EXTRACTION, MODIFICATION AND
CHARACTERIZATION OF PECTIN FROM BANANA PEEL
By
Maryam Rabbani
2014-GCUF-32196
Thesis submitted in partial fulfillment of
the requirements for the degree of
MASTER OF PHILOSOPHY
IN
ANALYTICAL CHEMISTRY
DEPARTMENT OF CHEMISTRY
GOVERNMENT COLLEGE UNIVERSITY, FAISALABAD
November, 2020
DEDICATION
Every challenging work needs self-efforts as well as guidance of elders especially those
who are very close to our heart. My humble effort I dedicate to my Worthy
Parents
Whose affection, love, encouragement and prayers of day and night make me able to get
such success and honor
Along with all hard working and respected Teachers
ⅰ
DECLARATION
The work reported in this thesis was carried out by me under the supervision of Dr.
Shahzad Ali Shahid Chatha, Associate Professor, Department of Chemistry,
Government College University Faisalabad, Pakistan.
I hereby declare that the title of thesis “Extraction, modification and
characterization of pectin from banana peel” and the contents of thesis are the product
of my own research and no part has been copied from any published source (except the
references, standard mathematical of genetic models/equations/formulas/protocols etc). I
further declare that this work has not been submitted for the award of any other
degree/diploma. The University may take action if the information provided is found
inaccurate at any stage.
__________________
Maryam Rabbani
2014-GCUF-32196
ⅰ
CERTIFICATE BY THE SCRUTINY COMMITTEE
It is certified that the thesis submitted by Maryam Rabbani, Registration No. 2014GCUF-32196 is according to prescribed format of the Government College University
Faisalabad.
1. Dr. Shahzad Ali Shahid Chatha /Associate Professor
____________
Convener of Scrutiny Committee
2. Dr. Atta ul Haq /Associate Professor
____________
Member of Scrutiny Committee
3. Dr. Fozia Anjum /Assistant Professor
Member of Scrutiny Committee
ⅰ
____________
CERTIFICATE BY THE RESEARCH SUPERVISORY COMMITTEE
It is certified that the contents and form of a thesis submitted by Maryam Rabbani,
Registration No. 2014-GCUF-32196 has been satisfactory and in accordance with the
prescribed format. We recommend it to be processed for the evaluation by the external
examiner for the award of a degree.
Supervisor
__________________________________
Dr. Shahzad Ali Shahid Chatha
Associate Professor
Department of Chemistry, GCUF
Member of supervisory committee
__________________________________
Dr.Syed Ali Raza Naqvi
Associate Professor
Department of Chemistry, GCUF
Member of supervisory committee
__________________________________
Prof. Dr. Shazia Anwar Bukhari
Professor
Department of Biochemistry, GCUF
Chairperson
__________________________________
Dr. Muhammad Zubair
Associate Professor, Chairman
Department of Chemistry, GCUF
Dean
__________________________________
Prof. Dr. Farhat Jabeen
Dean
Faculty of Physical Sciences
Government College University Faisalabad
ⅰ
TABLE OF CONTENTS
Contents
Page Number
Acknowledgments
i
List of abbreviations
ii
List of figures
iii
List of tables
v
Abstract
vi
Chapter 1 Introduction
1
Chapter 2 Review of Literature
5
2.1 Overview of pectin
5
2.2 Chemistry of pectin
6
2.3 Classification of pectin
8
2.3.1 Native pectin
8
2.3.2 Commercial pectin
9
2.3.3 High methoxy pectin
9
2.3.4 Low methoxy pectin
9
2.4 Properties of pectin
10
2.4.1 Gelling properties
10
2.4.2 Emulsifyning properties
10
2.4.3 Solubility
10
2.5 Applications of pectin
11
2.6 Biological properties of pectin
13
2.6.1 Antioxident activity of pectin
13
2.6.2 Antibacterial activity of pectin
15
2.6.3 Antifungal activity of pectin
15
2.6.4 Antitumor activity of pectin
16
2.6.5 Anti-inflammatory activity of pectin
17
2.6.6 Anticancer activity of pectin
17
2.7 Sources of pectin
18
2.8 Extraction methods of pectin
19
2.8.1 Acid extraction
19
2.8.2 Microwave assisted extraction
21
2.8.3 Ultrasound assisted extraction
22
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2.8.4 Thermo mechanical extraction
22
2.8.5 Enzymatic extraction
22
2.9 Drawbacks of native pectin
24
2.10 Modification methods of pectin
24
2.10.1 Chemical modification
25
2.10.2 Biological modification
28
2.10.3 Physical modification
29
2.11 Characterization techniques for pectin
31
2.11.1 Fourier transform infrared spectroscopy
31
2.11.2 Atomic force microscopy
31
2.11.3 Scanning electron microscopy
31
2.11.4 Thermal analysis
32
2.11.5 X-ray diffraction
32
2.11.6 High performance exclusion
32
chromatography
2.11.7 Mass spectroscopy
Chapter 3 Materials and Methods
33
35
3.1 Chemicals and reagents
35
3.2 Collection and pretreatment of plant material
36
3.3 Preparation of crude extract
36
3.4 Phytochemical screening of fruit aqueous extract
36
3.4.1 Carbohydrate
37
3.4.2 Proteins
37
3.4.3 Flavonoids
38
3.4.4 Phenols
38
3.4.5 Free amino acids
38
3.5 Extraction of pectin
38
3.6 Chemical analysis of pectin
40
3.6.1 Total phenolic contents
40
3.6.2 Total flavonoid contents
40
3.7 Modification of pectin
41
3.8 Characterization for pectin
41
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3.8.1 FTIR analysis
41
3.8.2 XRD analysis
41
3.10.3 SEM and EDX analysis
41
3.9 Antioxidant activities
42
3.9.1 DPPH radical antioxidant assay
42
3.9.2 Hydroxyl radical antioxidant assay
42
3.9.3 Ferric reducing antioxidant power
43
3.9.4 Phospho molybdenum phosphate assay
43
3.10 Statistical analysis
43
Chapter 4 Results and Discussion
44
4.1 Extraction yield
44
4.2 Phytochemical screening of fruit aqueous extract
44
4.2.1 Carbohydrate
44
4.2.2 Flavonoids
45
4.2.3 Phenols
45
4.2.4 Proteins
45
4.2.5 Free amino acids
45
4.3 Chemical analysis
46
4.3.1 total flavonoid content
46
4.3.2 Total phenolic content
47
4.4 Modification Yield
48
4.5 Characterization techniques
48
4.5.1 FTIR analysis
48
4.5.2 SEM analysis
49
4.5.3 EDX analysis
50
4.5.4 XRD analysis
51
4.6 Antioxidant activities
52
4.6.1 DPPH radical scavenging assay
52
4.6.2 Ferric reducing antioxidant power
54
4.6.3 Hydroxyl scavenging activity assay
55
4.6.4 Phosphomolybdenum antioxidant assay
57
Summary
59
References
60
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ACKNOWLEDGMENTS
I have only pearls of my eye to admire the blessing of the compassionate and omnipotent
because word are bound. Knowledge is limited and time is short to express His dignity,
ALMIGHTY ALLAH, the propitious, the benevolent and sovereign Whose Blessing and
Glory flourished my thoughts and thrived my ambitions, giving me affectionate parents,
talented teachers, sweet brother and unique friends.
Trembling lips and wet eye praise for HOLY PROPHET HAZRAT
MUHAMMAD (P.B.U.H) for enlightening our conscience with the essence of faith in
ALLAH, converging all His kindness and mercy upon him.
The work presented in this manuscript was accomplished under the sympathetic attitude,
scholarly criticism, cheering perspective and enlightening supervision of Prof. Dr.
Shahzad Ali Shahid Chatha, Associate Professor, Department of Chemistry, GC
University Faisalabad, Pakistan. I would like to express my gratitude to my research
supervisor for their guidance and advice given throughout the course of this work.
Without his kind help, the timely completion of my research work would have not
possible for me.
I expressed my deep sense of gratitude to Dr. Zubair, Chairman, Department of
chemistry, GC University, Faisalabad for his generous guidance, inspiring discipline,
sincere and unprecedented leadership as well as moral patronization.
Friends are there comrades of the battle, the battle to generate knowledge, sift
myths and fact and to remove ambiguity. Not least of all, I owe so much to my whole
family specially my parents and my beloved brother Engr. M. Abdullah Khan for my
success is really the fruit of their devoted prayers. Also for their undying support, their
unwavering belief that I can achieve. Unfortunately, I cannot thank everyone by name
because it would take a lifetime but, I just want you all to know that you count so much.
Had it not been for all your prayers and benedictions, were it not for your sincere love and
help, I would never have completed this thesis. So, thank you all.
(Maryam Rabbani)
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LIST OF ABBREVIATIONS
Symbol
Abbreviations
LMP
Low mthoxy pectin
HMP
High methoy pectin
DPPH
2,2-diphenyl -1picrylhydrazyl
FRAP
Ferric reducing antioxidant power
HPLC
High performance liquid chromatography
SEM
Scanning electron microscopy
XRD
X-ray diffractometry
EDX
Energy dispersive xray
ANOVA
Analysis of Variance
SD
Standard deviation
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LIST OF FIGURES
Figure
Page Number
Figure 2.1: Structure of pectin
5
Figure 3.1: Schematic diagram of methadology
35
Figure 3.2: Pretreatment of plant material
36
Figure 3.3: Schematic diagram of extracion
39
Figure 3.4: Chemical analysis of pectin
40
Figure 4.1: Standard curve of gallic acid
47
Figure 4.2: Standard curve of quercetin
47
Figure 4.3: FTIR spectra of native pectin
49
Figure 4.4: FTIR spectra of modified pectin
49
Figure 4.5: SEM spectra of native pectin
50
Figure 4.6: SEM spectra of modified pectin
50
Figure 4.7: EDX spectrum of native pectin
51
Figure 4.8: EDX spectrum of modified pectin
51
Figure 4.9: XRD spectra of native pectin
52
Figure 4.10: XRD spectra of modified pectin
52
Figure 4.11: scavenging actity of DPPH
54
Figure 4.12: Scavenging activity of FRAP
55
Figure 4.13: Scavenging activity of hydroxyl assay
56
Figure 4.14: Scavenging activity of Phosphomolybdenum assay
58
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LIST OF TABLES
Table
Page Number
Table 1: Different sources of extraction
19
Table 2: Different modes of extraction
23
Table 3: Yield of extracted pectin
44
Table 4: Phytochemical screening test results
46
Table 5: DPPH radical scavenging antioxidant assay
53
Table 6: FRAP radical scavenging antioxidant assay
55
Table 7: Hydroxyl scavenging antioxidant assay
56
Table 8: Phospho molybdenum assay
57
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ABSTRACT
Pectin having the properties like low cost, local availability, biodegradability and nontoxic behaviors gaining attention for biomedical applications. Banana peel is a rich source
of pectin. The present study was aimed to extract the pectin from banana peel. In this
study the pectin was extracted from the banana peel and modified by one of the chemical
methods. Pectin extraction was done by using acid based extraction method.
Characterization techniques such as FTIR was used to determine the functional groups on
the structure of pectin. The surface morphology, elemental composition and crystalline
structure will be determined by SEM, EDX and XRD. The biological applications like
anti-oxidant will be comparatively analyzed of native and modified pectin with the help
of reported methods. The modification of natural pectin extracted from banana peel
enhanced the anti-oxidant activities.
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Chapter 1
INTRODUCTION
Natural products obtained from plants, animals, microorganisms and marine have been
used for maintenance of natural life in both native as well as the modified form from the
beginning of human life (Wardropper et al., 2020). They play a vital role in the
sustainability of life by curing the several disorders either appeared in the environment or
in human and animal life (Arshad et al., 2011). Plants based natural products have been
gaining special attention to researchers, herbalists and industrialists. Especially, natural
products from medicinal plants have to turn out to be a natural source of medicine to treat
several kinds of human and animal diseases (Abbas et al., 2011; Ghasemzadeh et al.,
2010). Medicinal plants are appreciated for natural products due to their effectiveness,
sustainable, efficient, beneficial as well as green nature (Ahmad & Husain, 2008).
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Chapter 2
REVIEW OF LITERATURE
2.1 Polysaccharides
The polysaccharides are polymers usually formed from more than ten sugar units linked
to each other with the help of the glycosidic linkage (Xie et al., 2016). The concentration
range of polysaccharide occurrence in plants was reported from 1% to 90% (Ridley et al.,
2001). Chemical composition and structure of polysaccharides act as factors to classify
them.
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Chapter 3
MATERIALS AND METHODS
The research work described in this dissertation was performed according to standard
operating procedures and a piece of description in the form of flow sheet is elaborated in
figure 3.1.
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Chapter 4
RESULTS AND DISCUSSION
In this study hidden antioxidant potential of polysaccharides from Carissa carandas fruit
was evaluated. Hot water extraction was employed to make crude extract of Carissa
carandas fruit and this extract was allowed to perform phytochemical screening tests.
ⅰ
SUMMARY
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REFERENCES
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