coside was associated (P < 0.05) with the value of antioxidant
capacity determined by the three following tests: ACW PCL
(r = 0.924), ACL PCL (r = 0.916) and DPPH (r = 0.904). In case of
cyanidin-3-(feruloyl)(sinapoyl)-diglucoside-5-glucoside, its indi-
vidual content was positively and signicantly correlated
(P < 0.05) with the antioxidant capacity provided by until four
methods used (ACL PCL r = 0.926, DPPH r = 0.882, ACW PCL
r = 0.29, ORAC
FL
r = 0.828). A similar observation was noted for
cyanidin-3-(sinapoyl)(sinapoyl)-diglucoside-5-glucoside (ACL PCL
r = 0.944, ABTS r = 0.861, ORAC
FL
r = 0.836, DPPH r = 0.825). Our
data supported previous observations which demonstrated that
anthocyanins contribute to the antioxidant capacity (Sadilova
et al., 2007). Moreover, it has been recently exhibited that fermen-
tation and heat treatment of white cabbage increased the initial val-
ues of the antioxidant activity (Kusznierewicz, S
miechowska,
Bartoszek, & Namies nik, 2008). Conversely, in our study the antiox-
idant capacity of red cabbage products after fermentation, storage
and stewing was lower when compared to the fresh product. This
probably stems from the fact that since anthocyanins are strong
in vitro antioxidants and the antioxidant capacity of red cabbage
products studied in our experiments is positively and signicantly
correlated with anthocyanins concentration, the degradation of
anthocyanins during these processes resulted in a decrease of
antioxidant capacity of the products analysed. In addition, taking
into account the ndings indicating that anthocyanins acylated
with sinapic acid have the highest antioxidant capacity among
anthocyanins conjugated with others hydroxylcinnamic acids
(Wiczkowski et al., 2013) together with the assumptions derived
from this study that cyanidin-3-diglucoside-5-glucoside acylated
with sinapic acid show the highest rate of degradation, the
reduction of antioxidant capacity of red cabbage products is
observed justied.
4. Conclusion
The results indicated that all red cabbage processing applied
reduced the content of anthocyanins in the products obtained.
However, it is necessary to emphasise that the extent of respective
losses depended on the kind of the processing introduced. Among
individual red compounds found in red cabbage products, acylated
with sinapic acid derivatives of cyanidin-3-diglucoside-5-gluco-
side were characterised by the highest losses. Contrary, the lowest
rate of decrease was noted for nonacylated anthocyanins, however
these results might stem from the decomposition of acylated
anthocyanins. In case of antioxidant activity, fresh red cabbage
was characterised by a stronger ability to radical scavenge than
fermented, stored and stewed red cabbage where the diminution
of anthocyanins content entailed the reduction of antioxidant
capacity.
When it comes to the pro-health food, maximal retention of
bioactive compounds during processing is strongly desired since
benecial biological properties of food are to some extend attrib-
uted to their active constituents. On the basis of all the facts pre-
sented, red cabbage products in the fresh, fermented or short
time stewed form constitute valuable vegetables for daily con-
sumption as they remain rich in anthocyanins. What should also
be noted, fermented red cabbage stored up to 30 days is a signi-
cant source of anthocyanins, acting as strong antioxidants. Quanti-
cation of the losses and estimation of anthocyanin prole changes
allows for the consideration of optimisation possibilities as well as
the quest of processing techniques, manner of storage, and the raw
material in order to reduce losses and changes, as well as to
improve the pro-health potential of red cabbage products. Cur-
rently, further studies are needed to determine how anthocyanins
from different forms of red cabbage products behave after
consumption.
Acknowledgement
The research was supported by the National Science Centre
(Poland, project 1902/B/P01/2008/35).
122 W. Wiczkowski et al. / Food Chemistry 167 (2015) 115123
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