Manolopoulou and Varzakas J Food Nutr Disor 2014, 3:1

http://dx.doi.org/10.4172/2324-9323.1000131

Research Article

Application of Antibrowning
Agents in Minimally Processed
Cabbage
Eleni Manolopoulou1 and Theodoros Varzakas2*

Abstract
The aim of this study was to investigate the effect of storage at 0C
and 5C on the color and organoleptic quality, of fresh-cut cabbage
during the addition of ascorbic acid, citric acid and calcium chloride
and during Modified Atmosphere Packaging (MAP) with low 2
concentrations (1.5%) and high concentrations of C2 (17%).
Of the two studied temperatures (0 and 5C) the temperature of 0C
maintained better quality parameters (color, organoleptic quality)
and increased the shelf life of minimally processed cabbage.
Among the treatments (MAP, citric acid, ascorbic acid, calcium
chloride) MAP and citric acid showed the best results in terms of
color and total visual quality. The combination of low temperature
storage (0C) with MAP or citric acid treatment prolonged the shelf
life of minimally processed cabbage for 22 days, time sufficient for
acceptable marketing of the product.

Keywords
Antibrowning agents; Cabbage; MAP

Introduction
Fresh-cut produce is defined as any fresh fruit or vegetable or any
combination thereof that has been physically altered from its original
form, but remains in a fresh state. Regardless of commodity, it has
been trimmed, peeled, washed and cut into 100% usable product that
is subsequently bagged or packaged to offer consumers high nutrition,
convenience and value while still maintaining freshness (IFPA). In
recent years, consumers have become more health conscious in their
food choices, but have had less time to prepare healthful meals. As a
result the market demand for fresh-cut fruits and legumes has rapidly
increased.
t is well known that minimally processed fruits and vegetables
are generally more perishable than the original raw materials [1,2],
because the application of partial processing increases perishability
due to increased metabolic activities and decompartmentalization of
enzymes and substrates. This may cause browning, softening, and offflavour development [3].
Successful marketing of minimally processed fresh produce is
linked to several factors as the maintenance of high level of sensory
quality for an adequate duration subsequent to harvest [4], the
*Corresponding author: Theodoros Varzakas, Department of Food Technology,
School of Agricultural Technology, Technological Educational Institute of
Peloponnese, Greece, Tel: +30 2721045279; Fax: +3027210 45234; E-mail:
theovarzakas@yahoo.gr
Received: July 05, 2013 Accepted: January 09, 2014 Published: January 13,
2014

International Publisher of Science,

Technology and Medicine

Journal of Food &

Nutritional Disorders
a SciTechnol journal
harvest timing to achieve peak quality, quality control for postharvest
treatments and effective packaging.
Colour is an important quality attribute in the food and
bioprocess industries, both for its aesthetic value and for quality
judgement. Colour influences consumers choice and preferences.
An important consideration during fresh-cut fruit processing is the
preservation of normal tissue colour and control of discoloration or
surface browning.
Browning colouration is an important phenomenon in food
handling and processing it affects appearance quality. Enzymatic
browning involves the interaction of oxygen, phenolic compounds
and polyphenol oxidases (PPOs). To prevent browning at least one
component must be removed from the system. Browning can be
prevented by exclusion or removal of one or both of the substrates
(O2 and phenols), lowering the pH to 2 or more units below the
optimum, or adding compounds that inhibit PPO or prevent melanin
formation [5].
In leafy vegetables browning is considered as one of the most
important defects because it is easily noticeable. It does not occur
in intact plant cells since phenolic compounds in cell vacuoles are
separated from the PPO that is present in the cytoplasm. Once tissues
are damaged by slicing or cutting, the mixture of PPO and phenolic
compounds consequently results in a rapid browning reaction.
Enzymatic browning remains a costly concern for the fruit and vegetable
industry, as it is one of the primary causes of quality degradation.
Enzymatic browning may be controlled through the use of
physical and chemical methods and in most cases both are employed.
Physical methods may include reduction of temperature and /or
oxygen, use of MAP or edible coatings or treatment with gamma
irradiation or high pressure. Chemical methods utilize compounds
that act to inhibit the enzyme, remove its substrates (O2, phenolics)
or function as preferred substrate [6].
Among the compounds that have been shown to inhibit browning
are sulphites, and andioxidants which are grouped in accordance to
their mode of action (i.e., as acidulants- such as citric, malic, and
phosphoric acids-, reducing agents- such as ascorbic acid or the
erythrobate isomer of ascorbic acid-, chelating agents- such as sorbic
acid, polycarboxylic acids-,inorganic salts such as calcium, zinc and
sodium-, complexing, enzymes, and inhibitors) [7-18].
Temperature is the most important factor in the preservation of
fresh-cut fruit quality. The use of low temperatures during the storage,
handling and processing of minimal processing fruits and vegetables
can control the metabolic rates and the enzymatic activity that affect
quality. Refrigeration slows down both PPO and POD activities, since
low temperatures (0-4C) are far from optimal for these enzymes [19].
CA, MAP reduce oxygen and increase the carbon dioxide content
of the atmosphere surrounding the commodity and can produce
direct effects on phenolic composition. High levels of oxygen are
particularly effective in inhibiting enzymatic discoloration. It has been
hypothesized that high oxygen levels may cause substrate inhibition
of PPO, or alternatively, that high levels of colourless quinones
subsequently formed may cause feedback inhibition of PPO [20].

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Citation: Manolopoulou E, Varzakas T (2014) Application of Antibrowning Agents in Minimally Processed Cabbage. J Food Nutr Disor 3:1.

doi:http://dx.doi.org/10.4172/2324-9323.1000131

The objective of this study was to investigate the potential of

ascorbic acid, citric acid, calcium chloride and MAP, individually, to
extend the shelf life and maintain the quality of fresh-cut cabbage.

Materials and Methods

White Cabbage (Brassica oleracea L. var. capitata) var. Bunner
harvested on Mars and April from a farm located in Messinia
(Peloponnese, Greece) was minimally processed using the best
preparation techniques. The leaves were cut into quarters and
then sliced into 3 cm wide strips with a sharp stainless steel knife
sterilized in ethanol. The shredded cabbage was washed for 4 min in
chlorinated (100 ppm NaOCl) water at 5C (at a solid/ liquid ratio of
1:3) to remove released nutrients. Then the cabbage was rinsed with
tap water for 4 min and centrifuged for 1 min in a 23 cm diameter
basked rotating at 500 rpm.
The treatments were conducted in parallel and prepared for
the same batch of product. To minimize product heterogeneity,
processed cabbage was pooled, mixed and subsequently divided in
five equal parts. The first part was dipped in distilled water and was
used as control sample. The second portion was dipped in ascorbic
acid solution; the third was dipped in citric acid solution while the
fourth portion was dipped in calcium chloride solution. The fifth and
final portion was packaged in MAP, which was made by injection of a
mixture with 5% 2-10% C2 balanced with N2.
Dipping treatments with solutions of ascorbic acid
(Serva,Germany), citric acid (Merck, Germany) and calcium chloride
(Carlo Erba, Italy) were applied separately for 3 min, after draining.
All dipping solutions were made with distilled water at 5C and the
concentrations were 1% (w/v) in all cases.
One hundred 2 g of sliced cabbages treated with ascorbic acid,
citric acid, calcium chloride or distilled water (control) were placed
in clear trays of polystyrene (15.5 x 22.5 x 3 cm) covered with PVC
film (13 m) and stored at 0C and 5C and 90% H.R. in darkness.
The same quantity of sliced cabbage was placed in 30 m thick MDPE
packages (30 x 24 cm), (O2 permeability 6.170 730 mL m-2 d-1 bar-1
and CO2 permeability 41.520 1830 mL m-2 d-1 bar-1, S=PCO2/PO2=6.8
at 20C according to data provided by the film manufacturer) since
gas flushing was applied. Gas flushing was carried out using a gas
mixer (MAP Mix 9000, Denmark) and a vacuum compensation
chamber (Multivac A, 300/16, Germany). All tested packages
treatments mentioned above were stored at 0C and 5C (except
the MAP maintained only at 0C) and 90% relative humidity in the
dark for 22 days. Ten packages were prepared per sampling date and
treatment (ascorbic acid, citric acid, calcium chloride, control, MAP).
Measurements were taken during the 0, 7th, 14th, and 22rd day. Every
experiment was conducted in twice.
Volume 3 Issue 1 1000131

(O2)

(CO2)

20

21
18
15
12
9
6
3
0

16
12
8

CO2 %

O2%

Cabbage is an important member of cruciferous vegetables being

also an important dietary vegetable consumed as fresh cut salad or
cooked. Recently it was widely marketed as a minimally processed
product. Vegetables in the Brassica family are reported to possess
cancer-preventive properties [21] attributed to their glucosinolates
[22] and phenolic compounds [23,24]. Glucosinolates in Brassica
vegetables confer defence functions to the plants and provide a source
of bioactive compounds that are important to human nutrition
and health. Brassica vegetables are therefore of great interest for
their potential use in disease risk reduction and are considered as
functional foods [25]. Cabbage and broccoli, excellent sources of
indoles, dithiolthiones, isothiocyanates, and chlorophyllins, are also
effective in minimizing the risk of heart attack [26].

4
0
0

12 19 22

storage time (days)

Figure 1: Changes in O2 and CO2 of packaged shredded cabbage stored at
0C. (N=20) (I=LSD).

Parameters studied were: gas analysis (MAP) and color.

Organoleptic control was carried out for discoloration (browning) of
cut surfaces and total visual quality. The inpackage atmosphere (2,
C2) was measured with a headspace gas analyzer (CheckMate 9000,
PBI Densensor Co., Denmark) drawing up to 2 mL of air samples
Sampling took place with a hypodermic needle through a septum
pasted on the packaging Colour was measured with a Minolta CR300 chromameter (Minolta Corp. Japan) on CIE L*, a*, b* chromatic
space. The instrument was initially calibrated using a white ceramic tile
(Y=92.6 X=0.313 y=0.319). The L* chromatic variable, ranges from 0
(black) to 100 (white) and is an indicator of the lightening or darkening
due to the physicochemical changes taking place during storage. The a*
measures the degree of redness (+a*) and greenness (-a*) while the b*
the degree of yellowness (+b*) and blueness (-b*). Colour changes were
determined by parameters (L*), and hue angle [h=arctan-1(b*/a*)].
Quality evaluation (cut surface browning, and overall visual
acceptance) was assessed in 6 packs per treatment, by a panel of six
semi trained judges adopting a hedonic scale from 1 to 5, rating with
5=very fresh with no browning on the cut surfaces, 3=slight browning
on the cut surfaces with marginal marketability, 1=not to be consumed
[27]. A score of 3 was considered as the marketable limit.

Statistical analysis
The experiment was performed according to a full factorial design
(treatments x storage time) and subjected to analysis of variance
(ANOVA) using the Statgraphics Plus 5.1 (Statpoint Technologies,
Inc, VA, USA) statistical software. Mean values were subjected
to Fishers Least Significant Difference test (LSD) at p=0.05. This
statistical test is liberal with respect to the comparison wise error rate,
but is powerful for detecting true difference [28]. The presented data
in the figures are the mean value from 2 experimental series since no
significant difference between the series was found.

Results and Discussion

Gas composition
The composition of the atmosphere (O2 and CO2) inside the
package of shredded cabbage is presented in Figure 1. The O2
concentration was reduced during the first days, whereas C2
concentration increased.

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Citation: Manolopoulou E, Varzakas T (2014) Application of Antibrowning Agents in Minimally Processed Cabbage. J Food Nutr Disor 3:1.

doi:http://dx.doi.org/10.4172/2324-9323.1000131

CaCl2

control

80

ascorbic acid

0C
CC

70

CaCl2

control

60

50

50

40

40

30

30

20

20

10

10

5C

14

storage time (days)

22

CaCl2

control

MAP

14

22

storage time (days)

After 6 days of storage the composition of the atmosphere in the

packages was stabilized and remained at these levels until the end
of storage (22nd day). 2 concentration stabilized at 1.5% (from an
initial concentration of 5%) and C2 concentration remained at 17%
(from an initial concentration of 10%). This change in gases could
be attributed to the increase in respiratory activity of tissues due to
the cutting [29]. According to Hiroaki et al. [30] and Gorny [31],
the ideal gas concentration for shredded cabbage storage is 5-7.5%
2 and 15% C2, whereas Hu et al. [32] suggested a 2% 2 and 13%
C2 concentration. The sensitivity of minimally processed vegetables
to modified atmospheres might be quite different compared to the
whole product, withstanding more extreme concentrations of 2 and
C2 compared to whole vegetables and this is due to the fact that they
find fewer obstacles in the diffusion of gases [33].

Colour
The L* colour value and hue angle (h) were selected as the
most suitable parameters for discolouration of cut surfaces of
minimally processed fruits and vegetables [34]. When the colour
became darker, a decrease in L* and hue angle (h) was observed.
Hue angle (h), considered the qualitative attribute of colour. A
higher hue angle represents a lesser yellow character in the assays.
It has been extensively used in the evaluation of colour parameters
in green vegetables [35,36]. The ANOVA showed that: temperature,
treatments, storage time and their interaction had a significant effect
on L* and h (p=0.005).
The lightness of minimally processed cabbage decreased linearly
(Figure 2) from 70.94 6 to 63.8 8.5-61.3 8 for the chemical
treatments, to 60.28 7 for the MAP and to 59.4 6 for the control,
during 22 days at 0C.
At 5C the lightness decreased from 70.94 6 to 63.6 7.9-62.07
8.2 at all cases during 22 days of storage [37]. At the end of storage at
0C lightness change in treatments (ascorbic acid, citric acid, calcium
chloride, MAP) and control varied between 10-16.3%, whereas at 5C
this change ranged between 10-13%.
Among the treatments at 0C, citric acid proved to be the most
effective in maintaining the lightness in minimally processed cabbage
presenting the smallest change of L* value. At 5C no significant
difference in L* values between the control and treated samples was
observed after 22 days of storage. Temperature is the most influential
factor on pigment degradation. For many vegetables, shelf-life can

ascorbic acid

0C

105

100

100

95

citric acid

CaCl2

control

5C

110

105

95
90

85
0

Figure 2: Effect of chemical (ascorbic acid, citric acid, calcium chloride)

and physical (MAP) treatments on changes in lightness (L* ) of minimally
processed cabbage stored at 0C and 5C for 22 days (Mean values N=2
repetitions X 10 samples=20) (I=LSD).

Volume 3 Issue 1 1000131

citric acid

90

0
0

ascorbic acid

110

70

60

citric acid

80

L*

L*

MAP

h*(angle)

citric acid

h*( angle)

ascorbic acid

85
0

14

22

storage time (days)

14

22

storage time (days)

Figure 3: Changes in hue angle (h) of shredded cabbage stored at 0C and

5C which has undergone chemical and physical treatments (ean values
=2 repetitions 10 samples =20) (I=LSD).

be better extended by lowering the storage temperature to slightly

above freezing point of the tissue. Citric acid lowers the product pH,
helps control enzymatic browning [9], and maintains the lightness
of cabbage.
Hue angle values (Figure 3) during storage were significantly
affected by treatments (chemicals and physical) mainly at 0C. The
same observation was made by Manolopoulou et al. [37] for Hue
angle values and chemical treatments.
Hue angle control value decreased rapidly at 0C. Among the
treatments applied, MAP retained better the values of hue angle.
Modified atmosphere packaging and low temperature storage
have been used for many years to reduce respiration rates and the
deterioration of fresh-cut produce. MAP can decrease the rate of
browning reactions due to reduced O2 level and elevated CO2 level
in the surrounding atmosphere [38]. The decrease in hue according
to Bolin et al. [39] could be due to chlorophyll degradation from
the action of chlorophyllase. High concentrations in C2 keep
chlorophyll in cabbage [40] and reduce polyphenoloxidase and
phenolic compounds [41]. Among the chemical treatments citric acid
had the highest antioxidant capacity. Citric acid has been reported
extensively for its inhibitory activity on polyphenol oxidase, PPO
[42] and its antibrowning activity in minimally processed fruits and
vegetables [43]. Our results come in agreement with [27] who worked
on minimally processed Chinese cabbage and Kaur et al. [44] working
on minimally processed Indian cabbage. Ascorbic acid and calcium
chloride are intermediate inhibitors as seen by their hue angle values
(Figure 3).

Overall organoleptic quality

Freshness is the most important quality criterion that consumers
use to evaluate organoleptic quality of fruits and vegetables. Browning
of cut surfaces is one of the most limiting factors on the shelf life of
fresh-cut products and a significant factor of quality deterioration
of shredded cabbage [45]. Organoleptic quality evaluation with
quantifiable physical measurements, determine the shelf-life of fresh
cut vegetables. Changes in quality characteristics (overall acceptance
and browning of cut surfaces), are shown in Figures 4 and 5.
Overall acceptance together with cut surface browning
determined the shelf life of control samples which was 7 days at both
storage temperatures (Figures 4 and 5) using score 3 as the limit.
Among the different treatments (physical and chemical) shredded
cabbage packaged in MAP and stored at 0C, presented the highest
Page 3 of 5

Citation: Manolopoulou E, Varzakas T (2014) Application of Antibrowning Agents in Minimally Processed Cabbage. J Food Nutr Disor 3:1.

doi:http://dx.doi.org/10.4172/2324-9323.1000131

citric acid
0C
control

ascorbic acid

citric acid

CaCl2

control

overall acceptance

overall acceptance

acid ascorbic
CaCl2
MAP

4
3
2
1

5 C

4
3
2
1
0

0
0

10

15

20

25

10

15

20

25

storage time (days)

storage time (days)

Figure 4: Changes in overall acceptance of minimally processed cabbage

with different physical and chemical treatments during storage at 0C
and 5C (Mean values N=2 repetitions X 6 samples per repetition X 6
panellists=72) (I=LSD).

Conclusions
MAP with low 2 concentrations (1.5%) and high concentrations
of C2 (17%) retained the quality of shredded cabbage by reducing
browning of the cut surfaces and, retaining total visual quality. MAP
increased shelf life of shredded cabbage to 22 days at 0C making it
easily marketable. From chemical treatments citric acid better retains
the overall acceptance of shredded cabbage. It can reduce cut surface
browning. Citric acid treatment combined with low temperature
storage (0C) prolonged the shelf life of minimally processed cabbage
for 22 days.
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Author Affiliation

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Department of Plant Production, School of Agricultural Technology,

Technological Educational Institute of Peloponnese, Greece

Department of Food Technology, School of Agricultural Technology,

Technological Educational Institute of Peloponnese, Greece

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