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 a.mujtaba1987@gmail.com

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

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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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