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9 Mujtaba et al.
Int. J. Biosci. 2014
RESEARCH PAPER OPEN ACCESS
Potential role of calcium chloride, potassium permanganate and
boric acid on quality maintenance of tomato cv. Rio grandi at
ambient temperature
Ahmed Mujtaba1*, Tariq Masud1, Shahid Javed Butt2, Mudassar Ali Qazalbash1,
Wajiha Fareed1, Azka Shahid1
1Department of Food Science & Technology, PMAS-Arid Agriculture University Rawalpindi,
Pakistan
2Department of Horticulture, PMAS-Arid Agriculture University Rawalpindi, Pakistan
Key words: Lycopersicon esculentum, postharvest quality, calcium chloride, potassium chloride, boric acid.
http://dx.doi.org/10.12692/ijb/5.9.9-20
Article published on November 10, 2014
Abstract
Tomato (Lycopersicon esculentum Mill) fruits were harvested at turning stage and then treated with different
concentration of calcium chloride, potassium permanganate and boric acid. All these treatment were storage at
ambient temperature (27 oC ± 1) with relative humidity of 80-90 % for 60 days. The result indicated that 2%
calcium chloride and 800 ppm boric acid were effective in maintaining pH and titratable acidity as well as
lycopene and β-carotene respectively then all treated fruits and control. Similarly, higher values for total
phenolic content, ascorbic acid and total antioxidant content were also achieved for the same treatments as
compared to others. It was concluded that 2% calcium chloride and 800 ppm boric acid may be applied for
postharvest treatment of tomatoes for extending shelf life and maintaining functional attributes of tomato.
* Corresponding Author: Ahmed Mujtaba [email protected]
International Journal of Biosciences | IJB |
ISSN: 2220-6655 (Print) 2222-5234 (Online)
http://www.innspub.net
Vol. 5, No. 9, p. 9-20, 2014
10 Mujtaba et al.
Int. J. Biosci. 2014
Introduction
Tomato (Lycopersicon esculentum Mill) is one of the
most important and versatile food crop of world,
which is famous for its ability to be a potential
component of healthy diet. Besides their rich
nutritional composition, the important and valuable
phytochemical components make them highly favored
by the consumer worldwide (Tonucci et al., 1995).
Tomatoes are not only seasonal but highly perishable
after harvesting due to various chemical and physical
processes. The fruit losses its desired quality,
nutritional attributes and some could likely to result
in total waste (Znidarcic and Pozrl, 2006; Idha and
Aderibigbe, 2007). Tomato is climacteric in nature,
having a respiratory peak during ripening due to
release of ethylene (Sammi and Masud, 2007; Wills
and Ku, 2002). To extend the shelf life of tomato, the
respiratory metabolism must be hindered or slowed
down either by low temperature storage or storage in
a high carbon dioxide atmosphere (Kalt et al., 1999).
Various chemicals generally considered as safe
(GRAS) are widely used to improve shelf life of
perishable commodities. Among various permitted
chemicals, calcium delays the post-harvest ripening
controls development of physiological disorders,
improve quality and postharvest decay by improving
the strength of tissues and cell wall (Hong and Lee,
1999). It also reduces the rate of respiration weight
loss and solubilization of pectin hence maintains its
firmness during extended storage (Bhattarai and
Gautam, 2006; Luna-Guzman et al., 1999; Magee et
al., 2002; Conway et al., 1994).
Potassium permanganate can be used as ethylene
absorbent that plays a central role in fruit ripening
(Matsumoto and Ogawa, 1995). It degrades ethylene
into carbon dioxide and water that blocks the
synthesis of endogenous ethylene. In addition
potassium permanganate also has fungi static effect
against Botrytis Cinera that causes significant losses
of firmness in fruits (Bombelli and Wright, 2006).
The potassium permanganate has also significant
affect total soluble solid, titratable acidity and
ascorbic acid content due to its ethylene absorbent
capacity (Briceno et al., 1999).
Similarly, boric acid is also used to inhibit ethylene
production, ripening, and post-harvest disease
incidence and thus reduces postharvest decay by
minimizing microbial load in tomato and increase
shelf life (Wang and Morris, 1993; Prusky et al., 2001;
Nasrin et al., 2008).
For home consumption, tomatoes are usually
purchased when they are ripe; however, market
supplies are made at early ripe or turning stage. There
are enormous losses of tomato in the marketing chain
from harvesting to consumption. The reason is poor
handling and lack of postharvest technologies.
Furthermore, there is limited information about the
overall nutritional implications of storage on the
modern tomato cultivars. The objectives of this study
were to assess the effectiveness of post-harvest
treatment on shelf life of tomato and to determine the
effect of various treatments on their antioxidant
component during ambient storage.
Material and methods
Plant material
Tomato (Lycopersicon esculentum Mill) cv. Rio
Grandi was harvested at USDA stage III (turning
stage) from the fields of Usman Khattar, Taxila,
Pakistan. Fruits were selected with the consideration
that all tomatoes were of uniform size and maturity
level with absence of visual symptoms of any disease.
The fruits were immediately transported in bulk to
the post harvest laboratory of Department of Food
Technology, Pir Mehr Ali Shah Arid Agriculture
University Rawalpindi, Pakistan, where the study was
carried out.
Treatments
Tomato were washed with running water to remove
any dirt and dust and dried at room temperature with
forced air. Fruits were divided into 10 treatment
groups; containing 70 fruits in each and treatments
were applied in the following scheme:
T0 = Control, T1 = 1% CaCl2, T2 = 2% CaCl2, T3 = 3%
CaCl2, T4 = 400 ppm, KMnO4, T5 = 800 ppm, KMnO4,
T6 = Saturated KMnO4, T7 = 400 ppm, Boric acid, T8 =
800 ppm, Boric acid, T9 = 1000 ppm Boric acid
11 Mujtaba et al.
Int. J. Biosci. 2014
Application of chemicals
Fruits (T1, T2 and T3) were treated with calcium
chloride solution by dipped in 1, 2, 3%, respectively
for 1-2 minutes in each treatment, and then it was
dried in air. Sponge cubes of 1 cubic inch cutting (1
inches3) were dipped in 400, 800 ppm and saturated
solution of potassium permanganate. After that these
sponge cuttings were allowed to dry to the extent that
no drop of potassium permanganate falls from them.
Then one cutting of respective treatments (T4, T5 and
T6) was placed in polyethylene bag at one corner and
sealed the side to avoid contact the fruits. The
remaining three treatments (T7, T8 and T9) were
applied by dipping the fruits in 400, 800 and 1000
ppm boric acid solution for 30 to 60 seconds,
removed and dried in air. The following parameters
were evaluated at regular intervals of 15 days during
storage at ambient temperatures.
pH, titratable acidity and ascorbic acid
The pH values were measured by using electronic pH
meter (HANNA pH 210), titratable acidity was
determined by titration with 1 N NaOH and ascorbic
acid was determined by 2, 6-dicholorphenol
indophenols method as described in AOAC (2000).
Determination of total phenolic content
The Folin-Ciocalteau’s reagent was used to measure
the total phenolic content as described by Spanos and
Wrolstad (1990). The sample was extracted using 25
mL methanol and 5 grams tomato sample by shaking.
The methanolic extract was diluted with 6 ml of
double distilled water and 500 l of Folin-Ciocalteau’s
reagent. This reaction mixture was neutralized by
adding 1.5 ml of 20% w/v sodium carbonate, and
sample were vortexed for 20 sec. The samples were
incubated at 45oC for 15 min. and the absorbance was
measured at 765 nm using a CE-2021,
spectrophotometer (CECIL Instruments Cambridge,
England). The total phenolic content was expressed
as gallic acid equivalents (GAE) in mg per kg fresh
weight. A mixture of water and Folin-Ciocaleau’s was
used as a blank.
Determinatrion of antioxidant activity
The antioxidant activity was determined according to
the method described by Chang et al. (2006) with
some modification. Tomato fruits were cut into 10 x
10 x 10 mm3 cubes after cleaning. The sample was
dried at 70oC in a hot air oven until complete removal
of moisture. The sample was then ground to powder
and stored at 40oC until use. The free radical
scavenging activity of DPPH was detected according
to the method of Shimada et al. (1992) with some
modification. To 100L extracted sample (diluted 1:5
(v/v) with methanol), 3.9 ml of freshly prepared
DPPH methanolic solution was mixed and left it
stand for 30 min to react. The absorbance was
determined using a spectrophotometer (CE-2021,
2000 series CECIL Instruments Cambridge, England)
at 517 nm. The percentage DPPH scavenging activity
is expressed by:
%scavenging activity = [1- (Sample Absorbance/Blank
Sample Absorbance)] x 100%.
Determination of lycopene
Lycopene (mg/100g) was determined by the method
of Srivastava and Kumar (2004). Firstly 5-10 g fruit
sample was taken and crushed repeatedly in 5-10 mL
acetone. The acetone extract was transferred to a
separating funnel containing 10-15 mL petroleum
ether and was mixed gently to take up pigment into
petroleum phase. The lower phase was then
transferred to a 100 mL volumetric flask it was
extracted repeatedly with petroleum ether until
became colorless. The petroleum ether extracts were
combined and dried over a small quantity of
anhydrous sodium sulphate. Made the volume up to
100 mL with petroleum ether and measured the
optical density (OD) of solution at 503 nm by
spectrophotometer (CE-2021, 2000 series CECIL
Instruments Cambridge, England) using petroleum
ether as blank. lycopene content was calculated by
the following formula.
Lycopene (mg/100mL) = 3.1206 x O.D of sample x
volume make up x dilution/1 x wt of
sample x 100
Determination of β-carotene
12 Mujtaba et al.
Int. J. Biosci. 2014
β-carotene (mg/100g) was determined by the method
of Srivastava and Kumar (2004). Firstly 5-10 g fruit
sample was taken and crushed in acetone (5-10 mL)
by a pestle and mortor along with few crystal of
anhydrous sodium sulphate. The process was
repeated twice, the supernatant was combined and
transferred to a separating funnel and then 10-15 mL
petroleum ether were mixed thoroughly. Two layers
were separated out on standing. Discarded lower
layer and collected upper layers in a 100 mL
volumetric flask and then it is extracted with
petroleum ether until it became colorless. Make
volume up the 100 mL with petroleum ether and
measured optical density of the solution at 452 nm by
UV-spectrometer CE-2021 (CECIL Instruments
Cambridge, England) using petroleum ether blank.
Then the contents were calculated by the following
formula:
Β-carotene (mg/100 mL) = O.D x 13.9 x 104 x100/wt
of sample x 560 x1000
Statistical analysis
The data obtained was statistically analyzed by two
ways ANNOVA and Duncan’s multiple range tests for
comparison of mean as described by Steel et al.
(1997) using statistics 8.1 software.
Results and discussion
Fruit pH
pH is an important factor to measure the free acid
content in any commodity indicating the degradation
of organic acids into sugar (Bhattarai and Gautam,
2006). In the present study, pH of all treated fruit
and control were measured during 60 days of
ambient storage (Table. 1). All the treatments were
found to maintain the pH throughout storage interval
with gradual decrease in pH at the end of storage. The
lowest mean (after 60 day of storage) was recorded in
T1 (1% CaCl2) with the value of 4.71 indicating slowing
down the ripening process followed by T6 (Saturated
KMnO4) that is 4.79. The pH of all treatments that is
1, 2 and 3% CaCl2, 400, 800 ppm and saturated
KMnO4 and 400, 800 and 1000 ppm H3BO3 after 60
days of storage were 4.71, 4.96, 5.01, 4.81, 4,88, 4.79,
4.98, 4.90 and 5.03 respectively. These results are
coherent with the finding of Andrea et al. (1999) who
reported that the postharvest treatments reduces the
pH between the second and fourteenth day. It was
also observed that the increase in pH of control
sample is much slower than treated fruit except at day
30. The findings are in consistent with the finding of
Diaz-Sobac et al. (1996) and Pila et al. (2010) who
also revealed same pattern of result in mango samples
and tomato respectively. In comparison the pH of the
chemically treated fruit was found to be lower than
that of the pH in control set, which might be due to
the differences in atmosphere created by different
treatments. It is also confirmed from present study
that lower concentration of calcium has significant
effect on pH which are in line with the finding of
Andrea et al. (1999) and Pila et al. (2010).
Total Soluble Sugar (0Brix)
Changes in TSS contents were a natural phenomenon
occurred during ripening due to conservation of
starch into sugar (Kays, 1997). The sugar content
increase depending upon stage of ripeness at harvest
and storage interval (Jimenez et al., 1996). The TSS of
treated fruits were found relatively in higher range
(3.00-6.70 0Brix) as compare to fruits of control set
TSS (3.45-4.15 0Brix) during storage interval. The
treatments and their interactions had not a significant
(p>0.05) effect on TSS value (Table 1). The
interactions among treatments and storage intervals
showed that there is low TSS (oBrix) value in 60th days
in saturated potassium permaganate (T6) and 400
ppm potassium permanganate (T4) that is 3.80 and
3.90 respectively. Our results are in consistent with
the finding of Pila et al. (2010) and Rai et al. (2012),
who reported that the concentration of TSS
progressively increased with storage.
Titratable Acidity
Titratable acidity (TA) is directly related to the
concentration of organic acids present in the fruit as
free acid, anion or combined as a salt (Kays, 1997)
and is often related to maturity (Bhattacharya, 2004).
In the present investigation, titratable acidity
decreased with the passage of time at faster rate in
treated fruit (Table. 1) as organic acids usually
13 Mujtaba et al.
Int. J. Biosci. 2014
declines during ripening as they are respired or
converted to sugars (Tosun et al., 2008). The present
study demonstrate that T4 (400 ppm KMnO4)
exhibited highest percentage of acidity throughout the
storage. The reason is that the use of KMnO4
contributes to an increase in CO2 concentration as it
is a byproduct of ethylene degradation (Sammi and
Masud, 2007) and CO2 accumulation in the fruit
forms carbonic acid resulting in acidiosis (Carrillo et
al., 1995). CaCl2@2% (T2) and 400 ppm boric acid
(T7) proved to be better in decreasing acidity. The low
acidity at the end may be due to atmosphere created
by treatments as described by Kabir (2010) and Batu
and Thompson (1998). During the fruit ripening in
storage, fruit utilizes the acid for the production of
flavoring compounds (Bhattarai and Gautam, 2006).
This view has been further substantiated by Pila et al.
(2010) by citing the reasons that the change in
titratable acidity was mainly due to metabolic
activities during which depletion of organic acid takes
place. Furthermore, microorganisms may use citric
acid as a carbon source resulting in decrease of
titratable acidity (Sammi and Masud, 2007).
Table 1. Chemical evaluation of tomato treated with different post harvest treatment during 60 day at room
temperature.
Storage (Days) Treatment pH TSS (oBrix) TA (mg citric acid /100 ml) AA (mg/100 g)
0 5.00 3.45 0.13 4.00
15 1% CaCl2 4.85 3.80 0.15 3.90
2% CaCl2 4.70 3.30 0.16 2.50
3% CaCl2 4.84 3.00 0.14 2.90
400 ppm KMnO4 4.69 3.40 0.27 4.40
800 ppm KMnO4 4.88 3.30 0.30 3.40
Saturated KMnO4 4.74 3.30 0.17 4.60
400 ppm H3BO4 4.89 3.20 0.16 4.20
800 ppm H3BO4 4.88 3.30 0.17 3.40
1000 ppm H3BO4 4.77 3.10 0.26 3.70
Control 4.90 4.00 0.15 4.30
30 1% CaCl2 4.85 4.15 0.17 12.10
2% CaCl2 4.70 3.90 0.18 11.30
3% CaCl2 4.84 3.60 0.24 10.20
400 ppm KMnO4 4.69 4.20 0.32 3.70
800 ppm KMnO4 4.88 3.70 0.13 9.60
Saturated KMnO4 4.74 3.70 0.17 5.10
400 ppm H3BO4 4.89 3.40 0.13 8.70
800 ppm H3BO4 4.88 3.60 0.12 11.40
1000 ppm H3BO4 4.77 3.40 0.15 16.70
Control 4.70 4.15 0.18 18.00
45 1% CaCl2 4.69 4.20 0.19 21.30
2% CaCl2 4.81 4.70 0.18 18.30
3% CaCl2 4.88 4.00 0.22 12.70
400 ppm KMnO4 4.71 4.70 0.27 13.57
800 ppm KMnO4 4.86 4.20 0.28 10.36
Saturated KMnO4 4.76 3.90 0.15 14.64
400 ppm H3BO4 4.90 3.50 0.20 10.30
800 ppm H3BO4 4.65 3.80 0.15 18.60
1000 ppm H3BO4 4.90 4.50 0.16 13.60
Control 4.76 4.10 0.25 27.00
60 1% CaCl2 4.71 5.80 0.15 22.50
2% CaCl2 4.96 5.70 0.14 19.50
3% CaCl2 5.01 6.00 0.16 13.50
400 ppm KMnO4 4.81 3.90 0.19 14.57
800 ppm KMnO4 4.88 4.80 0.21 12.35
Saturated KMnO4 4.79 3.80 0.20 15.68
400 ppm H3BO4 4.98 5.00 0.13 11.00
800 ppm H3BO4 4.90 5.50 0.13 19.20
1000 ppm H3BO4 5.03 6.70 0.13 15.20
Control 0.00 0.00 0.00 0.00
Treatment (0.024) (0.060) (0.019) (0.185)
Storage interval (0.017) (0.043) (0.013) (0.131)
Interaction (0.053) (0.135) (0.042) (0.413)
Values are the means (n=3). L. S. D values are in bracket. P<0.05
14 Mujtaba et al.
Int. J. Biosci. 2014
Ascorbic Acid
Data pertaining to ascorbic acid contents of tomato
during storage is illustrated in Table 1. It is evident
that ascorbic acid contents increased significantly (P
< 0.05) in all treatments during the storage. The
highest ascorbic acid contents were observed in T1
(1% CaCl2) followed by T2 (2% CaCl2) at 60th day of
storage. The increase in ascorbic acid content is
thought to be indication that the fruit is still in
ripening process (Pila et al. 2010). The accumulation
of ascorbic acid in ripening stages of tomatoes was
also observed by Abushita et al. (1997), Giovanelli et
al. (1999), Lee and Kader (2000) and Kalt et al.
(1999). The high titratable acidity and phenolic
substances are responsible for stability and
accumulation of ascorbic acid (Miller and Evans,
1997). It can also be observed that fruit treated with
calcium chloride showed highest ascorbic acid
followed by boric acid treated lots. The increased in
the ascorbic content was evaluated by the Subbiah
and Perumal (1990). The high carbondioxide
atmosphere affected the ripening rate delaying
ascorbic acid synthesis.
Fig. 1. Effect of post-harvest treatments on storage
interval on the total phenolic (mg GAE/100g) content
of tomato fruit.
Total Phenolic Contents
The oxidation of phenolic contents in fruit and
vegetable are generally associated with the formation
of the brown substances (Lopez-Serrano and Ros
Barcelo, 1999). For this reason, phenolic contents
have been found to be closely related to the degree of
browning (Lee et al., 1990). The data shows that there
is general increase in total phenolic content in all
treated fruits except 1000 ppm H3BO4 (Fig. 1). The
high phenolic contents may be due to the libration of
phenolic compounds from the fruit matrix (Chism
and Haard, 1996). In the present study control set
showed lower values of phenolic contents as Rio
Grandi showed lowest phenolic contents as compared
to other (Hdider et al., 2013). It has also been
observed during storage period that the phenolic
contents varied significantly. These variations are
mainly dependent on ripening stages at the time of
harvest, environmental factors i.e. mainly light and
temperature (Dumas et al., 2003) and analytical
methodology. Moreover, contradictory results could
be attributed to different pattern of different classes
of phenolics during tomato fruit ripening as reported
by Raffo et al. (2002). There is no exact pattern of
change in phenolic content during ripening stages as
the highest phenolic content is observed in Kalvert at
orange red stage of maturity, while in high lycopene
varieties similar trend was found in green to green
orange stage (Hdider et al., 2013). In the present
study calcium chloride treated fruit showed lowest
change in phenolic content and similar results were
also observed by Hdider et al. (2013).
Fig. 2. Effect of post-harvest treatment on storage
interval on the antioxidant activity (%) on tomato
fruit.
Total Antioxidant Activity
The data illustrating the total antioxidant activity is
shown in Figure 2. The antioxidant activity of treated
fruits were found relatively in higher range (73.27 to
93.03 %) as compare to fruit of control set (78.17 to
92.37 %). There are different antioxidant compounds
in tomatoes which are carotenoids, ascorbic acid and
other polyphenols (Giovanelli et al., 1999). Increasing
antioxidant activity during storage may be due to the
15 Mujtaba et al.
Int. J. Biosci. 2014
ripening process and accumulation of phenolic
compounds (Cano et al., 2003). β-carotene was found
to be increased in 30th day of storage and then
decline. The phenolic compounds were found to be
varying during ripening period as some authors
observed that cholorgenic acid declined whereas no
change was observed in rutin (Naguib, 2000; Russo et
al., 2000). It is also found that at 30th day of storage
the lycopene and beta-carotene contents are lower
that is the main reasons for the decrease in total
antioxidant activity storage. The similar trends were
found in the findings of Cano et al. (2003) and
Javanmardi and Kubota (2006). Furthermore, biotic
and abiotic stress affect the pathways involved in
biosynthesis of terpenes, phenolics and nitrogen
containing compound (Javanmardi and Kubota,
2006; Leonardi et al., 2000). Control set showed the
lowest change in antioxidant activity in 40th day of
storage then other treated lots. The results are in
accordance with Cano et al. (2003) who found that
the hydrophilic activity is lower in pink stage. As
shown in the figure, 1% CaCl2 showed higher
scavenging activity, this was probably due to the
combined effect of phenolic compounds on various
concentrations (Chang et al., 2006).
Fig. 3. Effect of post-harvest treatment on storage
interval on the lycopene (mg/100ml) on tomato fruit.
Lycopene
The lycopene is said to be a good index to the level of
maturity (Gautier et al., 2008). In this study,
lycopene accumulation was observed in all treatments
throughout storage period (Fig 3). The figure shows
generally increasing pattern with significant variation
among different treatments. The variations in
lycopene during ripening are attributed by the factors
like plant nutrition, environment and genotype,
which can affect the biosynthesis of carotenoid
(Abushita et al., 2000; George et al., 2004). During
the onset of ripening and tomato development
destruction of food pigment (chlorophyll) occurs and
hence lycopene content increase (Toor and Savage,
2005). The reason for the slow development of
lycopene may be due to the formation of ethylene
which can be decreased by lower oxygen
concentration helpful in delaying lycopene
development (Shi and Maguer, 2000) to increase
shelf life. Tomatoes treated with 800 ppm potassium
permanganate delayed the lycopene biosynthesis.
Similar results have been reported by Nguyen (1999)
and Causse et al. (2002).
Fig. 4. Effect of post-harvest treatment on storage
interval on the β-carotene (mg/100ml) on tomato
fruit
β-Carotene
The accumulation of β-carotene is the index of
maturity due to red color (Abushita et al., 1997). The
current study describes that the β-carotene level in
tomato fruit increased during storage upto 30th day
after which it started to decline (Fig. 4). At the 30th
day tomatoes turned to dark pink color and the higher
values of β-carotene were obtained. Similar trends
were observed in all treatments however, control
showed a slower rate of increase in β-carotene value
(p > 0.05). Carotenoids are stable compounds that
are widely distributed in fruits and vegetable and
responsible for characteristic colour of tomatoes (Pila
et al., 2010; Fraser et al., 2001). The concentration of
-carotene increases in proportion to ripeness with
rapid accumulation of red pigment (Abushita et al.,
1997) which is in consistent with our finding that
there is sharp increase in -carotene during ripening.
16 Mujtaba et al.
Int. J. Biosci. 2014
During fruit ripening, maximum concentration of β-
carotene occurs at turning to breaking stage (Passam
et al., 2007). Tomatoes treated with calcium chloride
delayed the -carotene synthesis in the present study,
which has also been observed by Baloch et al. (2003)
where he also reported increased loss of carotenoid in
calcium chloride treated fruit.
Conclusion
The results of the above experiments indicates that,
relative to controls, all of the nine treatments
contributed to maintain the quality of tomatoes
delaying visual and quality defects as well as
increased the antioxidant capacity. Over all 2 %
calcium chloride and 800 ppm boric acid treatments
exhibited certain physiological and chemical roles for
post-harvest quality of tomatoes.
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