studies on bio-extract coatings and …...certificate-ii studies onthis is to certify that the...

STUDIES ON BIO-EXTRACT COATINGS

AND PACKAGING ON THE SHELF LIFE OF

POMEGRANATE FRUITS CULTIVAR

‘MRIDULA’

BY

SUCHISMITA JENA

(2015A45M)

Thesis submitted to the Chaudhary Charan Singh Haryana

Agricultural University, Hisar in the partial fulfillment of the

requirements for the degree of

MASTER OF SCIENCE

IN

HORTICULTURE- FRUIT SCIENCE

COLLEGE OF AGRICULTURE

CCS HARYANA AGRICULTURAL UNIVERSITY

HISAR- 125 004, HARYANA, INDIA

2017

CERTIFICATE-I

This is to certify that the thesis entitled “Studies on bio-extract coatings and

packaging on shelf life of pomegranate fruits cultivar Mridula” submitted in partial

fulfilment of the requirements for the award of Masters of Science in the subject of

Horticulture- Fruit Science to the Chaudhary Charan Singh, Haryana Agricultural

University, Hisar is a record of bonafide research work carried out by Suchismita Jena,

Admission No. 2015A45M, under my supervision and guidance and that no part of this thesis

has been submitted for any other degree.

The assistance and help received during the course of investigation have been fully

acknowledged.

Dr. R.K. Goyal Place: Hisar (Major Advisor)

Date: Professor

Department of Horticulture

CCS Haryana Agricultural University

Hisar- 125004 (Haryana), India

CERTIFICATE-II

This is to certify that the thesis entitled “Studies on bio-extract coatings and

packaging on shelf life of pomegranate fruits cultivar Mridula” submitted by

Suchismita Jena, Admission No. 2015A45M to the Chaudhary Charan Singh Haryana

Agricultural University, Hisar in partial fulfilment of the requirement for the Masters of

Science in the subject of Horticulture- Fruit Science, has been approved by the student‟s

advisory committee after an oral examination on the same.

MAJOR ADVISOR EXTERNAL EXAMINER

HEAD OF THE DEPARTMENT

DEAN, POST-GRADUATE STUDIES

ACKNOWLEDGEMENT

A precious debt as that of learning is the only debt, which is not only difficult but also impossible

to repay except perhaps through gratitude. The very idea of this work makes me amused in thanking to all

those who helped me in achieving this milestone of my academic pursuit. I would like to extend my

profound regards and deep sense of gratitude to my Major Advisor Dr. R.K. Goyal, Professor in the

Department of Horticulture. I am indebted to his advice, noble counseling and for providing necessary

facilities during my study. His every cordial behavior has imprinted everlasting impression on my mind. It

will not be an exaggeration to say that without his sincere efforts, the culmination of present research work

as well as my M.Sc. degree can’t be imagined.

With an overwhelming and genuine sense of obligations, I am thankful to the members of the

advisory committee Dr. Anil Kumar Godara, Principal Scientist (co-major advisor) who taught me how to

use sensors, and member of advisory committee Dr. J.K. Sandooja, Professor and Head, Department of

Plant Physiology, Dr. Yogesh Jindal, Asstt. Scientist, Department of Genetics and Plant Breeding and

Dean PGs nominee Dr. Pala Ram, Principal Scientist, Department of Entomology for their valuable

counselling and constructive suggestions that were much helpful throughout my research progress. I would

like to put on record my sincere thankfulness to Dr. Ashwini Kumar, Professor and Head, Department of

Horticulture for providing me the necessary facilities during my study programme. I also extend my sincere

thanks to Dr. S.K. Bhatia and all other faculties of the Department of Horticulture for their humanitarian

character and moral support for me during entire study period.

I avail this opportunity to express my deepest indebtedness to Dr. M.K. Rana, Department of

Vegetable Science, who sustained his incessant and exhilarating support to me.

I like to express my heart full thanks to my seniors, particularly Monalisha Hota, Ashish kumar

Panda, Reenu Kukshal, Anuradha Bishnoi and Vinita Rajput who have rendered their immense care and

support during my entire M.Sc. programme. My special thanks to my dear batch mates, Suchismita

Balabantaray, Bishnu, Abhilash, Biswabiplab, Palak, Ekkta, Shashi, Reetika, Komal, Neha, Rupakshi,

Priya, Gouravkant and Vikash who helped me a lot during my research as well as during my studies. I am

thankful to ICAR for providing me National Talent Fellowship during my study period.

At last, I owe this achievement to my father Shri Pramod Kumar Jena, my mother Smt. Sabita

Jena, my grandmother Smt. Saroja Rout and my younger brother Rajib Ranjan Jena along with all my

family members and well-wishers. Finally, I bow my head on the foot of “Sai Baba” for providing me

patience and strength without which it would not have been blossomed.

I convey my respect and gratitude to all my other near and dears who could not be

queued to find a mention here.

Place: HISAR

Dated: 29.05.2017 Suchismita Jena

CONTENTS

CHAPTER TITLE PAGE NO.

I INTRODUCTION 1-3

II REVIEW OF LITERATURE 4-13

III MATERIALS AND METHODS 14-21

IV RESULTS 22-42

V DISCUSSION 43-52

VI SUMMARY AND CONCLUSION 53-54

BIBLIOGRAPHY i-vi

APPENDIX I-II

LIST OF TABLES

Table Description Page

1. Physical characteristic of pomegranate cv. Mridula 22

2. Biochemical characteristic of pomegranate cv. Mridula 22

3. Effect of bio-extracts coating on physiological loss in weight (%) of pomegranate

cv. Mridula 23

4. Effect of bio-extracts coating on decay loss (%) of pomegranate cv. Mridula 24

5. Effect of bio-extracts coating on juice content (%) of pomegranate cv. Mridula 24

6. Effect of bio-extracts coating on total soluble solids (%) of pomegranate cv.

Mridula 25

7. Effect of bio-extracts coating on titratable acidity (%) of pomegranate cv.

Mridula 26

8. Effect of bio-extracts coating on total soluble solids to acid ratio (%) of

pomegranate cv. Mridula 26

9. Effect of bio-extracts coating on pH of pomegranate cv. Mridula 27

10. Effect of bio-extracts coating on ascorbic acid (mg/100g) of pomegranate cv.

Mridula 28

11. Effect of bio-extracts coating on anthocyanin content (mg/100 g) of pomegranate

cv. Mridula 28

12. Effect of bio-extracts coating on total sugars (%) of pomegranate cv. Mridula 29

13. Effect of bio-extracts coating on reducing sugars (%) of pomegranate cv.

Mridula 30

14. Effect of bio-extracts coating on non-reducing sugar (%) of pomegranate cv.

Mridula 31

15. Effect of bio-extracts coating on organoleptic rating of pomegranate cv. Mridula 31

16. Effect of packaging materials on physiological loss in weight (%) of


17. Effect of packaging materials on decay loss (%) of pomegranate cv. Mridula 33

18. Effect of packaging materials on juice content (%) of pomegranate cv. Mridula 33

19. Effect of packaging materials on total soluble solids (%) of pomegranate cv.

Mridula 34

20. Effect of packaging materials on titratable acidity (%) of pomegranate cv.

Mridula 34

21. Effect of packaging materials on total soluble solids to acid ratio (%) of


22. Effect of packaging materials on pH of pomegranate cv. Mridula 35

23. Effect of packaging materials on ascorbic acid (mg/100g) of pomegranate cv.

Mridula 36

24. Effect of packaging materials on anthocyanin content (mg/100 g) of pomegranate

cv. Mridula 37

25. Effect of packaging materials on total sugars (%) of pomegranate cv. Mridula 37

26. Effect of packaging materials on reducing sugars (%) of pomegranate cv.

Mridula 38

27. Effect of packaging materials on non-reducing sugars (%) of pomegranate cv.

Mridula 38

28. Effect of packaging materials on organoleptic rating of pomegranate cv. Mridula 39

List of Figures

List of Plates

Fig. Description

Page

1. Effect of various packaging film on the CO2 concentration (%) in modified

atmospheric packaging of pomegranate cv. Mridula 39

2. Effect of various packaging film on the O2 concentration (%) in modified

atmospheric packaging of pomegranate cv. Mridula 40

3. Effect of various packaging film on the temperature variation (⁰C) in

modified atmospheric packaging of pomegranate cv. Mridula 41

4. Effect of various packaging film on the RH (%) in modified atmospheric

packaging of pomegranate cv. Mridula 42

5. Effect of different bio-extracts coating on physiological loss in weight (%) in


6. Effect of different bio-extract coating on decay loss (%) in pomegranate cv.

Mridula 44

7. Effect of different packaging film on physiological loss in weight (%) in


8. Effect of different packaging film on the decay loss (%) in pomegranate cv.

Mridula 48

Plate Description

1. Pomegranate fruits procured from Mangiana in CFB boxes

2. Opened pomegranate fruit

3. Pomegranate arils

4. Fresh matured Aloe vera leaves

5. Separation of colourless hydro-parenchyma from outer rind

6. Prepared Aloe vera extract

7. Air drying of fruits after the coating on it

8. Peeling of ginger

9. Fruits dipped in ginger extract

10. Fresh mint leaves

11. Prepared mint extract

12. Pomegranate fruit in LDPE

13. Pomegranate fruit in PP

14. Pomegranate fruit wrapped in cling film

15. Fruits wrapped in cellophane paper

16. Use of sensors to evaluate the various components of MAP in pomegranate

LIST OF ABBREVIATIONS

% : Per cent

/ : Per

°Brix : Degree Brix

°C : Degree Celsius

°F : Degree Fahrenheit

µ : Micron

Cm : Centimetre

cv. : Cultivar

D Days

et al. : et alii (and others)

etc. : Etcetera

G : Gram

H Hours

i.e. : id est (that is)

Mg : Milligram

Ml : Millilitre

OD : Optical Density

var. : Variety

viz. : videlicet (namely)

1

CHAPTER-I

INTRODUCTION

Pomegranate (Punica granatum L.), a member of Punicaceae family with

chromosome number 16, is a fruit-bearing deciduous shrub. Punica is the only known genus

of Punicaceae family. Punica granatum has been classified into two subspecies, i.e.,

chlorocarpa and porphyrocarpa, which have been established based on ovary colour, a stable

feature retained even when they are reproduced by seeds. The subspecies chlorocarpa is found

mainly in the Transcaucasia, whereas, the second subspecies porphyrocarpa is distributed

chiefly in Central Asia. Pomegranate is one of the oldest known edible fruits popularly known

as Anar, a non-climacteric many seeded berry. It is one of the important fruit crops of tropical

and subtropical, arid and semiarid regions of the world. Its fruits cannot be sweet unless the

temperature is high for a sufficiently long period. Humid climate lowers the quality of fruits.

The versatile adaptability, hardy nature, low maintenance cost, fine table and therapeutic

values and possibilities to thrive in the rest period when irrigation potential is generally low

are its main features.

According to De-Candolle (1967), pomegranate is an ancient fruit, which has

originated from Southwest Asia, probably in Iran and some adjoining countries. Even though

it is native to Iran, it is extensively cultivated in Spain, Morocco, Egypt, Afghanistan and

Balauchistan. The cultivation has also been initiated on small scale in countries like United

States of America especially California and Florida, Mexico, Palestine, Israel, China, Japan,

Burma, USSR, Pakistan and many parts of India (Singh, 2000). While in India pomegranate is

found growing from Kanyakumari to Kashmir, but commercially, it is cultivated only in

Maharashtra, which is popularly known as pomegranate basket of India. Its plantation is also

seen on small scale in Gujarat, Rajasthan, Karnataka, Tamilnadu, Andhra Pradesh, Uttar

Pradesh, Punjab and Haryana. In India, the area under pomegranate is 0.193 million hector

and production is 2.198 million tonnes (Saxena, 2015).

The pomegranate fruit is balausta, having edible portion as the juicy outgrowth of

seed called arils. The fruits are generally harvested at full ripe stage with a waxy shiny surface

of reddish yellow or greenish red peel, depending on the cultivar. Fruits are an excellent

dietary source of organic acids, soluble solids, protein, fat, carbohydrates, tannin, vitamin C,

thiamine, riboflavin, niacin, nicotinic acid, agollic acid and minerals like calcium, iron,

2

phosphorus, and magnesium. Anthocyanins present in pomegranate fruit have shown

antioxidant activity higher than α-tocopherol, ascorbic acid and β-carotene. Along with

nutritive value, it also has anti-mutagenic, anti-inflammatory and anti-hypertension activities

(Bhowmik et al., 2013). It reduces liver injury, lowers the risk of heart diseases and helps to

fight against bacterial and fungal infections. The pomegranate fruit juice is highly beneficial

for leprosy patients. To highlight its importance, it was chosen as a symbol of 18th

International Horticultural congress held during 1970, showing pomegranate in a basket.

Post-harvest losses in fresh fruits was estimated approximately 10-20%. In India,

improper handling leads to post-harvest loss (6.70-15.88%) of fruits (Jha et al., 2015), thus,

reduces profit margin of the growers, therefore, it is quite essential to reduce these losses. In

countries like India where the horticultural infrastructures are lacking, the growers are liable

to depend upon other simple and low cost techniques to maintain its fruit quality attributes.

For the last few years, the use of various chemicals and waxing materials at pre- and post-

harvest stages is gaining importance among growers in order to enhance the shelf life of fruits

as well as meeting their quality standards but the application of these substances is believed to

be unsafe, as they are prone to objectionable residue on fruit surface, which may have direct

effect on human health. In order to overcome these short comings, there is an urgent need to

substitute the ecologically and economically unsafe substances with substances of biological

origin. The bio-extracts exhibiting growth regulating, fungicidal and insecticidal properties

can be exploited for retaining freshness and enhancing the shelf life of horticultural crops as

well as meeting their quality standards. Among the biological substances, botanical

formulations of Aloe vera, ginger and mint are the most definite alternative to overcome the

undesirable effects of chemicals (Chauhan, 2011), thus, herbal extracts are being increasingly

studied as additive in edible coating on fruits and vegetables, which would be an interesting

and innovative for commercial application and an alternative to the use of post-harvest

chemical treatment leading to the increment of shelf life of fruits and vegetables.

Although pomegranate is a non-climacteric fruit but it is subjected to continuous

physiological and biochemical changes after harvest with severe problems during post-harvest

handling, storage and marketing. Appearance, especially skin and aril colour, is an important

quality trait for the marketing of pomegranate. Many factors affect appearance including

bruising, water loss, decaying and the development of physiological disorders during storage.

The major cause limiting the storage potential of pomegranate is the development of decay,

which is often caused due to the presence of fungal infection especially in blossom end of the

fruit at harvest. Several post-harvest methods have been evaluated for long-term storage of

pomegranate, of which, the most successful method in reducing decay and physiological

disorders is the use of controlled atmosphere (CA) storage, however, CA storage facilities are

3

not always economical and available in many countries. The modified atmosphere packaging

(MAP) is a simple, economical and effective method for delaying post-harvest deterioration,

and maintaining quality of pomegranate (Selcuk and Erkan, 2016). Packaging material also

plays a significant role in attracting the consumers and prolonging the storage period of many

fruits and vegetables. It is often desirable to generate an atmosphere around the fruit low in

oxygen (O2) and/or high in carbon dioxide (CO2) to influence the metabolism of the packed

produce, which can result in reduction of respiratory activity, delaying softening, ripening,

senescencing and reducing incidence of physiological disorders and pathogenic infestations.

Therefore, the present experiment Studies on bio-extract coating and packaging on the shelf

life of the pomegranate fruits cultivar Mridula was planned with the following objectives:

1. To evaluate the effect of different bio-extracts coating on shelf life of pomegranate

2. To evaluate the effect of packaging on the shelf life of pomegranate

4

CHAPTER-II

REVIEW OF LITERATURE

In this chapter, an attempt has been made to review, update and collect literature on

research topic and thereby provide an overview of the subject. The perusal of literature

revealed not much published literature on Studies on bio-extract coatings and packaging on

the shelf life of pomegranate fruits cultivar Mridula.. Therefore, the relevant research work

done on these aspects of other fruit crops has been reviewed in this chapter under following

heads and subheads:

2.1 Effect of different bio-extract coatings

2.2 Effect of packaging

2.3 Effect of coating and packaging

2.1 Effect of different bio-extract coatings

The post-harvest losses in fresh fruits is estimated approximately 10-20%, therefore, it is quite

essential to reduce these losses. In countries like India where the horticultural infrastructures

are lacking, the growers are liable to depend upon other simple and low cost techniques to

maintain its fruit quality attributes. For the last few decades, the use of various chemicals and

waxing materials at pre- and post-harvest stages is gaining importance but the application of

these substances is believed to be unsafe, as they are prone to objectionable residue on fruit

surface. Hence, there is an urgent need to substitute the chemical substances with substances

of biological origin. The bio-extracts exhibiting growth regulating, fungicidal and insecticidal

properties can be exploited for retaining freshness and enhancing the shelf life of horticultural

crops as well as meeting their quality standards. Among the biological substances, botanical

formulations of Aloe vera, ginger and mint are the most definite alternative to overcome the

undesirable effects of chemicals (Chauhan, 2011). Aloe vera extract is used as anti-fungal,

anti-bacterial and anti-inflammatory (Serrano et al., 2005). Aloe vera gel has been proven one

of the best edible and biologically safe preservative coatings for different types of food

because of its film-forming properties, antimicrobial actions, biodegradability and

biochemical properties (Misir et al., 2014).

2.1.1 Physiological loss in weight (%)

In cold store, Aloe vera gel treatment was found significantly helpful in decreasing

dehydration and rapid weight loss in sweet cherries (Martinez-Romero et al., 2006). Aloe vera

5

gel (1:3) coating significantly reduced weight loss (9.99±2.1% compared to 13.79±0.13% in

control) in strawberry (Singh et al., 2011). Aloe vera gel coating (5 or 10%) was found

effective in suppressing weight loss in apple cv. Granny Smith during cold storage (Ergun and

Satici, 2012). Aloe vera gel applied as edible coating in sweet orange fruit reduced weight

loss during post-harvest storage (Adetunji et al., 2012). Aloe vera gel (33%) coating reduced

weight loss in sweet cherry (Asghari et al., 2013). The weight loss of papaya coated with Aloe

vera gel (100%) was recorded 7.93% against 22.5% in uncoated fruits at 25-29ºC temperature

and 82-84% relative humidity (Misir et al., 2014). Aloe Vera coating resulted in decreased

weight loss in oranges (Kumar and Bhatnagar, 2014). Aloe vera gel solution (1.5%) acts as a

physical barrier in reducing the weight loss in papaya during storage at room temperature

(Sharmin et al., 2015).

Cushioning with spearmint leaves reduced the physiological loss in weight due to

their effect on slowing down of physiological processes responsible for weight loss (Chauhan,

2011).

2.1.2 Decay loss (%)

Aloe vera gel applied as an edible coating has been found effective in reduction of microbial

spoilage of several fruits such as sweet cherry, table grape and nectarine (Valverde et al.,

2005). Aloe vera gel coating on sweet cherries was found significantly delaying stem

browning and decreasing microbial population during cold storage (Martinez-Romero et al.,

2006). Aloe vera gel coated sweet cherries and table grapes, which showed a reduction in the

population of mesophilic aerobic bacteria and yeast and mold during storage (Martínez-

Romero et al., 2006; Valverde et al., 2005). The population of mesophilic aerobic bacteria,

mould and yeast was significantly reduced on Aloe vera treated sweet cherries (Martinez-

Romero et al., 2006). Aloe vera gel prevented softening and oxidative browning and reduced

the risk of microbial contamination in fruits such as apple, banana, cherries, grapes and

papaya (Ahmad et al., 2009; Valverde et al., 2005; Martinez-Romero et al., 2006; Marpudi et

al., 2011). The decay incidence was significantly lower in sweet cherry treated with Aloe vera

gel and nitric oxide (NO) in combination than treated either with Aloe vera gel or nitric oxide

(Asghari et al., 2013). Aloe vera gel 100% + ascorbic acid 1% and citric acid 1% reduced

microbial spoilage in pomegranate arils for 12 days during storage in rigid polypropylene

boxes at 3°C temperature (Martinez-Romero et al., 2013). Aloe vera gel 50 and 75% as a

coating on mango fruits significantly reduced weight loss and improved post-harvest shelf life

at 15-22 and 13°C temperature (Sophia et al., 2015). Aloe vera gel coating effectively

controlled or inhibited microbial populations in strawberry fruits (Sogvar et al., 2016).

6

Red ginger and turmeric rhizome extracts 30% concentration inhibited 100 and 34%

growth of Thielaviopsis paradoxa fungus in Salak pondoh, respectively (Dharmaputra et al.,

2013).

Fruit treated with mint leaf extract 20% retained higher pectin content and proved to

be highly effective in reducing spoilage throughout storage (Chauhan, 2011). Edible coatings

with 0.5 or 1.0% mint essential oil showed in vitro antimicrobial activity as they either

reduced or inhibited the growth of E. coli and Salmonella in fresh-cut pineapple (Bitencourt et

al., 2014).

2.1.3 Quality retention

Aloe vera gel applied as an edible coating has been found effective in quality retention of

several fruits such as sweet cherry, table grape and nectarine (Valverde et al., 2005). Aloe

vera gel (1: 3 ratio) treatment in strawberries significantly maintained quality characteristics

when stored at 5ºC temperature and 95% relative humidity (Singh et al., 2011). Aloe vera gel

coating (5 or 10%) was effective in delaying the reduction of soluble solids and titratable

acidity in apple cv. Granny Smith during cold storage (Ergun and Satici, 2012). Aloe vera gel

coating delayed softening, ascorbic acid and TSS losses and maintained the quality of the

orange fruits (Adetunji et al., 2012). Coating of Aloe vera gel preserved the quality of apple

cv. Granny Smith (Ergun and Satici, 2012). The coating of peach and plum with Aloe vera or

Aloe arborescens gel significantly delayed the changes in quality parameters, such as colour

changes, reduction of acidity and increase in solid to acid ratio during post-harvest ripening at

20ºC temperature (Guillen et al., 2013). Aloe vera coating improved the quality of stored

kiwifruit slices at 4±1ºC (Benitez et al., 2013). Aloe arborescens gel was more effective than

Aloe vera gel for use as edible coating for preserving the quality of climacteric fruit (Guillen

et al., 2013). Aloe vera gel 50% as coating and storage temperature of 13ºC maintained the

quality of mango fruits (Sophia et al., 2015). Dipping in Aloe vera extract represented as a

natural tool for quality maintenance of minimally processed table grapes stored for 15 days at

4ºC temperature (Alberio et al., 2015)

Use of 10% wax in combination with 30% red ginger extract for coating salak

pondoh maintained fruit quality at room temperature (28.0±1.5°C) and relative humidity 65-

75% for 12 days against up to 9 days in control (Dharmaputra et al., 2013).

2.1.4 Total soluble solids (%)

Coating or wrapping in films significantly affects the total soluble solids of Citrus sinensis

(Adetunji et al., 2012). Aloe vera gel (100%) coated guava variety L-49 recorded maximum

total soluble solids (Verma et al., 2012). Aloe Vera coated oranges showed higher TSS as

compared to control fruits (Kumar and Bhatnagar, 2014). The lowest total soluble solids

7

(7.610%) was found in papaya preserved with 1.5% Aloe vera gel solution at 29-31°C

temperature (Sharmin et al., 2015).

2.1.5 Titratable acidity (%)

The maximum acidity (0.78%) was recorded in guava cv. L-49 coated with 100% Aloe vera

gel during storage (Verma et al., 2012). Treatment with either ascorbic acid, citric acid, or in

combination with Aloe vera gel maintained or increased titratable acidity in pomegranate arils

after 8 days of storage in rigid polypropylene boxes at 3°C temperature (Martinez-Romero et

al., 2013). Coating of Aloe vera increased the titratable acidity in oranges (Kumar and

Bhatnagar, 2014). Aloe vera gel 50 and 75% as a coating reduced the decrease in titratable

acidity of mango fruits at 13°C temperature (Sophia et al., 2015). Papaya treated with 1.5%

Aloe vera gel solution had comparatively higher titratable acidity (0.41%) at 12 days of

storage at 29-31°C temperature (Sharmin et al., 2015).

2.1.6 pH

Aloe vera gel treatment had no effect on apples cv. Granny Smith during storage at 2°C

temperature (Ergun and Satici, 2012). Aloe-pectin treatment showed lesser decrease in pH of

jujube fruits for 45 days at 5±2°C temperature (Padmaja and Bosco, 2014). Aloe vera gel

coating reduced the increase in pH of mango fruits at 13ºC temperature (Sophia et al., 2015).

2.1.7 Anthocyanin (µg/100 g)

Aloe vera gel 100% + ascorbic acid 1% and citric acid 1% coating on pomegranate arils

increased total anthocyanin after 8 days of storage in rigid polypropylene boxes at 3°C

temperature (Martinez-Romero et al., 2013).

2.1.8 Ascorbic acid (mg/100 g)

The maximum ascorbic acid content (299.6 mg/100 g) was recorded in 100% Aloe vera gel

coated guava cv. L-49 followed by whey protein and corn starch coated guava during storage

at room temperature (Verma et al., 2012). Aloe vera gel coating significantly reduced

ascorbic acid loss (6.48±0.50) in orange due to the low oxygen permeability of gel coating,

which lowered the activity of enzymes and prevented oxidation of ascorbic acid at ambient

room temperature (Adetunji et al., 2012). The coating on sweet cherry with Aloe vera gel

33% and nitric oxide (NO) 5μmol per litre in combination resulted higher vitamin C than

individual treatment (Asghari et al., 2013). Ascorbic acid content was higher in 1.5% Aloe

vera treated papaya fruits at room temperature (Sharmin et al., 2015).

2.1.9 Sugars (%)

Aloe vera gel coated sweet oranges showed the reducing sugars 7.61±0.45 as against

4.94±0.92 in uncoated fruits (Adetunji et al., 2012).

8

2.1.10 General appearance

Aloe vera gel imparted attractive natural shine to sweet cherry fruit quite close to the freshly

harvested fruits (Martinez-Romero et al., 2006). The sensory analysis reveals that sweet

cherry treated with Aloe vera gel showed a decrease in colour change and maintenance of

fruit visual aspect without any detrimental effect on taste, aroma and flavour (Martinez-

Romero et al., 2006). Aloe vera gel coating (5 or 10%) was effective in impeding changes in

appearance of apple cv. Granny Smith during cold storage (Ergun and Satici, 2012).

2.1.11 Overall acceptability

Guava cv. L-49 coated with 100% Aloe vera gel had maximum overall acceptability score

(Verma et al., 2012). Aloe vera 100% + ascorbic acid 1% and citric acids 1% treated

pomegranate arils perceived higher scores in sensory analysis by judges for flavour, texture,

aroma, colour and purchase decision during storage for 12 days at 3°C temperature (Martinez-

Romero et al., 2013).

2.1.12 Shelf life

Aloe vera gel significantly decreased respiration rate, accelerated softening and ripening of

sweet cherries, thereby extended storability in cold store (Martinez-Romero et al., 2006).

Martinez-Romero et al. (2006) confirmed the role of A. vera gel applied as a coating in

delaying post-harvest softening of sweet and sour cherry, table grape, strawberry, papaya and

nectarine fruits during ripening. Aloe vera gel (1: 3 ratio) coating extended the storability of

strawberry up to 16 days at 5°C temperature and 95% relative humidity (Singh et al., 2011).

Aloe vera gel applied as edible coating on sweet orange fruit had beneficial effects in

retarding the ripening process and offered possibility to extend the shelf life by providing a

semi-permeable barrier to gases and water vapour (Adetunji et al., 2012). The coating of

peach and plum with Aloe vera or Aloe arborescens gel significantly delayed ethylene

production, the effect being higher in plum, which had the highest ethylene production rates

(Guillen et al., 2013). The post-harvest treatment of table grapes with salicylic acid and Aloe

Vera gel extended the storage life and maintained their quality (Asghari et al., 2013). Coating

with Aloe vera gel significantly delayed the increase in respiration rate, rapid weight loss,

colour change, accelerated aging and ripening allowing the sour cherries storability extension

without any detrimental effect on taste, aroma and flavour (Ravanfar et al., 2014). Aloe vera

gel coating extended the shelf life of papaya fruits for a period of 15 days at low temperature

(Kumar and Bhatnagar, 2014). The storability of grapes coated with Aloe vera gel could be

extended up to 35 days at 1°C temperature (Kumar and Bhatnagar, 2014). Aloe vera gel 50

and 75% as coating on mango fruits significantly improved the post-harvest life at 15-22 and

13°C temperature (Sophia et al., 2015). Papaya coated with 1.5% Aloe vera gel solution could

be stored as long as 15 days at 29-31°C temperature (Sharmin et al., 2015).

9

2.2 Effect of packaging

Polymeric film packaging creates modified atmosphere around the produce inside the package

allowing lower degree of gases and interplays with physiological processes of commodity

resulting in reduced rate of respiration, transpiration and other metabolic processes of

strawberry fruits (Zagory and Kader, 1988). Storage in modified atmosphere packed with

perforated polypropylene affected the aroma development in strawberry cv. Korona fruits at

5°C temperature (Nielsen and Leufve, 2008).


Packaging of Blood Red orange in polyethylene film was most effective in reducing weight

loss during storage for five weeks at room temperature (Ahmad et al., 1989). Packaging of

Mango cv. Keitt individually in low and high density polyethylene films (LDPE and HDPE)

showed reduced weight loss and did not result any off-flavors for four weeks at 20ºC

temperature and 67% relative humidity (Gonzalez et al., 1990). Wrapping reduced weight

loss of avocado fruit during ripening more than waxing at 22°C temperature (Joyce et al.,

1995). Shrink wrapping was reported lower weight loss in pomegranate and papaya fruits

during storage (Nanda et al., 2001; Singh and Rao, 2005). Perforated polypropylene

packaging reduced weight loss of strawberries by preventing dehydration and also helped in

maintaining appearance of fruits for 10 days at 5°C temperature (Nielsen and Leufve, 2008).

Fruits of aonla cv. Kanchan packed in HDPE showed minimum physiological loss in weight

(6.36%) at room temperature (Vishwakarma et al., 2012). Shrink film wrapped pear cv.

Patharnakh fruits recorded lowest PLW (3.50%) as compared to unwrapped fruits, showing

highest PLW (6.20%) during four weeks of storage under super-market conditions, i.e., 20-

21ºC temperature and 85-90% relative humidity (Mahajan et al., 2013). The fruits of apple cv.

Royal Delicious packed in Cryovac heat shrinkable film showed least physiological loss in

weight for about 35 days at ambient room temperature (Sharma et al., 2013). The shrink film

packaging significantly reduced kinnow weight loss during storage under supermarket

conditions (Mahajan and Singh, 2014). Eva apple cultivar fruits in high density polyethylene

film packaging (HDPE) showed lowest ethylene production possibly because of the higher

concentrations of CO2 inside the package, subjected to lowest percentage of mass loss for 225

days at 0.5±0.5°C temperature (Fante et al., 2014). The minimum per cent loss in weight

(5.49%) was recorded in strawberry cv. Sweet Charlie fruits packed with LDPE 50 micron at

a temperature of 20±2°C and relative humidity 80-90% (Panda et al., 2016).


Loquat fruits wrapped in low density polyethylene (LDPE) showed minimum browning index

(Akhtar et al., 2012). The fungal infection in aonla cv. Kanchan fruits packed in HDPE film

was observed minimum under ambient room conditions (Vishwakarma et al., 2012). Apple

10

cv. Royal Delicious packed in cryovac heat shrinkable films showed least decay loss without

any adverse effect on quality for about 35 days under ambient room conditions (Sharma et al.,

2013). Modified atmosphere packaging prevented the occurrence of decay loss in fruits of

strawberry cv. Sweet Charlie only for a day at 20±2°C temperature and 80-90% relative

humidity (Panda et al., 2016).


Airtight polyethylene bags reduced moisture loss and polysaccharides hydrolysis, resulting in

less increase in loquat fruit TSS (Akhtar et al., 2012). The maximum TSS content was

observed in LDPE wrapped aonla cv. Kanchan fruits at ambient room temperature

(Vishwakarma et al., 2012). The TSS content of pear cv. Patharnak fruits packed in shrink

film increased slowly and steadily up to 21 days (13.25%), thereafter, declined gradually 28

days after storage (10.95%) at 20-21°C temperature and 85-90% relative humidity (Mahajan

et al., 2013). The shrink film wrapped kinnow fruits maintained 9.45% TSS after a storage

period of 5 days, which reached to a peak value of 12.30% after 20 days of storage and then

declined under supermarket conditions (Mahajan and Singh, 2014).


Slight increase in titratable acidity was detected in fruits of strawberry cv. Honeoye packed in

polypropylene during 10 days storage at 5°C temperature (Nielsen and Leufve, 2008). The

maximum acidity (0.52%) was recorded in peach fruits shrink wrapped in 50μ LDPE and 20μ

LLDPE at the end of 42 days storage at 5±1ºC temperature and 90-95% relative humidity

(Singh et al., 2009). Loquat fruit packed in perforated low density polyethylene (0.25%)

showed significantly lowest (0.32%) titratable acidity (Akhtar et al., 2012). Kinnow fruit

packed in shrink wrap film showed highest titratable acidity (0.54%) at 18-20°C temperature

and 80-85% relative humidity (Mahajan and Singh, 2014).


Ascorbic acid content of loquat decreased slightly in MAP as compared to control fruits

during storage of six weeks (Amaros et al., 2008). Fruits of aonla cv. Kanchan packed in

HDPE showed highest ascorbic acid (518.3 mg) at room temperature (Vishwakarma et al.,

2012). The maximum vitamin C content (19.18 mg) was observed in shrink film wrapped

kinnow fruits at 18-20°C temperature and 80-85% relative humidity (Mahajan and Singh,

2014). Fruits of strawberry cv. Sweet Charlie packed in LDPE 50 film bags showed better

level of ascorbic acid (31.56 mg/100 g) against control fruits stored at a temperature of

20±2ºC and relative humidity 80-90% (Panda et al., 2016).

11

2.2.6 Sugars (%)

Different packaging materials such as high density polyethylene, low density polyethylene,

newspaper, tissue paper and paddy straw increased the total and reducing sugars with the

advancement of storage in sand pear (Mohla et al., 2005). The sugar levels in fruits of

strawberry cv. Korona packed in perforated polypropylene dropped slightly during 10 days of

storage at 5ºC temperature (Nielsen and Leufve 2008). The pear cv. Patharnak fruits packed

with shrink film recorded maximum sugar content (8.36%) while lowest in control fruits

(7.80%) under super-market conditions, i.e., 20-21°C temperature and 85-90% relative

humidity (Mahajan et al., 2013).

2.2.7 General appearance

Polyethylene film packaging was most effective in retaining external characteristics of Blood

Red orange during storage for five weeks at room temperature (Ahmad et al., 1989). Apples

stored under modified atmosphere packaging had more appealing colour than those stored

under ambient cold storage conditions (Rocha et al., 2004). The 20μ linear low density

polyethylene (LLDPE) slightly checked the colour change in peach fruits during storage at

5±1°C temperature and 90-95% relative humidity (Singh et al., 2009).

2.2.8 Organoleptic rating

The organoleptic rating of Blood Red orange packed in polyethylene film was higher during

storage for five weeks at room temperature (Ahmad et al., 1989). The sensory attributes of

packed strawberry cv. Chandler fruits stored in micro-atmosphere with O2 <3%, CO2 >20%

levels and temperature 1ºC became less desirable (Shamaila et al., 1992). Micro-perforated

polypropylene packaging increased off-flavour compounds in fruits of strawberry cv.

Camarosa (Sanz et al., 1999). Perforated polypropylene packaging could be used for

maintaining strawberry quality during extended storage at 5°C temperature (Nielsen and

Leufve, 2008). The pear cv. Patharnak fruits wrapped in shrink film were rated as very much

desirable to moderately desirable after 3 and 4 weeks of storage under super-market

conditions, i.e., 20-21°C temperature and 85-90% relative humidity possibly due to creation

of favourable gaseous atmosphere (Mahajan et al., 2013). The heat shrink film wrapped

kinnow fruits showed gradual and steady increase in organoleptic rating up to 20 days at 18-

20°C temperature and 80-85% relative humidity (Mahajan and Singh, 2014). The fruits of

strawberry cv. Sweet Charlie packed in 50 micron LDPE and stored at a temperature of

20±2°C and relative humidity 80-90% were having better organoleptic ratings (Panda et al.,

2016).

2.2.9 Quality retention

Loquat fruit packed in perforated polyethylene retained their quality for 30 days at 1°C as

well as 5ºC temperature (Ding et al., 1998). Wrapping in heat shrinkable film maintained

12

acceptable appearance, flavour and overall eating quality of banana and kiwi fruits

(Kudachikar et al., 2007; Sharma et al., 2012). Modified atmosphere packaging especially

high density polyethylene film of 70μ thickness wrapping preserved the quality of apple cv.

Eva up to seven months during storage at 0.5°C temperature (Fante et al., 2014). The shrink

film packaging proved quite effective in maintaining the various qualities attributes of kinnow

fruit for 20 days under supermarket conditions (Mahajan and Singh, 2014).

2.2.10 Shelf life

Packaging of mango cv. Keitt individually in low and high density polyethylene films (LDPE

and HDPE) delayed fruit ripening up to 4 weeks at 20°C temperature and 67% relative

humidity (Gonzalez et al., 1990). Low density polyethylene packages maintained longer shelf

life of blueberry fruit under cold storage (Beaudry et al., 1992). Micro-perforated

polypropylene film packaging extended the shelf life of strawberry cv. Camarosa fruits (Sanz

et al., 1999). Individual heat shrink wrapping of peach fruits with 20μ linear low density

polyethylene (LLDPE) after post-harvest treatment with carbendazim 500 ppm retained better

quality and extended the shelf life up to 42 days at 5±1°C temperature and 90-95% relative

humidity (Singh et al., 2009). Perforated low density polyethylene (0.25%) retained

significantly highest firmness of loquat fruit and thereby increased the shelf life (Akhtar et al.,

2012). Among different packaging materials, HDPE proved to be the best for prolonging the

shelf life of aonla cv. Kanchan fruits up to 15 days at ambient room temperature

(Vishwakarma et al., 2012). Wrapping of individual fruit in 12 micron cling film was

effective in increasing the shelf life of fig cv. Daulatabad at room temperature (Sharma and

Singh, 2013). Shrink film packaging delayed the softening process in pear cv. Patharnakh

fruits and finally retained the desirable fruits firmness due to reduced transpiration loss and

respiration activity per se retained more turgidity in super-market conditions, i.e., 20-21°C

temperature and 85-90% relative humidity (Mahajan et al., 2013). The shelf life of

pomegranate arils coated with Aloe vera gel at 100% alone or with ascorbic acid 1% and

citric acid 1% and packed in rigid polypropylene boxes was extended up to 12 days at 3°C

temperature (Martinez-Romero et al., 2013). Shrink film improved the shelf life of kinnow

fruits for 20 days against 10 days in control at 18-20°C temperature and 80-85% relative

humidity (Mahajan and Singh, 2014).

2.3 Effect of coating and packaging

Wrapping and waxing both delayed softening, colour change and extended the shelf life of

harvested avocado fruit at 22°C temperature by about 50% as compared to untreated fruit

(Joyce et al., 1995). The modified atmosphere packaging (MAP) in combination with

essential oils improved the beneficial effect of MAP on maintaining sweet cherry fruit quality

during storage and extending shelf life (Serrano et al., 2005). Aloe vera gel + glycerol coating

13

combined with perforated plastic film packaging slowed damage to strawberry fruits (Arifin

et al., 2013).


The maximum TSS was recorded in mango cv. Langra coated with 4% CaCl2 and packed in

LDPE after 34 days of storage at 13±1°C temperature and 90-95% relative humidity (Singh

et al., 2013).


The fruits of mango cv. Langra coated with 2% CaCl2 and packed in LDPE showed highest

titratable acidity after 34 days of storage at 13±1°C temperature and 90-95% relative humidity

(Singh et al., 2013).

2.3.3 Ascorbic acid content (mg/100 g)

The maximum ascorbic acid was recorded in mango cv. Langra coated with 2% CaCl2 and

packed in LDPE after 34 days of storage at 13±1°C temperature and 90-95% relative

humidity (Singh et al., 2013).

2.3.4 Shelf life

Treatment with 2% CaCl2 + LDPE packaging maintained fruit firmness, inhibited senescence

changes and markedly extended the storage life of mango cv. Langra at 13±1°C temperature

and 90-95% relative humidity (Singh et al., 2013). Aloe vera gel coating (1:3) on jujube for 5

minutes and packed in LDPE film extended its shelf life up to 45 days at 5±2°C temperature

(Padmaja and Bosco, 2014).

2.3.5 Quality control

Aloe vera gel dipping (1:3) for 5 minutes effectively inhibited the undesirable

physicochemical and physiological changes during storage of jujube packaged in LDPE film,

for 45 days at 5±2°C temperature (Padmaja and Bosco, 2014).

2.3.6 Organolaptic rating

The quality attributes, i.e., visual aspect, firmness, crunchiness, juiciness, sweetness and

sourness, and overall acceptability of jujube fruits coated with Aloe vera and packaged in

LDPE film were highest for 45 days at 5±2°C temperature (Padmaja and Bosco, 2014).

14

CHAPTER-III

MATERIAL AND METHODS

The present experiment entitled Studies on bio-extract coatings and packaging on the

shelf life of pomegranate fruits cultivar Mridula was carried out in Post-harvest Technology

Laboratory of the Department of Horticulture, CCS Haryana Agricultural University, Hisar

during 2016. The material and methods used during the experimentation are given as under:

3.1 Experimental details

The fresh fruits of pomegranate cv. Mridula were procured from the Centre of Excellence for

Fruits, Mangiana (Haryana). The details of experiment are given below:

3.1.1 Bio-extract coating of fruits

Preparation of Aloe vera gel +(edible coatings)

The mature leaves of Aloe vera plant were harvested and washed with running tap water. Aloe

vera latex was then separated from the outer cortex of leave by keeping it for two hours in

vertical position and then the colourless hydro-parenchyma was ground in a blender. The

resulting mixture was filtered to remove the fibres. The liquid obtained constituted fresh Aloe

vera gel (100%). Then it was further diluted with distilled water in1:1 ratio (50% Aloe vera

extract) and in 3:1 ratio of Aloe gel and water (75% Aloe vera extract).

Preparation of ginger extract

Fresh mature rhizomes of gingers were bought from the vegetable market and washed

properly after peeling in running tap water to remove dirt, and then, they were cut into small

pieces with the help of stainless steel knife and were ground in a blender. Thereafter, the juice

was extracted by straining with muslin cloth. The liquid obtained constituted fresh ginger

extract. Then 10, 20 and 30 ml of above prepared extract were dissolved in 1 litre of distilled

water to prepare 1, 2 and 3% of ginger extract, respectively.

Preparation of mint extract

Fresh mint leaves were bought from the vegetable market and washed properly after peeling

in running tap water to remove dirt, and then, those were ground in a blender. Thereafter, the

juice was extracted by straining with muslin cloth. The liquid obtained constituted fresh mint

extract. Then 100, 200 and 300 ml of above prepared extract were dissolved in 1 litre of

distilled water to prepare 10, 20 and 30% of mint extract, respectively.

Plate 1: Pomegranate fruits procured from Mangiana in CFB boxes

Plate 2: Opened pomegranate fruit Plate 3: Pomegranate arils

Plate 4: Fresh matured Aloe vera leaves Plate 5: Separation of colourless hydro-

parenchyma from outer rind

15

T1: Aloe vera extract 50%



T4: Ginger extract 1%



T7: Mint extract 10%



T10: Control

Fruits were dipped in these prepared bio-extract solutions of different concentration

for three minutes, allowed to drain and then dried at room temperature to allow a thin film

layer to be formed on the fruits, and then kept them in corrugated fibre boxes (CFB).

3.1.2 Wrapping materials

T1: LDPE 25 micron

T2: Polypropylene 25 micron

T3: Cling film

T4: Cellophane paper

T5: Control

The individual fruits were wrapped in the above mentioned packaging films, and

thereafter, fruits were packed in CFB boxes.

3.1.3 Storage conditions

The fruits were stored at room temperature with maximum 29±2°C, minimum 12±2°C and

relative humidity 90±5%.

3.2 Observations recorded

The following observations were recorded at three days interval.


The pomegranate fruit weight of each box was recorded at three days interval and the

physiological loss in weight for each date of observations was calculated by using the

following formula:

Initial weight - Final weight

Loss in weight (%) = x 100

Initial weight

Plate 6: Prepared Aloe vera extract Plate 7: Air drying of fruits after the

coating on it

Plate 9: Peeling of ginger Plate 10: Fruits dipped in ginger extract

Plate 11: Fresh mint leaves

Plate 12: Prepared mint extract

16


The decay loss was calculated by using the following formula:

Weight of decayed fruits

Decay loss (%) = x 100

Initial weight of fruits

3.2.3 Juice content (%)

The pomegranate juice content was calculated by using the following formula:

Juice volume

Juice content (%) = x 100

Fruit weight


The total soluble solids of fruits were determined at room temperature by using hand

refractometer having a range of zero to 32 (ERMA) by putting a drop of pomegranate juice

and taking the readings. The refractometer was calibrated with distilled water after every use

and the values were expressed in %.


The titratable acidity was determined as per the method suggested by AOAC (1990). Five

millilitre of pomegranate juice was taken in a conical flask and diluted with a little quantity of

distilled water, and the final volume was made to 100 ml. A pinch of activated charcoal was

added for the disappearance of red colour. It was then filtered through a rough filter paper and

20 ml of aliquot was taken for titration. Two drops of phenolphthalein indicator was added to

the given aliquot and then titrated against N/10 NaOH solution. The end point was considered

as the appearance of pink colour. The acid content was calculated by using following

formulae and expressed as gram of citric acid per 100 ml of pomegranate juice.

0.0064 × titratable volume × total volume made

Acidity (%) = x 100

Volume of aliquot × volume of sample

3.2.6 pH

Freshly extracted pomegranate fruit juice from each sample was taken to determine the pH by

using digital pH meter, which was calibrated with buffer having pH= 4.0 and 9.2.

3.2.7 Anthocyanin content (mg/100 g)

Total anthocyanin was determined according to the pH differential spectroscopic method

(Cheng and Breen, 1991; Tonutare et al., 2014).

Reagents

pH 4.5 buffer: 400 ml of 1M sodium acetate (82 g/l) + 240 ml of 1N HCl (36.5 ml

concentrated HCl/l) + 360 ml distilled water

pH 1.0 buffer: 125 ml of 0.2 N KCl (14.9 g/l) + 385 ml of 0.2 N HCl

17

The pH of the buffer can be adjusted as required to obtain final pH values.

Procedure

To 20 ml of the sample 80 ml of buffer was added and mixed in a blender at full speed. The

order of dilution was such that the sample at pH 1.0 would have an absorbance of less than

1.0 and preferably in the range of 0.4-0.6. The dilution strength was same for both 1.0 and 4.5

samples.

For making the diluted samples clear and haze free, the sediments were removed by

centrifuging and filtering the samples. Thereafter, absorbance reading was taken at 700 nm

and 510 nm, and the absorbance was found zero at 700 nm.

Calculation

A = (A1-A2) – (A3-A4)

B = (A x MW x DF x 103)/ EL

Where,

A =Absorbance difference between pH 1.0 and 4.5

A1 =Absorbance at 510 at pH 1.0




B = Concentration (mg/100 g)

EL =Molar absorbance of the major anthocyanin (26.9)

MW =Molecular weight of the major anthocyanin (499.2)

DF = Dilution factor

103

= Factor for conversion of g to mg

3.2.8 Ascorbic acid content (mg/100 ml)

The ascorbic acid was estimated by using the procedure suggested by AOAC (1990).

Reagents

(a) Metaphosphoric acid solution (3%)

Metaphosphoric acid (HPO3) 15 g

Glacial acetic acid 40 ml

Final volume 500 ml

(b) 2,6-dichlorophenol indophenol dye

2,6-dichlorophenol indophenols dye 50 mg

18

Sodium bicarbonate 42 mg

Final volume 200 ml

(c) Standard ascorbic acid solution

50 mg of ascorbic acid was dissolved in metaphosphoric acid solution (3%) and the final

volume was made to 50 ml by adding metaphosphoric acid. One ml of standard ascorbic acid

solution was used to standardize the dye with appearance of pink colour as the end point, i.e.,

standard factor or dye factor.

Estimation

Five millilitre pomegranate juice was taken in a conical flask and a pinch of activated

charcoal was added for the disappearance of red colour. It was then filtered through filter

paper. The filtrate was added with 15 ml of three percent metaphosphoric acid as buffer. Two

ml aliquot was taken in conical flask and it was titrated against 2,6-dichlorophenol

indophenol dye. The endpoint was the appearance of pink colour persistent up to one minute.

The ascorbic acid content was calculated using the following formula and results were

expressed in mg of ascorbic acid per 100 ml of juice.

Titratable volume

Ascorbic acid = x 100

(mg/100 ml) Standard reading x ml of sample taken for titration

3.2.9 Sugars (%)

Sugars were estimated by using the method of Hulme and Narain (1931).

Reagents

1. Potassium ferricyanide solution: 8.25 g potassium ferricyanide and 10.6 g anhydrous

sodium carbonate were dissolved in 500 ml distill water, and the final volume was made to

one liter.

2. Potassium iodide solution: 12.5 g potassium iodide, 25 g zinc sulphate and 125 g sodium

chloride were dissolved in distill water, and the final volume was made to one liter.

3. 5 per cent acetic acid solution: 50 ml glacial acetic acid was dissolved with distill water,

and the final volume was made to one liter.

4. Sodium thiosulphate solution: 2.482 g sodium thiosulphate was dissolved in distill water,

and the final volume was made to one liter.

5. Starch solution indicator: Soluble starch 1 g and sodium chloride 20 g were dissolved in

distill water, and the final volume was made to 100 ml.

19

Extraction

Ten ml of distilled water was added to five millilitre of pomegranate fruit juice in each test

tube. The tubes were covered with aluminium foil and kept for 30 minutes in boiling water

bath. The material was filtered and same procedure was followed three times and the total

material was pooled in volumetric flask and the final volume was made with proper dilution.

Estimation

A) Reducing sugars (%)

Five ml of juice extract was taken in a test tube and five ml of potassium ferricyanide solution

was added to it. The tubes were covered with aluminum foil and kept for 15 minutes in

boiling water bath. After cooling under tap water, five ml of potassium iodide solution

followed by three ml of acetic acid solution was added in each test tube. The liberated iodine

was titrated with sodium thiosulphate solution using starch as an indicator. The end point was

the disappearance of blue and appearance of milky white colour. A blank was also run

simultaneously. The results were calculated by using the following formula and expressed in

percent.

X (mg sugars per 5 ml of juice extract)= [(ml of sodium thiosulphate used in blank ml

of sodium thiosulphate used for sample) + 0.05] x 0.338

X × dilution factor

Reducing sugars (%) = x 100

5×1000

B) Total sugars (%)

25 ml of diluted sugar extract was taken in flask, 4 ml concentrated HCl was added and kept

for 15 minutes in water bath at 68°C temperature. Acidity was neutralized by adding

anhydrous sodium carbonate until the effervescence stopped. Thereafter, the volume was

made to 50 ml and total sugars were then determined as described in reducing sugars.

C) Non-reducing sugar (%)

The non-reducing sugar was determined by subtracting the value of reducing sugars from the

estimated total sugars for each sample.


Three pomegranate fruits were selected at random from each of the replication and subjected

to sensory evaluation daily by a panel of five judges following the nine point hedonic scale as

described by Rangana (1977).

The scale was as follows:

Acceptability Marks

Extremely desirable 9

20

Very much desirable 8

Moderately desirable 7

Slightly desirable 6

Neither desirable nor undesirable 5

Slightly undesirable 4

Moderately undesirable 3

Very much undesirable 2

Extremely undesirable 1

3.3 Statistical Analysis

The data on different parameters were collected, and in order to evaluate comparative

performance of the various treatments, the data were analyzed by using the technique of

analysis of variance described by Fisher (1958). All the tests of significance were made at 5

per cent level of significance. To judge the significance of the difference between two means,

the critical difference (C.D.) was worked out by following the formula given as under:

C.D. at 5% level of significance = [(2 x error mean squares)1/2

] x „t‟ at 5%

Where,

C.D. = Critical difference

t = Value of t-distribution at 5 per cent level of significance at error

degrees of freedom

3.4 Use of sensors to study the permeability of various packaging films and components

of MAP

3.4.1. Sensors and the software

Data collection software: Logger Pro 3.11

Vernier Data Logger

Sensors: i) Vernier CO2 Gas Sensor

ii) Vernier O2 Gas Sensor

iii) Vernier Relative Humidity Sensor

iv) Vernier Stainless Steel Temperature Probe

3.4.2. Packaging materials used

T1: Low density polyethylene (LDPE) 25 micron

T2: Polypropylene (PP) 25 micron

T3: Cling film

T4: Cellophane paper

T5: Control

Plate 13 Pomegranate fruit in LDPE Plate 14: Pomegranate fruit in PP

Plate 15: Pomegranate fruit wrapped in

cling film

Plate 16: Fruits wrapped in cellophane

paper

Plate 17: Use of sensors to study the permeability of various packaging film

21

3.4.3. Preparation of samples and recording observations

The fresh fruits of pomegranate cv. Mridula were procured from the Centre of Excellence for

Fruits, Mangiana (Haryana). The fruits were washed properly in running tap water and dried

by wiping with the help of muslin cloth during the morning hours. Each of the packaging

films was wrapped around the individual fruits along with the sensor probe (4) and the

packages were made manually air tight immediately. Each of the sensors was connected to the

data logger (data collecting device), which was connected to the computer. The data

collecting software (Logger Pro 3.11) recorded the data immediately after making the

packages air tight and represented in graphical forms individually for each parameter with

respect to time scale.

22

CHAPTER-IV

EXPERIMENTAL RESULTS

The present experiment entitled Studies on bio-extracts coating and packaging on the

shelf life of pomegranate cv. Mridula fruits was conducted at Post-harvest Laboratory of the

Department of Horticulture, CCS Haryana Agricultural University, Hisar. The results so

obtained are presented in this chapter under the following heads:

Physical characteristics of pomegranate fruit cv. Mridula

The physical characteristics fruit length was 7.23 cm, fruit diameter 6.45 cm, fruit weight

135.67 g, rind weight 44.36 g, rag weight 12.63 g, aril weight 91.07 g, juice weight 64.45 g

and juice content 47.51% before the start of storage experiment on pomegranate cv. Mridula.

Table 1: Physical characteristics of pomegranate fruit cv. Mridula

Parameters Value

Fruit length (cm) 7.23

Fruit diameter (cm) 6.45

Fruit weight (g) 135.67

Rind weight (g) 44.36

Rag weight (g) 12.63

Aril weight (g) 91.07

Juice weight (g) 64.45

Juice content (%) 47.51

Biochemical characteristics of pomegranate fruit cv. Mridula

The biochemical characteristics total soluble solids were 13.37%, titratable acidity 0.43%, pH

3.48, ascorbic acid content 13.08 mg/100 g, anthocyanin content 13.86 mg/100 g, total sugars

Table 2: Biochemical characteristics of pomegranate fruit cv. Mridula

Parameters Value

Total soluble solids (%) 13.37

Titratable acidity (%) 0.43

Total soluble solids to acid ratio 31.09

pH 3.48

Ascorbic acid (mg/100 g) 13.08

Anthocyanin content (mg/100 g) 13.86

Total sugars (%) 9.34

Reducing sugars (%) 7.36

Non-reducing sugar (%) 1.98

23

9.34%, reducing sugars 7.36% and non-reducing sugar 1.98% before the start of storage

experiment on pomegranate cv. Mridula.

4.1. Experiment-1: Effect of various bio-extracts coating on physico-chemical

properties of pomegranate fruits during storage


The data presented in Table 3 clearly indicate that the bio-extract coating treatments

significantly affected the physiological loss in weight of pomegranate fruits. Under ambient

room conditions, the significantly minimum loss in weight on 3rd, 6th, 9th and 12th day of

storage was observed in pomegranate fruits treated with Aloe vera extract 100%, i.e., 1.85,

2.93, 4.70 and 6.32% as compared to all other treatments, followed by Aloe vera extract

75%, which was at par with Aloe vera extract 50% and the maximum loss in weight in

untreated pomegranate fruits, i.e., 5.12, 7.25, 10.31 and 12.61%, respectively (Table 1),

which was statistically at par with physiological loss in weight with ginger extract 1, 2 and

3% and mint extract 10, 20 and 30% coating.

Table 3: Effect of bio-extracts coating on physiological loss in weight (%) of pomegranate cv.

Mridula

Treatments Storage period (days)

3 6 9 12

Aloe vera extract 50% 3.32 4.90 6.58 8.56



Ginger extract 1% 4.77 6.85 9.79 11.92

Ginger extract 2% 4.53 6.56 9.58 11.87

Ginger extract 3% 4.24 6.36 9.49 11.71

Mint extract 10% 4.92 6.97 9.90 12.14

Mint extract 20% 4.59 6.60 9.61 11.98

Mint extract 30% 4.43 6.43 9.55 11.85

Control 5.12 7.25 10.31 12.61

C.D. at 5% level of significance 0.90 1.02 1.13 1.10


The data on decay loss given in Table 4 clearly show that the decay loss in pomegranate fruits

was significantly affected by the bio-extracts coating. In all the bio-extracts coating

treatments, no decay loss was found up to first eight days of storage under ambient room

conditions, while least decay loss, i.e., 0.63 and 9.65% was observed on 9th and 12th day of

storage in treatment ginger extract 3% and the most decay loss, i.e., 11.40 and 23.36% was

recorded in untreated fruits, respectively. The treatment Aloe vera extract 100% and ginger

extract 1%, Aloe vera extract 75% and mint extract 20% and Aloe vera extract 50% and mint

extract 10% on 9th day of storage, whereas, Aloe vera extract 100% and ginger extract 1%,

24

Aloe vera extract 75% and mint extract 30% and Aloe vera extract 50%, mint extract 10 and

20 % on 12th day of storage were found statistically at par with each other in respect of decay

loss.

Table 4: Effect of bio-extracts coating on decay loss (%) in pomegranate cv. Mridula


9 12

Aloe vera extract 50% 8.06 16.29



Ginger extract 1% 3.17 11.96



Mint extract 10% 8.18 16.43



Control 11.40 23.36

C.D. at 5% level of significance 0.98 1.03


The perusal of data on pomegranate juice content represented in Table 5 reveals that there

was a significant difference in juice content (%) of fruits coated with different bio-extracts

and also under different storage period. The maximum juice retention (47.17%) was found in

fruits coated with Aloe vera extract 100% and the minimum in control (45.56%), which was

statistically at par with juice content in fruits coated with mint extract 10% (45.75%).

Table 5: Effect of bio-extracts coating on juice content (%) of pomegranate cv. Mridula

Treatments Storage period (days) Mean

0 3 6 9 12

Aloe vera extract 50% 47.51 47.33 46.65 46.32 45.18 46.60



Ginger extract 1% 47.51 47.12 46.45 44.88 43.05 45.80

Ginger extract 2% 47.51 47.14 46.36 45.24 43.11 45.87

Ginger extract 3% 47.51 47.37 46.55 45.37 43.82 46.12

Mint extract 10% 47.51 46.98 45.99 45.07 43.2 45.75

Mint extract 20% 47.51 47.05 46.00 45.13 43.36 45.81

Mint extract 30% 47.51 47.09 46.01 45.22 43.35 45.84

Control 47.51 46.79 45.72 44.89 42.87 45.56

Mean 47.51 47.17 46.42 45.60 44.00

C.D. at 5% level of significance

Treatments (T) = 0.23

Storage period (S) = 0.28

Treatments × storage period = NS

25

With the advancement of storage period, the juice content in fruits decreased

gradually. The maximum juice content in pomegranate fruits (47.51%) was noticed on zero

day of storage and minimum on 12th day of storage (44.00%) under ambient room conditions.

The interaction of the bio-extracts coating and storage duration was found non-significant

with respect to juice content of pomegranate fruits.


The data given in Table 6 indicate that the storage period and the bio-extracts coating

showed significant effect on total soluble solids of pomegranate fruits, however, the

interaction between bio-extracts coating and storage duration had no significant effect on total

soluble solids of pomegranate fruits. The pomegranate fruits coated with Aloe vera extract

100% had minimum TSS (13.51%) and uncoated fruits had maximum TSS (13.89%), which

was statistically at par with fruits coated with ginger extract 1% and mint extract 10 and 20%,

while TSS of fruits coated with Aloe vera extract 75 and 50% were statistically at par with of

each other. With the increase in storage period, the TSS of fruit increased gradually. The

minimum TSS was recorded on zero day of storage (13.37%), which was statistically at par

with TSS on 3rd day of storage (13.44%) and maximum on 12th day (14.19%) of storage.

Table 6: Effect of bio-extracts coating on total soluble solids (%) of pomegranate cv.

Mridula


0 3 6 9 12

Aloe vera extract 50% 13.37 13.41 13.62 13.84 14 13.65



Ginger extract 1% 13.37 13.48 13.71 14.1 14.32 13.80

Ginger extract 2% 13.37 13.44 13.65 14.07 14.28 13.76

Ginger extract 3% 13.37 13.39 13.63 14.05 14.22 13.73

Mint extract 10% 13.37 13.51 13.8 14.19 14.4 13.85

Mint extract 20% 13.37 13.49 13.75 14.13 14.29 13.81

Mint extract 30% 13.37 13.44 13.69 14.08 14.24 13.76

Control 13.37 13.54 13.83 14.22 14.47 13.89

Mean 13.37 13.44 13.67 14.01 14.19






The data registered in Table 7 demonstrate that the different bio-extracts coating and the

interaction between different treatments and storage periods did not affect the titratable

acidity of pomegranate fruits significantly, however, over the storage period, significant

variation in titratable acidity was observed. The titratable acidity of fruits coated with

26

different bio-extracts went on decreasing with the advancement of storage period. Under

ambient room conditions, the maximum titratable acidity was noticed on zero day of storage

(0.43%) and minimum on 12th (0.40%) day of storage, while the titratable acidity on 3rd day

of storage (0.42%) and 6th (0.42%) day of storage was found statistically at par with each

other in respect of titratable acidity.

Table 7: Effect of bio-extracts coating on titratable acidity (%) of pomegranate cv. Mridula


0 3 6 9 12




Ginger extract 1% 0.43 0.42 0.42 0.41 0.40 0.41

Ginger extract 2% 0.43 0.42 0.42 0.41 0.40 0.42

Ginger extract 3% 0.43 0.43 0.42 0.41 0.41 0.42

Mint extract 10% 0.43 0.41 0.41 0.40 0.39 0.41

Mint extract 20% 0.43 0.42 0.42 0.41 0.40 0.41

Mint extract 30% 0.43 0.42 0.42 0.41 0.40 0.42

Control 0.43 0.41 0.41 0.40 0.39 0.41

Mean 0.43 0.42 0.42 0.41 0.40


Treatments (T) = NS



4.1.6 TSS to acid ratio (%)

The TSS to acid ratio of pomegranate fruits with different bio-extracts coating followed an

increasing trend with the advancement of storage period. Under ambient room conditions

Table 8: Effect of bio-extracts coating on TSS to acid ratio (%) of pomegranate cv. Mridula


0 3 6 9 12




Ginger extract 1% 31.09 32.10 32.64 34.39 35.80 33.66

Ginger extract 2% 31.09 32.00 32.50 34.32 35.70 32.76

Ginger extract 3% 31.09 31.14 32.45 34.27 34.68 32.69

Mint extract 10% 31.09 32.95 33.66 35.48 36.92 33.78

Mint extract 20% 31.09 32.12 32.74 34.46 35.73 33.68

Mint extract 30% 31.09 32.00 32.60 34.34 35.60 32.76

Control 31.09 33.02 33.73 35.55 37.10 33.88

Mean 31.09 32.00 32.55 34.17 35.48

27

(Table 6), the total soluble solids to acid ratio was noted minimum on the zero day (31.09%)

of storage and maximum on 12th (35.48%) day of storage. The total soluble solids to acid

ratio (33.88%) was found maximum in control fruits and the minimum TSS to acid ratio

(32.17%) in fruits treated with Aloe vera extract 100%.

4.1.7 pH

The pH of juice extracted from stored pomegranate fruits with different bio-extracts coating

recorded in Table 9 revealed statistically non-significant variation for different bio-extract

coating materials and the interaction between the bio-extracts coating and storage period,

however, significant variation was observed with respect to storage periods. Under ambient

room conditions, the minimum pH was recorded on zero day of storage (3.48) and the

maximum on 12th day (3.71) of storage.

Table 9: Effect of bio-extracts coating on pH of pomegranate cv. Mridula


0 3 6 9 12




Ginger extract 1% 3.48 3.59 3.60 3.66 3.73 3.72

Ginger extract 2% 3.48 3.58 3.59 3.64 3.71 3.70

Ginger extract 3% 3.48 3.56 3.56 3.63 3.69 3.68

Mint extract 10% 3.48 3.62 3.62 3.69 3.75 3.74

Mint extract 20% 3.48 3.60 3.60 3.68 3.74 3.73

Mint extract 30% 3.48 3.58 3.58 3.65 3.70 3.69

Control 3.48 3.61 3.64 3.71 3.77 3.76

Mean 3.48 3.57 3.58 3.65 3.71


Treatments (T) = NS




The data recorded in Table 10 reveal that the ascorbic acid content of pomegranate fruits

coated with different bio-extracts varied significantly with respect to both the period of

storage and the treatments, while the interaction between the treatments and storage periods

was found statistically not-significant. Under ambient room conditions, the maximum

ascorbic acid was recorded in fruits coated with Aloe vera extract 100% (12.82 mg/100 g).

The minimum ascorbic acid was recorded in fruits kept untreated (12.03 mg/100 g).

28

Table 10: Effect of bio-extracts coating on ascorbic acid (mg/100 g) of pomegranate cv. Mridula


0 3 6 9 12




Ginger extract 1% 13.08 12.80 12.03 11.79 11.63 12.27

Ginger extract 2% 13.08 12.83 12.15 11.86 11.74 12.33

Ginger extract 3% 13.08 12.88 12.51 11.99 11.83 12.46

Mint extract 10% 13.08 12.76 12.33 11.96 11.28 12.28

Mint extract 20% 13.08 12.77 12.41 12.24 11.55 12.41

Mint extract 30% 13.08 12.81 12.50 12.31 11.73 12.49

Control 13.08 12.84 12.09 11.43 10.70 12.03

Mean 13.08 12.85 12.46 12.11 11.68





With the advancement of storage period, the ascorbic acid decreased significantly. It

was recorded maximum on zero day of storage (13.08 mg/100 g), which was statistically at

par with ascorbic acid on 3rd day of storage (12.85 mg/100 g) and minimum on 12th day

(11.68 mg/100 g) of storage.


Table 11: Effect of bio-extracts coating on anthocyanin content (mg/100 g) of pomegranate cv.

Mridula


0 3 6 9 12



Aloe vera extract 100% 13.86 13.91 13.95 14 14.2 13.98

Ginger extract 1% 13.86 13.93 14.14 14.39 14.62 14.19

Ginger extract 2% 13.86 13.92 14.06 14.35 14.59 14.16

Ginger extract 3% 13.86 13.87 14 14.3 14.49 14.10

Mint extract 10% 13.86 13.96 14.26 14.42 14.68 14.24

Mint extract 20% 13.86 13.92 14.23 14.36 14.63 14.20

Mint extract 30% 13.86 13.86 14.07 14.31 14.6 14.14

Control 13.86 14 14.36 14.51 14.72 14.29

Mean 13.86 13.92 14.11 14.30 14.52





29

The experimental data pertaining to anthocyanin content of pomegranate fruits with different

bio-extracts coating in Table 11 during the study period revealed significant effect over the

storage period and different bio-extracts coating but the variation in anthocyanin content with

respect to interaction between packaging materials and storage conditions was statistically

non-significant. Fruits coated with Aloe vera extract 100% recorded least anthocyanin content

(13.98 mg/100 g) and highest in uncoated control fruits (14.29 mg/100 g). The fruits coated

with Aloe vera extract 50 and 75%, ginger extract 1 and 2% and mint extract 10 and 20%

were found statistically at par with anthocyanin content of each other. Under ambient room

conditions, anthocyanin content increased significantly with the increase in storage period,

where the minimum anthocyanin content was recorded on zero day or prior to storage (13.86

mg/100 g), which was statistically at par with anthocyanin content on 3rd day (13.92 mg/100

g) and the maximum on 12th day (14.52 mg/100 g), which was statistically at par with

anthocyanin content on 9th day (14.30 mg/100 g) of storage.

4.1.10 Total sugars (%)

The analysis of variance of total sugars of stored pomegranate fruits coated with different bio-

extracts presented in Table 12 followed significant variation over the storage period and

different bio-extracts coating, however, no significant variation was recorded with respect to

interaction between the coating materials and the period of storage. The total sugars were

recorded least in fruits coated with Aloe vera extract 100% (9.42%) and the highest in

uncoated control fruits (9.64%). The fruits coated with Aloe vera extract 50 and 75%, ginger

extract 1, 2 and 3% and mint extract 10, 20 and 30% were found statistically at par with total

sugars content of each other.

Table 12: Effect of bio-extracts coating on total sugars (%) of pomegranate cv. Mridula


0 3 6 9 12




Ginger extract 1% 9.34 9.43 9.57 9.71 9.96 9.60

Ginger extract 2% 9.34 9.42 9.56 9.67 9.89 9.58

Ginger extract 3% 9.34 9.39 9.5 9.65 9.83 9.54

Mint extract 10% 9.34 9.43 9.57 9.72 10.01 9.61

Mint extract 20% 9.34 9.42 9.53 9.7 9.99 9.60

Mint extract 30% 9.34 9.41 9.49 9.66 9.92 9.56

Control 9.34 9.45 9.6 9.73 10.09 9.64

Mean 9.34 9.41 9.51 9.64 9.87





30

Under ambient room conditions, the total sugars increased gradually with the

advancement of storage period. The minimum total sugars was observed on zero day (9.34%),

which was statistically at par with total sugars on 3rd day (9.41%) and maximum on 12th day

(9.87%) of storage.

4.1.11 Reducing sugars (%)

The perusal of data in Table 13 indicates that the reducing sugars in stored pomegranate fruits

coated with different bio-extracts revealed a significant variation with respect to the period of

storage and bio-extracts coating, however, the variation with respect to interaction between

bio-extracts coating and storage period was statistically non-significant. Fruits coated with

Aloe vera extract 100% recorded the least reducing sugars (7.41%) and the highest in

uncoated control fruits (7.57%), which was statistically at par with reducing sugars content of

fruits treated with ginger extract 1 and 2% and mint extract 10, 20 and 30%, whereas, the

fruits coated with Aloe vera extract 50, 75% and ginger extract 3% were found statistically at

par with reducing sugars content of each other. Under ambient room conditions, the minimum

amount of reducing sugars was recorded on zero day of storage (7.36%), which was

statistically at par with reducing sugars on 3rd day of storage (7.41%) and highest on 12th day

(7.75%) of storage.

Table 13: Effect of bio-extracts coating on reducing sugars (%) of pomegranate cv. Mridula


0 3 6 9 12




Ginger extract 1% 7.36 7.42 7.49 7.6 7.81 7.54

Ginger extract 2% 7.36 7.4 7.48 7.58 7.76 7.52

Ginger extract 3% 7.36 7.41 7.49 7.55 7.7 7.50

Mint extract 10% 7.36 7.43 7.52 7.6 7.85 7.55

Mint extract 20% 7.36 7.42 7.48 7.59 7.83 7.54

Mint extract 30% 7.36 7.4 7.45 7.58 7.8 7.52

Control 7.36 7.42 7.53 7.6 7.94 7.57

Mean 7.36 7.41 7.47 7.56 7.75





4.1.12 Non-reducing sugar (%)

The experimental data in Table 14 pertaining to non-reducing sugar in stored pomegranate

fruits coated with different bio-extracts illustrated an increasing trend with the advancement

of storage period. The lowest amount of non-reducing sugar was recorded on zero day of

31

storage (1.98%) and the highest on 12th day of storage (2.12%) under ambient room

conditions. The least non-reducing sugar was recorded in fruits coated with Aloe vera extract

100% (2.01%) and the highest in uncoated control fruits (2.07%).

Table 14: Effect of bio-extracts coating on non-reducing sugar (%) of pomegranate cv. Mridula


0 3 6 9 12




Ginger extract 1% 1.98 2.01 2.08 2.11 2.15 2.07

Ginger extract 2% 1.98 2.02 2.08 2.09 2.13 2.06

Ginger extract 3% 1.98 1.98 2.01 2.10 2.13 2.04

Mint extract 10% 1.98 2.00 2.05 2.12 2.16 2.06

Mint extract 20% 1.98 2.00 2.05 2.11 2.16 2.06

Mint extract 30% 1.98 2.01 2.04 2.08 2.12 2.05

Control 1.98 2.03 2.07 2.13 2.15 2.07

Mean 1.98 2.00 2.04 2.09 2.12


The data pertaining to organoleptic rating pomegranate fruit are presented in Table 15 the

organoleptic rating of pomegranate fruit differed with different bio-extracts coating and this

rating decreased gradually with the advancement of storage period. Under ambient room

conditions, the fruits coated with Aloe vera extract 100% illustrated the highest organoleptic

rating (8.0), while the least rating (7.1) was given to the fruits kept untreated. On zero day

(8.5), the pomegranate fruits had the maximum organoleptic rating and minimum on 12th day

(6.1) of storage.

Table 15: Effect of bio-extracts coating on Organoleptic rating of pomegranate cv. Mridula


0 3 6 9 12




Ginger extract 1% 8.5 8.1 7.5 6.7 6.0 7.4

Ginger extract 2% 8.5 8.1 7.6 6.9 6.0 7.4

Ginger extract 3% 8.5 8.2 7.8 7.0 6.1 7.5

Mint extract 10% 8.5 8.0 7.3 6.6 5.5 7.2

Mint extract 20% 8.5 8.0 7.5 6.7 5.7 7.3

Mint extract 30% 8.5 8.0 7.6 6.9 5.9 7.4

Control 8.5 7.8 7.0 6.5 5.5 7.1

Mean 8.5 8.1 7.7 7.0 6.1

32

4.2. Experiment-2: Effect of different packaging material on the physico-chemical



The data presented in Table 16 clearly indicate that the different packaging materials

significantly affected the physiological loss in weight of pomegranate fruits. On 3rd day, the

fruits packed in LDPE 25 micron had significantly least loss in weight (0.38%), which was

statistically at par with fruits packed in polypropylene 25 micron (1.06%), cling film (0.84%)

and cellophane paper (1.11%) in reducing the loss in weight. On 6th day, the least loss in

weight was observed in fruits packed in LDPE 25 micron (1.36%), which was statistically at

par with packaging material polypropylene 25 micron (1.87%) in respect to reducing the

physiological loss in weight, while on 9th and 12th day, the fruits wrapped in LDPE 25

micron illustrated least physiological loss in weight, i.e., 4.19 and 5.17%, respectively as

compared to other packaging materials under ambient room conditions. On 6th and 12th day,

the treatment cellophane paper and cling film, whereas, on 9th day the treatment

polypropylene 25 micron, cellophane paper and cling film were statistically at par with each

other in reducing the physiological loss in weight. The unwrapped fruits taken as control had

the highest physiological loss in weight, i.e., 5.12, 7.25, 10.31and 12.61% on 3rd, 6th, 9th and

12th day of storage period respectively.

Table 16: Effect of different packaging materials on physiological loss in weight (%) of

pomegranate cv. Mridula


3 6 9 12

LDPE 25 micron 0.38 1.36 4.19 5.17

Polypropylene 25 micron 1.06 1.87 5.97 7.33

Cling film 0.84 2.21 6.61 8.39

Cellophane paper 1.11 2.26 6.83 8.96

Control 5.12 7.25 10.31 12.61

C.D. at 5% level of significance 0.83 0.74 1.09 1.02


The perusal of data in Table 17 reveals that the different packaging materials significantly

affected the decay loss in pomegranate fruits. Under ambient room conditions, no decay loss

was found during first eight days of storage of pomegranate fruits, while the minimum decay

loss of 3.71 and 10.51% was recorded on 9th and 12th day of storage in fruits wrapped with

LDPE 25 micron packaging film and the maximum decay loss of 11.40 and 23.36% was

found in unwrapped fruits, respectively.

33

Table 17: Effect of different packaging materials on decay loss (%) of pomegranate cv. Mridula


9 12

LDPE 25 micron 3.71 10.51

Polypropylene 25 micron 4.43 11.22

Cling film 9.95 18.82

Cellophane paper 7.17 16.80

Control 11.40 23.36

C.D. at 5% level of significance 0.61 0.54


The data presented in Table 18 indicate that different packaging films and the storage period

had statistically significant effect on juice content of pomegranate fruits under ambient room

conditions, however, non-significant variation in juice content was found for the interaction

between packaging materials and storage period. The fruits wrapped in LDPE 25 micron

retained maximum juice content (46.94%) as compared to other packaging materials and the

minimum juice content (45.56%) was found in unwrapped control fruits, while the treatment

cellophane paper (45.97%) and cling film (45.99%) were statistically at par with each other in

respect of juice content of fruits. The juice retention in fruits showed a decreasing trend with

the advancement of the storage period, while in the early days of storage, the decrease in juice

content was slower but with increase in time, it became more rapid. The maximum juice

content in pomegranate fruits (47.51%) was noticed on zero day and the minimum on 12th

day (44.30%) of storage.

Table 18: Effect of different packaging materials on juice content (%) of pomegranate cv.

Mridula


0 3 6 9 12

LDPE 25 micron 47.51 47.26 46.91 46.73 46.28 46.94

Polypropylene 25 micron 47.51 47.12 46.63 45.98 44.85 46.42

Cling film 47.51 47.01 46.45 45.22 43.76 45.99

Cellophane paper 47.51 46.96 46.37 45.26 43.73 45.97

Control 47.51 46.79 45.72 44.89 42.87 45.56

Mean 47.51 47.03 46.42 45.62 44.30





4.2.4. Total soluble solids (%)

The total soluble solids of pomegranate fruits packed in different packaging materials

represented in Table 19 exhibited statistically significant variation with respect to the period

of storage but the packaging films and the interaction between packaging materials and

34

storage duration had no significant effect on total soluble solids of fruits. Under ambient room

conditions, the minimum total soluble solids were found on zero day (13.37%), which was

statistically at par with TSS on 3rd day of storage (13.46%) and the maximum TSS was found

on 12th day (14.24%) of storage.

Table 19: Effect of different packaging materials on total soluble solids (%) of



0 3 6 9 12

LDPE 25 micron 13.37 13.41 13.52 13.67 13.96 13.59


Cling film 13.37 13.44 13.6 13.87 14.25 13.71


Control 13.37 13.54 13.83 14.22 14.47 13.89

Mean 13.37 13.46 13.63 13.90 14.24


Treatments (T) = NS




The analysis of variance of the titratable acidity of stored pomegranate fruits packed with

different packaging films presented in Table 20 followed significant variation over the storage

period. However, no significant variation was recorded with respect to different packaging

films and the interaction between the packaging films and the period of storage.

Table 20: Effect of different packaging materials on titratable acidity (%) of



0 3 6 9 12

LDPE 25 micron 0.43 0.43 0.42 0.41 0.41 0.42


Cling film 0.43 0.42 0.41 0.41 0.39 0.41


Control 0.43 0.41 0.41 0.40 0.39 0.41

Mean 0.43 0.42 0.41 0.41 0.40


Treatments (T) = NS



The titratable acidity of pomegranate fruits packed in different packaging films went

on decreasing with the advancement of storage period. The titratable acidity was observed

maximum on zero day (0.43%) and minimum on 12th (0.40%) day of storage, whereas, the

35

titratable acidity on 6th (0.41%) and 9th day (0.41%) was found statistically at par with each

other under ambient room conditions.

4.2.6 TSS to acid ratio (%)

The TSS to acid ratio of pomegranate fruits packed in different packaging films presented in

Table 21 followed an increasing trend with the advancement of storage period. Under ambient

room conditions, the TSS to acid ratio was noticed minimum on zero day (31.09%) and

maximum on 12th (35.60%) day of storage, whereas, with respect to different packaging

films, the maximum TSS to acid ratio (33.88%) was observed in unwrapped fruits and the

minimum TSS to acid ratio (32.36%) in fruits packed with LDPE 25 micron.

Table 21: Effect of different packaging materials on TSS to acid ratio (%) of



0 3 6 9 12

LDPE 25 micron 31.09 31.19 32.19 33.34 34.05 32.36


Cling film 31.09 32.00 33.17 33.83 36.54 33.44


Control 31.09 33.02 33.73 35.55 37.10 33.88

Mean 31.09 32.05 33.24 33.90 35.60

4.2.7 pH

The data in Table 22 reveal statistically non-significant effect on pH of stored pomegranate

fruits packed in different packaging films. However, storage period affected the pH of fruits

significantly. Under ambient room conditions, the minimum pH was recorded on zero day

(3.48), which was statistically at par with pH on 3rd day of storage (3.57) and maximum on

12th day (3.71) of storage, which was statistically at par with pH on 9th day (3.67) of storage.

Table 22: Effect of different packaging materials on pH of pomegranate cv. Mridula


0 3 6 9 12

LDPE 25 micron 3.48 3.52 3.56 3.62 3.65 3.57


Cling film 3.48 3.58 3.61 3.67 3.73 3.61


Control 3.48 3.61 3.64 3.71 3.76 3.64

Mean 3.48 3.57 3.60 3.67 3.71


Treatments (T) = NS



36

No significant effect was found with respect to interaction between the packaging films and

storage period.


The data pertaining to ascorbic acid content of pomegranate fruits are presented in Table 23.

The perusal of data reveals that the ascorbic acid content of pomegranate fruits packed in

different packaging films varied significantly over the period of storage and with respect to

the packaging films, however, the interaction between the packaging materials and storage

period was found statistically non-significant. With the advancement of storage period, the

ascorbic acid content of fruits decreased significantly. It was recorded maximum on zero day

of storage (13.08 mg/100 g) and the minimum on 12th day (11.48 mg/100 g) of storage.

Under ambient room conditions, the maximum ascorbic acid was observed in fruits packed

with LDPE 25 micron packaging film (12.79 mg/100 g) and the minimum (12.03 mg/100 g)

in fruits kept unwrapped, whereas, the treatment cling film (12.38 mg/100 g) and cellophane

paper (12.27 mg/100 g) were statistically at par with each other when ascorbic acid content of

the fruit was concerned.

Table 23: Effect of different packaging materials on ascorbic acid (mg/100 g) of



0 3 6 9 12

LDPE 25 micron 13.08 12.94 12.81 12.67 12.45 12.79


Cling film 13.08 12.83 12.41 12.24 11.35 12.38


Control 13.08 12.84 12.09 11.43 10.70 12.03

Mean 13.08 12.86 12.47 12.15 11.48






The experimental results pertaining to anthocyanin content of pomegranate fruits packed in

different packaging films illustrated in Table 24. The storage period and different packaging

materials significantly affected the anthocyanin content of fruit, on the other hand,

statistically non-significant effect was found with respect to interaction between packaging

materials and storage period. The minimum anthocyanin content was recorded on zero day or

prior to storage (13.86 mg/100 g) and the maximum on 12th day of storage (14.44 mg/100 g),

which was statistically at par with anthocyanin content on 9th day (14.26 mg/100 g) of

storage under ambient room conditions.

37

Table 24: Effect of different packaging materials on anthocyanin content (mg/100 g) of



0 3 6 9 12

LDPE 25 micron 13.86 13.93 13.97 14.03 14.27 14.01


Cling film 13.86 13.73 14.23 14.27 14.41 14.10


Control 13.86 14.00 14.36 14.51 14.72 14.29

Mean 13.86 13.90 14.16 14.26 14.44


Treatments (T) = NS




The data pertaining to total sugars in stored pomegranate fruits packed in different packaging

materials in Table 25 revealed a significant variation with respect to period of storage,

however, no significant variation was recorded with respect to different packaging materials

and the interaction between the packaging materials and the storage period. Under ambient

room conditions, the total sugars increased with increasing storage period and were found

minimum on zero day (9.34%) of storage, which was statistically at par with total sugars on

3rd day of storage (9.42%) and the maximum total sugars was recorded on 12th day (9.90%)

of storage.

Table 25: Effect of different packaging materials on total sugars (%) of pomegranate

cv. Mridula


0 3 6 9 12

LDPE 25 micron 9.34 9.39 9.43 9.46 9.6 9.44


Cling film 9.34 9.43 9.53 9.66 9.98 9.59


Control 9.34 9.45 9.6 9.73 10.09 9.64

Mean 9.34 9.42 9.54 9.64 9.90


Treatments (T) = NS




The analysis of variance of reducing sugars of stored pomegranate fruits packed in different

packaging materials presented in Table 26 followed significant variation over the storage

period, however, statistically non-significant effect on reducing sugars was found with respect

to packaging materials and the interaction between packaging materials and period of storage.

38

Under ambient room conditions, the least amount of reducing sugars was recorded on zero

day of storage (7.36%), which was statistically at par with reducing sugars in pomegranate

fruit on 3rd day of storage (7.41%) and the highest on 12th day (7.77%) of storage.

Table 26: Effect of different packaging materials on reducing sugars (%) of



0 3 6 9 12

LDPE 25 micron 7.36 7.39 7.43 7.45 7.58 7.44


Cling film 7.36 7.42 7.51 7.59 7.8 7.54


Control 7.36 7.42 7.53 7.6 7.94 7.57

Mean 7.36 7.41 7.50 7.56 7.77


Treatments (T) = NS




The experimental results in Table 27 pertaining to non-reducing sugar in stored pomegranate

fruits packed in different packaging materials showed an increasing trend with the

advancement of storage period. The least amount of non-reducing sugar was recorded on zero

day of storage (1.98%) and the utmost amount of non-reducing sugar on 12th day of storage

(2.13%) under ambient room conditions.

Table 27: Effect of different packaging materials on non reducing sugar (%) of



0 3 6 9 12

LDPE 25 micron 1.98 2.00 2.00 2.01 2.02 2.00


Cling film 1.98 2.01 2.02 2.07 2.18 2.05


Control 1.98 2.03 2.07 2.13 2.15 2.07

Mean 1.98 2.01 2.04 2.08 2.13


The data on organoleptic rating are presented in Table 28. The pomegranate fruits packed in

different packaging materials varied from each other in respect of organoleptic rating. Under

ambient room conditions, the pomegranate fruits packed in LDPE 25 micron packaging film

illustrated the highest organoleptic rating (8.0), while the minimum organoleptic rating (7.2)

was given to the fruits kept unwrapped. On zero day of storage (8.5), the pomegranate fruits

39

had the maximum organoleptic rating and minimum organoleptic rating (6.6) on 12th day of

storage.

Table 28: Effect of different packaging materials on organoleptic rating of



0 3 6 9 12

LDPE 25 micron 8.5 8.4 8.2 7.9 7.2 8.0


Cling film 8.5 8.2 8.0 7.6 6.7 7.8


Control 8.5 8.0 7.2 6.6 5.7 7.2

Mean 8.5 8.2 7.9 7.4 6.6

Effect of packaging materials on the components of modified atmospheric packaging

The findings of this experiment recorded illustrated behavior of individual packaging film for

each of the major parameters playing a key role in modifying the in-package environment are

given under the following headings:

4.3.1 Effect of various packaging film on the carbon dioxide concentration

The data pertaining to permeability of carbon dioxide recorded immediately after keeping the

pomegranate fruit in different packaging materials and making the packages air tight are

Fig.1. Effect of various packaging film on the carbon dioxide concentration (%) in modified

atmospheric packaging of pomegranate cv. Mridula

presented in Fig.1, which reveals that in all the treatments, the carbon dioxide concentration

inside the package increased continuously but at differential rate. The control or unwrapped

fruits kept in CFB boxes had the lowest concentration of carbon dioxide around it, which was

almost equal to the concentration of outer atmosphere. In LDPE 25 micron packaging film,

0.00

0.20

0.40

0.60

0.80

1.00

1.20

1.40

1.60

1.80

2.00

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

LDPE 25

micron

PP 25 micron

CLING FILM

CELLOPHANE

PAPER

CONTROL

Time (hours)

CO

2co

nc.

(%)

40

the carbon dioxide concentration increased rapidly up to 6 hours (1.29%) and then gradually

increased at a slower rate up to 24 hours (1.77%). The rise of carbon dioxide concentration in

cling film could be illustrated in three different phases. Up to 4 hours (1.15%), a rapid

increase was noticed, which was followed by a comparative slower increase up to 13 hours

(1.30%) and after that stability in carbon dioxide concentration was found until 24 hours

(1.31%). In polypropylene and cellophane paper packaging, the carbon dioxide concentration

increased at a faster rate up to 6 hours 1.11 and 1.39% followed by the establishment of

equilibrium in carbon dioxide until 24 hours i.e., 1.18 and 1.40% respectively. This inferred

maximum permeability of carbon dioxide gas by the PP 25 micron film.

4.3.2 Effect of various packaging film on the oxygen concentration

The data presented in Fig. 2 depict a very clear view about the differential permeability for

oxygen of different packaging films. With the advancement of time, the oxygen concentration

followed a decreasing trend in all the treatments except for PP 25 micron film. The decrease

was at faster rate in initial hours, however, after reaching a certain level, the declining rate

was at reduced pace, and finally, it attained the constancy. Differential rate of the decreasing

permeability was due to differential permeability by various packaging materials. In LDPE 25

micron film, a sharp reduction in oxygen concentration was recorded up to 9 hours (10.27%)

Fig.2. Effect of various packaging film on the oxygen concentration (%) in modified atmospheric

packaging of pomegranate cv. Mridula

followed by comparatively slower decreasing rate until 24 hours (5.68%). A steady decrease

in oxygen concentration was recorded in cling film from 1 (20.65%) to 24 hours (13.54%). A

similar trend as that of cling film was recorded for that of cellophane paper (20.73-11.36%).

Among other packaging materials, PP 25 micron film maintained the maximum concentration

0.00

5.00

10.00

15.00

20.00

25.00

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

LDPE 25

micron

PP 25 micron

CLING FILM

CELLOPHANE

PAPER

CONTROL

O2

con

c. (

%)

Time (hours)

41

of oxygen throughout the observation period (20.48-17.71%), which indicated its maximal

permeability to oxygen. The fruits kept in CFB boxes as control had nearly constant rate of

oxygen concentration and was in equilibrium with atmospheric oxygen.

4.3.3 Effect of various packaging film on temperature (°C)

Temperature

The data in Fig. 3 represented the variation in temperature of pomegranate fruits kept inside

all the different packaging materials individually during the period of investigation. More or

less constant variation in temperature was recorded for all the packaging films. Almost all the

packaging films had a similar trend in variation of temperature inside the package. The fruits

kept as control in CFB boxes had a variation in temperature that might be due to the effect of

external atmospheric temperature, which showed an increasing trend in day time and

decreasing trend during night.

Fig.3. Effect of various packaging film on the temperature (°C) in modified atmospheric


4.3.4 Effect of various packaging film on relative humidity (%)

Data represented in Fig. 4 demonstrated the changes in relative humidity inside the package

of atmospherically modified packed pomegranate fruits. In LDPE 25 micron film packaging,

relative humidity depicted a sharp rise up to 3 hours (94.2%) and thence moved on constantly.

Exactly similar pattern was noticed in cling film. In PP, the relative humidity was almost

increased steadily in initial hours to 16 hours (89.3%) and then it followed the constant trend

until 24 hours (90.4%) and reached the equilibrium state. The maximum permeability was

observed in LDPE 25 micron film, in which, a steady increase was recorded up to 2 hours

(92.7%), and the maximum relative humidity was recorded on 24 hours (95.9%) and the

25

26

27

28

29

30

31

32

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

LDPE 25

micron

PP 25 micron

CLING FILM

CELLOPHANE

PAPER

CONTROL

Time (hours)

Tem

p.

(°C

)

42

minimum being on zero hours (85.4%). In cellophane paper, the sharp rise was up to 3 hours

(79.3%) and then gradually increased up to 17 hours (87.5%) and then followed a constant

trend until 24 hours (88.3%). The relative humidity around the unwrapped fruits was affected

by the external atmosphere.

Fig.4. Effect of various packaging film on the relative humidity (%) in modified atmospheric


0.0

20.0

40.0

60.0

80.0

100.0

120.0

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24

LDPE 25

micron

PP 25 micron

CLING FILM

CELLOPHAN

E PAPER

CONTROL

Time (hours)

RH

(%

)

43

CHAPTER-V

DISCUSSION

The present experiment entitled Studies on bio-extracts coating and packaging on

shelf life of pomegranate fruits cultivar Mridula was conducted in Laboratory of Post-harvest

Technology of the Department of Horticulture, CCS Haryana Agricultural University, Hisar

during the year 2016. In this chapter, the results of the experiment so obtained are discussed

under following heads in light of the findings of earlier research workers and efforts have

been made to establish causes and effects associated with the help of available evidences and

relevant literature:

5.1 Experiment-1: Effect of various bio-extracts coatings on physico-chemical properties

of pomegranate fruits

5.1.1. Physiological loss in weight (%)

The physiological loss in weight of pomegranate fruits varied with different bio-extracts

coating (Table 3). In this experiment, Aloe vera extract 100% coating was found very

effective in reducing the physiological loss in weight of pomegranate fruits. Fruit weight loss

occurred as a result of dehydration and loss of water from the fruit surface. Aloe vera gel

coating reduced weight loss in coated fruit because of hygroscopic properties that enabled the

formation of a barrier to water diffusion between fruit and environment, per se, avoiding its

external transference (Morillon et al., 2002).

Fig.5. Effect of different bio-extracts coating on physiological loss in weight (%) in pomegranate

cv. Mridula

0

2

4

6

8

10

12

14

Day 3

Day 6

Day 9

Day 12

Treatments

PL

W (

%)

44

The Aloe vera gel retarded the moisture loss and reduced respiration rates, these

effects being similar to those obtained with other edible coatings (Romanazzi et al., 2008).

Aloe vera gel coating significantly reduced weight loss during fruit ripening and low

temperature storage as compared to uncoated mango fruits (Sophia et al., 2015). Similar

reduction in weight loss has also been reported by Valverde et al. (2005), Martinez-Romero et

al. (2006), Marpudi (2011), Padmaja and Bosco (2014) in Aloe vera coated sweet cherry,

table grapes and jujube.

5.1.2. Decay loss (%)

Decay loss of pomegranate fruit was significantly controlled by different bio-extracts coating.

In this experiment, the least decay loss was found in fruits coated with ginger extract (Table

4). This might be due to the anti-fungal and antibacterial properties of ginger, which inhibited

the growth of different microorganisms. Ginger (Zingiber officinale) has long been used as

naturopathy due to their potential antimicrobial activity against different microbial pathogens

(Islam et al., 2014) and antibacterial properties (Obochi et al., 2009). These results are in

conformity with the findings of Dharmaputra et al. (2013) in Salak pondoh fruit by inhibiting

the growth of Thielaviopsis paradoxa during storage.

Fig.6. Effect of different bio-extract coating on decay loss (%) in pomegranate cv. Mridula

5.1.3. Juice content (%)

The juice content of pomegranate fruits was significantly affected by the bio-extracts coating

and the different storage period (Table 5). In initial days of storage, the decrease in juice

content was slower, but later, the juice reduction became more rapid. The less juice content in

pomegranate fruits at the end of storage period might be due to the excessive loss of moisture

from the fruits.

0

5

10

15

20

25

Day 9

Day 12

Treatments

Dec

ay

lo

ss (

%)

45


The total soluble solids (TSS) of pomegranate fruits coated with different bio-extracts coating

recorded significant variation with respect to the period of storage and different bio-extracts

coating. With the advancement of storage period, total soluble solids increased gradually. The

total soluble solids in Aloe vera extract coated pomegranate fruits was found least (Table 6) as

compared to other treatments, which might be due to lower evapo-transpirational losses. This

increase in total soluble solids prevented to some extent with the help of aloe-pectin edible

coating on fruits (Valverde et al., 2005). Similarly, the delayed and smaller increase in total

soluble solids has been reported by Valverde et al. (2005), Martinez-Romero et al. (2006),

Alberio et al. (2015) and Verma et al. (2012) in Aloe vera gel coated sweet cherry, table

grapes and guava cv. L-49 and in starch-coated strawberry fruits by Mali and Grossmann

(2003).

5.1.5. Titratable acidity (%)

The different bio-extracts coating reported non-significant differences in titratable acidity of

pomegranate fruits (Table 7). The acidity of pomegranate fruits declined as the storage period

advanced. These findings corroborate the findings of Arowora et al. (2013) in oranges and

Ergun and Satici (2012) in apples cv. Red Chief under cold storage conditions.

5.1.6. TSS to acid ratio (%)

The total soluble solids to acid ratio of pomegranate fruits with different bio-extracts coating

followed an increasing trend with the advancement of storage period (Table 8). The fruits

coated with Aloe vera extract were observed having minimum total soluble solids to acid ratio

as compared to other treatments.

5.1.7 pH

The experiment reveals that the variation in pH of juice from pomegranate fruits with

different bio-extracts coating was not significant; however, significant effect was found in

respect of storage period. With the increase in storage period, the pH of fruit increased (Table

9). This might be due to the high level of carbon dioxide, which caused the formation of

carbonic acids. The dissociation of carbonic acid in cytoplasm is likely to occur. Since the

vacuole is acidic, dissociation of carbonic acid would be slight and the buffering capacity of

the tissue is likely to absorb such changes. Alternatively, at vacuolar pH, bicarbonates could

form and increase the pH (Holcroft and Kader, 1999) in strawberry fruits.


In the present investigation, the bio-extracts coating and the storage period exhibited

significant influence on ascorbic acid content of pomegranate fruits, where Aloe vera coated

46

pomegranate fruits had higher ascorbic acid content (Table 10). This might be due to the low

oxygen permeability of coat, which delayed the deteriorative oxidation reaction of ascorbic

acid content (Ayranci et al., 2003). Srinu et al. (2012) reported that coating reduces

respiration of the fruits and retains ascorbic acid in the fruits. Brishti et al. (2013) found

higher ascorbic acid content in Aloe vera coated papaya fruits (86.55 mg) than the control

fruits (61.10 mg) during the storage at temperature 25-29°C and relative humidity 82-84%.

Similar results were obtained during storage in nectarines (Ahmed et al., 2009), guava cv. L-

49 (Verma et al., 2012), oranges (Arowora et al., 2013), longan (Duan et al., 2007) and jujube

(Zhu et al., 2009) fruits coated with Aloe gel.


Anthocyanin content of pomegranate fruits significantly increased with the advancement of

storage duration (Table 11). This might be due to higher loss of moisture through evapo-

transpiration with the increase in storage period. The increase in anthocyanin pigments during

post-harvest storage of pomegranate fruit is associated with the maturation process

(Mirdehghan et al., 2006; Sayyari et al., 2011). The modified atmosphere created by the Aloe

vera gel coating reduced moisture loss through evapo-transpiration, per se, anthocyanin

content increased very less. Similar results have also been reported by Cantos et al. (2002)

and Tripathi and Dubey (2004) in grapes, Brishti et al. (2013) in papaya and Martinez-

Romero et al. (2013) in pomegranate.


The different bio-extracts coating and the storage period significantly affected the total sugars

content of pomegranate fruits (Table 12). The fruits coated with Aloe vera extract had least

total sugars content, as the Aloe vera gel coating created modified atmosphere, which reduced

the moisture loss from the pomegranate fruits through evapo-transpiration. The total amount

of sugars did not increase but the percentage of sugars increased due to reduced water

content.


The different bio-extracts coating and the storage period significantly affected the reducing

sugars content of pomegranate fruits (Table 13). The minimum reducing sugars were

observed in fruits coated with Aloe vera extract, which reduced the moisture loss from the

pomegranate fruits through evapo-transpiration due to creation of modified atmosphere, thus,

the increase in reducing sugars content was very less.

47


The non-reducing sugar increased with the advancement of storage period (Table 14), where

the maximum non-reducing sugar was noticed on 12 day of storage period and minimum on

the very first day of storage. Aloe vera extract coated fruits had lowest non-reducing sugar as

compared to all other treatments, in this experiment.


In this experiment, the highest organoleptic rating given to pomegranate fruits coated with

Aloe vera extract since, the coated fruits had greater retention of bright red color than the

uncoated fruits (Table 15). During storage, the judging panel found that flavor was

satisfactory in Aloe vera coated fruits but it was unsatisfactory in control fruits. In table

grapes, the panelists also preferred the coated berries with A. vera gel, because of their

crunchiness, firmness, juiciness and visual aspects as compared to uncoated fruits (Valverde

et al., 2005), as well as in sweet cherry (Martinez-Romero et al., 2006), which looked shiny

and attractive. The results of present study are in agreement with the findings of Brishti et al.

(2013) in fruits of papaya and Tripathi and Dubey (2004) in table grapes.




Positive effects of packaging films were observed in reducing physiological loss in weight of

pomegranate fruits (Table 15). The fruits wrapped with LDPE 25 micron was found best

when physiological loss in weight of fruit was concerned, on the other hand, fruits retained

unwrapped exhibited the highest physiological loss in weight. Fruits packed in different

packaging films recorded lower weight loss, which was obvious due to their role in altering

Fig.7. Effect of different packaging film on physiological loss in weight (%) in pomegranate cv.

Mridula

0

2

4

6

8

10

12

14

LDPE 25

micron

PP 25 micron Cling film Cellophane

paper

Control

Day 3

Day 6

Day 9

Day 12

PL

W(%

)

Treatments

48

the CO2 and O2 concentration inside the packages and thereby, reducing the rate of

transpiration/respiration and maintaining higher humidity inside the wrappers (Ben, 1985).

These results are in conformity with the findings of Valero et al. (2006) in table grapes,

Maniwara et al. (2015) in purple passion fruit, Kumar and Nagpal (1996) in mango, Nielsen

and Leufven (2008) in strawberry, Siddiqui and Gupta (1997) in guava and Sonkar and

Ladaniya (1998) in nagpur mandarin.

5.2.2. Decay loss (%)

The decay loss of pomegranate fruits was significantly affected by the different packaging

material (Table 16). In this experiment, LDPE 25 micron was observed as the best packing

materials in terms of controlling decay loss, where no decay loss was found during first eight

days of storage. This might be due to the property of packaging films to retain a higher level

of CO2 inside the package. Higher CO2 level in atmosphere showed fungi-static effect (Li and

Kader, 1989). The results are in line with the findings of Ozkaya et al. (2009) who reported

that the modified atmosphere packaging resulted in lower decay loss in strawberry fruits than

the control fruits.

Fig.8. Effect of different packaging film on the decay loss (%) in pomegranate cv. Mridula

5.2.3. Juice content (%)

The packaging material and the storage period had significant effect on juice content of

pomegranate fruits (Table 17). The juice content decreased with respect to storage period.

LDPE 25 micron found to be very effective packing material in controlling reduction in juice

content of fruits. Earlier, similar results in Kinnow mandarin were reported by Mahajan et al.

(2006).


In this experiment, the storage period significantly affected the total soluble solids content of

pomegranate fruits. However, packaging had non-significant effect on this (Table 18). The

0

5

10

15

20

25

LDPE 25

micron

PP 25

micron

Cling film Cellophane

paper

Control

Day 9

Day 12

Treatments

Dec

ay

loss

(%

)

49

increase in TSS during storage period could be attributed to the water loss and hydrolysis of

starch and other polysaccharides to soluble form of sugar (Wills et al., 1980). The gradual

increase in TSS over a longer period in film wrapped peach fruits was possibly due to

retarded ripening and senescence processes, which simultaneously delayed the conversion of

starch into sugars (Pongener et al., 2010). The results of present study corroborate the

findings of O‟Grady et al. (2014) of pomegranate arils.

5.2.5. Titratable acidity (%)

The titratable acidity of pomegranate fruits was affected significantly by storage period only,

not by the packaging material (Table 19). Most of the polyethylene bags retained higher

acidity content of fruits as compared to control fruits. This might be due to the development

of modified atmosphere around the fruits stored in polyethylene bags, which might have

slowed down various metabolic processes, resulting in lower utilization of acids in respiration

(Wavlah and Athale,1988). Variability in titratable acidity could be attributed to the effect of

increased CO₂ solubility inside the packages (Caleb et al., 2013). Similar results were also

reported by O‟Grady et al. (2014) in pomegranate arils, McCollum et al. (1992) in mango and

Nielsen and Leufven (2008) in strawberry fruits.

5.2.6. TSS to acid ratio (%)

The total soluble solids to acid ratio were recorded maximum in unwrapped fruit and the

minimum in fruits wrapped in LDPE 25 micron packaging film (Table 20). This ratio

increased gradually with the advancement of storage period, hence, the TSS to acid ratio was

found minimum on the very first day and the maximum on 12th day of storage period. This

might be due to the increase in total soluble solids and decrease in titratable acid content in

the fruits with the increase in storage duration.

5.2.7. pH

Different bio-extracts coating has non-significant effect on pH of pomegranate fruit (Table

21), however, storage period had significant effect. The pH of fruit increased with the

increase in storage period and this might be due to the reduction of titratable acidity and

increase in TSS of fruits. Variability in pH values in the studies could be attributed to the

effect of increased CO2 solubility inside the packages (Caleb et al., 2013). The results of

present study are in line with the findings of O‟Grady et al. (2014) in pomegranate.


The ascorbic acid content of pomegranate fruits packed in different packaging films varied

significantly over the period of storage. In the present investigation, the packaging materials

illustrated significant influence on ascorbic acid content of the fruits (Table 22). Such results

might be attributed to better modification of the atmosphere inside the package by reducing

50

O2 concentration, which concomitantly decreased the enzymatic oxidation of ascorbic acid

(Agrahari et al., 2001). Over the prolongation of storage period, the ascorbic acid content was

on a decreasing trend. This might be due to the oxidation and irreversible conversion of

ascorbic acid to dehydro-ascorbic acid in the presence of enzyme ascorbinase. Similar results

were also obtained by Sood et al. (2012) in strawberry.


Anthocyanin content of the fruits significantly increased with the prolongation of storage

period (Table 23). In the present study, the variation in anthocyanin content of pomegranate

fruit as a function of various packaging materials was not significant. This is in close

agreement with the findings of Nielsen and Leufven (2008) in strawberry fruits, where the

packaged strawberries maintained their colour and lustre much better than the unpackaged

samples.


The study revealed that the variation in total sugars in juice extracted from the stored

pomegranate fruits packed in different packaging film materials was non-significant.

However, the storage period had significant effect total sugars in juice of pomegranate fruit

(Table 24). The minor rise in sugar concentration in juice of packed fruit was probably due to

water loss at slower rate. The delayed increase in the sugar content under film packaging

might be attributed to the inherent property of films in delaying the metabolic activities of

fruits during storage due to delay in ethylene production and respiration rate (Abeles et al.,

1992). The results of present study are in accord with the findings of Nielsen and Leufven

(2008) in strawberry and Mohla et al. (2005) in pear. Bhullar (1983) in kagzi lime observed

that conversion of cell wall materials like hemicellulose and pectin from insoluble to soluble

fraction might have also added towards rise in sugars during later period of storage.


Significant variation in reducing sugars of stored pomegranate fruits was observed with

respect to the period of storage, however, there was no such significant variation with respect

to packaging materials. The slight increase in reducing sugars was possibly due to the loss of

moisture from the fruits with the increase in storage period. The results are in accordance with

the reports of Kahlon and Bajwa (1991) in litchi and Waskar et al. (1999) in sapota who have

reported an increase in reducing sugars content during storage.


The non-reducing sugar content of pomegranate fruit found minimum in LDPE 25 micron

coated fruits and the maximum in unwrapped fruits. With the prolongation of storage period,

51

the non-reducing sugar went on increasing trend, where the highest non-reducing sugar was

found on 12th day and the least on 0th day of storage.


The sensory quality or organoleptic rating of stored pomegranate fruits packed in different

packaging materials, differed with respect to the type of packaging materials as well as under

various storage period. The fruits got the maximum rating on the 0th day (8.25) and the

minimum on the 12th day (5.47) of storage. Among the different packaging materials, LDPE

25 micron depicted the best organoleptic rating to stored fruits. This characteristic feature of

LDPE film, having a proper balance for the permeability of carbon dioxide and oxygen and

relative humidity, maintained better overall sensory quality in strawberry fruits (Panda and

Goyal, 2016). It is expected that the altered atmosphere might disturb the aroma profile but

there was no loss of aroma in strawberries cv. Honeoye (Nielsen and Leufven, 2008).

Effect of permeability of packaging materials on the components of MAP

Carbon dioxide (%)

In all the packaging treatments, the carbon dioxide concentration followed an increasing trend

in the early phases. Such steady rise in carbon dioxide was because of the very high

respiratory behavior of pomegranate fruits. Inside the packaging material, the respiration rate

declined after some time, hence, the carbon dioxide concentration could not increase so

rapidly. Similar trend of carbon dioxide concentration was reported by Nielsen and Leufve,

(2008) in strawberry fruits in modified atmosphere packaging. In films having a greater

permeability of carbon dioxide, the time taken for reaching constancy was quite high than that

of the films having lesser permeability. The rise in carbon dioxide concentration in the PP

film was very slow, which might be due to its maximum permeability for carbon dioxide.

Oxygen (%)

Because of the utilization of limited amount of oxygen inside the packages during the process

of respiration, the oxygen concentration in all the packages went on decreasing. In films

having a lesser permeability to oxygen, the constant decreasing trend was achieved than that

of the film having a greater permeability. The oxygen concentration was maintained at a

higher level in PP 25 micron film than that of the other films. In most of the packaging

materials except PP 25 micron, similar trend of decreasing oxygen concentration was

recorded. The results of present study corroborate the findings of Nielsen and Leufve (2008)

in strawberry fruits.

Temperature (°C)

The variation in temperature was not affected by the modification inside the package rather it

might be due to the variation in atmospheric temperature. Such variation affected the

52

modified atmosphere by regulating the relative humidity level along with various metabolic

activities directly. No such strong evidence was found for temperature, which could be

correlated with the permeability of various packaging films.

Relative Humidity (%)

The relative humidity inside the packaging material as a measure of the moisture level is

prominently affected by the transpiration and other metabolic processes of the cells of any

fruit. During early phase of storage, a rapid increase in relative humidity might be attributed

to the rapid metabolic processes. Later, as the moisture loss from the fruits is checked, the

relative humidity inside the package reached a certain peak. Hence, a constant trend in

relative humidity was observed after a certain time. As the temperature increased, an increase

in relative humidity was noticed. This might be attributed to the increased moisture holding

capacity of the package because of the expansion of volume as a consequence of increased

temperature.

53

CHAPTER-VI

SUMMERY AND CONCLUSION

The present experiment entitled Studies on bio-extracts coating and packaging on the

shelf life of pomegranate fruits cv. Mridula was conducted at Post-harvest Laboratory of the

Department of Horticulture, CCS Haryana Agricultural University, Hisar to evaluate the best

bio-extract coating material or the most effective packaging film for prolonging the shelf life

and maintaining physico-chemical characteristics of high valued pomegranate fruits. The

salient findings of the study are briefly summarized below:

6.1. Experiment-1: Effect of various bio-extracts coating on physico-chemical


In this experiment, the loss in weight of fruits was observed from 3rd day onwards under

ambient room conditions, where Aloe vera extract 100% proved as the most effective one in

reducing the physiological loss in weight of pomegranate fruit.

Bio-extracts coating prevented the decaying of pomegranate fruits during first eight

days of storage under ambient room conditions. Among all the treatments, fruits coated with

ginger extract 3% had least decay loss.

The quality parameters such as juice content, ascorbic acid and titratable acidity

decreased with the advancement of storage period. Variation in such parameters played very

crucial role in determining the optimum storage quality of pomegranate fruits. The maximum

juice content, ascorbic acid and titratable acidity was observed in fruits coated with Aloe vera

extract 100% and on the very first days with respect to the storage period.

In pomegranate fruits, the total soluble solids, pH, total soluble solids to acid ratio,

anthocyanin contents, total sugars, reducing sugars and non-reducing sugars increased with

the prolongation of storage period and noticed highest on 12 day of storage. Among the

treatments, Aloe vera extract 100% coated pomegranate fruit had least content of TSS and

sugars, highest anthocyanin content but pH remain unaffected.

Under storage conditions, the overall acceptability of pomegranate fruits was found

best in pomegranate fruits coated with Aloe vera extract 100%. During the period of storage,

the organoleptic rating of fruits declined, where the highest rating was given on the very first

day of storage and the least on 12 day of storage.

54



In this experiment, the loss in weight of fruits was affected by the different packaging

materials, where LDPE 25 micron packaging film proved the best in controlling physiological

loss in weight of pomegranate fruit. This loss in weight was observed from 3rd day onwards

under ambient room conditions.

The different packaging materials significantly affected the decay loss in

pomegranate fruits. No decaying was noticed in fruits during first eight days of storage under

ambient room conditions. Fruits wrapped in LDPE 25 micron had least decay loss, among all

the treatments.

Variation in juice content, ascorbic acid content and titratable acidity are very

important in determining the optimum storage quality of pomegranate fruits. These contents

decreased with the advancement of storage period and the minimum was observed on 12th

day of storage. The fruits packed in LDPE 25 micron had the highest juice retention and

ascorbic acid content, whereas, the titratable acidity of the fruit was not affected by the

treatments.

Pomegranate fruits had the highest total soluble solids, pH, total soluble solids to acid

ratio, anthocyanin contents, total sugars, reducing sugars and non-reducing sugar on 12th day

of storage, as these increased with the advancement of storage period but the above

mentioned quality parameters were not affected by the treatments.

The pomegranate fruits wrapped in LDPE 25 micron packaging film had higher

overall acceptability or organoleptic rating under storage conditions. This rating declined

where the maximum organoleptic rating was given on very first day of storage and the

minimum on 12th day of storage.

Among the different packaging materials, LDPE 25 micron recorded the maximum

rise in CO2 concentration with sharp reduction in oxygen concentration and highest increase

in relative humidity, while PP 25 micron was found to have the lowest concentration of CO2,

highest concentration of O2 and the lowest rise in relative humidity through sensors. The

cellophane paper and cling films showed intermediate result for most of the components of

MAP. From the study, it is concluded that LDPE 25 micron was the best packaging material

for the modified atmosphere packaging of pomegranate fruits.

i

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I

APPENDIX-I

(9 POINT HEDONIC RATING SCALE)

Name …………….. Date……………

Fruit……………. Time ………….

INSTRUCTIONS: Taste the given samples and check how much you each like or dislike.

Use appropriate scale to show your attitude by assigning points that best describe your

feelings about the sample. An honest expression of yours will help us to evaluate on the basis

of the following scale.

Score Preference Code

Extremely desirable 9

Very much desirable 8

Moderately desirable 7

Slightly desirable 6

Neither desirable nor undesirable 5

Slightly undesirable 4

Moderately undesirable 3

Very much undesirable 2

Extremely undesirable 1

Sample

code

Colour and

appearance

Flavour Taste Overall

acceptability

Remarks

Signature

II

APPENDIX- II

Minimum and maximum temperature during the month of September and October 2016

Morning and evening Relative Humidity (%) during the month of September and October, 2016

Source: Department of Agricultural Metrology, CCS Haryana Agricultural University, Hisar

0.0

5.0

10.0

15.0

20.0

25.0

30.0

35.0

40.0

1 S

ept.

16

4 S

ept.

16

7 S

ept.

16

10

Sep

t. 1

6

13

Sep

t. 1

6

16

Sep

t. 1

6

19

Sep

t. 1

6

22

Sep

t. 1

6

25

Sep

t. 1

6

28

Sep

t. 1

6

1 O

ct. 1

6

4 O

ct. 1

6

7 O

ct. 1

6

10

Oct

. 16

13

Oct

. 16

16

Oct

. 16

19

Oct

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ABSTRACT

1. Title of Thesis : Studies on bio-extract coatings and packaging on the

shelf life of pomegranate fruits cultivar Mridula

2. Full Name of Degree Holder : Ms. Suchismita Jena

3. Admission Number : 2015A45M

4. Title of the Degree : Masters of Science

5. Name of Discipline : Horticulture- Fruit science

6. Name and Address of : Dr. R.K. Goyal Major Advisor Professor, Department of Horticulture

CCS HAU, Hisar-125004 (Haryana), India

7. Degree Awarding University : CCS Haryana Agricultural University

Hisar-125004 (Haryana), India

8. Year of Award of Degree : 2017

9. Major Subject : Horticulture- Fruit Science

10. Total No. of Pages in Thesis : 54 + vi + II

11. Number of Words in Abstract : 439

Keywords: Bio-extract coating, packaging, shelf life, quality, sensors, storage, pomegranate, Mridula

The experiment-I comprising of Aloe vera extract 50, 75 and 100%, ginger extract 1, 2 and

3% and mint extract 10, 20 and 30% and experiment-II comprising of LDPE 25 micron, PP 25 micron,

cling film and cellophane paper were conducted at Post-harvest Laboratory of the Department of

Horticulture, CCS Haryana Agricultural University, Hisar to evaluate the best bio-extract coating

material and the most effective packaging film for prolonging shelf life and maintaining physico-

chemical characteristics of high valued pomegranate fruits. In experiment-I, coating with Aloe vera

extract 100% was most effective in reducing physiological loss in weight and ginger extract 3% in

reducing decay loss. Different quality attributes like total soluble solids, pH, total soluble solids to acid

ratio, anthocyanin, total sugars, reducing sugars and non-reducing sugar of fruits increased, while juice

content, ascorbic acid and titratable acidity decreased with the advancement of storage period under

ambient room conditions. Among the treatments, Aloe vera extract 100% coated pomegranate fruit had

least content of total soluble solids and sugars and significantly highest content of juice, anthocyanin

and ascorbic acid but had no effect on titratable acidity and pH of the fruits. Hence, the highest

organoleptic rating was given to pomegranate fruits coated with Aloe vera extract 100%.

In experiment-II, the pomegranate fruits packed in LDPE 25 micron film had the least

physiological loss in weight and decay loss and highest juice and ascorbic acid content. The different

packaging material had no significant effect on total soluble solids, titratable acidity, pH, total soluble

solids to acid ratio, anthocyanin contents, total sugars, reducing sugars and non-reducing sugar,

however, storage period had significant effect on them and the significantly maximum total soluble

solids, sugars and anthocyanin content and pH was observed on 12th day of storage, as these increased

with the passage of time, on the other hand, the juice content, ascorbic acid and titratable acidity were

found maximum on the very first day but they decreased with the prolongation of storage period. The

pomegranate fruits wrapped in LDPE 25 micron packaging film had higher organoleptic rating and

overall acceptability under storage conditions. Among the different packaging materials, LDPE 25

micron recorded the maximum rise in CO2 concentration with sharp reduction in oxygen concentration

and highest increase in relative humidity, while PP 25 micron was found to have the lowest

concentration of CO2, highest concentration of O2 and the lowest rise in relative humidity through

sensors. The cellophane paper and cling films showed intermediate result for most of the components

of MAP. From the study, it is concluded that LDPE 25 micron was the best packaging material for the

modified atmosphere packaging of pomegranate fruits.

Major Advisor Head of the Department Signature of the Student

CURRICULUM VITAE

1. Name : Ms. Suchismita Jena

2. Date of Birth : January 25th, 1994

3. Place of Birth : Buguda, Ganjam (Odisha)

4. Mother’s Name : Mrs. Sabita Jena

5. Father’s Name : Mr. Pramod Kumar Jena

6. Permanent Address : Aurobindo Nagar 2nd

lane,

Buguda, Ganjam

Odisha- 761118

7. Mobile : +91-9812110226

+91-9178589847

8. E-mail : [email protected]

9. Academic Qualifications

Degree University Year of

Passing

Percentage

of marks

Subjects

M.Sc.

(Horticulture) CCS HAU, Hisar 2017 8.76/10

Major: Horticulture

Minor: Plant Physiology

B.Sc. (Hons.) Agri. OUAT, Bhubaneswar 2015 8.50/10 Agriculture

+2 Science People‟s College,

Buguda, Ganjam 2011 74.00

Odia, English, Physics,

Chemistry, Math., Biology

10th B.N. Girls High School,

Buguda, Ganjam, Odisha 2009 84.67

FLO, SLE, TLS, MTH, SCP,

SCL, SSG, SSH

10. Co-curricular activities

Keen interest in paintings, singing and dancing.

Participated in Duet Dance competition of Inter College Youth Festival, 2016 at

CCSHAU, Hisar.

Participated in “National Conference on Trends in Nanobiotechnology” at CCSHAU,

Hisar (November 29-30, 2016).

Participated in “State Level Workshop on Nursery Raising in Protected Conditions” at

CCSHAU, Hisar (March 5-6, 2016).

Participated in “Bakery and Confectionary Training Programme” at CCSHAU, Hisar

(February 6-18, 2017)

Participated in “Training Programme in Protected Cultivation of Horticultural Crops” at

CCSHAU, Hisar (December 21-01, 2016).

11. Medals/ Honours received

Awarded with “Bidya-bhushan Award” by Amul Company, during higher secondary

(12th).

Awarded by Odisha Government Prerna Scholarship throughout the Bachelor‟s degree

programme.

Secured AIR-43 in ICAR‟s 20th All India Entrance Examination (AIEEA-PG-2015),

and awarded with National Talent Scholarship by ICAR throughout the Master‟s degree

programme.

1st position in duet dance competition of Inter College Youth Festival, 2016 at

CCSHAU, Hisar.

12. Publications

Anuradha and Suchismita Jena, 2017. Citrus fruits - A source of antioxidant. Indian

Farmer, 4(3): 263-264.

UNDERTAKING OF COPY RIGHT

I, Suchismita Jena, Admission No. 2015A45M, hereby undertake that I give the full

copyrights of my thesis entitled Studies on bio-extract coatings and packaging on the shelf

life of pomegranate fruits cultivar Mridula to the Chaudhary Charan Singh Haryana

Agricultural University, Hisar.

I also undertake that the patent, if any, arising out of the research work conducted

during the programme shall be filed by me only with due permission with the competent

authority of Chaudhary Charan Singh Haryana Agricultural University, Hisar.

SIGNATURE OF STUDENT

studies on bio-extract coatings and …...certificate-ii studies onthis is to certify that the...

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