42
STUDIES ON PRODUCTION OF PECTIN FROM FRUIT WASTES
AVAILABLE IN PAKISTAN. IT’S BIO- CHARACTERIZATION
AND UTILIZATION IN THE DEVELOPMENT OF
PHARMACEUTICAL AND FOOD PRODUCTS
BY
NAUSHEEN HAMEED SIDDIQUI
B-Pharm., M-Pharm.
Thesis submitted for the partial fulfillment of the degree of
DOCTOR OF PHILOSOPHY
DEPARTMENT OF PHARMACOGNOSY
FACULTY OF PHARMACY AND PHARMACEUTICAL SCIENCES
UNIVERSITY OF KARACHI
KARACHI 75250
PAKISTAN
2016
43
DEDICATION
Dedicated to my parents for their continuous support
To my husband for be my strength and
to my priceless loving children
Maryam, Mysha and Ayaan
who are source of inspiration for me.
44
TABLE OF CONTENTS
List of Tables vii
List of Figures xvi
List of Abbreviations xix
Acknowledgements xxii
Abstract xxv
Urdu Translation xix
PART - I
INTRODUCTION 1
REVIEW OF LITERATURE 14
History of pectin 14
Chemistry and type of pectin 15
Biosynthesis of pectin 16
Classification of Pectin 17
Extraction and purification of pectin 18
Traditional method of extraction 22
Microwave- assisted extraction 23
Enzymatic Extraction of pectin 25
Purification of Pectin 27
Uses and application of pectin 34
Large scale production of pectin 39
45
PART - II
MATERIALS AND METHODS 42
Equipment 42
List of chemicals and reagents used 45
List of apparatus 48
List of solutions used 49
Extraction of pectin 53
Processing and preparation of peel 53
Screening of seasonal fruits and their wastes for the presence of pectin
contents
53
Extraction of pectin from selected fruit wastes and their comparative
study
54
Mechanical, pH and heating factors used during the study 55
Effects of organic acid and inorganic acids and their strength on pectin
yield from sapodilla
58
Extraction with 0.1N HCl, using 5 mechanical procedures, 5 pH, two
boiling methods and varying time of boiling (10, 20, 40 and 60 min)
58
Extraction using 1N HCl, using 5 mechanical procedure, 5 pH, two
boiling methods and varying time of boiling (10, 20, 40 and 60 min)
58
Extraction using organic acids 59
Extraction with different strengths of inorganic acid 59
Physical and biochemical characterization of pectin from Sapodilla 59
46
Percent yield 59
Identification tests for pectin 59
Stiff gel test 60
Test with 95% ethanol 60
Test with potassium hydroxide (KOH) 60
Iodine test 60
Biochemical characterization 61
Preparation of sample 61
Qualitative test for ammonia 61
Detection of moisture, ash, methoxy content and equivalent weight 61
Moisture 61
Ash 61
Equivalent weight 62
Methoxyl content 62
Anhydrouronic Acid 62
Grading of pectin 63
Galacturonic acid content 64
Water holding capacity (WHC) 64
Water binding capacity (WBC) 65
Fat binding capacity (FBC) 66
Fourier Transform Infrared Spectroscopy of selected pectin 66
47
Dynamic Light Scattering studies of selected pectin 66
Optimization of the yield of pectin from sapodilla peel thorough
Response Surface Methodology (RSM)
67
Pharmaceutical preparation 68
Formulation of tablet using extracted pectin from sapodilla peel 68
Evaluation of granules 71
Angle of repose (α) 71
Bulk density (ρb) 71
Tapped density (ρt) 72
Compressibility Index 72
Hausner’s ratio 72
Loss on drying (LOD) 72
Tablet compression 73
Tablets testing 73
Weight variation 74
Tablet thickness and diameter 74
Tablet hardness 74
In-Vitro dissolution studies 74
Formulation of antidiarrheal preparation (suspension) from extracted
sapodilla pectin.
75
Evaluation of Suspension 76
48
Color, odor and taste 76
Viscosity 76
Sedimentation Volume 76
Redispersibility 77
Food preparation 79
Preparation of Sapodilla jam 79
Physico-chemical analysis of jam 80
Determination of pH 80
Determination of viscosity 80
Determination of moisture and ash contents 80
Determination of total titratable acidity (TTA) 80
Determination of total soluble sugar 80
Determination of total solids 80
Determination of vitamin C (Ascorbic Acid) 81
Sensory Evaluation 81
Preparation of pudding from extracted sapodilla pectin 83
PART - III
RESULTS 86
Extraction and purification of pectin from the selected fruit waste. 86
Physical and Biochemical characterization of the purified pectin 88
Optimization of the yield of pectin from sapodilla peel thorough 88
49
response surface methodology
Utilization of the purified pectin for the development of pharmaceutical
and food products
89
PART – IV
DISCUSSION AND CONCLUSION 169
DISSCUSSION 169
CONCLUSION 206
REFERENCES 212
ANNEXURES
PUBLICATIONS
50
LIST OF TABLES
Table 1 Showing the uses of different types of pectin in various types of
food products
19
Table 2 Showing some investigated sources of pectin with their applied
mode of extractions
20
Table 3 Showing different uses and properties of pectin. (Source: J.F.
Hydrocolloids)
37
Table 4 Composition of Paracetamol tablet with different concentrations of
pectin used
69
Table 5 Composition of Ibuprufen tablet with different concentrations of
pectin used
70
Table 6 Composition of antidiarrheal preparation using extracted sapodilla
pectin.
78
Table 7 Formulation of the sapodilla Jam (1KG) with different concentration
of pectin
82
Table 8 Showing different pectin concentration used for each selected
extracted fruit pectin
85
Table 9 List of fruits used in the screening study for the presence of pectin 91
Table 10 Showing percent yield of pectin from five selected fruits 92
Table 11 Analysis of variance (mean squares) of yield for five selected fruits 93
Table 12 Means comparison of yield for sapodilla fruit peel (Mechanical
procedure pH interaction mean±SE )
94
Table 13 Means comparison of yield for sapodilla fruit peel (Mechanical
procedure boiling method interaction mean±SE)
94
Table 14 Means comparison of yield for sapodilla fruit peel (Mechanical
procedure pH boiling method interaction mean±SE)
95
51
Table 15 Means comparison of yield for banana fruit peel (Mechanical
procedure pH interaction mean±SE)
96
Table 16 Means comparison of yield for banana fruit peel (Mechanical
procedure boiling method interaction mean±SE)
96
Table 17 Means comparison of yield for banana fruit peel (Mechanical
procedure pH boiling method interaction mean±SE)
97
Table 18 Means comparison of yield for muskmelon fruit peel (Mechanical
procedure pH interaction mean±SE)
98
Table 19 Means comparison of yield for muskmelon fruit peel (Mechanical
procedure boiling method interaction mean±SE)
98
Table 20 Means comparison of yield for muskmelon fruit peel (Mechanical
procedure pH boiling method interaction mean±SE)
99
Table 21 Means comparison of yield for apple fruit peel (Mechanical
procedure pH interaction mean±SE)
100
Table 22 Means comparison of yield for apple fruit peel (Mechanical
procedure boiling method interaction mean±SE)
100
Table 23 Means comparison of yield for apple fruit peel (Mechanical
procedure pH boiling method interaction mean±SE)
101
Table 24 Means comparison of yield for orange fruit peel (Mechanical
procedure pH interaction mean±SE)
102
Table 25 Means comparison of yield for orange fruit peel (Mechanical
procedure boiling method interaction mean±SE)
102
Table 26 Means comparison of yield for orange fruit peel (Mechanical
procedure pH boiling method interaction mean±SE)
103
Table 27 Percentage yield of pectin from sapodilla fruit peel after using
different physicomechanical procedures (pH, mechanical procedure,
104
52
boiling method and time of boling) with 0.1N HCl
Table 28 Analysis of variance (mean squares) of yield of pectin from
sapodilla fruit peel after using different physicomechanical
procedure different fruits with 0.1N HCl
105
Table 29 Means comparison of yield for sapodilla fruit peel after 10 min of
boiling using 0.1N HCl (Mechanical procedure pH interaction
mean±SE)
106
Table 30 Means comparison of yield for sapodilla fruit peel after 10 min of
boiling using 0.1N HCl (Mechanical procedure boiling method
interaction mean±SE)
106
Table 31 Means comparison of yield for sapodilla fruit peel after 10 min of
boiling using 0.1N HCl (Mechanical procedure pH boiling
method interaction mean±SE)
107
Table 32 Means comparison of yield for sapodilla fruit peel after 20 min of
boiling using 0.1N HCl (Mechanical procedure pH interaction
mean±SE)
108
Table 33 Means comparison of yield for sapodilla fruit peel after 20 min of
boiling using 0.1N HCl (Mechanical procedure boiling method
interaction mean±SE)
108
Table 34 Means comparison of yield for sapodilla fruit peel after 20 min of
boiling using 0.1N HCl ( Mechanical procedure pH boiling
method interaction mean±SE)
109
Table 35 Means comparison of yield for sapodilla fruit peel after 40 min of
boiling using 0.1N HCl (Mechanical procedure pH interaction
mean±SE)
110
Table 36 Means comparison of yield for sapodilla fruit peel after 40 min of
boiling using 0.1N HCl ( Mechanical procedure boiling method
interaction mean±SE)
110
53
Table 37 Means comparison of yield for sapodilla fruit peel after 40 min of
boiling using 0.1N HCl (Mechanical procedure pH boiling
method interaction mean±SE)
111
Table 38 Means comparison of yield for sapodilla fruit peel after 60 min of
boiling using 0.1N HCl Mechanical procedure pH interaction
mean±SE)
112
Table 39 Means comparison of yield for sapodilla fruit peel after 60 min of
boiling using 0.1N HCl (Mechanical procedure boiling method
interaction mean±SE)
112
Table 40 Means comparison of yield for sapodilla fruit peel after 60 min of
boiling using 0.1N HCl (Mechanical procedure pH boiling
method interaction mean±SE)
113
Table 41 Percentage yield of pectin from sapodilla fruit peel after using
different physicomechanical procedures (pH, mechanical procedure,
boiling method and time of boling) with IN HCl
114
Table 42 Analysis of variance (mean squares) of yield of pectin from
sapodilla fruit peel after using different physicomechanical
procedure different fruits with 1N HCl
115
Table 43 Means comparison of yield for sapodilla fruit peel after 10 min of
boiling using 1N HCl ( Mechanical procedure pH interaction
mean±SE)
116
Table 44 Means comparison of yield for sapodilla fruit peel after 10 min of
boiling using 1N HCl ( Mechanical procedure boiling method
interaction mean±SE)
116
Table 45 Means comparison of yield for sapodilla fruit peel after 10 min of
boiling using 1N HCl (Mechanical procedure pH boiling method
interaction mean±SE)
117
54
Table 46 Means comparison of yield for sapodilla fruit peel after 20 min of
boiling using 1N HCl (Mechanical procedure pH interaction
mean±SE)
118
Table 47 Means comparison of yield for sapodilla fruit peel after 20 min of
boiling using 1N HCl ( Mechanical procedure boiling method
interaction mean±SE)
119
Table 48 Means comparison of yield for sapodilla fruit peel after 20 min of
boiling using 1N HCl (Mechanical procedure pH boiling method
interaction mean±SE)
120
Table 49 Means comparison of yield for sapodilla fruit peel after 40 min of
boiling using 1N HCl (Mechanical procedure pH interaction
mean±SE)
121
Table 50 Means comparison of yield for sapodilla fruit peel after 40 min of
boiling using 1N HCl (Mechanical procedure boiling method
interaction mean±SE)
121
Table 51 Means comparison of yield for sapodilla fruit peel after 40 min of
boiling using 1N HCl ( Mechanical procedure pH boiling
method interaction mean±SE)
122
Table 52 Means comparison of yield for sapodilla fruit peel after 60 min of
boiling using 1N HCl (Mechanical procedure pH interaction
mean±SE)
123
Table 53 Means comparison of yield for sapodilla fruit peel after 60 min of
boiling using 1N HCl (Mechanical procedure boiling method
interaction mean±SE)
123
Table 54 Means comparison of yield for sapodilla fruit peel after 60 min of
boiling using 1N HCl ( Mechanical procedure pH boiling
method interaction mean±SE)
124
55
Table 55 Means comparison of yield for sapodilla fruit peel after using
different inorganic acid
125
Table 56 Analysis of variance (mean squares) of yield of pectin from
sapodilla fruit peel after using different inorganic acid
126
Table 57 Means comparison of yield for sapodilla fruit peel after using citric
acid (Boiling method x mechanical procedure interaction mean±SE)
126
Table 58 Means comparison of yield for sapodilla fruit peel after using citric
acid (Acid % x boiling method interaction mean±SE)
127
Table 59 Means comparison of yield for sapodilla fruit peel after using citric
acid (Boiling method x Acid% x mechanical procedure interaction
mean±SE)
127
Table 60 Means comparison of yield for sapodilla fruit peel after using oxalic
acid (Boiling method x mechanical procedure interaction mean±SE)
128
Table 61 Means comparison of yield for sapodilla fruit peel after using oxalic
acid (Acid% x boiling method interaction mean±SE
128
Table 62 Means comparison of yield for sapodilla fruit peel after using oxalic
acid (Boiling method x Acid% x mechanical procedure interaction
mean±SE)
129
Table 63 Means comparison of yield for sapodilla fruit peel after using
tartaric acid (Boiling method x mechanical procedure interaction
mean±SE)
130
Table 64 Means comparison of yield for sapodilla fruit peel after using
tartaric acid (Acid% x boiling method interaction mean±SE)
130
Table 65 Means comparison of yield for sapodilla fruit peel after using
tartaric acid( Boiling method x Acid% x mechanical procedure
interaction mean±SE)
131
Table 66 Percentage yield of pectin from sapodilla fruit peel using different 132
56
strengths of same inorganic acid.
Table 67 Analysis of variance (mean squares) of yield of pectin from
sapodilla fruit peel after using different strength of same inorganic
acid
133
Table 68 Means comparison for yield of pectin from sapodilla fruit peel after
using 0.1N HCl (Mechanical procedure pH interaction mean±SE )
134
Table 69 Means comparison for yield of pectin from sapodilla fruit peel after
using 0.1N HCl (Mechanical procedure boiling method interaction
mean±SE)
134
Table 70 Means comparison for yield of pectin from sapodilla fruit peel after
using 0.1N HCl (Mechanical procedure pH boiling method
interaction mean±SE)
135
Table 71 Means comparison for yield of pectin from sapodilla fruit peel after
using 0.5N HCl (Mechanical procedure pH interaction mean±SE)
136
Table 72 Means comparison for yield of pectin from sapodilla fruit peel after
using 0.5N HCl (Mechanical procedure boiling method interaction
mean±SE)
136
Table 73 Means comparison for yield of pectin from sapodilla fruit peel after
using 0.5N HCl (Mechanical procedure pH boiling method
interaction mean±SE)
137
Table 74 Means comparison for yield of pectin from sapodilla fruit peel after
using 1N HCl (Mechanical procedure pH interaction mean±SE)
138
Table 75 Means comparison for yield of pectin from sapodilla fruit peel after
using 1N HCl (Mechanical procedure boiling method interaction
mean±SE)
138
Table 76 Means comparison for yield of pectin from sapodilla fruit peel after
using 1N HCl (Mechanical procedure pH boiling method
139
57
interaction mean±SE)
Table 77 Identification tests for presence of pectin from three selected fruits 140
Table 78 Biochemical characterization of the purified pectin extracted from
sapodilla pectin at different pH
140
Table 79 Water holding, water binding and fat binding capacity of sapodilla
pectin
141
Table 80 Showing FTIR spectral values of sample and standard pectin along
with associated functional groups.
141
Table 81 Summary of the physical characterization of standard pectin and the
different pectin extracted from various source and at different pH by
DLS studies.
144
Table 82 Box-Bechen experimental design and levels of factors used for
optimization of pectin yield
150
Table 83 Box-Behnken experimental design and corresponding results for
responses
150
Table 84 Analysis of Variance for Yield (Box-Bechen experimental design,
response surface methodology)
151
Table 85 Estimated Regression Coefficients for Yield (Box-Bechen
experimental design, response surface methodology)
152
Table 86 Predicted values of yield. (Box-Bechen experimental design,
response surface methodology)
152
Table 87 Flow properties of granules made for paracetamol tablets 157
Table 88 Pharmaceutical characteristics of compressed formulation of
paracetamol tablet
158
Table 89 Dissolution studies of paracetamol tablet 159
Table 90 Flow properties of granules made for ibuprofen tablets 163
58
Table 91 Pharmaceutical characteristics of compressed formulation of
ibuprofen tablet
163
Table 92 Dissolution studies of ibuprufen tablets 164
Table 93 Basic evaluation test of antidiarrheal formulation prepared from
sapodilla pectin
166
Table 94 Effect of different concentration of sapodilla pectin on the chemical
properties of the jam samples
167
Table 95 Scores for sensory parameters of jam as judged by twenty (20)
panelists.
167
59
LIST OF FIGURES
Fig. 1 Structure of plant cell showing location of pectin in the cell wall 1
Fig. 2 Showing colour and texture of pectin 2
Fig. 3 (A) Showing formation of viscous gel and (B) showing increased bulk
volume of stool by Pectin
3
Fig. 4 Showing effect of retention of NFM by pectin on outer and inner
stratum corneum of the skin. The improved hydration of the
epidermis leads of enhanced mechanical stability of the stratum
granulosum, stratum spinosum and stratum basle
4
Fig. 5 Showing glycoside linkage 5
Fig. 6 α-4-linked galactosyluronic acid (GaIA) 6
Fig. 7 Schematic representations of the conventional (A) and alternative
(B) structures of pectin.
7
Fig. 8 Showing major sources of pectin used arround the world 10
Fig. 9 Showing The overall application and uses of pectin in
pharmaceutical, food, cosmetics and chemical industries
10
Fig. 10 Showing major pectin producers and their market share 11
Fig. 11 Showing the global pectin market 12
Fig. 12 Showing current and growing market size of hydrocolloids in the
different regions of the world
13
Fig. 13 Schematic diagram for biosynthesis of pectin 17
Fig. 14 Extraction of pectin from soy hull. 28
Fig. 15 Extraction of pectin from leaves of Nephrolepis biserrata 29
Fig. 16 Extraction of pectin from raw papaya peel 30
Fig. 17 Commercial extraction of pectin ( Ensymm) 31
60
Fig. 18 Commercial extraction of pectin (Cargillfood) 32
Fig. 19 Commercial extraction of pectin (GENU) 33
Fig. 20 Showing marketed of pectin in Middle east and Australia. 38
Fig. 21 Flow chart showing general procedure employed for the extraction
of pectin on commercial scale.
41
Fig. 22 Showing flowchart of extraction of pectin followed during the
current study
57
Fig. 23 FTIR spectra of food grade pectin 142
Fig. 24 FTIR spectra of sapodilla peel pectin extracted at pH 5 142
Fig. 25 FTIR spectra of sapodilla peel pectin extracted at pH 3 143
Fig. 26 FTIR spectra of sapodilla peel pectin extracted at pH 1 143
Fig. 27 Physical characterization of standard pectin by DLS studies 145
Fig. 28 Physical characterization of apple pectin extracted at pH5.0 by
DLS studies
146
Fig. 29 Physical characterization orange pectin extracted at pH5.0 by DLS
studies
147
Fig. 30 Physical characterization of sapodilla pectin extracted at pH3.0 by
DLS studies.
148
Fig. 31 Physical characterization of sapodilla pectin extracted at pH5.0 by
DLS studies
149
Fig. 32 Showing the optimal conditions for the extraction of pectin from
sapodilla fruit peel(Box-Bechen experimental design, response
surface methodology)
153
Fig. 33 Response surface graph and contour plot of effect of pH and
temperature on yield of pectin at constant time
154
Fig. 34 Response surface graph and contour plot of effect of pH and time 155
Boiling for 5 min
61
on yield of pectin at constant temperature
Fig. 35 Response surface graph and contour plot of effect of temperature
and time on yield of pectin at constant pH
156
Fig. 36 Formulated paracetamol tablest ( F1, F2, F3) 160
Fig. 37 Formulated paracetamol tablest ( F4, F5, F6) 161
Fig. 38 Formulated paracetamol tablest ( F7, F8, F9) 162
Fig. 39 Formulated ipubrufen tablets ( R1, R2, R3, R4) 165
Fig. 40 Formulated antidiarrheal preparation 166
Fig. 41 Formulation of Jam made from extracted pectin 168
Fig. 42 Formulation of pudding made from extracted pectin. 168
62
Abbreviation
Abbreviation Full Form
AGA Apigalacturonan
AOAC Association of Official Analytical Chemists
ADI Accepted Daily Intake
AUA Anhydrouronic acid
ANOVA Analysis of Variance
ACF Autocorrealtion function
API Active Pharmaceutical ingredients
Ca++ Calcium ion
CAS Chemical abstract service
CDTA Cyclohexanediaminetetraacetic acid
CFR Code of Federal Regulations
CeO Cerium oxide
CP Centipoise
Coef Coefficient
DLS Dynamic light scattering
DM Degree of methoxylation
DE Degree of esterification
DI Deionized water
EDTA Ethylenediaminetetraacetic Acid
FTIR Fourier transform infrared spectroscopy
63
FDA Food and Drug Administration
FAO Food and Agriculture Organization of the United Nations
FGP
g
Food grade pectin
Gram
GA Galacturonic acid
GI Gastrointestinal
GC-MS Gas chromatography–mass spectrometry
Gal A Galacturonic acid
GRAS Generally Recognized as Safe
HG Homogalacturonan
HGA Homogalacturonan
H2SO4 Sulphuric acid
HPLC High Performance Liquid Chromatography
HCL Hydrochloric acid
HM High methoxyl pectin
H2O Water
HIV Human immunodeficiency virus
IMS Industrial methylated spirit
JECFA The Joint FAO/WHO Expert Committee on Food Additives
KOH Potassium hydrochloride
Kg Kilogram
KP Kilopond
64
LM Low methoxyl pectin
MT Metric ton
MW Microwave
MAE Microwave assisted extraction
ml Millilitre
mg Milligram
mW Milliwatt
MOH Ministry of health
mM Millimolar
N Normal
NFM Natural moisturizing factor
NaOH Sodium hydroxide
nm Nanometer
NH4OH Ammonium hydroxide
NaN3 Sodium azide
NaCl Sodium chloride
%PD Perrcent polydispersity
PCSIR Pakistan Council of Scientific and Industrial Research
ppm Parts pur million
RG-I Rhamnogalacturonan I
RG-II Rhamnogalacturonan II
RSM Response surface methodology
65
RH Radius of hydration
Rpm
s
Revolutions per minute
second
SE Standard error
SD Standard deviation
TSS Total soluble solids
TTA Total titratable acidity
TS Total solids
USP 36/NF United States Pharmacopeia (USP) and the National Formulary
VIFS Variance influence factor
Vit C Vitamin C
W Watt
w/v Weight by volume
WBC water binding capacity
WHC Water holding capacity
WHO World health organization
XGA Xylogalacturonan
66
ACKNOWLEDGEMENT
All praises goes to Allah Almighty because without his countless blessing and love this
work would have been possible.
I would like to express my deep and sincere gratitude to my supervisor, Professor Dr
Iqbal Azhar, Dean Faculty of Pharmacy and Pharmaceutical Sciences, University of
Karachi for his kind support, advices, productive and useful opinions and suggestions
throughout my work. His constant encouragement and attitude has always been a moral
support for me. His continuous help and confidence in my abilities have been an integral
part for my understanding I will always be grateful.
I owe a lot of gratitude to my second supervisor Dr Zafar Alam Mahmood, Country
Manager, Colorcon Limited-England. It is my pride and privilege to express my sincere
thanks and deep sense of gratitude for his motivation, earnestness and immense
knowledge. His preeminent guidance supported me during the time of research and
writing of this thesis. His tutoring and keen interest made my research both enjoyable and
exciting. It is my great honor to study and do research under his talented supervision.
In addition, I would in particular like to thank Dr. M. Mohtasheemul Hasan (Chairman),
Prof Dr. Waseemuddin Ahmed, Professor Dr Mansoor Ahmed, Prof. Dr. Ghazala H.
Rizwan, (former Dean of Faculty of Pharmacy), Ms Farah Mazhar (Assistant prof), Mr.
Salman Ahmed and Mrs. Safia Abdi (Lecturer) Department of Pharmacognosy
University of Karachi and Prof. Dr. M. Shaiq Ali (Assistant professor International
Centre for Chemical & Biological Sciences, ICCBS, HEJ Research Institute of
Chemistry, University of Karachi) for their special supports. I also give thanks to my
67
colleagues and workers at Department of Pharmacognosy Faculty of Pharmacy for their
kind support. I am very grateful to Dr. Abid Ali (Assistant professor International Centre
for Chemical & Biological Sciences, ICCBS, HEJ Research Institute of Chemistry,
University of Karachi) for providing me support in the DLS studies and the important
extensive discussions and contributions concerning my work
I owe loving thanks to my husband, Tariq for his amazing love and support especially
through the hard times. His soothing words always made me feel better and urged me on.
I thank him for his sacrifices and supports throughout the course of my PhD.
I feel particularly indebted to my parents for their ever-lasting love, understanding and
encouragement. It was their unshakable faith in me that has always helped me to proceed
further. I cannot express my love and gratitude I have for them. Thank you for
encouraging me and supporting me in every step so that I can fulfill my dreams.
I wish to thank my brothers and sisters and their spouses who were always there for me in
my difficulties, without their encouragement, understanding, support and prayers it would
have been impossible for me to finish this work. Thanks to my in-laws specially my
Father in law and friends for their constant encouragement when I needed it. I feel short
of words to express my most heartfelt and cordial thanks to my children, who have
always stood by my side at the toughest times their unconditional love kept me going.
Finally, I wish to extend warm thanks to my aunt Mrs Shamsunisa Siddiqui, my sister
Ambreen, nieces Romina and Adina, cousins Marya and Nida and to everybody involved
directly or indirectly with my work.
Nausheen Hameed Siddiqui
68
ABSTRACT
With the rising awareness and use of plant based functional fibers there is a growing
market demand of pectin throughout the world. However, this is mainly linked with the
price and quality of the products availability in the market for different purposes. The
market price and current production of pectin has been influenced by many factors
among which the provision of raw material for the extraction of pectin is of main
concern. The purpose of this research investigations was not only to explore new source
of pectin which can aid production of pectin but also to extract it in a simpler way as to
minimize or reduce the cost of production. A systematic literature search was also carried
out to evaluate the present findings comprising the earlier results in term of its application
in developing food and pharmaceutical products.
This thesis is based on work to find out the effective extraction of pectin from various
fruit wastes available in Pakistan, its extraction in a most simple and effective way and
then after extracting its usage in both pharmaceutical and food product. For this purpose
numerous seasonal fruits available in Pakistan were studied in their respective time of
availability as a screening program.The present research was focused on the isolation,
physicochemical characterization and functional properties of pectin extracted from a
new source. The extraction process for effective extraction was developed after using
different solvents (organic and inorganic acids of different strengths), five extracting pH
(1,3 ,5 6 and 7), five mechanical procedure (homogenizing , grinding, hammering, cutting
and chopping) , two different boiling techniques, (Bunsen burner and microwave heating)
. Sapodilla fruits when selected finally was subjected to other affecting parameters for
better yield, like time of boiling (10, 20, 40 and 60 min respectively) with two different
69
strength of acid (0.1 and 1N HCl) strength of inorganic acid (0.1, 0.5 and 1N HCl
respectively), and effect of organic acid (critic, tartaric and acetic acid)
Initially five fruits, sapodilla, banana, muskmelon, apple and orange were selected for
studying the physic mechanical effects among which two fruits (Apple and orange) were
purely used for comparative study purpose. After this study three fruits (banana sapodilla
and muskmelon) were selected for further evaluation. . Out of three fruits, highest pectin
yield was recorded from banana (10.5%) followed by sapodilla (4.7%) and muskmelon
(4.4%) respectively. In comparison orange peel indicated 22.7% and apple peel 4.85%.
Identification and jelly grade tests were performed which supported sapodilla as most
appropriate against banana and muskmelon pectin for further investigation to find out a
new source of pectin which can be chemically stable, easily available, and
pharmaceutically useful in nature. Hence sapodilla peel was used to extract pectin after
using different phyico-mechanical processes. It was observed that the best extraction
parameter to obtain maximum yield of pectin was after 10 min of boiling of chopped
peels of sapodilla maintaining the pH with 1NHCl to be at pH 5.
Present research was also concerned with the bio characterization of extracted sapodilla
pectin. With the equivalent weight of 1700, degree of esterification 73.63% and jelly
grade 100, sapodilla pectin was evaluated as high methoxyl pectin. Water binding, water
holding and fat binding capacity was also assessed which also showed promising results.
FTIR spectroscopy performed found that the pectin at pH5 was of best quality as
compared to the other extracted pectin at different pH. Dynamic light scattering studies
(DLS) were also performed to determine the partial side and molecular weight of
extracted pectin and was compared with the commercial food grade pectin. The DLS
70
studies showed similarity in the particle size and molecular weight between standard and
extracted pectin which also proved that the extraction performed was effective.
Optimization of process of extraction of pectin was a studied after applying statistical
software Box Behnken design. According to response surface methodology the best yield
of pectin (3.7%) from sapodilla fruits can be achieved by keeping the pH 5 at 61.11 oC
for 90 min of heating time. The verification of predicted model also gave similar results,
3.5% pectin was extracted on the predicted pH, temperature and time of boiling.
The extracted pectin was also used in the formulation of two types of solid
pharmaceutical dosage form and an oral liquid preparation (suspension). The tablets were
first tested for its micromeritics properties of the granules. After the formulation of the
desired tablets the tablets were compressed and were tested for hardness, thickness, loss
on drying and dissolution properties. It was found that the concentration of added pectin
has influence on both the micrometric properties of granules as well as on the dissolution
profile of formulated tablets. Both the tablets showed increased in hardness and lowering
of dissolution rate with the addition of increased amount of pectin. However the best
formulation for paracetamol was F4 and F5 with 40 and 50 mg of pectin respectively
while for ibuprofen R1 with 50mg of pectin concentration was best. The antidiarrheal
preparation also exibited similar results in terms of its evaluation as suspension.
The extracted sapodilla pectin was then used in preparation of food and pharmaceutical
products. Two types of food preparation, jam and pudding were made using the extracted
sapodilla pectin. The jam was also evaluated for its chemical and sensory attributes and
found sapodilla pectin can be used in making of jam with a slightly higher concentration
(10mg) than the food grade pectin while pudding was evaluated for its textural properties
71
mainly and found that the addition of sapodilla pectin (10 and 15g) has no significant
impact on the textural properties of pudding and the pudding was equally acceptable as
pudding made from equal amount of food grade pectin
72
73
74
INTRODUCTION
Considerable focus, both on global and regional level has been given during last decade
towards the systematic research and application of the various findings or outcomes to
support the pharmaceutical, food and cosmetic industries in developing useful
formulations or products. This idea has created a great impact and geared up the research
activity in the field of both primary and secondary metabolites of different biochemical
groups, especially the high molecular weight compounds produced naturally and with
biodegradable property. Perhaps, it will not be exaggerated to interpret that the scientific
knowledge of phytochemicals or phytopharmaceuticals with the current and future
resources will have significant impact in the process development for the introduction of
varieties of industrial or commercial products from renewable plants, fruits and
vegetables. Among various biologically active groups, one of the leading groups of
natural product, which has been given especial attention, is the polysaccharide moiety.
Figure – 1: Structure of plant cell showing location of pectin in the cell wall.
(Source: Molecular expressions, 2016)
75
The polysaccharides are of tremendous importance with regard to their application in
pharmaceutical, nutraceuticals, cosmeceuticals and chemical industry and of best
example which can be quoted here is “Pectin” which is the most structurally complicated.
Polysaccharide in the plant cell wall (Figure – 1) with vast applications in these
commercial organizations with respect to health, food and personal care productsIn plant
cells, pectin is linked to cellulose and hemicellulose. The initial form is protopectin
which is a water insoluble pectic substance and are present as an important parts of
middle lamella among the cells as calcium pectate and magnesium pectate (Figure -1).
The cellulose to which pectin in linked in the cell, provides sufficient support to the
tissues in maintaining their rigidity while the pectic component makes the plant flexible
in nature. Pectin is present as protopectin in unripe fruits which is transformed when the
fruit ripens into pectin, which when mixed with water has a tendency of forming gel and
also among sources of water soluble fibres. However, the jellying capacity of pectin
diminishes when in over ripe fruits pectin is converted into pectic acid. Comes from the
Greek word meaning congealed or curdled, pectin was first isolated and reported in 1790
by Vauquelin, followed by its characterization 1825 Barconnot from the cell wall of fruits
and vegetables. At present, commercial pectin is mainly obtained from citrus fruits and is
Figure – 2: Showing colour and texture of pectin. (Source: Wikipedia, 2016)
76
a white to light brown powder (Figure - 2). However, production methods (isolation,
extraction and purification) used by different manufactures are highly confidential and
has patent protection.
The role of pectin is quite diversified in pharmaceuticals. These include both therapeutic
as well as excipient. In view of its properties to increase the viscosity and volume of stool
it has its applications both in diarrhoea and constipation. Though, FDA has discontinued
pectin in 1992, it is still use in other countries. It is also extensively used in Alternative
System of Medicine (such as Ayurvedic and Unani system of medicine) alone or in
combination with certain herbs or minerals as a compound or poly herbal formulations.
Pectin has the property to absorb water and can swell more than 40 times in volume to
form a viscous gel that helps to lubricate the lining of the intestine for ease of defecation.
It also increases bulk volume.
Figure – 3 (A) Showing formation of viscous gel and (B) showing increased bulk
volume of stool by Pectin. (Source: Anonymous)
The viscous gel that is formed softens and adds bulk to the stool to accelerate and
regulate the movement of food through the intestine leading to overall bowel cleansing
and detoxification. Although a bactericidal action of pectin has been proposed to explain
A B
77
the effectiveness of pectin treating diarrhoea, most experimental results do not support
this theory. However, some evidence suggests that under certain in-vitro conditions,
pectin may have a light antimicrobial action toward Echerichia coli (Thakur et al., 1997).
Pectin promisingly affects cholesterol levels in blood and is reported to reduce blood
cholesterol in a wide range of subjects and experimental conditions (Ginter et al., 1979;
Miettinen and Tarpila, 1977; Sriamornsak, 2001; Sundar et al., 2012). As excipients, the
functionalities include, binding and gelling agent. In the medical device, application of
pectin has mostly been focused in wound healing or dressing and as an adhesive, such as
colostomy devices. Ostomy care products.
Figure – 4: Showing effect of retention of NFM by pectin on outer and inner stratum
corneum of the skin. The improved hydration of the epidermis leads of enhanced
Outer stratum
corneum
Inner stratum
corneum
Stratum
granulosum
Stratum
spinosum
Stratum basale
78
mechanical stability of the stratum granulosum, stratum spinosum and stratum basale.
(Source: Anonymous)
When used in personal care products, pectin forms a layer of moisture on the surface of
the skin (Figure - 4) which helps optimise the fluid balance of the epidermis. This not
only supports the skin to retain and maintain its own natural moisturising factor (NMF)
but skin becomes smoother, firmer and full of moisture.
The family of polysaccharides containing α-(1-4)-linked D-galactopyranosyl uronic acid
residue is commonly known as pectin (Yeh et al., 2011). The primary cell wall is mainly
comprised of polysaccharide apart of cellulose and hemicellulose pectin, is one of the
class of polysaccharide which constitutes around 34% portion of call wall. (Darvill et al.,
1980). Pectin is a complex polysaccharide, made up of D--galacturonic acid in which
few are naturally present free carboxyl group are methylated ester while rest can join
with calcium and magnesium ions (Jain et al., 2005). The basic structure of pectin
polysaccharide is D-galacturonic acid, joined by glycoside linkages ( Figure-
5) (Abbaszadeh., 2008).
79
Figure - 5: Showing : Showing glycoside linkageglycoside linkage. (
Source : Study blue, 2016)
Structurally the pectic polysaccharides are partitioned into homogalacturonan (HG),
Xylogalacturonan(XGA), apiogalacturonan(AGA), rhamnogalacturonan II(RG-II), and
rhamnogalacturonan I(RG-I) (Caffall & Mohnen, 2009). While during extraction of
pectin two types of pectins are isolated. The difference in types depends upon the degree
of methoxylation (DM). The pectin obtained with a DM more than 50% require high
amount of sucrose and low pH values to form gels while pectins with lower DM values
than 50% need a divalent cation for ex Ca++ to form gels (Carr et al., 1995). One another
group which is also low-methoxYl pectin known as amidated pectin are extracted in the
presence of ammonia. In these types of pectin polymers, amide groups are formed with
ester and acids moieties (May, 1990).
A general structural pattern of pectin polysaccharides is described by many scientists
(Willats, 2001; Caffall and Mohnen, 2009; Ridley et al., 2001). Mainly the structure of
pectin consist of two regions the linear and ramified region. The linear region of
homogalacturonan composed of 1,4-linked D- galactopyranosyluronic acid residues.
These regions are linked by L-rhamnopyranose residues one or two in numbers,
which are involved in the linear chain by a 1,2-linkage. Most of the pectins has this very
structure as their backbone, the difference among them is the length of the chain. The
ramified region composed of three auxiliary units: RG-I, arabinogalactan, and
xylogalacturonan, which can be found in different ratios (Ovodov, 2009).
80
Due to the effect of the isolation procedure, storage and processing of plant material. It is
difficult to describe the exact structure of pectin (Sriamornsak, 2002). However it is
presumed that RG-I and RG-II are covalently linked to HGA domains but the direct
linkage between RG-I and RG-II does not have a proper proof. HGA is a
linear homopolymer of (1→4)-αlinked- D-galacturonic acid and is thought to include
some100–200 Galacturonic acid (GalA) residues. HGA is the most common and
frequently found part of pectin which is synthesized in Golgi. RG-I composed of about
100 repeats of the disaccharide (1→2)-α-L-rhamnose-(1→4)-α-D-galacturonic acid is the
acidic pectin domain/part. Here the chains are mainly consisting of of arabinan
andgalactan. RG-II sounds similar to RG-I due to its name but it has no structural
relationship with RG-I. It is a part of pectin which has numerous branches and a major
part of HGA. RG-II consists of almost 9 GalA residues which are connected by a (1→4)-
α-linkage which is substituted by 4 heteropolymeric side chains of known and consistent
lengths. There are eleven types of sugars attached with the side chains such as, 2-keto-3-
deoxy-D manno-octulosonic acid, apiose and acetic acid. (Figure-6) (Abbaszadeh, 2008)
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Figure – 6: α-4-linked galactosyluronic acid (GaIA) (Ridley et al., 2001)
In foods, including confectionery, beverage and canning industries, the application is
mainly confined as thickening and gelling agent while in cosmetic formulations it is
incorporated for its gelling, thickening and stabilizing properties (May, 1990;
Sriamornsak, 2001 and 2002 and Srivastava and Malviya, 2011). The food additive code
number for pectin is 440 or E440. The applications of pectin in solid oral dosage form
Figure-7: Schematic representations of the conventional (A) and alternative (B)
structures of pectin. Willats et al. (2006).
82
have been comprehensively represented in a review article (Sriamornsak, 2001). As a
therapeutic agent, it has two major applications, which include its application in
antidiarrheal formulations for infants and children as well as a dislipidemia, especially to
reduce elevated blood cholesterol (Sriamornsak, 2001). However, it requires a very high
intake of pectin, above 6 g/day which may reduce around 13% of serum cholesterol in
two week time (Ginter et al., 1979; Mietinnen and Tarplia, 1977). In addition, pectin has
been reported to be useful in elimination of lead and mercury from respiratory organs and
gastrointestinal tract (Kohn, 1982). Another application to treat disorders relating to
overeating has also been reported (Di Lorenzo et al., 1988). As an excipient, pectin act as
binder and compressibility enhancer (Slany et al., 1981a, 1981b; Kim et al., 1998) as well
as in the development of modified release dosage form, such as controlled release matrix
formulations (Krusteve et al., 1990; Naggar et al., 1992; Sungthongjeen et al., 1999),
pectin beads as well as insoluble hydrophilic coating for sustained release drug delivery
system (Aydin and Akbuga, 1996; Sriamornsak, 1996; Sriamornsak et al., 1997a and
1997b), and above all its application as a carrier material in colon-specific drug delivery
system due to its insolubility and resistant property against intestinal enzymes but
sensitivity against colonic pectinolytic enzymes (Ashford et al., 1993; Rubinstein et al.,
1993; Englyst et al., 1987; Sandberg et al., 1983 and Rubinstein et al., 1992)
Pectin provides a natural structure to various cosmetic preparations, such as pastes,
creams and ointments, while its thickening and stabilizing action is useful in applying
hair tonics, body lotions, shampoos and conditioners (Ref: Cargill). When used in skin
formulations, pectin forms a layer of moisture on the surface of the skin. The major
83
source as reported by IMR international (2009) for the production of pectin is citrus fruits
(Lemon, lime or oranges), followed by apple and sugar beet (Figure-8). The industrial
applications of pectin include use in plasticizers, edible films, foams and paper substitute.
The overall application and uses of pectin in pharmaceutical, food, cosmetics and
chemical industries have been graphically presented in Figure –9
Figure-8: Showing major sources of pectin used arround the world. ( Source: IMR
International 2016)
84
Figure-9: Showing the overall application and uses of pectin in pharmaceutical,
food, cosmetics and chemical industries. (Source: IMR International 2016)
The major pectin manufacturing companies include: Danisco, , CP Kelco, Herbstreith &
Fox (Germany), Cargill and Yantal. And according an estimate (IMR 2009) 42,000 MT
of pectin was produced worldwide with a positive growth between 3 to 6%. CP Kelco is
the largest pectin producer in the world (Figure-10).
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.
Figure-10: Showing major pectin producers and their market share. ( Source: IMR
International 2016)
In view the importance and diversified applications of pectin which categorises it as an
exciting and promising component for the pharmaceuticals, nutraceuticals and
cosmeceuticals, the search of new sources for its production demands a comprehensive
studies by applying both pure and applied knowledge. Based on this approach, present
study was taken to investigate the possibility of having a better quality of pectin from
seasonal fruits available in Pakistan and to explore and synchronize the physic-chemical
parameters for increased yield on lab scale basis. This was followed by the application of
the crude as well as refined pectin in some pharmaceutical oral and liquid dosage form
and to calculate the economics of overall process. According to the most recent survey
86
available on internet (Vision, 2015) the size of the global pectin market would be of $1 B
by 2017 and the compound annual growth rate will be 8% during the year 2012-2017
(Figure-11)
Figure -11: Showing the global pectin market. (Source: Vision, 2015)
87
Figure-12: Showing current and growing market size of hydrocolloids in the
different regions of the world (Source: Markets and Markets, 2016)
Hydrocolloid market in general is on constant growth which can be seen in figure-12.
According to this market survey report hydrocolloid market is growing fastest in Asia-
Pacific while due to the changing lifestyle and growing understanding of healthy and
nutritional foods the demand of hydrocolloids, of which pectin is a part, also increasing in
south-east Asia (Markets and Markets).
88
REVIEW OF LITERATURE
History of pectin
The pectic substances were first discovered by the French Chemist Louis Vauquelin in
1790, while its properties were first reported by Henri Braconnot another French chemist
in 1825 (Georgiev et al.,2012). Pectin has a key action in the maintenance of the
structural integrity of plant by providing firmness to the plant by composing a hydrated
cross linked three dimensional networks (Shi et al., 2002). Pectin not only has a role in
ripening and texture building of fresh fruits in plants but it is an important agent in
processing industry for its characteristics property of jelling and thickening agent in
canned and processed food products (Nurdjanah,2013).The traditional use of pectin was
due to its gelling property and it was used in making jam jellies and marmalades and
other bakery products (Jameson et al., 1923; Nanji et al 1934).Pectin was also used as
emulsifying agent in many food products containing oils and fats. It was presumed that
pectin, due to its gelling property will not only help in stabilizing a product under regular
climatic conditions but also helps in pasteurizing the product (Douglas and Loesch,
1927). Other food items were also investigated to improve the quality of food products in
which pectin was used. Breakfast foods and cereals were studied to improve the texture
and quality of this food product (Spalding and Conn, 1936).
Numerous application of pectin has been studies over the years which include its
traditional uses to modern usage in pharmaceutical application. Pectin was also studied
for its other medicinal properties like hemostasis (Kertesz, 1951), lipid lowering (Ershoff
and Wells, 1962), activity on enzymes and hormones (Morgan et al., 1979) and
cholesterol lowering agent (Ink and Hurt, 1987). Pectin also got under investigation in the
89
management of both types of diabetes mellitus and showed that it helps in lowering blood
glucose and insulin levels (Jenkins et al., 1976). Due to its ability to absorb water it is
also used in weight reduction (Holt et al., 1979). The addition of pectin in pharmaceutical
preparation due to its properties like delaying absorption of drug (Kertesz, 1951). It has
been used in combination with many other agents in treatment of diarrhea (Tompkins,
1938; Bennett, 1958). Pectin was also used in the prophylaxes and management of heavy
metal toxicity (Paskins- Hurlburt et al., 1977). Studies were also found for the use of
pectin in dentistry and skin care products (Lochridge, 1951).Other interesting uses
reported in literature were the use of pectin in therapeutic gels (Raudnitz, 1979).
Chemistry and Type of Pectin
In search for naturally occurring compounds plants are being studied with great interest.
Likewise pectin has also been studies because of its complex structures and vast number
of uses (Cosgrove and Jarvis, 2012).Although pectin is being considered as most complex
and heterogeneous naturally originated structure and its being studied upon for more than
180 years, it is astonishing to learn that due to different physiological changes occurring
in call wall the actual molecular structure of pectin are still studied so as to understand
the basic structure of pectin molecule (Schols et al., 2009; Cybulska et al., 2015).
Numerous techniques and apparatus are being used in order to understand the actual
chemical structure of pectin such as atomic force microscopy and force microscopy
(Morris et al., 2011).Studies on the structure of pectin are important as the properties and
functions of pectin depend upon its chemical configuration. (Funami et al., 2011).The
studies on structure of pectin further extend the possibility of its uses and application in a
90
more scientific way both in food and pharmaceutical properties (Sila et al., 2009).The
structure of pectin is mainly consist of D-galacturonic acid (Gal A) units joined in chains
by means of alpha-(1-4) glycosidic linkage (Srivastavaand Malviya, 2011).The
proportion of occurrence among HG, XGA, RGI, and RGII is also variable; typically HG
is the most common polysaccharide, composing around 65% of the pectin, while RGI
makes 20% to 35%. XGA and RGII are other lessor segments, each constituting less than
10 % (Acton, 2013).
Biosynthesis of pectin
The parts of the plants involved in the biosynthesis of pectin are mainly Golgi vesicles
however it is believed that some preliminary steps occurs in the endoplasmic reticulum. A
large numbers of enzymes are also involved in synthesis of these complex polysaccharides
which are mostly located in the Golgi apparatus. There are almost 67 enzymes which have
been identified which are required during the process which include glycosyltransferases,
methyltransferases and acetyltransferases (Harholt et al., 2010). The initiation of the
formation of pectic polysaccharides is still not known and the involvement of lipids and
protein donors are also not established. The pectin biosynthesis investigation has been a
tough task for researchers with very passive growth. Figure -13 shows a diagrammatic
representation of biosynthesis of pectin. According to this predictive model the pectin
biosynthesis mainly rely on nucleotide-sugars which are formed by nucleotide-sugar
transformation reactions which takes place in cytosyolic side of Golgi. A specialized
nucleotide-sugar which is called nucleoside monophosphate antiporters are responsible for
the movement of nucleotide-sugar into the Golgi (Ridley et al., 2001)
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Figure- 13: Schematic diagram for biosynthesis of Pectin. (Ridley et al., 2001)
Classification of Pectin
Commercially pectin is divided into different types on the basis of its gel forming
property. The gel forming property of pectin depends upon the degree of esterification.
The backbone structure of pectin which is partially methyl-esterified is used for
classification of pectin. The degree of esterification (DE) less than 50% is low DE pectin
while DE more than 50% is high DE pectin (Thirawong et al., 2007). It was also reported
that lower the methoxyl content present in pectin reduces is its gelling property
(Srivastava and Malviya, 2011).Activities like cholesterol lowering properties, also
depends upon the natural chemical properties of pectin (Brouns et al.,2012).
The types of pectin also effect drug permeability when used as films (Hagesaether et al.,
2009). It also has an effect on food grade of pectin, when pectin used in jams it effects the
92
texture and color of the jam (Kopjar et al., 2009). Other effects like mucoadhesive,
(Hagesaether and Sande, 2007) and the relationship between pectin and mucin by mucin-
particle method are also under investigation (Sriamornsak et al., 2010).Little influence
has been of the degree of esterification (DE) of the pectin molecules on chemical abstract
service (CAS) emulsion stability at different pH.(Surh et al., 2006).Pectin used in making
different food items and the type of pectin used are presented in Table – 1.
Extraction and purification of pectin
Kertesz in 1951 suggested that the extraction of pectin is a complex physico-chemical
process which may involve solubilization, extraction and DE polymerization of pectin
complex molecules from plant tissues. In lab-scale extraction, hot water extraction is used for
high esterified pectin. While the low-esterified pectin is not extracted efficiently when
extracted with the same procedure for them, chelating agents like Ethylenediaminetetraacetic
acid (EDTA) and Cyclohexylenedinitrilotetraacetic acid (CDTA) are used (QI et al., 2002).
While commercially, pectin is extracted through hot dilute mineral acid pH around 2 with
varying length of time. The time depends upon the type of pectin required and from one
manufacturer to another (May, 1990).Various factors can affect the extraction of pectin. The
extracting conditions may affect the probable structure of pectin. Studies on, the use of
different pH levels, reveals that it has a profound effect on both the extractability and in the
breakdown of pectin molecule (Kaya et al., 2014).). Some of the recent referenced natural
sources of pectin with respective method of extraction have been highlighted in the Table-2.
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Table - 1: Showing the uses of different types of pectin in various types of food
products
Food item Type of pectin Reference
Jams, marmalades, fruit
jellies and fruit spreads
HM Pectin Rababah et al., (2014)
Beverages HM pectin Walsh et al., (2014)
Fruit preparations for
yougurt
LM pectin
LM amidated pectin
Rinaldia et al., (2014)
Fruit preparations for
bakery products
LM pectin Wuestenberg, (2014)
Confectionery HM pectin Wuestenberg, (2014)
Glazes LM pectin
LM amidated pectin
Wuestenberg, (2014)
Deserts LM pectin
LM amidated pectin
Thakur., et al (1997)
Acidified dairy drinks HM pectin Bonnaillie., et al (2014).
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Table - 2: Showing some investigated sources of pectin with their applied mode of
extractions.
SOME SOURCES OF PECTIN
Source (Common Names) Mode of extraction Scientist/scientists
Gold kiwifruit Acid, water, enzymetic Yuliarti et al., (2015)
Water Melon Microwave Assisted Maran et al., (2014)
Grape fruit Ultrasound-assisted heating
extraction Xu et al., (2014)
Passion fruit peel Microwave-induced heating Seixas et al., (2014)
Grape Pomace Ultrasound-assisted
extraction
Minjares-Fuentes et al.,
(2014)
Dragon Fruit Citric Acid Muhammad et al., (2014)
Apple pomace and citrus
peel Subcritical water Wang et al., (2014)
Pumpkin Enzymatic extraction Cui and Chang., ( 2014)
Citrus limon Alcohol precipitation
method Kanmani et al., (2014)
Pomelo Microwave Tan et al., (2014)
Lemon pomace Different solvents Azad et al., (2014)
Mango Chelating agents Kermani et al., (2014)
Sisal waste Aqueous extraction Santos et al,. (2013)
lime peel High hydrostatic pressure
treatment Naghshineh et al., (2013)
Sugar beet Organic acids Ma et al., (2013)
Orange
Ultra-high pressure,
microwave or traditional heating
Guo et al., (2012)
Dragon Fruit Extraction using ammonium Ismail et al., (2012)
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SOME SOURCES OF PECTIN
Source (Common Names) Mode of extraction Scientist/scientists
oxalate/oxalic acid
Sugar Beet Microwave-assisted Li et al., (2012)
Sweet potato Disodium phosphate
solution Zhang and Mu., (2012)
Sugar Palm Microwave assisted Rungrodnimitchai., ( 2011)
Apple pomace Hydrochloric acid and citric
acid
Kumar and Chauhan., (
2010)
Red Dragon Fruit Extraction using citric acid Woo et al., ( 2010)
Mulberry Use of Hydrocholoric acid Liu et al., (2010)
Passion fruit Mixture of acids Kliemann et al., (2009)
Taiwan tangerine HCl extraction Tamaki et al., (2008)
Passion Fruit Peel Citric acid Pinheiro et al., (2008)
Pumpkin Microbial enzymes Ptichkina et al., (2008)
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Traditional method of extraction
Extraction with hot water is by far the easiest and old method of extraction of pectin and
its related compounds substances (Huang, 1973) but the study of literature also revealed
the use of organic and dilute inorganic acids such as sulfuric acid aids in hydrolysis of
pectin thus giving good yield of pectin in less time (Jameson et al., 1924; Nanji et al
1927). Traditionally for commercial purposes the pectin was extracted with the help of
different mineral acids like sulfuric, hydrochloric and nitric acids. Sometimes
combination of hydrochloric acid with ion exchange resins were also used in the
commercial production of pectin (Rouse and Crandall, 1978).The main emphasis during
extraction of pectin historically was to improve the grade of pectin as jellying was only
considered the important parameter of pectin later it was found that there are other factors
which are also important to study as they affect in the jellying property of pectin.
Techniques like HPLC were used to determine the degree of methoxylation,
anhydrogalauronic acid and degree of acetylation (Voragen et al., 1986).
Different mode of extraction was also studied in order to understand the factors affecting
the yield of pectin. It was learned that temperature influenced the production of pectin in
the most positive manner and factors like rate of extraction and diffusion coefficient
investigated further. Although commercially pectin was extracted with the help of hot
diluted mineral acids like hydrochloric acid and sulfuric acid however the factors
influencing the yield were dependent on the type of source used to extract pectin and also
on the production methods of company’s manufacturing pectin.(May , 1990). The
industrial extraction of pectin by the use of mineral acids aroused numerous problems
97
faced during the production of pectin which lead to the investigation of new process to
overcome those challenges. Enzyme extraction was one of the methods used so that the
problems like pulp maceration, difficult residual filtration and corrosion of the used
equipment were few of them (Sakamoto et al., 1995).
Physicochemical properties of pectin are also very important to understand and
investigate. The physicochemical properties of pectin depend upon many factors like
source from which pectin is extracted and also the process used for extraction. There are
others factors also which influence the extraction and yield of pectin are pH, temperature,
extraction time, type of solvents selected and the involvement of chelating agents like
EDTA and CDTA (Yeoh et al., 2008).Different techniques were used to determine the
actual structure of pectin in which spectroscopic techniques are used effectively.
Microwave was also investigated for better extraction of pectin for both in terms of yield
and quality. Image study of microwave extracted pectin was carried out to understand the
difference in quality or quantity of pectin extracted by using microwave (Zhongdong et
al., 2006).The method for the extraction of pectin has to be carefully designed in order to
get maximum yield with good quality of pectin. For this, numerous methods have been
studied and applied like direct boiling and microwave heating.
Microwave- assisted extraction
The traditional or conventional method of extraction of pectin is time consuming, it is
important to investigate for other methods for the extraction of pectin with good yield in
less time. For this purpose microwave heating is used presently (Koh et al.,
2014).Extraction with the help of microwave heating has given a desirable and positive
98
effect in getting good quantity of pectin (Maran et al., 2014).The good quality of pectin
includes low moisture and ash contents (Koh et al., 2014).The use of microwave (MW)
also helps in collecting high-quality antioxidant extracts from different plant materials.
This is because the fast and efficient heating of MW transmits heat straight to the
material under consideration releasing the phytochemical compounds readily and rapidly.
It also requires lesser amounts of solvent and time (Dorta et al., 2013).To optimize the
yield of pectin, extraction with numerous parts of the peel with different types of methods
has also been studied like flevedo and albedo of orange peels (Liu et al., 2006). A number
of other factors are equally important to consider while studying the yield, they are effect
of pH, extractant to peel ratio and temperature (Kulkarni and Vijayanand, 2010)
(Zhongdong et al., 2006). The yield obtained from microwave extraction was also
studies by applying statistical tools for the optimization of yield from the novel source of
pectin like apple (Wang et al., 2007). Not only apple but orange was also studied for its
effect of microwave heating on quality and quantity of pectin extracted from it. It was
noted that extraction with the help of microwave increased yield of pectin due to the
rupturing of parenchymal cells which assisted the release of pectin from the tissues.
(Kratchanova et al., 2004). Same source was used to determine the effect on yield of
pectin when pressure was applied along with microwave heating. The study also revealed
that molar mass, size and intrinsic viscosity of extracted pectin also increased as a result
of microwave heating during extraction of pectin from orange peel. This was confirmed
by size-exclusion chromatography (Fishman et al., 1999). Microwave pretreatment was
another method studied for its effect on the quality of novel sources of pectin like orange
lemon and apple (Kratchanova et al., 1996; Kratchanova et al., 1994).
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Enzymatic extraction of pectin
For the augmentation in the yield and to find convenient, less time consuming and
efficient process of extraction of pectin, different types of methods are studied and
investigated. Enzymatic extraction is one of them; use of enzymatic catalyst gave
competitive results (Dominiaket al.,2014). Currently large scale production of pectin
involves potent acids and elevated temperatures, Pectinolytic enzymes also help in the
degradation of pectin and hence used alternate methods for the extraction of pectin
(Mikami et al., 1987). The method is attaining popularity because it is eco-friendly.
Enzymes are excellent catalysts which can break or agitate cell walls and membranes
which aids in extracting considerable amount of functional compounds (Puri et al., 2012).
Polygalacturonase, which is acidic depolymerase, can also be used effectively in the
effective extraction of pectin (Franchi et al., 2014).
The enzymes mode of extraction of pectin involves breaking of pectin linkages lowering
the viscosity of the solution which in turns aids in the process of filtration and
centrifugation. More over enzymatic extraction is eco-friendly as it minimized water
pollution, the main drawback with this type of extraction can be the difficult and
expensive production of enzymes used in this process. The enzymatic extraction should
be monitored with great precision and care so as to stop the degradation of pectin at a
proper time otherwise it may affect the gelling property of pectin (Munarin et
al.,2012).The concentration of enzyme affects the yield of pectin as well as the molar
mass of pectin however no effect on its GalA concentration (Yuliarti et al.,
2011).Celluclast, Econase and Viscoferm are few examples of enzymes which gave good
results (Wikiera et al., 2015).Other new methods like application of high hydrostatic
100
pressure technology for enzymatic extraction has also been reported (Naghshineh et al.,
2013). A study done using microwave assisted flash extraction under control pressures
and temperature claims that microwave assisted extraction (MAE) extracts provides high
molar mass, moderate viscosity pectin in less time using low temperature when
compared to conventional heating (Fishman et al., 2008). Additional methods include
use of microbial enzymes is also reported (Ptichkina et al., 2008).
A number of solvents have been used by different worker to extract pectin, however, the
best results was reported from ethanol. Oxalic acid based extraction also gives good
result when Microwave assisted extraction is used(Tan etal., 2014).The Quantity of
solvent used also has an effect on yield of pectin (Chan and Choo,2013) the ratio of citric
acid required to make the solution pH 2.5 gave better results than with the ratio which
made the solution pH 4.
The use of ionic liquids are also found in past researches and has proved themselves as
an effective alternate solvents in extraction of biologically active compounds with the
help of microwave. They are not only effective but also environmental friendly (Guolin et
al.,2012;Du et al., 2007).The ionic liquids are a combination of organic cations and
inorganic or organic anions (Du,et al., 2007). Whereas the use of new solvents such as
ammonium oxalate (Ismail et al., 2012)disodium phosphate are also investigated (Zhang
and Mu, 2011). Optimization with the help of organic acids like nitric acid has also been
conducted (Vriesmann et al., 2011).
101
Purification of Pectin
For the purification of pectin different methods have been reported, Yapo, and Koffi,
(2014), first purification pectin using 95% ethanol and thus washed pectin precipitates
two-times with 70% ethanol, followed by 95% ethanol and acetone before drying them in
oven. In another study, Shaha et al (2013) washed the precipitated pectin through 95%
ethanol, followed by 55% and then with 70% ethanol. Isopropanol and acetone were also
used for washing pectin for further purification (Ismail et al., 2012) .Different methods
have also been discussed and compared by Sotanaphun, (2011).The authors used dialysis,
Amberlite XAD-16 polystyrene resins for the purification of pectin. Some of the very old
methods used for the purification of pectin include treatment with bromine, chlorine
water or iodine (Krishnamurti andGiri 1949) (Nanji and Chinoy, 1934).Celite also used
for purification since long. It was used in the extraction of pectin to remove impurities
and color that may be present in extracting solution (AOAC, 2012)Patents have also been
granted for celite to be used for the purification of pectin. Celite was used to filter the
solution containing soluble pectin before final precipitation with isopropanol. The celite
was used on a nylon cloth and the solution was filtered on a sieve and also as a filter aid
during vacuum filtration. (Buchholt et al., 2005). Recently a research is being conducted
in which celite 545 is used in the purification of pectin (Dominiaket al., 2014). Figure -13
to 15 shows different lab scale extraction of pectin while Figure- 16 to 18 shows different
industrial scale production of pectin.
102
Figure -14: Extraction of pectin from soy hull (Monsoor and Proctor, 2001).
Soy hull (400 g)
Extraction with 0.05 N HCl
Collect supernatant
Add equal volume of isopropanol
Adjust pH to 3.5
Collect Precipitate
Wash two times with isopropanol
Final wash with 70% isopropanol
Collect precipitate
Disperse with deionized water
Spray dry
103
Figure-15: Extraction of pectin from leaves of Nephrolepis biserrata (Pagarra et al 2014)
Nephrolepis biserrata leaves
Drying of sample
Add aquadest adjusted to 1;40 w/v
Add 0.5 H2SO4
Sample solution at pH 1.5
60; 90; 120 minutes of boiling at 80OC
Filtiration
Add 95% Ethanol (1:1 v/v)
Precipitation, 1 day
Washing with 95% Ethanol
Precipitized
Dry oven 40˚C
Yield of Pectin
104
Figure- 16: Extraction of pectin from raw papaya peel (Boonrod et al., 2006)
Raw papaya peel 15g
Treated material
Pectin solution
Pectin precipitate Pectin
Precipitate
Ethanol-insoluble
pectin
Aluminum-insoluble
pectin
Route
1
Route
2 1\2
Mincing
pH adjustment to 4-4.5
Precipitation with 95% ethanol
for 2 hr
Filtration
Acid-extraction in boiling water bath
with 0.06 M HCl 3 times at volume ratio
of 2:1, 1:5:1 and 1:5:1
Centrifugation at 4,500xg 30
pH adjustment to 3.5
Precipitation with 1.2%
aluminum chloride, pH 4.0 for 2
hr. Filtration
Washing with 70%
ethanol
Oven-drying at 37˚C
Washing with 70%
ethanol
Oven-drying at 37˚C
105
Figure-17: The commercial production of pectin ( Source: ENSYMM)
Citrus Waste
Hydrolysis Reactor
Expansion Tank
Filter
Condense/Decanter
Biogas Digester Precipitator Fermenter
Dryer
Distillation
Sulfuric
Acid Water
Steam
Limestone
Hydrolysat
e Soli
d
Biogas Pectin
Dried Pectin
Ethanol
Stilage
Liquid
Pectin
Depleted
Residue
106
Figure- 18: Commercial production of pectin (Source: Cargill foods)
107
Figure-19: Commercial production of pectin (Source: GENU)
Uses and application of pectin
The family of polysaccharides containing α-(1-4)-linked D-galactopyranosyluronic acid
residue is commonly known as pectin (Yeh et al., 2011). Pectin, a part from cellulose and
hemicellulose is one of the class of polysaccharides involve in the formation of primary
cell wall (Besson et al., 2013). Well known and traditional use of pectin is in food
industries, yet several pharmaceutical uses and applications have also been reported. Use
of Pectin in the production of jams and jellies has provided its additional application in
the manufacturing of many other types of food products, which include dairy products,
desserts and soft drinks (Munarin et al., 2012). Pharmaceutical uses include cholesterol
lowering formulations (Brouns et al., 2011), pectin-derived oligosaccharides considering
as a new source of prebiotics (Gullon et al., 2013), Ophthalmic and nasal formulations
(Mittal and Kaur, 2014 and Morris et al., 2010) and colon specific delivery systems
Pre treatment
of raw material Extraction Filtrations
Water + acid Filter aids
FRUIT
Insoluble material
ion-exchange
(calcium
removal)
Evaporative Concentration
de-
esterficiation
water acid
Precipitation de-esterficiation
and/ or amidation alcohol
washing
drying and
milling blending and
standardization
alkali or
ammonia alcohol
alcohol
alcohol
recover
y Solid waste( alcohol
soluble)
Standardized
Pectin Sugar and other pectin
preparations
Optional
process
Process
aids
Waste/re
cycle
Raw
materials/
products
Standardizing
agents
108
(Kushwaha et al., 2011). Recently, studies on its cosmoceutical applications have also
been undertaken (Suh et al., 2014). New approaches include development of hydrogels in
presence of proteins which can be used in the formulation of low calorie food products
(Wu et al., 2014).
Among many current known pharmaceutical uses pectin is under constant consideration
for finding even more important and beneficial uses. It is studied that both, pectin source
and physico-chemical properties of pectin renders its effect in lowering of cholesterol in
humans. The physico-chemical properties include molecular weight (MW), viscosity and
degree of esterification (DE) (Brouns et al., 2011).Among the advancements in new drug
delivery systems,pecsys is a new technology in which the pectin-based formulation, gels
when it comes in contact with mucosal surface. Such formulations gives rapid absorption,
controlling Cmax, reduces runoff and high bioavailability (Archimedes Pharma, 2011).
Pectin is also studied for the formulation of microencapsulated drug delivery systems in
which it is used in the form of biopolymer particles and are used in both food and
pharmaceutical preparations (Jones et al., 2010).The said drug–delivery systems are
under close observation through clinical trials in order to understand the functionality and
side effects (Watts and Smith, 2009).Pectin are important part of soluble dietary fibers;
these fibers are essential for proper physiological well-being of humans and are used to
treat dyslipidemias and other diseases (Dhingra et al., 2011).The source of pectin used for
pharmaceutical purpose are citrus peels and apple pomace. Different formulations have
been made and studied in connection with the colon-specific drug delivery systems.
Pectin microspheres have been under in-vitro investigations and the drug release from
microspheres are being understood in different conditions which may affect the release of
109
the drug (Kushwaha et al., 2011). The diversified properties of pectin can be associated
with its complex structure that’s the reason pectin is studied for its anti-tumor activities
too (Maxwell et al., 2012). But this association of pectin structure with its bioactivity
cannot be fully explained in literary works as the relationship with origin of pectin and its
possible chemical modification which results in molecular fragmentation are not fully
expressed (Leclere et al., 2013). Recently a work has been done on nanocomposite Pectin
scaffolds which will provide localized therapy in ovarian cancer (Chandran et al., 2013).
Pectin is still in use as antidiarrheal agent in formulations with kaolin and other
ingredients. Some currently marketed products are shown in Figure-20 with their
country/region of availability.
Pectin is commonly used as gelling, thickening and stabilizing agents in foods and to a
certain extent in pharmaceuticals as well. Pectin is mostly used to construct the desired
texture of products which result in controlling the moisture or water in the product
(Imason , 2010). Traditionally pectin was used mainly in food and food preparations, due
to its thickening and jelling properties, jams and jellies are few examples. Pectin is
considered very well tolerated and safe among numerous food additives therefore it is
categorized and recognized in “Acceptable Daily Intake (ADI) levels of “not specified”
products by the FAO/WHO Joint Expert Committee on Food Additives (JECFA). (Food
and Agriculture Organization of the United Nations 2009; Endreb and Christensen,
2009). The stature is equivalent in European Union where Pectin E440 (i) and Amidated
Pectin E440 (ii), both have been accustomed as ADI "not specified" by the Scientific
Committee for Food Specifications. Where as in United States pectin is designated as
GRAS (generally recognized as safe) by the FDA (U S Food and Drug Administration;
110
Endreb and Christensen, 2009). Moreover, it is permissible to use pectin in all non-
standardized foods and in accordance with the Code of Federal Regulations (21CFR
184.1588) Table-3 shows different uses of pectin in food industry. Due to the trust of
consumers and regulatory authorities on inertness and safe nature, pectin is gaining
popularity in making of cosmetics and personal care products. CP Kelco has number of
patents in this field such as GENU pHreshTM pectin which restores skin and maintain a
skin friendly pH has been presented (CP Kelco). Trudsoe et al (2014) describes use of
acidified pectins in personal care products that can be good for human skin. Personal care
products may comprise of a styling gel, moisturizer, lotion, deodorant, toothpaste, body
wash, bath gel, body gel, hand sanitizer, shampoo, conditioner, or combinations.
111
Table - 3: Showing different uses and properties of pectin.
(Source: J.F. Hydrocolloids)
Application/uses of pectin Role of pectin
1. Meat and Poultry
Provides neutral flavor and pH.
Helps in absorption of oil
Provides high content of dietary fiber.
Have more water retention capabilities
2. Fruit and Based (inc. fruit‐based
beverages)
Different types of pectin used having different
setting time
Provides nice and delicate texture.
Enhances body and mouthfeel.
Regulates setting time and temperature.
Helps in flavor release.
Viscosity of beverages is maintained
3. Dairy (inc. dairy‐based beverages) Provides optimum stability of heat treated
dairy drink.
Protects whey separation and casein
precipitation.
Enhances shelf‐life.
Smoother mouthfeel.
Helps flavor release
4. Confectionary Provides nice and delicate texture.
Regulates setting time and temperature.
Helps in flavor release.
5. Bakery Provides delicate and nice texture.
Regulates setting time and temperature.
Enhances flavor release.
112
A
B
c
Figure – 20: A and B available in Middle East while C is available in Australia.
113
Large scale production of pectin
Commercial pectins are largely extracted with acid extraction method from citrus peels
and apple pomace and the yield is usually 25 and 12% pectin, respectively (Ismail Sah et
al., 2012). As far as the structure of commercial pectin is concern they are less complex
as the processes of extraction and purification in industry removes almost all the neutral
sugars in pectin (Seymour, 2002). The pectic skeleton is made up of homoplymer of
galaturonic acid binded with (1→4) linkage with partial methyl esterified carboxyl
groups. According to United Nation’s Food and Agriculture Organization [FAO] and
European Union [EU] commercial pectin must be composed of atleast 65% of
galacturonicacid (Calderon, 2012).Flow chart shows a general procedure of commercially
extracted pectin. Apple pomace and citrus peels are the major raw materials which are the
main source of industrial pectin. But the supply of raw material also dependent upon
available local crop sources. Therefore some other types of raw materials are also used in
different parts of the world which includes potatoes pulps, sugar beet pulp, sunflower
heads etc. although the industrial process for the extraction of pectin is explained by
many scientists but detailed process differ among companies, but the general process is as
follows.
The materials used for the extraction of pectin are heated using reflux technique with
dilute mineral acid for 1 -10 hours at a pH approximately equal to 2 and at temperature of
60-100°C. If the source is apple or peach pomace after separating the hot pectin extract
from solid residue, to hydrolyze starch pectinase-free α-amylase is mixed. The obtained
extract is reduced in volume under vacuum to an approximately 4% pectin material and
114
then it is precipitated with 2- propanol. The precipitated pectin is then washed, dried and
reduced in particle size after grinding. For different types of pectin different mediums are
used for example for low DE values acidic treatments are used and when ammonia is
used amidated pectin were produced. The process is summarized in Figure- 21.
115
Figure -21: Flow chart showing general procedure employed for the extraction of
pectin on commercial scale. Turakhozhaev and Khodzhaev (1993).
Preliminary treatment of raw
material
Protopectin hydrolysis and
pectin extraction
Filtration and concentration
Precipitation and purification
Drying and grinding
Standardization
116
3. MATERIALS AND METHODS
Equipment
The equipment used in this study are as under
Analytical balance
OHAUS (PA 214) USA, Sartorious GmbH; type A 6801
Water bath
Thermostatic Grant Type JB2 water bath. USA
Homogenizer
Panasonic MX-J120P mechanical Blender and grinder.Japan
pH meter
Jenway 3510 pH meter. UK
Microwave oven
National (IEC-705) 700 W microwave. Japan
Freeze Dryer
Tro,science Co, Limited freeze dryer, model number TR-FD-BT-50 Japan
117
Spectrophotometer
A Nicolet Evolution 300, Thermo Electron Corporation, USA,UV-Vis spectrometer 1800
(Shimadzu, Kyoto, Japan)
FT-IR spectrometer
Thermo Nicolet 380, FTIR Instrument. USA
Centrifuge
Laboratory centrifuge shanghai China, model number 800 , Heltich Universal 32R
Tablet Punch Machine
Korsch, Erweka, Germany
Oven
Memmert, Germany
Furnace
Gallen Kamp Scientific Tech. UK
Hardness Tester
Fujiwara, Seisukusho Corporation, Japan
Friabilator
H.Jurgens and Co-Gmbh, D2800, Germany
118
Disintegration apparatus
Eeweka, ZT2, Heusenstamm Germany
Dissolution apparatus
Erweka DT, Heusenstamm, Germany
Refractometer
Hand held ATAGO refractometer, Japan
Viscometer
Brookfield viscometer
Laser spectroscatter
Laser spectroscatter-201 system , RiNA GmbH Berlin, Germany
Mixer
Eisco scientific
119
LIST OF CHEMICALS AND REAGENTS USED
BDH-VWR Laboratories
Industrial methylated spirit, ammonium hydroxide, celite, sodium hexameta- phosphate,
charcol, tartaric acid, phosphorus pentaoxide, sodium tetraborate.
Fisher Scientific
Ammonia, ethylenediamine tetraacetic acid (EDTA).
J T Baker
Hydrochloric acid, sulfuric acid.
Merck
Acetic acid, ethanol, nitric acid, sodium carbonate, sodium chloride, sodium citrate,
sodium hydroxide, m-hydroxydiphenyl, magnesium sulphate, oxalic acid, citric acid,
phenophthelene, phenol red, magnesium sulphate, potassium dihydrogen phosphate,
methanol, lactose,Propyl Paraben (Sodium),
Sigma- Aldrich
Galacturonic acid, pectin, sodium azide, bronopol, Microcrystalline cellulose
Mallinckrodt Speciality Chem Co
Paracetamol, kaolin
120
Fauji Pakistan
Starch
Vijai Minerals
Talc
Taihei Chemical Industrial Co. Japan
Magnesium stearate
Sensient Colors UK
Erythrocine
BASF Aktiengesellschaft, Germany
Bronopol, Ibuprufen
Asian Chemical Company Limited, Japan.
Sodium Saccharin
Nippon Chemical Trading Co. Japan
Sodium carboxy methyl cellulose
Lever Brothers Ltd. – Pakistan
Glycerine
121
Vanderbilt Minerals, LLC
Veegum
Local preparation
Distilled water
Deionized water
122
LIST OF APPARATUS
Knives ( IKEA) , motor and pestle( porceline), kitchen steak hammer (IKEA), kitchen
hand chopper (IKEA), muslin ( local market), burner( made in China) , sieve (80 mesh
sizes), metal dish (5 cm in diameter with lid), sieve ( number 16, 20 and 40) , stainless
steel pan ( local market) , wooden spoon ( Local market) , litmus paper, distillation
assembly , jelly vessels, ice box, burette, pipette ,buchner funnel (porceline), vacuum
pump, porcelain crucible (Yancheng Yongkang Lab Instruments Factory, Jiangsu,
China), reflux condenser (Shanghai Heqi Glass ware Co., Ltd., China), diaphragm
vacuum pump. vernier caliper in mm (CD-6, CSX, Mitutoyo, Japan), brookfield DV-E
viscometer, laboratory glass wares beakers (100ml, 500ml, 1000ml, 3000ml ), glass rods,
glass funnels, test tubes, right arm flask , conical flask (250ml) , volumetric flask (
100ml, 250ml), glass jars with lids, measuring cylinder( 100ml)] (Pyrex, England),
porcelain crucible (Yancheng Yongkang Lab Instruments Factory, Jiangsu, China),
desiccator (Duran, Germany),tripod stand. Centrifuge tubes (30ml), sintered glass
crucible, filter (0.22 mm Millipore, USA), quartz SUPRASIL ®.
123
LIST OF SOLUTIONS USED
Hydrochloric acid (HCl) solutions
0.1N: The solution was prepared by slowly adding 8.212 ml of concentrated HCl
to 250 ml deionized water. The final volume was adjusted to 1000 ml with deionized
water.
1N: The solution was prepared by slowly adding 82.117 ml of concentrated HCl
to 250 ml deionized water. The final volume was adjusted to 1000 ml with deionized
water.
Sodium hydroxide (NaOH) solutions
0.5N: The solution was prepared by dissolving 22 g of NaOH in 1000 ml of deionized
water.
0.1N: The solution was prepared by dissolving 4.5 g of NaOH in 1000 ml of deionized
water.
Sulphuric acid solutions
0.5N: The solution was prepared by slowly adding 14.027 ml of concentrated sulphuric
acid to 250 ml deionized water. The final volume was adjusted to 1000 ml with deionized
water.
124
1N: The solution was prepared by slowly adding 28.024 ml of concentrated sulphuric
to 250 ml deionized water. The final volume was adjusted to 1000 ml with deionized
water.
Nitric acid solution
1N: The solution was prepared by slowly adding 63.704 ml of concentrated nitric acid
to 250 ml deionized water. The final volume was adjusted to 1000 ml with deionized
water.
Oxalic acid solutions
1%: To make 1 % solution, 10 g of oxalic acid was dissolved in 1000 ml of deionized
water.
10%: To make 10% solution, 100 g of oxalic acid was dissolved into 1000 ml of
deionized water.
Tartaric acid solutions
1%: To make 1 % solution, 10 g of tartaric acid was dissolved in 1000 ml of deionized
water.
10%: To make 10% solution, 100 g of tartaric acid was dissolved into 1000 ml of
deionized water.
125
Citric acid solutions
1%: To make 1 % solution, of 10 g of citric acid was dissolved in 1000 ml of deionized
water.
10%: To make 10% solution, 100 g of citric acid was dissolved into 1000 ml of
deionized water.
Citric acid solution for jelly grade
50 g of citric acid was dissolved in distilled water and diluted to 100 ml with deionized
water.
Sodium citrate solution
25 g of sodium citrate was dissolved in distilled water and dilute to 100 ml with distilled
water.
Galacturonic acid stock solution
Galacturonic acid stock solution was prepared by dissolving 100 mg dry galacturonic
acid powder in 100 ml distilled water. This makes the concentration of the solution to be
1mg/ml. The solution was refrigerated. Fresh solution was prepared if the sample is kept
longer than 4 weeks.
M/80 Sodium tetraborate in sulphuric acid (0.0125M solution)
1.192 g of sodium tetraborate was weighed and transferred added into a 250 ml
volumetric flask and the volume was adjusted with concentrated sulphuric acid.
126
Sodium hydroxide (5%)
5 g of sodium hydroxide was transferred in a 100 ml volumetric flask and the volume
adjusted with deionized water.
m-hydroxydiphenyl solution (0.15%)
0.15 g of m-hydroxydiphenyl was weighed and transferred into a 100 ml flask and the
volume was adjusted with the help of 0.5% sodium hydroxide solution and refrigerated.
Phosphate buffer saline pH 7
1.20 g of sodium dihydrogen phosphate and 0.885 g of disodium hydrogen phosphate
were dissolved in 1 liter volume distilled water.
Tris saline buffer
20 mM Tris–HCl buffer was taken in a beaker the pH was adjusted to 7.5with 150mM
sodium chloride (NaCl) and 0.01% sodium azide (NaN3).
127
Extraction of pectin
Processing and preparation of peel
Fruits were purchased from the local market, Karachi, Pakistan and identified. Their
voucher specimen numbers were deposited in the department of Pharmacognosy, Faculty
of Pharmacy and Pharmaceutical Sciences, University of Karachi, Pakistan. All fruits
were washed with distilled water properly before subjecting to any procedure. Excess
water was drained and after air drying, the fruits were peeled off using sharp knife. The
respective peels thus obtained from different fruits were cut into thin slices of few mm in
thickness, weighed at a constant weight and used for extraction.
Screening of seasonal fruits and their wastes for the presence of pectin
contents
The method of extraction of pectin used was originally described by Patel et al (2012).
The modified method used for extraction in the present study is presented in Figure-20.
20 g of thin sliced peels of each fruit were dipped in 100 ml of boiling industrial
methylated sprit (IMS) and each content were boiled for 5 min on a water bath. The IMS
was then carefully decanted in a separate container. The peels obtained after boiling in
IMS were transferred in the jar of a mechanical blender and after adding small quantity of
DI water the contents were grinded for about 30 sec and then transferred the whole
content in a beaker to form loose slurry.
The slurry obtained from mechanical procedure of each fruit peels were decanted
separately into beakers and code numbers were assigned. The volume was raised to 60ml
128
for each test performed. The contents were boiled for 10 min using bunsen burner. The
mixture was then brought to room temperature and DI water was added in each beaker to
adjust the volume, if evaporated during boiling. The pH of the each content was recorded
and adjusted to desired pH 6 ±0.5, either using NH4OH or 0.1N HCl. The solids of each
content were separated, first using a Buchner funnel, followed by centrifugation at 4000
rpm. The supernatants, thus obtained from each content were then further processed by
adding ethanol (95%) in 1:4 ratios which is approximately 240ml. The addition of ethanol
lead to precipitation of pectin in the liquid phase, which is then separated with the help of
centrifugation, freeze dried, weighed and percentage yield was calculated. All freeze
dried materials in glass vials were kept in refrigerator for further study. The list of fruits
which were screened is shown in annexure 1.
Extraction of pectin from selected fruit wastes and their comparative
study
Materials and methods are same as described for the screening of seasonal fruits and their
wastes for the presence of pectin contents. Three fruits were selected in view of screening
results for the identification of new pectin source, ease of availability and cost. Fruits
selected were “Banana”, “Sapodilla” and “Muskmelon”. The fruits were screened for
their maximum pectin release, subjecting them to different mechanical procedure at
different pH using two different heating methods. Same procedure was adopted for
standard (apple and orange) fruits to improve yield in order to get clear yield comparison
between mechanical procedures, pH and heating methods. (Figure-22). A pictorial
representation is given in annexure 2.
129
Mechanical, pH and heating factors used during the study
The extraction procedure for all five fruits was kept similar as used and described in
screening study with slight modification. The peels of the selected fruits after boiling in
IMS were subjected to different mechanical tools, such as, homogenization, grinding,
cutting, chopping and hammering.
Homogenization Grinding Cutting Chopping Hammering
For each mechanical procedure, following techniques were applied:
Homogenization: The peels obtained after boiling in IMS were transferred in the jar of a
mechanical blender and after adding small quantity of DI water the content was grinded
for about 30 sec. and then transferred the whole content in a beaker to form loose slurry.
Grinding: The peels obtained after boiling in IMS were transferred in the jar of a
mechanical grinder and grinded without adding DI water for 30 sec. The grinded peels
were then transferred into a beaker containing required amount of water to form loose
slurry.
Cutting: The peels obtained after boiling in IMS were transferred in the container of a
hand chopper and sized / reduced into small slices by pressing the hand chopper 200
130
times. The processed peels were then transferred into a beaker containing required
amount of water to form loose slurry.
Chopping: The peels obtained after boiling in IMS were transferred in mortar and the
peels were chopped by using a pestle for about 200 times. The processed peels were then
transferred into a beaker containing required amount of water to form loose slurry.
Hammering: The peels obtained after boiling in IMS were transferred on a cutting
board and hammered for about 200 times. The processed peels were then transferred into
a beaker containing required amount of water to form loose slurry.
pH: Five strengths of pH were selected .The pH of the each slurry was maintained at
1,3,5,6 and 7 respectively. The slurry was checked for the initial pH and then HCl (0.1N)
or NH4OH were used to adjust the pH.
Heating methods and time. Two heating methods were used during this procedure
which were microwave heating and bunsen burner. The mixture was boiled for 10 min
using both the heating modules.
131
Figure-22: Showing flowchart of extraction of pectin followed during the current
Fruit peels 20 gms
100 ml IMS
60 ml H₂O
Adjustment of pH with HCL/ NH₄OH
Purification by passing through celite
Filtrate ------------> Add Ethanol 1:4 ratio
Centrifuge at 4000 rpm
Freeze Dryer
Percent Yeild
Boiling for 5 min
Boil for 10 min
Filter with cheeze cloth
132
study
Effects of organic acid and inorganic acids and their strength on pectin
yield from sapodilla
To determine the best extraction procedure, sapodilla fruit was subjected to different
types of extraction procedures using organic and inorganic acids, maintaing pH around
4±0.5and NH4OH to observe the yield at pH 6 to 7. The basic method used for extraction
of pectin was kept similar as used in screening of seasonal fruits and their waste for the
presence of pectin contents. The difference in the type, and concentration, strength of
organic and inorganic acids used are described below.
Extraction with 0.1N HCl, using 5 mechanical procedures, 5 different
pH, two boiling methods and varying time of boiling (10, 20, 40 and
60min)
In this procedure peels of sapodilla were selected and were subjected to the same
extraction procedure as mentioned in preceding text with the exception that the peels
were boiled for 10, 20, 40, and 60 min using both burner and microwave. For the
maintenance of pH 0.1N HCl was used.
Extraction using 1N HCl, using 5 mechanical procedure, 5 different pH,
two boiling methods and varying time of boiling (10, 20, 40 and 60min)
Aforementioned procedure is followed except that for the maintenance of pH 1N HCl
was used.
133
134
Extraction using organic acids
The same foregoing procedure is used except that for pH maintenance three different
organic acids oxalic, citric and tartaric acid were used in 1% and 10% concentrations.
Extraction with different strengths of inorganic acid
In this procedure after following the same extraction method described earlier the pH
was maintained with three different strengths (0.1N, 0.5N, and 1N) of same inorganic
acid (Hydrochloric acid).
Statistical Evaluations
Three factor factorial completely randomized design (CRD) was applied and mean
comparison was done by using Tukey HSD (Steel et al., 1997) at 5% level of significance.
Statistical analysis was performed using SPSS13 and Minitab13.1.
Physical and biochemical characterization of pectin
Percent yield
The percent yield of the dried pectin was calculated as follows
% Yield =Weight of vial with pectin
Weight of empty vial× 100
Identification tests for pectin
135
Identification tests were carried out according to the procedures performed by Sharma et
al (2013).
Stiff gel test
1g of pectin with 9 ml of distilled water was heated on a water bath to get a solution and
cooled
Test with 95% ethanol
To a 1% w/v solution equal volume of ethanol (95%) was added.
Test with potassium hydroxide (KOH)
5ml of 1% w/v solution of pectin sample was prepared and 2% w/v solution of KOH was
added, the solution was left to stand for 15 min.
Iodine test
To a freshly boiled 5ml solution of 2% w/v pectin, 0.15ml of iodine solution was added.
136
Biochemical characterization
Preparation of sample
The dried pectin was stored in a cool dry place. The sample is oven dried and used for all
the tests performed.
Qualitative test for ammonia
To 0.5g of dried pectin, 1ml of 0.1N NaOH was added and the sample was heated to
observe the release of ammonia if any (Ranganna, 1986).
Detection of Moisture, ash, methoxy content and equivalent weight
Moisture, ash, equivalent weight and methoxy content of pectin was determined by the
procedure used by Aina et al (2012) and Ranganna (1986) with slight modifications.
Moisture
1g of sample was weighed in a tared metal dish. The sample was dried at 100OC for four
hours after which the dish was cooled in a dessicator.
Ash
1 g of pectin was taken and weighed in an empty tarred crucible. The crucible was heated
at 600OC in a furnace for 24 hrs. The crucible was then taken out of the furnace and set
aside to cool in a desiccators. After cooling the crucible was weighed again and the ash
was calculated with the help of following formula:
137
Ash content (%) = (Weight of ash / Weight of sample) × 100
Equivalent weight
0.5 g of pectin was taken into a 250ml conical flask. The pectin was soaked with 5ml
ethanol and 1.0 g of NaCl was added, 100ml of distilled water and a few drops of phenol
red indicator was also added to the flask to completely dissolve the content. The solution
was then titrated with 0.1M NaOH to a pink color endpoint. Equivalent weight was
calculated using the equation below:
Equivalent weight = (Weight of sample / Volume of alkali (ml) × Molarity of alkali) ×
100%
Methoxyl content
A previously neutralized solution (obtained after the equivalent weight determination)
was used to determine methoxy content. 25ml of 0.25 N sodium hydroxide was added,
shaked vigorously and allowed to stand for 30 min at room temperature. The mixture was
then titrated with 0.1N NaOH to the same end point as used in equivalent weight. The
methoxyl content was calculated with the equation given below:
Methoxyl content % = (Volume of alkali (ml) × Normality of alkali × 3.1/ Weight of
sample (mg)) × 100
Anhydrouronic Acid.
The anhydouronic acid (AUA) content was calculated by the method suggested by Azad,
et al., 2014. The AUA was determined by the following equation:
138
% of AUA= 176x0.1zx100 + 176x 0.1y x100
w x1000 w x1000
When molecular unit of AUA (1 unit) = 176 g
Where,
z = ml (titre) of NaOH from equivalent weight determination.
y = ml (titre) of NaOH from methoxyl content determination.
w = weight of sample
Grading of pectin
Grade of jelly was determined by the method described by Ranganna (1986). 320 ml of
water was taken separately in a kettle, 500 g of sugar was mixed with 3.3 g of pectin
assuming the grade of pectin is 150. To the water in kettle 0.5ml of the citric acid
solution and 1 ml of sodium citrate solution was added. The pectin sugar mixture was
also added later and stirred thoroughly. The mixture was heated rapidly with constant
stirring. The mixture was removed from flame occasionally to avoid the excessive
evaporation of water. The mixture was heated till the weight of solution became 770g.
When the solution reached at the desired weight it was removed from the flame and
allowed to cool for 30 sec at room temperature. 2 ml of citric acid and 0.5ml of sodium
citrate solution was then added. The jelly was transferred into a vessel or any other
desired container and allowed to stand for at least 18 hours at 260 C. After 18 hours the
jelly was checked for overall firmness and compared with standard jelly prepared with
same procedure. The process is repeated with the increased amount of pectin if there is a
difference between the consistencies of standard and sample jellies.
139
Galacturonic acid content
Pectin assay using m-hydroxydiphenyl
Pectin Assay using m-hydroxydiphenyl was used to determination of pectic substance in
the desired samples. The colorimetric assay using m-hyrdoxydiphenyl was used for its
specificity for uronic acids as it can tolerate the presence of sucrose upto 1000 ppm.
Preparation and measurement of samples:
2 ml solution from the galacturonic acid stock solution was taken into a 100 ml
volumetric flask and the volume was made up with deionized water to make 20
microgram / ml of galacturonic acid. Similarly different concentration (40, 60 and 80
micrograms / ml) of galacturonic acid in the sample were also made. Sixteen test tubes
were placed in an ice box to cool before the experiment started. Three tubes were used
for each sample (2 for samples and one is used for blank determination). The sulphuric
acid/ sodium tetraborate solution was also kept in an ice bath throughout the experiment.
1.0 ml of all the test material was then filled in each of the three respective labeled cold
test tubes and was left to cool for a minute. About 6 ml of sodium teteraborate solution
was then added to the test tubes and mixed thoroughly with the help of a test tube stirrer.
The tubes were kept in the ice box until all the samples were prepared. The tubes were
then heated for about 6 minutes after which the tubes were placed again in the ice bath
for cooling. 0.1 ml of m-hydroxydiphenyl was then added to the first tube to develop
color; it was also mixed thoroughly with the stirrer. For blank 0.1ml of 0.5% sodium
hydroxide was taken in the third tube. It was also mix thoroughly and was allowed to
stand for 15–20 min at room temperature to dissipate any bubbles formed during stirring.
140
The absorbance was measured at 520 nm on a spectrophotometer by reading sample
against corresponding blank tube with 0.5% sodium hydroxide. The total absorbance for
each sample was obtained by subtracting absorbance due to m-hydroxydiphenyl and the
absorbance for sample blank. A calibration curve of Absorbance (y-axis) was plotted
against concentration of galacturonic (x-axis). This curve was use for standardization.
Water holding capacity (WHC)
The method used for water holding capacity was measured using Daou and Zhang,
(2011) method with slight modification as the sample used here was dried pectin. 1g
sample was accurately weighed in a graduated test tube. The sample was hydrated with
0.02% sodium azide dissolved in 30ml of deionized water for 18hours. After the assigned
period the supernatant was removed and the sample was drained into already weighed
filter paper which was dried to a constant weight in a forced-air oven at 110OC and
weighed again. WHC was expressed as the amount of water retained per gram dry sample
(g/g dry weight).
WHC (g/g) = (Hydrated residue weight-Dry residue weight)/ Dry residue weight
Water binding capacity (WBC)
The water binding capacity was also determined after slight modification of the method
described by Daou and Zhang (2011). 1g sample was taken in a graduated centrifuge
tube, 0.02% sodium azide dissolved in 30ml of deionized water was added and left at
room temperature for 18 hours. The tubes were centrifuged (3000 rpm) for 20min, the
supernanatent was separated by passing through a sintered funnel under vacuum. The
141
weight of the hydrated sample was noted and the sample was dried at 105OC for 24 hours
to get a constant weight. The value was expressed as the amount of water retained per
gram dry sample
WBC (g/g) = Residue hydrated weight after centrifugation -Residue dry weight / Residue
dry weight
Fat binding capacity (FBC)
The method used for fat binding capacity was previously used by Daou and Zhang,
(2011). 5g sample was taken and added to 20 ml soybean oil in a 50 ml centrifuged tube.
The mixture was then stirred after every 5 min for 30 sec and after 30 min the tubes were
centrifuged at 1600 rpm for 25 min. The free oil was discarded and the absorbed oil was
determined by weight difference and was expressed as ml (oil)/gram sample
FBC (ml/g) = (precipitation weight-Dry weight)/Dry weight
FTIR spectroscopy of selected pectin
FT-IR spectrometer was employed to investigate the characteristic spectra of the
extracted pectin obtained through different variations in extraction procedures. Dried
sample (1 g) was ground and analyzed through Smart Arc attachment, thereafter it was
scanned within the range of 4000-400 cm-1 (Park et al., 2006).
Dynamic Light Scattering (DLS) Analysis
In order to find some basic information regarding the particle size and polydispersity of
pectin. DLS studies were performed as described by Hameed et al., 2014. A laser
142
spectroscatter-201 system with a He–Ne laser providing a 690 nm light source and an
output power in the range of 10–50 mW was used in the study. An auto piloted run of 50
measurements at every 20 s, with a wait time of 1 s was used at 25 C was used for all
measurements. Standard and all other pectin samples was prepared (2% w/v) in Tris–
saline buffer .Samples (20 µl), reagents and buffers were filtered using 0.22 mm filter
(Millipore, USA) before filling into a special quartz SUPRASIL ® cell (light path 1.5
mm) for measurement. The scattered light was recorded at a constant scattering angle of
90o. The autocorrelation functions were evaluated using the CONTIN program to obtain
hydrodynamic radius (RH) distributions. The RH is associated with the diffusion
coefficient by the Einstein–Stokes equation. The data were analyzed using Xtal Concepts
software (XtalConcepts GmbH, Hamburg Germany) present with the instrument.
Optimization of the yield of pectin from sapodilla peel thorough
Response Surface Methodology (RSM).
The sapodilla peel was further studied in order to establish the optimum conditions of
extraction to get the maximum yield of pectin. To determine the optimum extraction
conditions for the extraction of pectin Box-Behnken design was used. A response surface
experimental design, with three independent variables was used to extract pectin from
sapodilla fruit peel. The variables used were time of boiling, temperature at which the
extraction was carried out and pH of the extracting medium. The design of experiment
and regression analysis was performed for response surface methodology by Minitab
(ver. 14) program (Minitab Inc. Quality Plaza, 1829 Pine Hall Rd. State College. PA.
16801. United States).The level of each variable was selected
143
Pharmaceutical preparation
Formulation of tablet using extracted pectin from sapodilla peel
Nine formulations for paracetamol and four formulations for Ibuprufen were designed by
the method described by Menon et al (2011) with slight modifications for direct
compression. The composition of 700mg tablet made by wet granulation method is
mentioned in Table 5 for paracetamol tablet and Table 6 for ibuprofen tablets. The
amount of active ingredient (Paracetamol / Ibuprofen), binder, disintegrant and diluent
were mixed after weighing each amount according to the size of batch. All the
ingredients were mixed and passed through sieve # 40 separately. The ingredients were
then mixed with distilled water as granulating agent and a dough is formed. The dough
was then passed through sieve #16 to get coarse granules which were then dried on 450C
for almost 2 hours. The dried granules were then passed through sieve # 20 to obtain
equal size granules. The obtained granules were then mixed with the weighed amount of
glident and lubricant. The mixture was mixed evenly through tumbler movement. The
finally obtained granules were then compressed through single punch machine to make
tablets of required weight and hardness. A pictorial representation of the process is given
in annexure 3. The comparative effect of binding property of pectin was determined,
control tablets were made with the addition of starch instead of pectin as binding agent.
144
Table - 4: Composition of Paracetamol tablet with different concentrations of pectin used
S.NO Ingredients S1
(mg/tab)
F1
(mg/tab)
F2
(mg/tab)
F3
(mg/tab)
F4
(mg/tab)
F5
(mg/tab)
F6
(mg/tab)
F7
(mg/tab)
F8
(mg/tab)
F9
(mg/tab)
1 Paracetamol 500 500 500 500 500 500 500 500 500 500
2 Microcrystalline
cellulose
30 30 30 30 30 30 30 30 30 30
3 Pectin 10 20 30 40 50 60 80 100 120
4 Starch 10
5 Lactose 153 153 143 133 123 113 103 83 63 43
6 Talc 5 5 5 5 5 5 5 5 5 5
7 Magnesium Sterate 2 2 2 2 2 2 2 2 2 2
Where F1= test formulation one, F2= test formulation two, F3=test formulation three, F4 = test formulation four, F5=test formulation
5, F6=test formulation six, F7 test formulation seven, F8= test formulation eight, F9=test formulation nine
145
Table - 5: Composition of Ibuprufen tablet with different concentrations of pectin used
S.NO Ingredients S2
(mg/tab)
R1
(mg/tab)
R2
(mg/tab)
R3
(mg/tab)
R4
(mg/tab)
1 Ibuprofen 500 500 500 500 500
2 Microcrystalline cellulose 30 30 30 30 30
3 Pectin 50 75 100 125
4 Starch 10
5 Lactose 153 113 88 63 38
6 Talc 5 5 5 5 5
7 Magnesium sterate 2 2 2 2 2
Where R1= TrRal formulation one, R2 = test formulation two, R3 =test formulation three , R4 = test formulation four
91
Evaluation of granules
The granules were evaluated for the flowabilty according to the method given in USP 36/
NF 31, 2013 guidelines, these are as under:
Angle of repose (α)
Angle of repose (α) was determined by funnel method.It is defined as angle possible
between the surface of a pile of the powder and the horizontal plane. (Banerjee and
Singh, 2013).The mixture of granules was carefully passed through a glass funnel. The
granules were passed carefully under the force of gravity on a piece of graph paper or
butter paper. The height of heap or cone (h) was calculated while the radius of the heap
was calculated as (r). The angle of repose was calculated as under:
𝛼 = tan−1 ( ℎ
𝑟)
Bulk density (ρb)
Bulk density of the granules was measured by weighing small amount of granules and
transferring it into a measuring cylinder. The volume of granules occupied in the cylinder
was carefully noted and is called bulk volume (Vb). Weight of the granules is denoted by
(M). The bulk density was calculated as under:
ρb = M/ Vb
92
Tapped density (ρt)
The tapped density was measured by tapping weighed amount of granules in a measuring
cylinder for a specific time. The minimum volume (Vt) occupied by the granules and
weight (M) was used to measure tapped density as under:
ρt = M/ Vt
Compressibility Index
It is measured in % compressibility (C). It can be explained as the measurement of the
free flow property through which a materials flow characteristics can be predicted. It was
measured as under:
C = (ρt –ρb) / ρt * 100
Hausner’s ratio
It can aslo be explined as the easiness of flow of granules and is calculated as under
Hausner’s ratio = ρt / ρb
Loss on drying (LOD)
1 to 2 g of test sample was weighed and kept in a stoppered bottle. The stoppered bottle
was previously weighed and dried under the same conditions for the test sample and
cooled in a desiccator. The test specimen was accurately weighed with stopper and was
93
put in the drying chamber without stoppers. Drying was done till a constant weight was
achieved.
% LOD = D-A x 100
C
Where A = Bottle with cover weight
B = Sample + Bottle with Cover (Initial weight)
C = Sample weight
D = Sample + Bottle with Cover (Final weight.)
Tablet compression
The granules obtained were compressed using a single punch machine in which a convex
shaped punch of diameter 12.38 mm was fitted to get oval shaped tablets each weighing
about 700 mg (±5%). The hardness was set between 6-7 kg compressions ranges. The
compression was done at room temperature and a minimum of fifty tablets were
compressed for each batch.
Tablets testing
The finally compressed tablets were evaluated for quality parameters following the
guidelines of USP 36/ NF 31 2013 and some non- pharmacopeial methods which are
described below
94
Weight variation
The variation in weight of the test formulations and reference tablets were studied by
taking weight of each 20 tablets individually on a Type 1 balance. Mean weight and
standard deviation were calculated
Tablet thickness and diameter
The diameter and thickness of 20 tablets, were determined by a Vernier caliper in mm
Tablet hardness
It is calculated by randomly selected 20 tablets of the test formulations using a hardness
tester .
In-Vitro dissolution studies
The dissolution studies of the test and standard tablets performed by following method
given in USP 32/NF 27, 2009 guidelines by using a USP apparatus II. It was carried out
in 900 ml of phosphate buffer pH 5.8 at 37±5 °C at 50 rpm. An aliquot of 10 ml of
solvent was taken out from vessels at 5, 10, 15, 20, 25, 30, 45, 60, 90 and 120 minutes
and volume was made up again by fresh medium. Spectrometer was used to calculate
drug concentration at 278 nm with dissolution medium taken as blank. The dissolution
profile was also established in distilled water, 0.1 N HCl of pH 1.2 and phosphate buffers
at pH 4.5, 6.8 using the same sampling times as described above to evaluate the release of
drug in the new formulations. Each experiment was repeated in triplicate.
95
Formulation of antidiarrheal preparation (suspension) from extracted
sapodilla pectin
In a 3000 ml container 620 ml distill water was taken. Erythrosine, bronopol, propyl
paraben and sodium saccharin were added one by one. In the next step pectin is added,
followed by veegum, sodium carboxymethyl cellulose. The mixture was stirred and
soaked overnight. More distill water was added with glycerin and sodium citrate at this
stage. The mixture was then continuously stirred and kaolin was added gradually. The
mixture was further mixed thoroughly for an hour and the final volume was adjusted
using DI water. Lastly vanilla flavor was added. The formulation ingredients with their
respective amounts are given in Table - 6
96
Evaluation of Suspension
Color, odor and taste
Organoleptic evaluation of suspension was performed for its color, odor and taste
properties
pH
The pH of the suspension was measured by using pH meter.
Viscosity
The viscosity of suspension was evaluated using Oswald viscometer. The suspension is
filled in the required volume very carefully with the help of a pipette into the viscometer.
The viscometer was then kept into a water bath to achieve the desired temperature. When
the temperature reached to a constant temperature the volume in the viscometer was
adjusted again carefully using pipette. The pressure is then released and the time is noted
for the suspension to reach from one point to the other.
Sedimentation Volume
25ml of suspension was taken in a 50ml stoppered graduated measuring cylinder. The
suspension was then moved upside down two three times and then allowed to settle for
three minutes and the volume of sediment was noted which was considered as the
original volume (H0). The cylinder was then kept stationary for 7 days and the volume of
sediment was noted at 7hrs and 24hrs for consecutive 7 days. This was considered as the
final volume (Hu). The sedimentation volume was calculated as
Sedimentation Volume (F) = Hu/ Ho
97
The height of the solid phase after settling relies on the particle size and the concentration
of solid. For a desirable suspension F should be 0.9 for 1 hr
Redispersibility
For the measurement of redispersity a fixed volume of suspension was kept in a
stoppered cylinder at room temperature for 7 days. The suspension in the cylinder was
moved upside down at regular intervals to remove any sediments present at the bottom of
the cylinder.
98
Table - 6: Composition of antidiarrheal preparation nusing extracted sapodilla
pectin.
S.No Ingredient 1litre 1000 ml
1 Deionized water 0.62 lit 620ml
2 Erythrosine 0.0000248kg 24.8mg
3 Bronopol 0.0001kg 100mg
4 Propylparaben (Sodium) 0.00055kg 550mg
5 Sodium saccharin 0.00075kg 750mg
6 Pectin 0.00434kg 4340mg
7 Veegum 0.004kg 4000 mg
8 Sodium carboxy methyl cellulose 0.0016kg 1600mg
9 Water 0.02 litre 20ml
10 Sodium citrate 0.002kg 2000mg
11 Glycerine 0.1lit 100ml
12 Kaolin 0.2kg 200000mg
13 Flavours 0.004lit 4 ml
Make up the volume to 1 L 1000 ml
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FOOD PREPARATIONS
Preparation of Sapodilla jam
Sapodilla jam was prepared from its fruit and pectin extracted from its peel was used as a
gelling agent. 450g of fruit pulp, 550g of sugar, 5g of citric acid and 5g of pectin was
used in the preparation of jam (Ahmed et al., 2011)
The pulp of sapodilla fruit and sugar was mixed first in a stainless steel pan and cooked
on a medium heat on fire. Pectin was added and the mixture was left for heating with
constant stirring till boiling. The mixture was boiled until the required consistency of jam
was achieved which was checked with the help of spoon test. To check the consistency
stainless steel spoons were kept in refrigerator before the test, a spoon when required was
withdrawn from the refrigerator and little jam was taken out from it. The spoon with the
jam was left aside for a min after which the spoon was held in an upright positon. If the
jam spills from the spoon it means the jam required more quickly or else jam was
removed from the fire. After the cooking the jam were transferred into glass jars and
capped. The bottles were then boiled for 10min in an upside down position in a separate
pan containing water; this process is called processing of jam. The bottles were then
cooled and stored at room temperature for further study. Evaluation of the jam cooked
with the three different concentration and standard pectin was performed and the results
were compared. The ingredients and their amounts are shown in Table-7
100
Physico-chemical analysis of jam
Determination of pH
The pH of jam was observed by the method described in AOAC
Determination of viscosity
The viscosity of jam was recorded at 25oC by viscometer using t-shaped spindle at 20rpm
Determination of moisture and ash content
The ash and moisture contents were determined following the methods described by
Nwosu et al., (2014).
Determination of total titratable acidity (TTA)
10g of sample was diluted to 100ml with distilled water and titrated against 0.1N NaOH
solution using phenolphthalein as indicator to a pink color endpoint.(AOAC)
Determination of total soluble sugar
The Reducing and non-reducing sucrose was estimated by lane Eynon method. (Islam et
al., 2012)
Determination of total solids
Total solids were determined by subtracting percentage moisture from hundred as
followed by Shahnawaz et al. (2009)
101
% total solids = 100 - % moisture.
Determination of vitamin C (Ascorbic Acid)
Titrimetric estimation of ascorbic acid was performed by method described by Hussain et
al., (2010)
Sensory Evaluation
The sensory evaluation of jam prepared from extracted and standard pectin was
commenced at PCSIR (Pakistan Council for Scientific and Industrial Research, Karachi -
Pakistan). The jam was judged by 20 panelists which were randomly picked from the
institution. The panelist after tasting the jam marked the different parameters of sensory
evaluation. The jam was assessed on a 9-point hedonic scale, where 1 represented
extremely disliked and 9 extremely liked (Stone and Sidel, 2004)
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Table - 7: Formulation of the sapodilla Jam (1KG) with different
concentration of pectin
Ingredients S
(g)
F1
(g)
F2
(g)
F3
(g)
Fruit pulp 450 450 450 450
Sugar 550 550 550 550
Pectin 5 5 7 10
Citric acid 5 5 5 5
WhereS = Jam with standard pectin, F1 = Jam with 5g pectin, F2 = Jam with 7g pectin,
F3 = Jam with 10g pectin
103
Preparation of pudding from extracted sapodilla pectin
Sample puddings with three different concentration of pectin were prepared. Two
pudding samples were also made as a reference from food grade pectin which has the
same concentration as two among the three sample puddings while one of the puddings
has less quantity of pectin than the reference pudding
Samples of milk pudding were prepared by using different concentration of extracted and
a standard concentration of food grade pectin. The pudding was prepared after mixing
required amount of extracted pectin with 100 g of sugar, 1 egg and approximately 990 ml
of milk (Table- 8) in a saucepan. The mixture was stirred to dissolve sugar and pectin
into the mixture, after 10 min of stirring the mixture was heated till boil. After the
required time of boiling the mixture was kept aside from fire and then poured into the
desired containers for cooling. The cooled pudding mixtures were then kept in
refrigerator for 10 hrs till further analysis. Sensory evaluation of puddings were
performed by the 20 panellist specified for the job.
The sensory evaluation of pudding was performed in by 20 panellist .The testing
procedure chosen for the study was Duo-trio test (Kim et al., 2015). Three milk pudding
samples were provided to the panellist among which one was the reference pudding
sample while among the remaining two one of the sample pudding was same in
concentration as reference pudding in terms of pectin. Among the remaining two samples
one of the sample was same as reference sample while the other was different. The
pudding made from standard pectin was used as reference for the first 10 panellist while
pudding made with extracted pectin in the same quantity as reference for the remaining
104
10 panellist. To avoid any biasness the samples were coded. An evaluation form was
given to each participant to rate the sensory attributes. The example of evaluation form is
given in Annexure 4. Same procedure was followed for the remaining pudding samples.
105
Table-8: Showing different pectin concentration used for each selected extracted
fruit pectin
Material P1 P2 P3 Ps1 Ps2
Pectin ( gram) 5 10 15 10 15
Sugar (gram) 100 100 100 100 100
Milk (ml) 990
(approx.)
990
(approx.)
990
(approx.)
990
(approx.)
990
(approx.)
Egg 1 1 1 1 1
P1, P2 and P3 are sapodilla pectin pudding samples while Ps1 and Ps2 and reference samples while approx is approximately
106
RESULTS
4.1 EXTRACTION AND PURIFICATION OF PECTIN FROM THE SELECTED
FRUIT WASTE
Screening results relating to the determination of pectin content of different seasonal
fruits have been presented in Table - 7. In total of nineteen fruits wastes (peels + exocarp)
were evaluated in the present study to determine the pectin content. A simple standard
procedure based on homogenization of peels followed by extraction with DI water at a
pH maintained between 5-6.5, followed by precipitation of pectin with ethanol was used
in the screening study. The whole process has been shown in Figure -20. Out of nineteen,
twelve fruits indicated presence of pectin while insignificant quantity of pectin was noted
in seven fruits. Among twelve the highest content of pectin (20%) was noted in orange
followed by apple 6%, sapodilla (5.5%) and guava (4%). Rest of the fruits indicated
between 2-3.5% of pectin.
After the screening process three fruits (Sapodilla, Banana and Muskmelon) were
selected for further evaluation while two fruits apple and orange were used for comparing
the extraction process and to observe and compare the difference in the yield of pectin.
Table - 8 represents the yield of pectin from five fruits using different method of
extraction. The highest yield obtained from banana, sapodilla and muskmelon were
10.5%, 4.75% and 2.65% respectively when microwave was used as a heating medium
while when Bunsen burner was used the yield came slightly lesser 8.85% for banana, 4%
for sapodilla and 2.45% for muskmelon. Before further analysis and study, the pectin
obtained from fruits (Banana, Sapodilla and Muskmelon) were subjected to the
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preliminary identification test of pectin which revealed that pectin obtained from
muskmelon and banana fails to form stiff gel which is characteristic of pectin. Jelly grade
test was also performed which further proved that the quality of pectin obtained from
banana was of week nature while muskmelon failed to form any jell. The jelly grade of
banana came out between 80 to 85 while sapodilla jelly grade came higher 100-110
which makes it favorable for further analysis and to figure out best extraction method.
Two strengths of inorganic acids (0.1N and 1N) was selected to check the influence of
the concentration of acids. Table – 25 shows the yield of pectin obtained from 0.1N HCl
while Table - 39 shows results acquired from the use of 1NHCl. The time of boiling was
also taken into account along with difference in pH, mechanical method and mode of
boiling. Among the four different time of boiling the best yield was obtained from 0.1N
HCl after 20 min of boiling (6.35% yield at pH5) using microwave as the mode of boiling
and hammering as the mechanical tool. While almost same results (6.7%) were obtained
after only 10 min of boiling when 1NHCl was used for the extraction of pectin at same
pH (pH5) using microwave as the boiling method but the mechanical method chopping
was proved effective.In quest for understanding the integrated effects of different
variables on the yield of pectin extraction with organic acid was also carried out (Table -
54). The organic acids were used in two strengths (1% and 10%). As in our prior studies
we found that pH between 3-5 was effective for extraction so the pH was maintained in a
range of 3 to 5. 1% oxalic acid gave best result (4.95%) using microwave as the heating
method and homogenizing as mechanical tool. While heating through Bunsen burner
gave best average result also with homogenizing succeeding over the other mechanical
methods gave 3.95% yield with 1% oxalic acid.The last study under the section of
108
extraction was done to learn the effect of the different strength of acids (Table – 65). For
this purpose three different strengths (0.1N, 0.5N and 1N) of same inorganic acid (HCl)
was used. The yield of pectin came out to be best when the strength of acid was
increased, 6.7% of pectin yield was obtained when the strength of acid was increased to
1N HCl. The chopping method was useful in all the strengths of acid while heating with
microwave gave almost good yield in every strength.
4.2 PHYSICAL AND BIOCHEMICAL CHARACTERIZATION OF THE
PURIFIED PECTIN
The pectin isolated from the peels of banana, sapodilla and muskmelon was analyzed for
quality. For this purpose preliminary identification tests were carried which are shown in
Table-75. Banana and sapodilla showed positive results while muskmelon failed to show
desired results hence it was omitted for further investigation. The basic characterization
tests was performed on banana and sapodilla peels and shown in Table- 76 and found that
the pectin obtained from sapodilla peel was better in quality than banana. Hence the rest
of the tests were performed on sapodilla peel pectin only. Water holding, water binding
and fat binding properties of pectin was also assessed and shown in Table-77. The
sapodilla pectin was further tested by subjecting it to FTIR spectroscopy (Table-78)
4.3 OPTIMIZATION OF THE YIELD OF PECTIN FROM SAPODILLA PEEL
THOROUGH RESPONSE SURFACE METHODOLOGY
Response surface methodology with Box Bechen design was applied for determination
of optimal condition employed to get maximum yield. The factors studied were time of
boiling, temperature and pH at the time of extraction and the response observed was yield
109
(Table-79). The experimental design was consisted of fifteen experiments (Table-80).
The analysis of variance (ANOVA) was performed which is used to validate the process
variables for the optimization model. It is also used to investigate and understand the
response interactions of the variables, quality of the fit of the polynomial model
(coefficient of determination (R2), adjusted coefficient of determination (adj-R2 ) and
predicted coefficient of determination (pre-R2) and optimization of process condition
were obtained(Table-81).
4.4 UTILIZATION OF THE PURIFIED PECTIN FOR THE DEVELOPMENT OF
PHARMACEUTICAL AND FOOD PRODUCTS
Sapodilla peel pectin, used as a binder in formulation of paracetamol and ibuprofen tablet
was evaluated for pre compression, micrometrics and dissolution properties. Table-84
and 87 represents the micrometrics properties of granules formulated with different
concentrations of binder of paracetamol and ibuprofen tablets respectively. The weight of
both the types (paracetamol and ibuprofen) remained under the limit of ± 5%. The
diameter and thickness of all the tablets also didn’t exceeded from the required level
(Table- 85and 88).
In paracetamol tablets,10 and 20 mg of pectin were found not suitable to achieve desired
hardness granules when compressed into tablets were soft and thus further concentration
of pectin was increased. When the concentration increased from 30mg/tab desired
hardness was achieved( Table-86) however interestingly as the concentration of pectin
was increased,dissolution was noted to be decreased. The best hardness and dissolution
was achieved with the formulation F4 and F5 when the concentration of 40mg /tab and
110
50 mg/tab were used respectively. Dissolution was recorded as 80.43% and 86.35%
respectively (Table -87). Further increased in pectin from 60 to 120mg not only increased
the hardness of tablet but also noted to decrease the dissolution significantly. While the
ibuprofen tablets showed a similar type of results it is also noted that the higher
concentrations of pectin had detrimental effect on dissolution properties of tablets.
Antidiarrheal preparation formulated from extracted pectin also showed similar results as
compared to the marketed product (Table 93).
The jam was prepared using different concentrations of pectin are shown in Fig.1. The
jam due to different concentrations of pectin appeared to possess different types of
physical and chemical attributes Table- 90. The pH of the jam showed to be in range
from 3.00 to 3.30 .The TTA came out to be in between 1.00 to 1.14. All the formulations
except F3 showed the value of TSS nearer to the desirable level. The Vit C content of jam
came in between 17.6 to 18.3. The moisture found in these formulations was same as that
of previously studied jam. The three formulated jams with different concentration of
pectin were subjected to hedonic testing the results of the sensory evaluation of the three
formulations compared with standards are presented in Table-91
The result of the sensory evaluation of pudding made from extracted pectin through duo
trio test indicated among the 20, 9 correctly identified the difference in sample while 10
failed to identify any difference. To constitute significant differentiation among the
samples 15 correct judgement among 20 were required. The result showed that the
panelist as a whole did not found any significant difference (P> 0.05) between the
pudding made from standard and extracted pectin.
111
Table - 9: List of fruits used in the screening study for the presence of pectin.
(a) is Munarin et al., 2012; (b) is Bhat and Singh, 2014; (c) is Baker, 1997; (d) is Baissise
et al., 2010. (e) is Rasheed, 2008; (f) is Aina et al., 2012.
S no
Fruit
%Yield Reported
amount Common name Parts
used
1 Orange peel 20 6-26% a
2 Apple exocarp 6 2-19%a
3 Grape Fruit peel 3.5 13-32%a
4 Sapodilla peel 5.5 Notfound
5 Muskmelon peel 2 Not found
6 Guava (unripe) exocarp 0 Not found
7 Guava(ripe) exocarp 4 16.8% b
8 Chinese apple (Jungle Fruit) exocarp 0 Notfound
9 Strawberry exocarp 0 0.35–0.44%.C
10 Sweet lime peel 3 6-26%a
11 Peach exocarp 0 4-18%a
12 Apricot exocarp 0 4.97%d
13 Canteloup peel 3 Not found
14 Plum exocarp 0 Not found
15 Watermelon peel 2.35 15.70% e
16 Mango(Unripe) peel 0 Not found
17 Mango (ripe) peel 3.5 9-29%a
18 Banana peel 3.5 2-15%a
19 Lemon peel 3.5 2.7% f
112
Table - 10: Showing percent yield of pectin from five selected fruits
Mechanical
procedure
p
H
Sapodilla Banana Muskmelo
n Apple Orange
B M B M B M B M B M
Homogenizin
g
1 0.5 1.0
5
3.2
5 5.6 0.55 0.55 0.5 1
16.1
5
15.2
5
3 0.7
5 1.6
6.8
5
4.3
5 2 1
1.9
5
2.7
5 18.2 19.8
5 1.2 1.5 8.8
5 8.4 1.45 2.3
2.3
5 3.7 19.4 22.7
6 1.5 2 5.8
5
9.7
5 1.15 1 1.5 1.4
13.9
5 2.85
7 0.5 0.5 5.4
5 3.1 0.55 1.8
0.0
5
0.3
5 21.7 7.5
Grinding
1 0.5
5 0.8
2.4
5 3.2 2.25 2.65
2.4
5 2.5 2.6 3.95
3 4 3.5 4.2 6.9 1.4 1.8 0.4
5
1.1
5 7.8 5.3
5 2.4
5 2.9 3.5 4.5 1.95 2.4 1 1.1 2.3 7.1
6 2 2 3.6
5
2.6
5 0.95 1.6 2.4
2.8
5 6.45 6.6
7 1.9
5 2.3
1.2
5
0.2
5 0.55 1
0.0
5
0.5
5 0.6 0.2
Cutting
1 0.8 0.9 0 2.5 0.5 0.5 2.8
5 3.9 8.1 7
3 1.1 1.5 1.9
5 1.2 2.45 2.55
2.9
5 4.5 8.5 12
5 0.4 1 5 5.5 1 1 2.7 3.7 11.8
5
20.6
5
6 1 1.2 1 5.4 0.7 0.95 1.1
5 2.9 6.95 10.2
7 0.7 1 3 6.5 1.3 2.2 0.0
5 0.7 4 7.4
Chopping
1 2.7 4.7 1.6
5 2.1 0.05 0.7
2.8
5
3.3
5 2.6 3.35
3 3.9 3.4
5 4 3.3 0.05 0.5
1.2
5
2.7
5 7.95
10.0
5
5 3.9
5
4.0
5
5.9
5 6.6 1.7 2.1
0.0
5
1.7
5 10.4 15.8
6 2.3 2.4
5
5.8
5
6.3
5 2.25 1.75 0.1 0.9
15.2
5
17.1
5
7 3.9 2.4 1.8 1.6 0.5 1.05 0.2 0.6 5.35 9.35
113
5 5 5
Hammering
1 1.5 2.0
5 4.5 7 0.65 1.25 1.5 1.7
13.5
5
16.6
5
3 0.6 1.5
5 5.5 5.5 0.5 0.8 2.7 3.5 7.2 8.35
5 0.8 1.9
5 8.5
10.
5 0.65 0.3
3.0
5
4.8
5 7.4
12.1
5
6 0.8 1.4 5 5.6 0.95 0.5 1.2
5
2.0
5 5.55 10.7
7 0.5 0 0.5 0.5 0.45 0.05 1.7 2.0
5 5.5 8.85
B= Boiling on burner, M= heating in microwave
114
Table - 11: Analysis of variance (mean squares) of yield for five selected fruits
Source of variation
Degrees
of
freedom
Mean squares
Sapodilla Banana Muskmelon Apple Orange
Mech. procedure (MP)
pH level
Boiling method (BM)
MP pH
MP BM
pH BM
MP pH x BM
Error
Total
4
4
1
16
4
4
16
100
149
32.6575**
3.3917**
3.2413**
2.5462**
0.3392**
1.0112**
0.6258**
0.0266
51.245**
84.818**
22.349**
16.774**
4.640**
6.332**
4.698**
0.191
4.3869**
1.5751**
2.0184**
2.7688**
0.2814**
0.4112**
0.4141**
0.0112
9.5920**
17.3200**
22.6981**
5.6967**
0.8120**
0.8975**
0.2720**
0.0314
493.729**
142.586**
59.914**
88.941**
81.797**
44.377**
21.750**
1.0100
NS = Non-significant (P>0.05); * = Significant (P<0.05); ** = Highly significant (P<0.01) , Mech = mechanical
115
Table – 12: Means comparison of yield for sapodilla fruit peel (Mechanical procedure pH interaction mean±SE )
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 0.78 ± 0.13ij 0.68 ± 0.06j 0.85 ± 0.03hij 3.70 ± 0.47a 1.78 ± 0.14ef 1.56 ± 0.23C
3 1.18 ± 0.19gh 3.75 ± 0.15a 1.30 ± 0.09g 3.68 ± 0.13a 1.08 ± 0.21ghi 2.20 ± 0.24A
5 1.35 ± 0.08g 2.68 ± 0.11c 0.70 ± 0.14j 4.00 ± 0.12a 1.38 ± 0.26g 2.02 ± 0.23B
6 1.75 ± 0.12f 2.00 ± 0.06ef 1.10 ± 0.05ghi 2.38 ± 0.10cd 1.10 ± 0.14ghi 1.67 ± 0.10C
7 0.50 ± 0.01jk 2.13 ± 0.10de 0.85 ± 0.07hij 3.18 ± 0.35b 0.25 ± 0.11k 1.38 ± 0.22D
Mean 1.11 ± 0.10C 2.25 ± 0.19B 0.96 ± 0.05D 3.39 ± 0.16A 1.12 ± 0.12C
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 13: Means comparison of yield for sapodilla fruit peel (Mechanical procedure boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 0.89 ± 0.11e 2.19 ± 0.30b 0.80 ± 0.07e 3.36 ± 0.20a 0.84 ± 0.09e 1.62 ± 0.14B
M 1.33 ± 0.14c 2.30 ± 0.25b 1.12 ± 0.06d 3.41 ± 0.25a 1.39 ± 0.20c 1.91 ± 0.13A
B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean.
Table - 14: Means comparison of yield for sapodilla fruit peel (Mechanical procedure pH boiling method
interaction mean ± SE)
BM pH Mechanical procedure BM pH
116
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 0.50 ± 0.03qrs 0.55 ± 0.02pqr 0.80 ± 0.04n-r 2.70 ± 0.14ef 1.50 ± 0.06i-l 1.21 ± 0.22E
B 3 0.75 ± 0.03n-r 4.00 ± 0.15bc 1.10 ± 0.03j-o 3.90 ± 0.14bcd 0.60 ± 0.03o-r 2.07 ± 0.41B
B 5 1.20 ± 0.07j-n 2.45 ± 0.10efg 0.40 ± 0.02rs 3.95 ± 0.22bcd 0.80 ± 0.03n-r 1.76 ± 0.35C
B 6 1.50 ± 0.07i-l 2.00 ± 0.10ghi 1.00 ± 0.04l-q 2.30 ± 0.13fg 0.80 ± 0.03n-r 1.52 ± 0.16D
B 7 0.50 ± 0.02qrs 1.95 ± 0.08ghi 0.70 ± 0.03n-r 3.95 ± 0.14bcd 0.50 ± 0.01qrs 1.52 ± 0.36D
M 1 1.05 ± 0.06k-p 0.80 ± 0.03n-r 0.90 ± 0.03m-r 4.70 ± 0.26a 2.05 ± 0.14gh 1.90 ± 0.40BC
M 3 1.60 ± 0.08hij 3.50 ± 0.16cd 1.50 ± 0.06i-l 3.45 ± 0.14d 1.55 ± 0.05h-k 2.32 ± 0.26A
M 5 1.50 ± 0.04i-l 2.90 ± 0.06e 1.00 ± 0.03l-q 4.05 ± 0.14b 1.95 ± 0.11ghi 2.28 ± 0.29A
M 6 2.00 ± 0.09ghi 2.00 ± 0.08ghi 1.20 ± 0.04j-n 2.45 ± 0.16efg 1.40 ± 0.02j-m 1.81 ± 0.13C
M 7 0.50 ± 0.02qrs 2.30 ± 0.12fg 1.00 ± 0.04l-q 2.40 ± 0.08efg 0.00 ± 0.00s 1.24 ± 0.26E
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
.
117
Table – 15: Means comparison of yield for banana fruit peel (Mechanical procedure pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 4.43 ± 0.55fgh 2.83 ± 0.19j 1.25 ± 0.56klm 1.88 ± 0.12k 5.75 ± 0.60cd 3.23 ± 0.36D
3 5.60 ± 0.57cde 5.55 ± 0.64cde 1.58 ± 0.17kl 3.65 ± 0.19hij 5.50 ± 0.18cde 4.38 ± 0.34C
5 8.63 ± 0.31ab 4.00 ± 0.25ghi 5.25 ± 0.21def 6.28 ± 0.27c 9.50 ± 0.53a 6.73 ± 0.41A
6 7.80 ± 0.90b 3.15 ± 0.26ij 3.20 ± 0.99ij 6.10 ± 0.22cd 5.30 ± 0.19def 5.11 ± 0.42B
7 4.28 ± 0.54gh 0.75 ± 0.23lm 4.75 ± 0.80efg 1.73 ± 0.07k 0.50 ± 0.01m 2.40 ± 0.38E
Mean 6.15 ± 0.41A 3.26 ± 0.33D 3.21 ± 0.40D 3.93 ± 0.37C 5.31 ± 0.55B
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 16: Means comparison of yield for banana fruit peel (Mechanical procedure boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 6.05 ± 0.50a 3.01 ± 0.29e 2.19 ± 0.46f 3.86 ± 0.50cd 4.80 ± 0.69b 3.98 ± 0.27B
M 6.24 ± 0.67a 3.50 ± 0.59de 4.22 ± 0.55c 3.99 ± 0.57cd 5.82 ± 0.87a 4.75 ± 0.31A
B= Boiling on burner , M= heating in microwave In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
118
Table - 17: Means comparison of yield for banana fruit peel (Mechanical procedure pH boiling method interaction
mean ± SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 3.25 ± 0.19l-q 2.45 ± 0.11o-u 0.00 ± 0.00x 1.65 ± 0.12r-w 4.50 ± 0.16h-l 2.37 ± 0.41D
B 3 6.85 ± 0.24def 4.20 ± 0.19i-m 1.95 ± 0.06q-v 4.00 ± 0.19j-n 5.50 ± 0.27e-i 4.50 ± 0.44C
B 5 8.85 ± 0.51b 3.50 ± 0.09l-p 5.00 ± 0.31g-k 5.95 ± 0.22d-h 8.50 ± 0.29b 6.36 ± 0.56B
B 6 5.85 ± 0.26d-h 3.65 ± 0.24k-o 1.00 ± 0.03u-x 5.85 ± 0.31d-h 5.00 ± 0.21g-k 4.27 ± 0.49C
B 7 5.45 ± 0.30e-j 1.25 ± 0.07t-x 3.00 ± 0.12m-s 1.85 ± 0.08q-v 0.50 ± 0.03vwx 2.41 ± 0.46D
M 1 5.60 ± 0.30d-i 3.20 ± 0.16l-q 2.50 ± 0.14o-t 2.10 ± 0.09p-u 7.00 ± 0.43cd 4.08 ± 0.52C
M 3 4.35 ± 0.12i-m 6.90 ± 0.40de 1.20 ± 0.07t-x 3.30 ± 0.15l-q 5.50 ± 0.31e-i 4.25 ± 0.53C
M 5 8.40 ± 0.43bc 4.50 ± 0.23h-l 5.50 ± 0.26e-i 6.60 ± 0.46def 10.50 ± 0.57a 7.10 ± 0.59A
M 6 9.75 ± 0.46ab 2.65 ± 0.14n-t 5.40 ± 0.27f-j 6.35 ± 0.29d-g 5.60 ± 0.21d-i 5.95 ± 0.62B
M 7 3.10 ± 0.09l-r 0.25 ± 0.01wx 6.50 ± 0.32def 1.60 ± 0.07s-w 0.50 ± 0.02vwx 2.39 ± 0.61D
B= Boiling on burner , M= heating in microwave; BM= boiling method
. In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table -18: Means comparison of yield for muskmelon fruit peel (Mechanical procedure pH interaction mean±SE)
pH level Mechanical procedure Mean
119
Homogenizing Grinding Cutting Chopping Hammering
1 0.55 ± 0.02m-p 2.45 ± 0.12a 0.50 ± 0.01n-q 0.38 ± 0.15pqr 0.95 ± 0.14ijk 0.97 ± 0.15D
3 1.50 ± 0.23fg 1.60 ± 0.10ef 2.50 ± 0.05a 0.28 ± 0.10qr 0.65 ± 0.07l-o 1.31 ± 0.15B
5 1.88 ± 0.20cd 2.18 ± 0.13b 1.00 ± 0.02ij 1.90 ± 0.10cd 0.48 ± 0.08o-r 1.49 ± 0.13A
6 1.08 ± 0.05hi 1.28 ± 0.15gh 0.83 ± 0.06jkl 2.00 ± 0.12bc 0.73 ± 0.10k-n 1.18 ± 0.09C
7 1.18 ± 0.28hi 0.78 ± 0.10j-m 1.75 ± 0.21de 0.78 ± 0.13j-m 0.25 ± 0.09r 0.95 ± 0.12D
Mean 1.24 ± 0.11C 1.66 ± 0.12A 1.32 ± 0.14B 1.07 ± 0.15D 0.61 ± 0.06E
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 19: Means comparison of yield for muskmelon fruit peel (Mechanical procedure boiling method interaction
mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 1.14 ± 0.15d 1.42 ± 0.17b 1.19 ± 0.18d 0.91 ± 0.24e 0.64 ± 0.05f 1.06 ± 0.08B
M 1.33 ± 0.17bc 1.89 ± 0.16a 1.44 ± 0.21b 1.22 ± 0.16cd 0.58 ± 0.11f 1.29 ± 0.09A
B= Boiling on burner , M= heating in microwave;
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table -20: Means comparison of yield for muskmelon fruit peel (Mechanical procedure pH boiling method interaction
mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 0.55 ± 0.03qr 2.25 ± 0.13bcd 0.50 ± 0.03qr 0.05 ± 0.00s 0.65 ± 0.03pqr 0.80 ± 0.20F
120
B 3 2.00 ± 0.04def 1.40 ± 0.05i-l 2.45 ± 0.06abc 0.05 ± 0.00s 0.50 ± 0.02qr 1.28 ± 0.24BCD
B 5 1.45 ± 0.05h-k 1.95 ± 0.10d-g 1.00 ± 0.04m-p 1.70 ± 0.05f-i 0.65 ± 0.03pqr 1.35 ± 0.13B
B 6 1.15 ± 0.06k-n 0.95 ± 0.03m-p 0.70 ± 0.05opq 2.25 ± 0.10bcd 0.95 ± 0.04m-p 1.20 ± 0.15DE
B 7 0.55 ± 0.02qr 0.55 ± 0.01qr 1.30 ± 0.07j-m 0.50 ± 0.03qr 0.45 ± 0.02qr 0.67 ± 0.09G
M 1 0.55 ± 0.03qr 2.65 ± 0.12a 0.50 ± 0.02qr 0.70 ± 0.04opq 1.25 ± 0.05j-m 1.13 ± 0.22E
M 3 1.00 ± 0.03m-p 1.80 ± 0.09e-h 2.55 ± 0.09ab 0.50 ± 0.02qr 0.80 ± 0.06n-q 1.33 ± 0.20BC
M 5 2.30 ± 0.09a-d 2.40 ± 0.14abc 1.00 ± 0.03m-p 2.10 ± 0.08cde 0.30 ± 0.01rs 1.62 ± 0.22A
M 6 1.00 ± 0.05m-p 1.60 ± 0.08g-j 0.95 ± 0.05m-p 1.75 ± 0.04e-i 0.50 ± 0.02qr 1.16 ± 0.12DE
M 7 1.80 ± 0.11e-h 1.00 ± 0.04m-p 2.20 ± 0.09bcd 1.05 ± 0.07l-o 0.05 ± 0.00s 1.22 ± 0.20CDE
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
121
Table -21: Means comparison of yield for apple fruit peel (Mechanical procedure pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 0.75 ± 0.12ijk 2.48 ± 0.06d 3.38 ± 0.26bc 3.10 ± 0.15c 1.60 ± 0.05gh 2.26 ± 0.19B
3 2.35 ± 0.19de 0.80 ± 0.16ij 3.73 ± 0.36ab 2.00 ± 0.34ef 3.10 ± 0.20c 2.40 ± 0.22A
5 3.03 ± 0.31c 1.05 ± 0.04i 3.20 ± 0.24c 0.90 ± 0.38i 3.95 ± 0.42a 2.43 ± 0.26A
6 1.45 ± 0.05h 2.63 ± 0.12d 2.03 ± 0.39ef 0.50 ± 0.18jkl 1.65 ± 0.18fgh 1.65 ± 0.16C
7 0.20 ± 0.07l 0.30 ± 0.11l 0.38 ± 0.15kl 0.43 ± 0.08jkl 1.88 ± 0.10fg 0.64 ± 0.12D
Mean 1.56 ± 0.20B 1.45 ± 0.18BC 2.54 ± 0.26A 1.39 ± 0.22C 2.44 ± 0.20A
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 22: Means comparison of yield for apple fruit peel (Mechanical procedure boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 1.27 ± 0.23e 1.27 ± 0.27e 1.94 ± 0.31c 0.90 ± 0.29f 2.04 ± 0.19c 1.48 ± 0.12B
M 1.84 ± 0.33c 1.63 ± 0.24d 3.14 ± 0.36a 1.87 ± 0.28c 2.83 ± 0.32b 2.26 ± 0.15A
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 23: Means comparison of yield for apple fruit peel (Mechanical procedure pH boiling method interaction
mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
122
B 1 0.50 ± 0.02r-u 2.45 ± 0.09fgh 2.85 ± 0.16d-g 2.85 ± 0.13d-g 1.50 ± 0.05i-m 2.03 ± 0.25C
B 3 1.95 ± 0.06hij 0.45 ± 0.02r-u 2.95 ± 0.14c-f 1.25 ± 0.08k-o 2.70 ± 0.15efg 1.86 ± 0.25C
B 5 2.35 ± 0.10gh 1.00 ± 0.05m-r 2.70 ± 0.05efg 0.05 ± 0.00u 3.05 ± 0.15cde 1.83 ± 0.30C
B 6 1.50 ± 0.08i-m 2.40 ± 0.07fgh 1.15 ± 0.06l-p 0.10 ± 0.01u 1.25 ± 0.09k-o 1.28 ± 0.20D
B 7 0.05 ± 0.00u 0.05 ± 0.00u 0.05 ± 0.00u 0.25 ± 0.00tu 1.70 ± 0.09i-l 0.42 ± 0.17F
M 1 1.00 ± 0.06m-r 2.50 ± 0.10e-h 3.90 ± 0.19b 3.35 ± 0.20bcd 1.70 ± 0.04i-l 2.49 ± 0.29B
M 3 2.75 ± 0.16efg 1.15 ± 0.04l-p 4.50 ± 0.21a 2.75 ± 0.14efg 3.50 ± 0.10bc 2.93 ± 0.30A
M 5 3.70 ± 0.15b 1.10 ± 0.04m-q 3.70 ± 0.18b 1.75 ± 0.09ijk 4.85 ± 0.23a 3.02 ± 0.37A
M 6 1.40 ± 0.07j-n 2.85 ± 0.11d-g 2.90 ± 0.06d-g 0.90 ± 0.04n-s 2.05 ± 0.05hi 2.02 ± 0.21C
M 7 0.35 ± 0.02stu 0.55 ± 0.02q-u 0.70 ± 0.04o-t 0.60 ± 0.02p-u 2.05 ± 0.12hi 0.85 ± 0.16E
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
123
Table -24: Means comparison of yield for orange fruit peel (Mechanical procedure pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 15.70 ± 0.45b 3.28 ± 0.32j 7.55 ± 0.30fgh 2.98 ± 0.19j 15.10 ± 0.88bc 8.92 ± 1.05C
3 19.00 ± 0.91a 6.55 ± 0.58ghi 10.25 ± 0.81d 9.00 ± 0.54def 7.78 ± 0.33e-h 10.52 ± 0.87B
5 21.05 ± 1.02a 4.70 ± 1.09ij 16.25 ± 2.07b 13.10 ± 1.27c 9.78 ± 1.11de 12.98 ± 1.18A
6 8.40 ± 2.51d-g 6.53 ± 0.21ghi 8.58 ± 0.81d-g 16.20 ± 0.61b 8.13 ± 1.17d-g 9.57 ± 0.84C
7 14.60 ± 3.20bc 0.40 ± 0.09k 5.70 ± 0.78hi 7.35 ± 0.92fgh 7.18 ± 0.78fgh 7.05 ± 1.07D
Mean 15.75 ± 1.14A 4.29 ± 0.49C 9.67 ± 0.82B 9.73 ± 0.91B 9.59 ± 0.65B
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table -25: Means comparison of yield for orange fruit peel (Mechanical procedure boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 17.88 ± 0.78a 3.95 ± 0.73e 7.88 ± 0.69d 8.31 ± 1.17d 7.84 ± 0.80d 9.17 ± 0.65B
M 13.62 ± 2.02b 4.63 ± 0.67e 11.45 ± 1.35c 11.14 ± 1.34c 11.34 ± 0.83c 10.44 ± 0.68A
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
124
Table -26: Means comparison of yield for orange fruit peel (Mechanical procedure pH boiling method interaction
mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 16.15 ± 0.44d-g 2.60 ± 0.14uvw 8.10 ± 0.33m-r 2.60 ± 0.11uvw 13.55 ± 0.48g-j 8.60 ± 1.49DE
B 3 18.20 ± 1.36b-e 7.80 ± 0.21m-r 8.50 ± 0.20l-r 7.95 ± 0.33m-r 7.20 ± 0.41n-s 9.93 ± 1.14CD
B 5 19.40 ± 0.80a-d 2.30 ± 0.10uvw 11.85 ± 0.64i-l 10.40 ± 0.66j-n 7.40 ± 0.20m-r 10.27 ± 1.52BC
B 6 13.95 ± 0.76f-i 6.45 ± 0.40p-t 6.95 ± 0.41o-s 15.25 ± 0.72e-h 5.55 ± 0.24q-u 9.63 ± 1.12B
B 7 21.70 ± 0.72a 0.60 ± 0.03vw 4.00 ± 0.21stu 5.35 ± 0.20r-u 5.50 ± 0.18q-u 7.43 ± 1.97BC
M 1 15.25 ± 0.80e-h 3.95 ± 0.16s-v 7.00 ± 0.17o-s 3.35 ± 0.14t-w 16.65 ± 1.12c-g 9.24 ± 1.52A
M 3 19.80 ± 1.29abc 5.30 ± 0.28r-u 12.00 ± 0.46h-k 10.05 ± 0.51k-o 8.35 ± 0.22m-r 11.10 ± 1.33CD
M 5 22.70 ± 1.36a 7.10 ± 0.39n-s 20.65 ± 1.25ab 15.80 ± 0.54efg 12.15 ± 0.66h-k 15.68 ± 1.55CD
M 6 2.85 ± 0.19uvw 6.60 ± 0.22p-t 10.20 ± 0.66j-o 17.15 ± 0.67c-f 10.70 ± 0.46i-m 9.50 ± 1.28EF
M 7 7.50 ± 0.35m-r 0.20 ± 0.01w 7.40 ± 0.35m-r 9.35 ± 0.43k-p 8.85 ± 0.48k-q 6.66 ± 0.90F
Means sharing similar letter in a row or in a column are statistically non-significant (P>0.05). Small letters represent comparison among interaction means and
capital letters are used for overall mean
125
Table - 27: Percentage yield of pectin from sapodilla fruit peel after using different
physicomechanical procedures (pH, mechanical procedure, boiling method and time of
boiling) with 0.1N HCl
Mechanical procedure
pH %yield 10 Min %yield 20min %yield 40 min %yield 60min
B M B M B M B M
Ham
mer
ing
1 1.5 2.05 0.5 1.1 0.05 0.05 1.2 1.8
3 0.6 1.55 2.2 4.85 0.05 0.1 4.1 3.15
5 0.8 1.95 5.5 6.35 0.05 0.05 4.5 4.6
6 0.8 1.4 3.5 5.2 1.65 1.65 3 3.45
7 0.5 0 2.6 4.35 0.05 0.95 1.75 3.85
Gri
nd
ing
1 0.55 0.8 0.75 1.6 0.1 1.15 0.25 0.35
3 4 3.5 3.55 4 1 1.35 0.05 0.05
5 2.45 2.9 2.05 3.8 2.4 2.75 0.05 2.1
6 2 2 3.4 2.7 2.1 3.1 0.05 0.05
7 1.95 2.3 2.85 3.3 0.95 1.75 0.05 2.3
Cu
ttin
g
1 0.8 0.9 1.45 1.5 0.05 0.05 0.05 0.1
3 1.1 1.5 1.95 3.15 0.45 0.05 0.4 1.4
5 0.4 1 1.5 2.2 1.2 3.95 0.05 0.5
6 1 1.2 1.45 1.85 0.65 1 0.1 0.9
7 0.7 1 1.35 1.8 0.1 1.5 0.05 1.05
Ho
mo
gen
izin
g
1 0.5 1.05 0.05 0.15 0.05 0.05 0.05 0.05
3 0.75 1.6 0.05 1 2.5 1.3 2.75 2.8
5 1.2 1.5 0.05 0.1 3 3.2 2.7 6
6 1.5 2 0.05 0.05 2.25 2.45 2.7 4.5
7 0.5 0.5 0.1 0.05 2.45 2.8 0.15 2.65
Ch
op
pin
g
(mort
ar/
pes
tle
1 2.7 4.7 0.05 0.05 0.05 0.05 1.85 2.75
3 3.9 3.45 4.75 6.5 1.6 1.35 1.25 3
5 3.95 4.05 2.25 3.05 2.05 2.35 1.95 5.6
6 2.3 2.45 2.5 2.8 2.7 3.6 3.25 5
7 3.95 2.4 0.1 0.7 1 2 3.6 3.65
B= Boiling on burner , M= heating in microwave
126
Table -28: Analysis of variance (mean squares) of yield of pectin from sapodilla fruit peel after using different
physicomechanical procedure different fruits with 0.1N HCl
Source of
variation
Degrees of
freedom
Mean squares
Yield 10 min Yield 20 min Yield 40 min Yield 60 min
MP
pH
BM
MP x pH
MP x BM
pH x BM
MP x pH x BM
Error
Total
4
4
1
16
4
4
16
100
149
32.6575**
3.3917**
3.2414**
2.5462**
0.3392**
1.0112**
0.6258**
0.0081
49.7412**
27.4284**
18.6772**
7.5601**
1.7538**
1.4724**
0.4729**
0.0129
12.1446**
20.1921**
6.1206**
2.3355**
1.0438**
1.6176**
0.5655**
0.0070
55.6906**
15.5938**
39.7837**
4.3065**
2.0295**
3.7639**
1.5634**
0.0122
NS = Non-significant (P>0.05); * = Significant (P<0.05); ** = Highly significant (P<0.01). Mech = Mechanical
Table – 29: Means comparison of yield for sapodilla fruit peel after 10 min of boiling using 0.1N HCl (Mechanical procedure
pH interaction mean±SE)
127
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 1.78 ± 0.13g 0.68 ± 0.06kl 0.85 ± 0.02k 3.70 ± 0.45b 0.78 ± 0.12k 1.56 ± 0.23D
3 1.08 ± 0.21j 3.75 ± 0.14b 1.30 ± 0.09hi 3.68 ± 0.11b 1.18 ± 0.19ij 2.20 ± 0.24A
5 1.38 ± 0.26h 2.68 ± 0.10d 0.70 ± 0.13k 4.00 ± 0.07a 1.35 ± 0.07hi 2.02 ± 0.23B
6 1.10 ± 0.13j 2.00 ± 0.04f 1.10 ± 0.05j 2.38 ± 0.04e 1.75 ± 0.12g 1.67 ± 0.10C
7 0.25 ± 0.11m 2.13 ± 0.08f 0.85 ± 0.07k 3.18 ± 0.35c 0.50 ± 0.01l 1.38 ± 0.22E
Mean 1.12 ± 0.12C 2.25 ± 0.19B 0.96 ± 0.05D 3.39 ± 0.15A 1.11 ± 0.09C
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 30: Means comparison of yield for sapodilla fruit peel after 10 min of boiling using 0.1N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 0.84 ± 0.09f 2.19 ± 0.30c 0.80 ± 0.07f 3.36 ± 0.19a 0.89 ± 0.11f 1.62 ± 0.14B
M 1.39 ± 0.20d 2.30 ± 0.24b 1.12 ± 0.06e 3.41 ± 0.24a 1.33 ± 0.14d 1.91 ± 0.13A B= Boiling on burner , M= heating in microwave; BM= boiling method In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table -31: Means comparison of yield for sapodilla fruit peel after 10 min of boiling using 0.1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 1.50 ± 0.02ij 0.55 ± 0.01qr 0.80 ± 0.02m-q 2.70 ± 0.05de 0.50 ± 0.01qr 1.21 ± 0.22F
B 3 0.60 ± 0.02pqr 4.00 ± 0.10b 1.10 ± 0.02klm 3.90 ± 0.09b 0.75 ± 0.02n-q 2.07 ± 0.41B
B 5 0.80 ± 0.01m-q 2.45 ± 0.03ef 0.40 ± 0.01r 3.95 ± 0.12b 1.20 ± 0.03jkl 1.76 ± 0.35D
128
B 6 0.80 ± 0.01m-q 2.00 ± 0.05gh 1.00 ± 0.02l-o 2.30 ± 0.05fg 1.50 ± 0.04ij 1.52 ± 0.15E
B 7 0.50 ± 0.02qr 1.95 ± 0.06h 0.70 ± 0.02o-r 3.95 ± 0.11b 0.50 ± 0.01qr 1.52 ± 0.36E
M 1 2.05 ± 0.05gh 0.80 ± 0.02m-q 0.90 ± 0.02l-p 4.70 ± 0.10a 1.05 ± 0.01lmn 1.90 ± 0.39C
M 3 1.55 ± 0.05i 3.50 ± 0.14c 1.50 ± 0.03ij 3.45 ± 0.02c 1.60 ± 0.04i 2.32 ± 0.25A
M 5 1.95 ± 0.05h 2.90 ± 0.05d 1.00 ± 0.01l-o 4.05 ± 0.10b 1.50 ± 0.05ij 2.28 ± 0.29A
M 6 1.40 ± 0.03ijk 2.00 ± 0.08gh 1.20 ± 0.03jkl 2.45 ± 0.04ef 2.00 ± 0.05gh 1.81 ± 0.12CD
M 7 0.00 ± 0.00s 2.30 ± 0.02fg 1.00 ± 0.02l-o 2.40 ± 0.06ef 0.50 ± 0.01qr 1.24 ± 0.26F
B= Boiling on burner , M= heating in microwave; BM= boiling method In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table -32: Means comparison of yield for sapodilla fruit peel after 20 min of boiling using 0.1N HCl (Mechanical procedure
pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 0.80 ± 0.13k 1.18 ± 0.19j 1.48 ± 0.03i 0.05 ± 0.00m 0.10 ± 0.02m 0.72 ± 0.11E
3 3.53 ± 0.60e 3.78 ± 0.12d 2.55 ± 0.27g 5.63 ± 0.40b 0.53 ± 0.21l 3.20 ± 0.34A
5 5.93 ± 0.19a 2.93 ± 0.40f 1.85 ± 0.16h 2.65 ± 0.19g 0.08 ± 0.01m 2.69 ± 0.37B
6 4.35 ± 0.38c 3.05 ± 0.17f 1.65 ± 0.09hi 2.65 ± 0.08g 0.05 ± 0.00m 2.35 ± 0.28C
7 3.48 ± 0.40e 3.08 ± 0.12f 1.58 ± 0.10i 0.40 ± 0.13l 0.08 ± 0.01m 1.72 ± 0.27D
Mean 3.62 ± 0.35A 2.80 ± 0.19B 1.82 ± 0.10D 2.27 ± 0.38C 0.17 ± 0.05E In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 33: Means comparison of yield for sapodilla fruit peel after 20 min of boiling using 0.1N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
129
B 2.86 ± 0.44c 2.52 ± 0.28d 1.54 ± 0.06g 1.93 ± 0.47f 0.06 ± 0.01i 1.78 ± 0.18B
M 4.37 ± 0.47a 3.08 ± 0.23b 2.10 ± 0.15e 2.62 ± 0.61d 0.27 ± 0.10h 2.49 ± 0.22A B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 34: Means comparison of yield for sapodilla fruit peel after 20 min of boiling using 0.1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 0.50 ± 0.01ab 0.75 ± 0.01YZa 1.45 ± 0.03UVW 0.05 ± 0.00c 0.05 ± 0.00c 0.56 ± 0.14H
B 3 2.20 ± 0.04OPQ 3.55 ± 0.10GH 1.95 ± 0.01P-S 4.75 ± 0.09D 0.05 ± 0.00c 2.50 ± 0.42C
B 5 5.50 ± 0.07B 2.05 ± 0.03PQR 1.50 ± 0.05TUV 2.25 ± 0.06NOP 0.05 ± 0.00c 2.27 ± 0.48D
B 6 3.50 ± 0.05GHI 3.40 ± 0.13HIJ 1.45 ± 0.03UVW 2.50 ± 0.06MNO 0.05 ± 0.00c 2.18 ± 0.35D
B 7 2.60 ± 0.02MN 2.85 ± 0.09KLM 1.35 ± 0.02VWX 0.10 ± 0.01c 0.10 ± 0.01c 1.40 ± 0.31F
M 1 1.10 ± 0.01WXY 1.60 ± 0.03S-V 1.50 ± 0.05TUV 0.05 ± 0.00c 0.15 ± 0.01bc 0.88 ± 0.18G
M 3 4.85 ± 0.13CD 4.00 ± 0.12EF 3.15 ± 0.06IJK 6.50 ± 0.17A 1.00 ± 0.03XYZ 3.90 ± 0.49A
M 5 6.35 ± 0.05A 3.80 ± 0.12FG 2.20 ± 0.03OPQ 3.05 ± 0.10JKL 0.10 ± 0.01c 3.10 ± 0.55B
M 6 5.20 ± 0.04BC 2.70 ± 0.08LM 1.85 ± 0.05Q-T 2.79 ± 0.10KLM 0.05 ± 0.01c 2.52 ± 0.45C
M 7 4.35 ± 0.14E 3.30 ± 0.10HIJ 1.80 ± 0.05R-U 0.70 ± 0.02Za 0.05 ± 0.00c 2.04 ± 0.43E
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 35: Means comparison of yield for sapodilla fruit peel after 40 min of boiling using 0.1N HCl (Mechanical procedure
pH interaction mean±SE)
130
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 0.05 ± 0.00l 0.63 ± 0.23ij 0.05 ± 0.00l 0.05 ± 0.00l 0.05 ± 0.00l 0.17 ± 0.06D
3 0.08 ± 0.01kl 1.18 ± 0.08g 0.25 ± 0.09k 1.48 ± 0.06ef 1.90 ± 0.27d 0.98 ± 0.14C
5 0.05 ± 0.00l 2.58 ± 0.08b 2.58 ± 0.62b 2.20 ± 0.08c 3.10 ± 0.09a 2.10 ± 0.23A
6 1.65 ± 0.02e 2.60 ± 0.23b 0.83 ± 0.08h 3.15 ± 0.20a 2.35 ± 0.07c 2.12 ± 0.16A
7 0.50 ± 0.20j 1.35 ± 0.18fg 0.80 ± 0.31hi 1.50 ± 0.23ef 2.63 ± 0.09b 1.36 ± 0.16B
Mean 0.47 ± 0.12D 1.67 ± 0.16B 0.90 ± 0.21C 1.68 ± 0.20B 2.01 ± 0.20A
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 36: Means comparison of yield for sapodilla fruit peel after 40 min of boiling using 0.1N HCl ( Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 0.37 ± 0.17f 1.31 ± 0.22d 0.49 ± 0.11e 1.48 ± 0.24c 2.05 ± 0.28a 1.14 ± 0.12B
M 0.56 ± 0.17e 2.02 ± 0.21a 1.31 ± 0.38d 1.87 ± 0.31b 1.96 ± 0.31ab 1.54 ± 0.14A
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 37: Means comparison of yield for sapodilla fruit peel after 40 min of boiling using 0.1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 0.05 ± 0.00r 0.10 ± 0.00r 0.05 ± 0.00r 0.05 ± 0.00r 0.05 ± 0.01r 0.06 ± 0.01H
B 3 0.05 ± 0.00r 1.00 ± 0.01p 0.45 ± 0.01q 1.60 ± 0.05lm 2.50 ± 0.09fgh 1.12 ± 0.23E
B 5 0.05 ± 0.00r 2.40 ± 0.05h 1.20 ± 0.01op 2.05 ± 0.05j 3.00 ± 0.11cd 1.74 ± 0.28D
131
B 6 1.65 ± 0.02l 2.10 ± 0.02ij 0.65 ± 0.01q 2.70 ± 0.03efg 2.25 ± 0.08hij 1.87 ± 0.19C
B 7 0.05 ± 0.00r 0.95 ± 0.01p 0.10 ± 0.01r 1.00 ± 0.06p 2.45 ± 0.05gh 0.91 ± 0.23F
M 1 0.05 ± 0.00r 1.15 ± 0.01op 0.05 ± 0.00r 0.05 ± 0.01r 0.05 ± 0.00r 0.27 ± 0.12G
M 3 0.10 ± 0.00r 1.35 ± 0.02mno 0.05 ± 0.00r 1.35 ± 0.05mno 1.30 ± 0.04no 0.83 ± 0.17F
M 5 0.05 ± 0.00r 2.75 ± 0.06def 3.95 ± 0.10a 2.35 ± 0.09hi 3.20 ± 0.13c 2.46 ± 0.35A
M 6 1.65 ± 0.03l 3.10 ± 0.09c 1.00 ± 0.01p 3.60 ± 0.08b 2.45 ± 0.07gh 2.36 ± 0.25B
M 7 0.95 ± 0.01p 1.75 ± 0.03kl 1.50 ± 0.03lmn 2.00 ± 0.06jk 2.80 ± 0.07de 1.80 ± 0.16CD
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
132
Table - 38: Means comparison of yield for sapodilla fruit peel after 60 min of boiling using 0.1N HCl Mechanical procedure
pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 1.50 ± 0.13g 0.30 ± 0.02lm 0.08 ± 0.01mn 2.30 ± 0.21f 0.05 ± 0.00n 0.85 ± 0.17D
3 3.63 ± 0.21c 0.05 ± 0.00n 0.90 ± 0.22j 2.13 ± 0.40f 2.78 ± 0.04e 1.90 ± 0.26C
5 4.55 ± 0.06a 1.08 ± 0.46ij 0.28 ± 0.10lmn 3.78 ± 0.82c 4.35 ± 0.74ab 2.81 ± 0.40A
6 3.23 ± 0.10d 0.05 ± 0.00n 0.50 ± 0.18kl 4.13 ± 0.40b 3.60 ± 0.41c 2.30 ± 0.33B
7 2.80 ± 0.47e 1.18 ± 0.50hi 0.55 ± 0.22k 3.63 ± 0.04c 1.40 ± 0.56gh 1.91 ± 0.27C
Mean 3.14 ± 0.21A 0.53 ± 0.16C 0.46 ± 0.09C 3.19 ± 0.24A 2.44 ± 0.34B In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 39: Means comparison of yield for sapodilla fruit peel after 60 min of boiling using 0.1N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 2.91 ± 0.34d 0.09 ± 0.02i 0.13 ± 0.04i 2.38 ± 0.24e 1.67 ± 0.34f 1.44 ± 0.17B
M 3.37 ± 0.25b 0.97 ± 0.27g 0.79 ± 0.12h 4.00 ± 0.30a 3.20 ± 0.54c 2.47 ± 0.21A B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 40: Means comparison of yield for sapodilla fruit peel after 60 min of boiling using 0.1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 1.20 ± 0.01qr 0.25 ± 0.01st 0.05 ± 0.00t 1.85 ± 0.05o 0.05 ± 0.00t 0.68 ± 0.19G
B 3 4.10 ± 0.03e 0.05 ± 0.00t 0.40 ± 0.01st 1.25 ± 0.03qr 2.75 ± 0.05l 1.71 ± 0.41E
133
B 5 4.50 ± 0.04d 0.05 ± 0.00t 0.05 ± 0.00t 1.95 ± 0.03no 2.70 ± 0.02l 1.85 ± 0.45D
B 6 3.00 ± 0.06jkl 0.05 ± 0.00t 0.10 ± 0.01t 3.25 ± 0.09hij 2.70 ± 0.10l 1.82 ± 0.38DE
B 7 1.75 ± 0.03op 0.05 ± 0.01t 0.05 ± 0.01t 3.60 ± 0.04fgh 0.15 ± 0.01st 1.12 ± 0.37F
M 1 1.80 ± 0.02o 0.35 ± 0.00st 0.10 ± 0.01t 2.75 ± 0.09l 0.05 ± 0.01t 1.01 ± 0.29F
M 3 3.15 ± 0.05ijk 0.05 ± 0.00t 1.40 ± 0.03pq 3.00 ± 0.12jkl 2.80 ± 0.08kl 2.08 ± 0.32C
M 5 4.60 ± 0.11d 2.10 ± 0.04no 0.50 ± 0.02s 5.60 ± 0.07b 6.00 ± 0.21a 3.76 ± 0.57A
M 6 3.45 ± 0.03ghi 0.05 ± 0.00t 0.90 ± 0.02r 5.00 ± 0.17c 4.50 ± 0.14d 2.78 ± 0.53B
M 7 3.85 ± 0.06ef 2.30 ± 0.08mn 1.05 ± 0.02qr 3.65 ± 0.07fg 2.65 ± 0.11lm 2.70 ± 0.27B
B= Boiling on burner , M= heating in microwave; BM= boiling method In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
134
Table - 41: Percentage yield of pectin from sapodilla fruit peel after using different
physicomechanical procedures (pH, mechanical procedure, boiling method and time of
boling) with IN HCl.
Mechanical
procedure pH
% yield 10 min %Yield 20 min %yield 40 min % yield 60 min
B M B M B M B M
Hammering
1 2.65 3.15 1.5 2.2 3.5 3 0.3 2
3 5.65 6.55 3.45 5.45 4.5 4.5 0.4 2.85
5 2.3 4.6 4.4 4.5 4.2 5.25 0.15 2.3
6 2.15 2.95 2.45 4.3 2 3.4 3.45 2.6
7 2.5 3.1 2.4 2.45 2 2.55 3.5 0.05
Grinding
1 3.1 4.15 1.55 1.65 1.4 3.7 1.4 3
3 5.6 4.8 2.55 5.4 3.25 5.15 3.15 5
5 1.05 6.1 4.2 5.1 3.8 5.5 1 3.3
6 1.45 3.55 4.3 3.65 3.5 3.3 0.8 1.2
7 0.35 2.75 1.3 2.5 2.15 2.6 1.65 1.6
Cutting
1 0.05 0.05 0.5 1.4 3.65 4 0.65 0.05
3 0.05 1.6 1 2.35 2.5 3.5 0.75 0.1
5 2.95 5.3 2.05 2.7 4.1 5.5 0.1 0.05
6 0.15 3.5 3.35 3.7 1.5 3 0 0.6
7 0.05 1.2 2.6 1.8 2 3.25 0.1 0.05
Homogenizing
1 1.55 4.95 2.1 3.6 3.4 4.6 0.6 0
3 2.4 5.35 3.55 5.25 3.65 4.55 0.05 0.15
5 1.7 4.85 3.55 5.9 2.7 3 0.15 3.4
6 0.6 1.5 3.8 3.15 3.9 3.5 0.05 0.25
7 1.2 2.55 1.25 2.2 3.35 3.3 0.1 0.6
Chopping 1 3 5.5 2.5 5.9 1.95 3.2 0.05 3.55
(mortor/pastle 3 3.7 5.4 1.4 6 1.15 4.85 0.2 0.1
5 4.1 6.7 1.25 6.1 4.5 6.5 0.45 0.5
6 3.75 3.8 1.2 4.9 2.5 3.1 0 1
7 0.05 0.25 1.8 2.5 1 3.95 0.1 0.6
B= Boiling on burner , M= heating in microwave
135
Table - 42: Analysis of variance (mean squares) of yield of pectin from sapodilla fruit peel after using different
physicomechanical procedure different fruits with 1N HCl
Source of
variation
Degrees of
freedom
Mean squares
Yield 10 min_NHCl Yield 20 min_NHCl Yield 40 min_NHCl Yield 60 min_NHCl
MP
pH
BM
MP x pH
MP x BM
pH x BM
MP x pH x BM
Error
Total
4
4
1
16
4
4
16
100
149
23.648**
38.736**
106.361**
8.396**
1.944**
4.779**
2.360**
0.022
8.5850**
21.8147**
71.7604**
3.9668**
10.4560**
4.7719**
1.5394**
0.0173
0.5439**
16.3404**
42.5601**
3.5228**
3.5189**
0.9204**
1.2315**
0.0168
21.8563**
0.8645**
14.8838**
4.0399**
2.1424**
4.7014**
3.3080**
0.0054
NS = Non-significant (P>0.05); * = Significant (P<0.05); ** = Highly significant (P<0.01); Mech= mechanical
136
Table - 43: Means comparison of yield for sapodilla fruit peel after 10 min of boiling using 1N HCl ( Mechanical procedure
pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 2.90 ± 0.12j 3.63 ± 0.24fg 0.05 ± 0.00p 4.25 ± 0.56cd 3.25 ± 0.76i 2.82 ± 0.33C
3 6.10 ± 0.21a 5.20 ± 0.19b 0.83 ± 0.35no 4.55 ± 0.39c 3.88 ± 0.66ef 4.11 ± 0.37A
5 3.45 ± 0.51ghi 3.58 ± 1.13fgh 4.13 ± 0.53de 5.40 ± 0.59b 3.28 ± 0.71hi 3.97 ± 0.34B
6 2.55 ± 0.18k 2.50 ± 0.47k 1.83 ± 0.75lm 3.78 ± 0.07f 1.05 ± 0.20n 2.34 ± 0.24D
7 2.80 ± 0.14jk 1.55 ± 0.54m 0.63 ± 0.26o 0.15 ± 0.05p 1.88 ± 0.30l 1.40 ± 0.22E
Mean 3.56 ± 0.27A 3.29 ± 0.34B 1.49 ± 0.33D 3.63 ± 0.38A 2.67 ± 0.31C In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 44: Means comparison of yield for sapodilla fruit peel after 10 min of boiling using 1N HCl ( Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 3.05 ± 0.35d 2.31 ± 0.50e 0.65 ± 0.31g 2.92 ± 0.40d 1.49 ± 0.16f 2.08 ± 0.19B
M 4.07 ± 0.37b 4.27 ± 0.31a 2.33 ± 0.50e 4.33 ± 0.60a 3.84 ± 0.41c 3.77 ± 0.21A
B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 45: Means comparison of yield for sapodilla fruit peel after 10 min of boiling using 1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 2.65 ± 0.03n-q 3.10 ± 0.05lmn 0.05 ± 0.01w 3.00 ± 0.06mno 1.55 ± 0.03st 2.07 ± 0.31F
137
B 3 5.65 ± 0.12cd 5.60 ± 0.07cd 0.05 ± 0.01w 3.70 ± 0.10jk 2.40 ± 0.08pq 3.48 ± 0.56C
B 5 2.30 ± 0.05pq 1.05 ± 0.03tu 2.95 ± 0.11no 4.10 ± 0.15ij 1.70 ± 0.04rs 2.42 ± 0.28E
B 6 2.15 ± 0.04qr 1.45 ± 0.01st 0.15 ± 0.01vw 3.75 ± 0.08jk 0.60 ± 0.01uv 1.62 ± 0.34G
B 7 2.50 ± 0.05opq 0.35 ± 0.01vw 0.05 ± 0.01w 0.05 ± 0.01w 1.20 ± 0.03st 0.83 ± 0.25H
M 1 3.15 ± 0.09lmn 4.15 ± 0.05ij 0.05 ± 0.00w 5.50 ± 0.08d 4.95 ± 0.12e-h 3.56 ± 0.52C
M 3 6.55 ± 0.08ab 4.80 ± 0.13gh 1.60 ± 0.05s 5.40 ± 0.20de 5.35 ± 0.10def 4.74 ± 0.45B
M 5 4.60 ± 0.02hi 6.10 ± 0.20bc 5.30 ± 0.07d-g 6.70 ± 0.24a 4.85 ± 0.16fgh 5.51 ± 0.22A
M 6 2.95 ± 0.09no 3.55 ± 0.05kl 3.50 ± 0.05klm 3.80 ± 0.13jk 1.50 ± 0.05st 3.06 ± 0.22D
M 7 3.10 ± 0.01lmn 2.75 ± 0.09nop 1.20 ± 0.03st 0.25 ± 0.02vw 2.55 ± 0.05opq 1.97 ± 0.29F
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 46: Means comparison of yield for sapodilla fruit peel after 20 min of boiling using 1N HCl Mechanical procedure
pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 1.82 ± 0.15kl 1.60 ± 0.03l 0.95 ± 0.20m 4.20 ± 0.77cd 2.85 ± 0.34h 2.28 ± 0.27D
3 4.45 ± 0.45abc 3.98 ± 0.64de 1.68 ± 0.30kl 3.70 ± 1.03ef 4.40 ± 0.39bc 3.64 ± 0.32B
5 4.45 ± 0.06abc 4.65 ± 0.21ab 2.38 ± 0.15i 3.68 ± 1.09f 4.73 ± 0.53a 3.98 ± 0.28A
6 3.38 ± 0.41g 3.98 ± 0.15de 3.53 ± 0.10fg 3.05 ± 0.83h 3.48 ± 0.16fg 3.48 ± 0.19C
7 2.43 ± 0.03i 1.90 ± 0.27jk 2.20 ± 0.18i 2.15 ± 0.16ij 1.73 ± 0.21kl 2.08 ± 0.09E
Mean 3.30 ± 0.23BC 3.22 ± 0.27C 2.15 ± 0.18D 3.36 ± 0.37AB 3.44 ± 0.25A
138
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
139
Table - 47: Means comparison of yield for sapodilla fruit peel after 20 min of boiling using 1N HCl ( Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 2.84 ± 0.27d 2.78 ± 0.34d 1.90 ± 0.28f 1.63 ± 0.13g 2.85 ± 0.27d 2.40 ± 0.13B
M 3.77 ± 0.34c 3.66 ± 0.39c 2.39 ± 0.21e 5.08 ± 0.37a 4.02 ± 0.37b 3.78 ± 0.18A
B= Boiling on burner , M= heating in microwave In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
140
Table - 48: Means comparison of yield for sapodilla fruit peel after 20 min of boiling using 1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 1.50 ± 0.02qrs 1.55 ± 0.03qrs 0.50 ± 0.01u 2.50 ± 0.06j-m 2.10 ± 0.05mno 1.63 ± 0.18F
B 3 3.45 ± 0.02ghi 2.55 ± 0.05jkl 1.00 ± 0.02t 1.40 ± 0.03q-t 3.55 ± 0.06ghi 2.39 ± 0.28D
B 5 4.40 ± 0.06e 4.20 ± 0.07ef 2.05 ± 0.03nop 1.25 ± 0.02rst 3.55 ± 0.05ghi 3.09 ± 0.33C
B 6 2.45 ± 0.04j-n 4.30 ± 0.04e 3.35 ± 0.05hi 1.20 ± 0.03st 3.80 ± 0.10fg 3.02 ± 0.29C
B 7 2.40 ± 0.05j-n 1.30 ± 0.01rst 2.60 ± 0.09jk 1.80 ± 0.05opq 1.25 ± 0.01rst 1.87 ± 0.15E
M 1 2.13 ± 0.07l-o 1.65 ± 0.01pqr 1.40 ± 0.01q-t 5.90 ± 0.19a 3.60 ± 0.08gh 2.94 ± 0.45C
M 3 5.45 ± 0.08b 5.40 ± 0.13b 2.35 ± 0.02j-n 6.00 ± 0.15a 5.25 ± 0.14bc 4.89 ± 0.35A
M 5 4.50 ± 0.10de 5.10 ± 0.10bc 2.70 ± 0.06j 6.10 ± 0.08a 5.90 ± 0.17a 4.86 ± 0.33A
M 6 4.30 ± 0.03e 3.65 ± 0.06gh 3.70 ± 0.14gh 4.90 ± 0.11cd 3.15 ± 0.07i 3.94 ± 0.16B
M 7 2.45 ± 0.04j-n 2.50 ± 0.08j-m 1.80 ± 0.04opq 2.50 ± 0.05j-m 2.20 ± 0.06k-o 2.29 ± 0.08D
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
141
Table - 49: Means comparison of yield for sapodilla fruit peel after 40 min of boiling using 1N HCl (Mechanical procedure
pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 3.25 ± 0.12gh 2.55 ± 0.51k-n 3.83 ± 0.09ef 2.58 ± 0.28j-m 4.00 ± 0.28de 3.24 ± 0.17C
3 4.50 ± 0.05c 4.20 ± 0.43d 3.00 ± 0.23hi 3.00 ± 0.83hi 4.10 ± 0.21de 3.76 ± 0.22B
5 4.73 ± 0.24bc 4.65 ± 0.38bc 4.80 ± 0.32b 5.50 ± 0.46a 2.85 ± 0.08ij 4.51 ± 0.21A
6 2.70 ± 0.32jkl 3.38 ± 0.04g 2.25 ± 0.34o 2.80 ± 0.14ijk 3.70 ± 0.10f 2.97 ± 0.13D
7 2.28 ± 0.13no 2.38 ± 0.10mno 2.63 ± 0.28j-m 2.48 ± 0.66l-o 3.33 ± 0.04g 2.62 ± 0.15E
Mean 3.49 ± 0.20B 3.43 ± 0.22B 3.30 ± 0.20C 3.27 ± 0.31C 3.60 ± 0.11A
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 50: Means comparison of yield for sapodilla fruit peel after 40 min of boiling using 1N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 3.24 ± 0.29e 2.81 ± 0.24f 2.75 ± 0.26f 2.22 ± 0.34g 3.40 ± 0.11d 2.88 ± 0.12B
M 3.74 ± 0.27c 4.05 ± 0.30b 3.85 ± 0.24c 4.32 ± 0.34a 3.79 ± 0.18c 3.95 ± 0.12A
B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 51: Means comparison of yield for sapodilla fruit peel after 40 min of boiling using 1N HCl ( Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 3.50 ± 0.05j-n 1.40 ± 0.02uv 3.65 ± 0.05i-m 1.95 ± 0.04t 3.40 ± 0.10k-o 2.78 ± 0.25G
142
B 3 4.50 ± 0.11def 3.25 ± 0.04mno 2.50 ± 0.05rs 1.15 ± 0.02uv 3.65 ± 0.12i-m 3.01 ± 0.30F
B 5 4.20 ± 0.04efg 3.80 ± 0.11g-k 4.10 ± 0.09fgh 4.50 ± 0.04def 2.70 ± 0.10pqr 3.86 ± 0.17C
B 6 2.00 ± 0.04t 3.47 ± 0.03j-n 1.50 ± 0.01u 2.50 ± 0.03rs 3.90 ± 0.05g-j 2.67 ± 0.24G
B 7 2.00 ± 0.04t 2.15 ± 0.03st 2.00 ± 0.03t 1.00 ± 0.03v 3.35 ± 0.08l-o 2.10 ± 0.20H
M 1 3.00 ± 0.11opq 3.70 ± 0.05h-l 4.00 ± 0.10ghi 3.20 ± 0.05no 4.60 ± 0.09de 3.70 ± 0.16D
M 3 4.50 ± 0.04def 5.15 ± 0.14bc 3.50 ± 0.03j-n 4.85 ± 0.09cd 4.55 ± 0.09de 4.51 ± 0.15B
M 5 5.25 ± 0.09bc 5.50 ± 0.07b 5.50 ± 0.08b 6.50 ± 0.19a 3.00 ± 0.05opq 5.15 ± 0.31A
M 6 3.40 ± 0.10k-o 3.30 ± 0.03l-o 3.00 ± 0.04opq 3.10 ± 0.06nop 3.50 ± 0.08j-n 3.26 ± 0.06E
M 7 2.55 ± 0.07rs 2.60 ± 0.05qr 3.25 ± 0.07mno 3.95 ± 0.14ghi 3.30 ± 0.05l-o 3.13 ± 0.14EF
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 52: Means comparison of yield for sapodilla fruit peel after 60 min of boiling using 1N HCl (Mechanical procedure
pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 1.15 ± 0.38fg 2.20 ± 0.36c 0.35 ± 0.13hi 1.80 ± 0.78d 0.30 ± 0.13ij 1.16 ± 0.23B
3 1.63 ± 0.55e 4.08 ± 0.42a 0.43 ± 0.15hi 0.15 ± 0.02jk 0.10 ± 0.02k 1.28 ± 0.31A
5 1.23 ± 0.48f 2.15 ± 0.52c 0.08 ± 0.01k 0.48 ± 0.02h 1.78 ± 0.73de 1.14 ± 0.24B
6 3.03 ± 0.20b 1.00 ± 0.09g 0.30 ± 0.13ij 0.50 ± 0.22h 0.15 ± 0.05jk 1.00 ± 0.21C
7 1.78 ± 0.77de 1.63 ± 0.03e 0.08 ± 0.01k 0.35 ± 0.11hi 0.35 ± 0.11hi 0.84 ± 0.20D
Mean 1.76 ± 0.25B 2.21 ± 0.24A 0.25 ± 0.05E 0.66 ± 0.19C 0.54 ± 0.18D In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 53: Means comparison of yield for sapodilla fruit peel after 60 min of boiling using 1N HCl (Mechanical procedure
boiling method interaction mean±SE)
143
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 1.56 ± 0.42c 1.60 ± 0.22c 0.32 ± 0.08f 0.16 ± 0.04g 0.19 ± 0.06g 0.77 ± 0.12B
M 1.96 ± 0.27b 2.82 ± 0.36a 0.17 ± 0.06g 1.15 ± 0.33d 0.88 ± 0.34e 1.40 ± 0.17A
B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 54: Means comparison of yield for sapodilla fruit peel after 60 min of boiling using 1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 0.30 ± 0.01r-u 1.40 ± 0.05kl 0.65 ± 0.03op 0.05 ± 0.01vw 0.60 ± 0.02opq 0.60 ± 0.12E
B 3 0.40 ± 0.02q-t 3.15 ± 0.07de 0.75 ± 0.03o 0.20 ± 0.01t-w 0.05 ± 0.00vw 0.91 ± 0.31D
B 5 0.15 ± 0.01uvw 1.00 ± 0.03mn 0.10 ± 0.01uvw 0.45 ± 0.01p-s 0.15 ± 0.01uvw 0.37 ± 0.09F
B 6 3.45 ± 0.09bc 0.80 ± 0.03no 0.00 ± 0.00w 0.00 ± 0.00w 0.05 ± 0.01vw 0.86 ± 0.36D
B 7 3.50 ± 0.08bc 1.65 ± 0.03j 0.10 ± 0.01uvw 0.10 ± 0.01uvw 0.10 ± 0.01uvw 1.09 ± 0.36C
M 1 2.00 ± 0.05i 3.00 ± 0.08ef 0.05 ± 0.00vw 3.55 ± 0.07b 0.00 ± 0.00w 1.72 ± 0.39B
M 3 2.85 ± 0.05f 5.00 ± 0.10a 0.10 ± 0.01uvw 0.10 ± 0.01uvw 0.15 ± 0.01uvw 1.64 ± 0.53B
M 5 2.30 ± 0.09h 3.30 ± 0.12cd 0.05 ± 0.00vw 0.50 ± 0.02pqr 3.40 ± 0.05bc 1.91 ± 0.37A
M 6 2.60 ± 0.07g 1.20 ± 0.04lm 0.60 ± 0.03opq 1.00 ± 0.05mn 0.25 ± 0.02s-v 1.13 ± 0.22C
M 7 0.05 ± 0.01vw 1.60 ± 0.05jk 0.05 ± 0.00vw 0.60 ± 0.02opq 0.60 ± 0.02opq 0.58 ± 0.15E B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
144
Table - 55: Percentage yield of pectin from sapodilla fruit peel after using different
organic acid
Mechanical
procedure Inorganic acid
% of inorganic
acid pH
% yield
B M h
om
og
en
izin
g
Citric acid 1%
3 to 5
2.55 3.25
10% 2.55 4.24
Oxalic acid 1% 3.95 4.95
10% 0.5 3.7
Tartaric acid 1% 1.8 3.25
10% 2.5 2.55
ch
op
pin
g
Citric acid 1% 2.4 4.3
10% 0.05 0.1
Oxalic acid 1% 0.05 1.5
10% 0.15 1.85
Tartaric acid 1% 0.75 1.25
10% 0.35 3.2
Grin
din
g
Citric acid 1% 1.35 2.15
10% 0.05 1.2
Oxalic acid 1% 0.1 0.3
10% 1.25 1.05
Tartaric acid 1% 1.8 2.43
10% 1.65 3.15
Cu
ttin
g
Citric acid 1% 1.4 2.64
10% 0.1 1.1
Oxalic acid 1% 0.05 1.2
10% 0.13 1.32
Tartaric acid 1% 0.2 1.87
10% 0.32 1.45
Ha
mm
erin
g
Citric acid 1% 1.6 1.8
10% 0.54 1.67
Oxalic acid 1% 0.1 1.3
10% 0.43 2
Tartaric acid 1% 0.33 1.4
10% 0.44 2.1
B= Boiling on burner , M= heating in microwave
145
Table - 56 : Analysis of variance (mean squares) of yield of pectin from sapodilla
fruit peel after using different organic acid
Source of
variation
Degrees of
freedom
Mean squares
Yield (citric acid) Yield (oxalic
acid)
Yield (tartaric
acid)
BM
MP
Acid%
BM x MP
BM x Acid%
MP x Acid%
BM x MP x Acid%
Error
Total
1
4
1
4
1
4
4
40
59
14.5829**
7.7555**
21.0278**
0.1237**
0.0049NS
5.6981**
1.0202**
0.0126
23.2877**
14.9075**
0.1882**
1.8114**
0.9077**
5.0161**
0.7483**
0.0090
23.4750**
6.0553**
1.0375**
0.3754**
0.5245**
0.3919**
1.5337**
0.0101
NS = Non-significant (P>0.05); * = Significant (P<0.05); ** = Highly significant (P<0.01) BM= Boiling method. MP= Mechanical procedure
Table - 57: Means comparison of yield for sapodilla fruit peel after using citric acid
(Boiling method x mechanical procedure interaction mean±SE)
Mechanical
procedure
Boiling Method Mean
B M
Homogenizing 2.55 ± 0.05b 3.75 ± 0.23a 3.15 ± 0.21A
Chopping 1.23 ± 0.53e 2.20 ± 0.94c 1.71 ± 0.54B
Grinding 0.70 ± 0.29f 1.68 ± 0.21d 1.19 ± 0.23D
Cutting 0.75 ± 0.29f 1.87 ± 0.34d 1.31 ± 0.27CD
Hammering 1.07 ± 0.24e 1.74 ± 0.04d 1.40 ± 0.15C
Mean 1.26 ± 0.18B 2.25 ± 0.24A
B= Boiling on burner , M= heating in microwave.In a row and column statistically non-significant (P>0.05)
represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 58: Means comparison of yield for sapodilla fruit peel after using citric acid
Acid (% x boiling method interaction mean±SE)
Inorganic Boiling Method Mean
146
acid B M
1% 1.86 ± 0.14b 2.83 ± 0.24a 2.34 ± 0.16A
10% 0.66 ± 0.26d 1.66 ± 0.37c 1.16 ± 0.24B B= Boiling on burner , M= heating in microwave Among the two concentrations of acid 1% has found more effective with microwave as heating procedure
Table - 59: Means comparison of yield for sapodilla fruit peel after using citric acid
(Boiling method x Acid% x mechanical procedure interaction mean±SE)
Acid% x MP Boiling Method
Mean B M
1% x MP1 2.55 ± 0.08c 3.25 ± 0.10b 2.90 ± 0.17B
1% x MP2 2.40 ± 0.09cd 4.30 ± 0.16a 3.35 ± 0.43A
1% x MP3 1.35 ± 0.04fg 2.15 ± 0.04d 1.75 ± 0.18D
1% x MP4 1.40 ± 0.02fg 2.64 ± 0.04c 2.02 ± 0.28C
1% x MP5 1.60 ± 0.04ef 1.80 ± 0.05e 1.70 ± 0.05D
10% x MP1 2.55 ± 0.08c 4.24 ± 0.12a 3.40 ± 0.38A
10% x MP2 0.05 ± 0.00i 0.10 ± 0.00i 0.08 ± 0.01G
10% x MP3 0.05 ± 0.00i 1.20 ± 0.02g 0.63 ± 0.26F
10% x MP4 0.10 ± 0.00i 1.10 ± 0.01g 0.60 ± 0.22F
10% x MP5 0.54 ± 0.01h 1.67 ± 0.05ef 1.11 ± 0.25E
BM= Boiling method. MP= Mechanical procedure; B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by
similar letters. Comparison among interaction means are represented by lower case alphabets while capital
alphabets are used for overall mean
Table - 60: Means comparison of yield for sapodilla fruit peel after using oxalic acid
(Boiling method x mechanical procedure interaction mean±SE)
Mechanical
procedure
Boiling Method Mean
B M
Homogenizing 2.23 ± 0.77b 4.33 ± 0.29a 3.28 ± 0.51A
Chopping 0.10 ± 0.02f 1.68 ± 0.08c 0.89 ± 0.24B
Grinding 0.68 ± 0.26e 0.68 ± 0.17e 0.68 ± 0.15C
Cutting 0.09 ± 0.02f 1.26 ± 0.03d 0.68 ± 0.18C
Hammering 0.27 ± 0.07f 1.65 ± 0.16c 0.96 ± 0.22B
Mean 0.67 ± 0.21B 1.92 ± 0.24A
147
B= Boiling on burner , M= heating in microwave; B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by
similar letters. Comparison among interaction means are represented by lower case alphabets while capital
alphabets are used for overall mean
Table - 61: Means comparison of yield for sapodilla fruit peel after using oxalic acid
Acid% x boiling method interaction mean±SE
Inorganic
acid
Boiling Method Mean
B M
1% 0.85 ± 0.42 1.85 ± 0.43 1.35 ± 0.31A
10% 0.49 ± 0.11 1.98 ± 0.25 1.24 ± 0.19B
B= Boiling on burner , M= heating in microwave
Among the two concentrations of acid 1% has found more effective with microwave as
heating procedure
Table - 62: Means comparison of yield for sapodilla fruit peel after using oxalic acid
(Boiling method x Acid% x mechanical procedure interaction mean±SE)
Acid% x MP Boiling Method
Mean B M
1% x MP1 3.95 ± 0.15b 4.95 ± 0.14a 4.45 ± 0.24A
1% x MP2 0.05 ± 0.00h 1.50 ± 0.02d 0.78 ± 0.32E
1% x MP3 0.10 ± 0.01h 0.30 ± 0.01fgh 0.20 ± 0.05F
1% x MP4 0.05 ± 0.00h 1.20 ± 0.01e 0.63 ± 0.26E
1% x MP5 0.10 ± 0.01h 1.30 ± 0.02de 0.70 ± 0.27E
10% x MP1 0.50 ± 0.02f 3.70 ± 0.10b 2.10 ± 0.72B
10% x MP2 0.15 ± 0.01gh 1.85 ± 0.03c 1.00 ± 0.38D
10% x MP3 1.25 ± 0.04de 1.05 ± 0.02e 1.15 ± 0.05CD
10% x MP4 0.13 ± 0.01h 1.32 ± 0.02de 0.73 ± 0.27E
10% x MP5 0.43 ± 0.02fg 2.00 ± 0.05c 1.22 ± 0.35C
BM= Boiling method. MP= Mechanical procedure ; B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by
similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
148
Table - 63: Means comparison of yield for sapodilla fruit peel after using tartaric
acid ( Boiling method x mechanical procedure interaction mean±SE)
Mechanical
procedure
Boiling Method Mean
B M
homogenizing 2.15 ± 0.16b 2.90 ± 0.17a 2.53 ± 0.16A
Chopping 0.55 ± 0.09d 2.23 ± 0.44b 1.39 ± 0.33C
Grinding 1.73 ± 0.04c 2.79 ± 0.17a 2.26 ± 0.18B
Cutting 0.26 ± 0.03e 1.66 ± 0.10c 0.96 ± 0.22D
Hammering 0.39 ± 0.03de 1.75 ± 0.16c 1.07 ± 0.22D
Mean 1.01 ± 0.15B 2.27 ± 0.14A
B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by
similar letters. Comparison among interaction means are represented by lower case alphabets while capital
alphabets are used for overall mean
Table - 64: Means comparison of yield for sapodilla fruit peel after using tartaric
acid (Acid% x boiling method interaction mean±SE)
Inorganic
acid
Boiling Method Mean
B M
1% 0.98 ± 0.19c 2.04 ± 0.20b 1.51 ± 0.17B
10% 1.05 ± 0.24c 2.49 ± 0.18a 1.77 ± 0.20A
B= Boiling on burner , M= heating in microwave
Among the two concentrations of acid 1% has found more effective with microwave as heating procedure
149
Table - 65: Means comparison of yield for sapodilla fruit peel after using tartaric
acid( Boiling method x Acid% x mechanical procedure interaction mean±SE)
Acid% x MP Boiling Method
Mean B M
1% x PM1 1.80 ± 0.03cd 3.25 ± 0.08a 2.53 ± 0.33A
1% x PM2 0.75 ± 0.02g 1.25 ± 0.02f 1.00 ± 0.11E
1% x PM3 1.80 ± 0.05cd 2.43 ± 0.09b 2.12 ± 0.15B
1% x PM4 0.20 ± 0.02h 1.87 ± 0.08cd 1.04 ± 0.38E
1% x PM5 0.33 ± 0.02h 1.40 ± 0.02ef 0.87 ± 0.24E
10% x PM1 2.50 ± 0.07b 2.55 ± 0.12b 2.53 ± 0.06A
10% x PM2 0.35 ± 0.02h 3.20 ± 0.08a 1.78 ± 0.64C
10% x PM3 1.65 ± 0.03de 3.15 ± 0.10a 2.40 ± 0.34A
10% x PM4 0.32 ± 0.02h 1.45 ± 0.02ef 0.89 ± 0.25E
10% x PM5 0.44 ± 0.03h 2.10 ± 0.05c 1.27 ± 0.37D
B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by
similar letters. Comparison among interaction means are represented by lower case alphabets while capital
alphabets are used for overall mean
150
Table - 66: percentage yield of pectin from sapodilla fruit peel using different
strengths of same inorganic acid.
Mechanical
procedure pH
0.1N HCl 0.5N HCl 1N HCl
B M B M B M
Hammering
1 1.5 2.05 0.65 1 2.65 3.15
3 0.6 1.55 0.05 3.3 5.65 6.55
5 0.8 1.95 0.3 2.2 2.3 4.6
6 0.8 1.4 0.45 1.65 2.15 2.95
7 0.5 0 0.25 0.05 2.5 3.1
Grinding
1 0.55 0.8 0.3 4.6 3.1 4.15
3 4 3.5 0.1 2.85 5.6 4.8
5 2.45 2.9 0.1 1 1.05 6.1
6 2 2 3.15 4.05 1.45 3.55
7 1.95 2.3 2.5 2.8 0.35 2.75
Cutting
1 0.8 0.9 3.65 4.2 0.05 0.05
3 1.1 1.5 4 2.25 0.05 1.6
5 0.4 1 4.35 3.75 2.95 5.3
6 1 1.2 0.55 4.75 0.15 3.5
7 0.7 1 1.25 0.5 0.05 1.2
Homogenizing
1 0.5 1.05 4.15 4.95 1.55 4.95
3 0.75 1.6 3.2 4.95 2.4 5.35
5 1.2 1.5 0.15 4.55 1.7 4.85
6 1.5 2 1.8 0.3 0.6 1.5
7 0.5 0.5 3.6 0.6 1.2 2.55
Chopping 1 2.7 4.7 4.4 4.65 3 5.5
(mortor/pastle 3 3.9 3.45 4.8 4.95 3.7 5.4
5 3.95 4.05 2.25 2.65 4.1 6.7
6 2.3 2.45 1.5 0.65 3.75 3.8
7 3.95 2.4 1 0.1 0.05 0.25
BM= Boiling method. MP= Mechanical procedure B= Boiling on burner , M= heating in microwave
133
Table - 67 : Analysis of variance (mean squares) of yield of pectin from sapodilla fruit peel after using different strength of
same inorganic acid
Source of
variation
Degrees of
freedom
Mean squares
Yield_0.1N HCl Yield_0.5N HCl Yield_1N HCl
MP
pH
BM
MP x pH
MP x BM
pH x BM
MP x pH x BM
Error
Total
4
4
1
16
4
4
16
100
149
32.6575**
3.3917**
3.2413**
2.5462**
0.3392**
1.0112**
0.6258**
0.0093
19.2084**
20.5359**
21.2064**
8.7942**
4.8684**
6.8626**
5.2626**
0.0104
24.375**
37.244**
109.739**
7.934**
1.832**
4.535**
2.303**
0.047
NS = Non-significant (P>0.05); * = Significant (P<0.05); ** = Highly significant (P<0.01) B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure
134
Table - 68: Means comparison for yield of pectin from sapodilla fruit peel after using 0.1N HCl ( Mechanical procedure pH
interaction mean±SE )
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 0.78 ± 0.12k 0.68 ± 0.06kl 0.85 ± 0.03k 3.70 ± 0.45b 1.78 ± 0.12g 1.56 ± 0.23D
3 1.18 ± 0.19hij 3.75 ± 0.13b 1.30 ± 0.09hi 3.68 ± 0.12b 1.08 ± 0.21j 2.20 ± 0.24A
5 1.35 ± 0.07h 2.68 ± 0.10d 0.70 ± 0.13kl 4.00 ± 0.09a 1.38 ± 0.26h 2.02 ± 0.23B
6 1.75 ± 0.11g 2.00 ± 0.04f 1.10 ± 0.05ij 2.38 ± 0.05e 1.10 ± 0.14ij 1.67 ± 0.10C
7 0.50 ± 0.01l 2.13 ± 0.09f 0.85 ± 0.07k 3.18 ± 0.35c 0.25 ± 0.11m 1.38 ± 0.22E
Mean 1.11 ± 0.09C 2.25 ± 0.19B 0.96 ± 0.05D 3.39 ± 0.15A 1.12 ± 0.12C
B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 69: Means comparison for yield of pectin from sapodilla fruit peel after using 0.1N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 0.89 ± 0.11e 2.19 ± 0.30b 0.80 ± 0.07e 3.36 ± 0.19a 0.84 ± 0.09e 1.62 ± 0.14B
M 1.33 ± 0.14c 2.30 ± 0.24b 1.12 ± 0.06d 3.41 ± 0.24a 1.39 ± 0.20c 1.91 ± 0.13A
B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 70: Means comparison for yield of pectin from sapodilla fruit peel after using 0.1N HCl (Mechanical procedure pH
boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 0.50 ± 0.02qr 0.55 ± 0.01qr 0.80 ± 0.02m-q 2.70 ± 0.05de 1.50 ± 0.04ij 1.21 ± 0.22F
135
B 3 0.75 ± 0.02n-q 4.00 ± 0.10b 1.10 ± 0.03klm 3.90 ± 0.10b 0.60 ± 0.03pqr 2.07 ± 0.41B
B 5 1.20 ± 0.03jkl 2.45 ± 0.04ef 0.40 ± 0.01r 3.95 ± 0.11b 0.80 ± 0.04m-q 1.76 ± 0.35D
B 6 1.50 ± 0.02ij 2.00 ± 0.05gh 1.00 ± 0.03l-o 2.30 ± 0.05fg 0.80 ± 0.03m-q 1.52 ± 0.15E
B 7 0.50 ± 0.01qr 1.95 ± 0.08h 0.70 ± 0.01o-r 3.95 ± 0.10b 0.50 ± 0.01qr 1.52 ± 0.36E
M 1 1.05 ± 0.03lmn 0.80 ± 0.01m-q 0.90 ± 0.02l-p 4.70 ± 0.10a 2.05 ± 0.03gh 1.90 ± 0.39C
M 3 1.60 ± 0.02i 3.50 ± 0.10c 1.50 ± 0.02ij 3.45 ± 0.08c 1.55 ± 0.05i 2.32 ± 0.25A
M 5 1.50 ± 0.03ij 2.90 ± 0.05d 1.00 ± 0.03l-o 4.05 ± 0.15b 1.95 ± 0.03h 2.28 ± 0.29A
M 6 2.00 ± 0.03gh 2.00 ± 0.08gh 1.20 ± 0.02jkl 2.45 ± 0.08ef 1.40 ± 0.05ijk 1.81 ± 0.12CD
M 7 0.50 ± 0.01qr 2.30 ± 0.08fg 1.00 ± 0.01l-o 2.40 ± 0.05ef 0.00 ± 0.00s 1.24 ± 0.26F
B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
136
Table - 71:Means comparison for yield of pectin from sapodilla fruit peel after using 0.5N HCl (Mechanical procedure pH
interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 4.55 ± 0.19b 2.45 ± 0.96fg 3.93 ± 0.13c 4.53 ± 0.08b 0.83 ± 0.08l 3.26 ± 0.32A
3 4.08 ± 0.39c 1.48 ± 0.62i 3.13 ± 0.39e 4.88 ± 0.08a 1.68 ± 0.73i 3.05 ± 0.32B
5 2.35 ± 0.99g 0.55 ± 0.20m 4.05 ± 0.14c 2.45 ± 0.10fg 1.25 ± 0.43j 2.13 ± 0.30C
6 1.05 ± 0.34jk 3.60 ± 0.21d 2.65 ± 0.94f 1.08 ± 0.19jk 1.05 ± 0.27jk 1.89 ± 0.28D
7 2.10 ± 0.67h 2.65 ± 0.07f 0.88 ± 0.17kl 0.55 ± 0.20m 0.15 ± 0.05n 1.27 ± 0.22E
Mean 2.83 ± 0.34B 2.15 ± 0.29D 2.93 ± 0.29A 2.70 ± 0.33C 0.99 ± 0.19E
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 72: Means comparison for yield of pectin from sapodilla fruit peel after using 0.5N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 2.58 ± 0.39c 1.23 ± 0.35e 2.76 ± 0.41b 2.79 ± 0.41b 0.34 ± 0.05f 1.94 ± 0.19B
M 3.07 ± 0.57a 3.06 ± 0.33a 3.09 ± 0.41a 2.60 ± 0.53c 1.64 ± 0.29d 2.69 ± 0.20A B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 73: Means comparison for yield of pectin from sapodilla fruit peel after using 0.5N HCl (Mechanical procedure pH
boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 4.15 ± 0.09def 0.30 ± 0.02s-v 3.65 ± 0.07h 4.40 ± 0.05cd 0.65 ± 0.03r 2.63 ± 0.48D
B 3 3.20 ± 0.02j 0.10 ± 0.01v 4.00 ± 0.03fg 4.80 ± 0.10ab 0.05 ± 0.01v 2.43 ± 0.53E
B 5 0.15 ± 0.00uv 0.10 ± 0.01v 4.35 ± 0.10cde 2.25 ± 0.05n 0.30 ± 0.01s-v 1.43 ± 0.45H
B 6 1.80 ± 0.02o 3.15 ± 0.07jk 0.55 ± 0.00rst 1.50 ± 0.04op 0.45 ± 0.02r-u 1.49 ± 0.26H
137
B 7 3.60 ± 0.08hi 2.50 ± 0.05mn 1.25 ± 0.03pq 1.00 ± 0.02q 0.25 ± 0.02tuv 1.72 ± 0.32G
M 1 4.95 ± 0.12a 4.60 ± 0.05bc 4.20 ± 0.05def 4.65 ± 0.11abc 1.00 ± 0.03q 3.88 ± 0.39A
M 3 4.95 ± 0.05a 2.85 ± 0.06kl 2.25 ± 0.03n 4.95 ± 0.13a 3.30 ± 0.11ij 3.66 ± 0.30B
M 5 4.55 ± 0.12bc 1.00 ± 0.02q 3.75 ± 0.05gh 2.65 ± 0.08lm 2.20 ± 0.08n 2.83 ± 0.33C
M 6 0.30 ± 0.01s-v 4.05 ± 0.11efg 4.75 ± 0.05ab 0.65 ± 0.03r 1.65 ± 0.03o 2.28 ± 0.48F
M 7 0.60 ± 0.02rs 2.80 ± 0.05lm 0.50 ± 0.01rst 0.10 ± 0.01v 0.05 ± 0.01v 0.81 ± 0.27I B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
138
Table - 74: Means comparison for yield of pectin from sapodilla fruit peel after using 1N HCl (Mechanical procedure pH
interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 3.25 ± 0.76ghi 3.63 ± 0.24fg 0.05 ± 0.00n 4.25 ± 0.56cd 2.90 ± 0.12hij 2.82 ± 0.33B
3 3.88 ± 0.66def 5.20 ± 0.19b 0.83 ± 0.35lm 4.55 ± 0.39c 6.10 ± 0.23a 4.11 ± 0.37A
5 3.28 ± 0.71gh 3.58 ± 1.13fg 4.13 ± 0.53cde 5.40 ± 0.59b 3.45 ± 0.52fg 3.97 ± 0.34A
6 1.05 ± 0.20l 2.50 ± 0.47j 1.83 ± 0.75k 3.78 ± 0.04ef 2.55 ± 0.19j 2.34 ± 0.24C
7 1.88 ± 0.30k 1.55 ± 0.54k 0.63 ± 0.26lm 0.48 ± 0.35mn 2.80 ± 0.14ij 1.47 ± 0.21D
Mean 2.67 ± 0.31C 3.29 ± 0.34B 1.49 ± 0.33D 3.69 ± 0.36A 3.56 ± 0.27A In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 75: Means comparison for yield of pectin from sapodilla fruit peel after using 1N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 1.49 ± 0.16f 2.31 ± 0.50e 0.65 ± 0.31g 2.92 ± 0.40d 3.05 ± 0.35d 2.08 ± 0.19B
M 3.84 ± 0.41c 4.27 ± 0.31ab 2.33 ± 0.50e 4.46 ± 0.55a 4.07 ± 0.37bc 3.79 ± 0.21A
B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 76: Means comparison for yield of pectin from sapodilla fruit peel after using 1N HCl (Mechanical procedure pH
boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 1.55 ± 0.05rst 3.10 ± 0.03i-m 0.05 ± 0.01w 3.00 ± 0.08j-n 2.65 ± 0.05l-o 2.07 ± 0.31F
139
B 3 2.40 ± 0.02m-p 5.60 ± 0.13bc 0.05 ± 0.01w 3.70 ± 0.10hij 5.65 ± 0.15bc 3.48 ± 0.56C
B 5 1.70 ± 0.03p-s 1.05 ± 0.02s-v 2.95 ± 0.04k-n 4.10 ± 0.09gh 2.30 ± 0.03n-q 2.42 ± 0.28E
B 6 0.60 ± 0.01uvw 1.45 ± 0.03rst 0.15 ± 0.01w 3.75 ± 0.05hi 2.15 ± 0.07o-r 1.62 ± 0.34G
B 7 1.20 ± 0.01stu 0.35 ± 0.01vw 0.05 ± 0.00w 0.05 ± 0.01w 2.50 ± 0.10l-o 0.83 ± 0.25H
M 1 4.95 ± 0.12cde 4.15 ± 0.10fgh 0.05 ± 0.00w 5.50 ± 0.10bcd 3.15 ± 0.08i-l 3.56 ± 0.52C
M 3 5.35 ± 0.12cd 4.80 ± 0.08d-g 1.60 ± 0.05q-t 5.40 ± 0.10bcd 6.55 ± 0.20a 4.74 ± 0.45B
M 5 4.85 ± 0.13def 6.10 ± 0.12ab 5.30 ± 0.10cde 6.70 ± 0.18a 4.60 ± 0.12efg 5.51 ± 0.22A
M 6 1.50 ± 0.03rst 3.55 ± 0.09h-k 3.50 ± 0.09h-k 3.80 ± 0.08hi 2.95 ± 0.10k-n 3.06 ± 0.22D
M 7 2.55 ± 0.09l-o 2.75 ± 0.05l-o 1.20 ± 0.03stu 0.92 ± 0.66tuv 3.10 ± 0.06i-m 2.10 ± 0.26F
B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
140
Table - 77: Identification tests for the presence of pectin from three selected fruits
Test Name Banana Sapodilla Muskmelon
Stiff gel test - + -
Test with 95% Ethanol + + Not clear
Test with Potassium Hydroxide (KOH) + + +
Iodine test + + +
Table - 78: Biochemical Characterization of the purified pectin extracted from
sapodilla pectin at different pH
Test Name Sapodilla
( pH5)
Sapodilla
(pH3)
Sapodilla
(pH1)
FGP*
Quantitative test for ammonia. No
ammonia
No
ammonia
No
ammonia
No ammonia
Moisture (%) 5.29 6.02 5.52 7.02
Ash (%) 5.11 4.3 4.89 1.16
Equivalent weight 1700 2680 2290 1271
Methoxyl Content (%) 5.1 4.4 4.9 8.16
Anhydrouronic Acid. (%) 39.33 31.78 34.56 70.50
Jelly Grade 100 99 98 150
Galacturonic Acid Content (%) 77.7 65.8 60.5 76.19
Protien 1.71 3.36 4.04
Degree of esterificatoion (%) 73.63 72.1 70.2 76.41
*FGP = Food grade pectin
Table - 79: Water holding, water binding and fat binding capacity of sapodilla
pectin
141
Test Name Sapodilla peel
pectin
Apple
pectin
Orange
pectin
Fibers
Apple Orange Grape
Fruit
WBC g/g 7.6 - - 1.62 1.65 2.09
WHC g/g 6.99 16.51 28.07 - - -
FBC g/g 2.2 - - 0.95 1.81 1.52
Table – 80: Showing FTIR spectral values of sample and standard pectin along with
associated functional groups.
Functional groups FOOD
GRADE
Sapodilla
(pH5)
Sapodilla
(pH3)
Sapodilla
(pH1)
O-H stretching 3319.2 3287.3 3271.1 3289.2
C-H stretching , symmetric,
asymmetric 2932.4 2923.6 2929.6 2930.2
C=O esterified 1733.3 1735.6 1738.6 1737.8
COO- asymmetric stretching
1622.2 1611.2 1612.2
COO- symmetric stretching 1426.4 1424.3 1423.3 1425.8
C-H bending 1335.4 1362.0 1363.0
C=O stretching 1234.9 1234.1 1234.5 1228.6
142
Figure – 23: FTIR spectra of food grade pectin
Figure – 24: FTIR spectra of sapodilla peel pectin extracted at pH 5
Figure – 25: FTIR spectra of sapodilla peel pectin extracted at pH 3
403.
3
467.
1
523.
2
679.
9
858.
2
909.
2
995.
2
1052
.6
1234
.9
1273
.913
35.4
1426
.4
1733
.3
1996
.0
2068
.9
2349
.0
2932
.4
3319
.2
3554
.1
*Pectin f ood grade
62
64
66
68
70
72
74
76
78
80
82
84
86
88
90
92
94
96
98
100
102
%T
500 1000 1500 2000 2500 3000 3500 4000
Wavenumbers (cm-1)
143
Figure – 26: FTIR spectra of sapodilla peel pectin extracted at pH 1
DLS studies
Table – 81: Summary the physical characterization of standard pectin and the
different pectin extracted from various source and at different pH by DLS.
144
Sample Extraction at
pH
Time of
extraction RH (nm)
PD (%)
(Đ)
Standard -- -- 73.7-1979.92 8.6-37.9
Apple 5.0 10 94.37-1357 18.4-47.8
Orange 5.0 10 473.07-482.92 36.8-36.9
Sapodilla 3.0 10 464.37-548.53 29.3-30.1
Sapodilla 5.0 10 390.21-421.17 28.1-29.3
RH= Radius of hydration ; PD = polydispersity
145
Standard
Figure – 27:.Physical characterization of standard pecti.by DLS. (A) DLS results of
sstandard pectin.illustrating the experimental conditions i.e., the mean autocorrelation
function (a), monodispersity and radius plot (b-c), respectively. (B) Comparative
corresponding radius distribution of pectin standard (a) and standardpectin (b). All
experiments were performed with an auto–piloted run of 50 measurements at every 20 s,
with a wait time 1 s (at 25 °C).
Figure- 28:.Physical characterization of apple pectin extracted at pH5.0. by DLS. (A)
DLS results of apple pectin extracted at pH5.0. illustrating the experimental conditions
i.e., the mean autocorrelation function (a), monodispersity and radius plot (b-c),
respectively. (B) Comparative corresponding radius distribution of pectin standard (a)
and apple pectin extracted at pH5.0 (b). All experiments were performed with an auto–
piloted run of 50 measurements at every 20 s, with a wait time 1 s (at 25 °C).
146
Figure – 29: Physical characterization of Orange pectin extracted at pH5.0. by DLS. (A)
DLS results of Orange pectin extracted at pH5.0. illustrating the experimental conditions
i.e., the mean autocorrelation function (a), monodispersity and radius plot (b-c),
respectively. (B) Comparative corresponding radius distribution of pectin standard (a)
and Orange pectin extracted at pH5.0 (b). All experiments were performed with an auto–
piloted run of 50 measurements at every 20 s, with a wait time 1 s (at 25 °C).
147
Figure – 30: Physical characterization of sapodilla pectin extracted at pH3.0. by DLS.
(A) DLS results of sapodilla pectin extracted at pH3.0. illustrating the experimental
conditions i.e., the mean autocorrelation function (a), monodispersity and radius plot (b-
c), respectively. (B) Comparative corresponding radius distribution of pectin standard (a)
and sapodilla pectin extracted at pH3.0 (b). All experiments were performed with an
auto–piloted run of 50 measurements at every 20 s, with a wait time 1 s (at 25 °C).
Figure – 31: Physical characterization of sapodilla pectin extracted at pH5.0. by DLS.
(A) DLS results of sapodilla pectin extracted at pH5.0. illustrating the experimental
conditions i.e., the mean autocorrelation function (a), monodispersity and radius plot (b-
c), respectively. (B) Comparative corresponding radius distribution of pectin standard (a)
and sapodilla pectin extracted at pH5.0 (b). All experiments were performed with an
148
auto–piloted run of 50 measurements at every 20 s, with a wait time 1 s (at 25 °C). Also
see Table 5 for details.
Response surface methodology
Table - 82: Box-Bechen experimental design and levels of factors used for
optimization of pectin yield
Variables Symbol Low High
pH X1 1 5
Temperature(oC) X2 50 100
Time ( minutes) X3 10 90
Table - 83: Box-Behnken experimental design and corresponding results for
responses
Std Order Run Order Pt Type Blocks pH Temperature Time Yield
10 1 2 1 3 100 10 1.6
3 2 2 1 1 100 50 1.7
14 3 0 1 3 75 50 1.5
1 4 2 1 1 50 50 1.6
2 5 2 1 5 50 50 3.5
6 6 2 1 5 75 10 2.6
9 7 2 1 3 50 10 1.5
12 8 2 1 3 100 90 1.65
4 9 2 1 5 100 50 2.5
8 10 2 1 5 75 90 3.5
11 11 2 1 3 50 90 1.4
13 12 0 1 3 75 50 2
5 13 2 1 1 75 10 3
15 14 0 1 3 75 50 1.8
149
7 15 2 1 1 75 90 1.5
Table - 84: Analysis of Variance for Yield ( Response surface methodology)
Source DF Adj SS Adj MS F-value Prob.
Model
Linear
pH
Temperature
Time
Square
pH*pH
Temperature*Temperature
Time*Time
2-Way Interaction
pH*Temperature
pH*Time
Temperature*Time
Error
Lack-of-Fit
Pure Error
Total
9
3
1
1
1
3
1
1
1
3
1
1
1
5
3
2
14
7.15996
2.40188
2.31125
0.03781
0.05281
3.00996
2.57694
0.28348
0.00848
1.74813
0.30250
1.44000
0.00563
0.51104
0.38438
0.12667
7.67100
0.79555
0.80063
2.31125
0.03781
0.05281
1.00332
2.57694
0.28348
0.00848
0.58271
0.30250
1.44000
0.00563
0.10221
0.12813
0.06333
7.78*
7.83*
22.61**
0.37
0.52
9.82*
25.21**
2.77
0.08
5.70*
2.96
14.09*
0.06
2.02
0.018
0.025
0.005
0.570
0.504
0.015
0.004
0.157
0.785
0.045
0.146
0.013
0.824
0.348
* = Significant (P<0.05); ** = Highly significant (P<0.01)
S = 0.3197
R² = 93.34%
R²(adj) = 81.35%
R²(pred) = 16.11%
150
Table - 85: Estimated Regression Coefficients for Yield (Box-Bechen experimental
design, response surface methodology)
Term Effect Coef. SE(Coef.) t-value Prob. VIF
Constant
pH
Temperature
Time
pH*pH
Temperature*Temperature
Time*Time
pH*Temperature
pH*Time
Temperature*Time
1.075
-0.137
-0.163
1.671
-0.554
0.096
-0.550
1.200
0.075
1.767
0.538
-0.069
-0.081
0.835
-0.277
0.048
-0.275
0.600
0.038
0.185
0.113
0.113
0.113
0.166
0.166
0.166
0.160
0.160
0.160
9.57**
4.76**
-0.61
-0.72
5.02**
-1.67
0.29
-1.72
3.75*
0.23
0.000
0.005
0.570
0.504
0.004
0.157
0.785
0.146
0.013
0.824
1.00
1.00
1.00
1.01
1.01
1.01
1.00
1.00
1.00
* = Significant (P<0.05); ** = Highly significant (P<0.01)
Yield = 0.76 - 0.947 pH + 0.0784 Temperature - 0.0303 Time + 0.2089 pH² -
0.000443 Temperature² + 0.000030 Time² - 0.00550 pH*Temperature
+ 0.00750 pH*Time + 0.000037 Temperature*Time
Table - 86: Predicted values of yield. (Box-Bechen experimental design, response
surface methodology)
Yield Composite
Solution pH Temperature Time Fit Desirability
1 5 61.1111 90 3.79087 1.00000
2 5 85.2577 86.8718 3.48497 0.99284
3 5 67.7177 17.6639 2.83699 0.68428
151
4 5 67.7177 17.6639 2.83699 0.68428
5 1 90.6592 10 2.79074 0.66226
Figure – 32: Showing the optimal conditions for the extraction of pectin from
sapodilla fruit peel (Box-Bechen experimental design, response surface
methodology)
152
Figure – 33: Response surface graph and contour plot of effect of pH and
temperature on yield of pectin at constant time
Yield = -0.6969-0.5663*x+0.0811*y+0.2079*x*x-0.0055*x*y-0.0004*y*y
> 3.5 < 3.5 < 3 < 2.5 < 2 < 1.5
Yield = Distance Weighted Least Squares
> 4 < 4 < 3.5 < 3 < 2.5 < 2 < 1.5 < 1 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5
pH
40
50
60
70
80
90
100
110
Tem
pe
ratu
re
153
Figure- 34: Response surface graph and contour plot of effect of pH and time on
yield of pectin at constant temperature
Yield = 4.0523-1.3913*x-0.0289*y+0.2142*x*x+0.0075*x*y+4.3269E-5*y*y
> 4.5 < 4.5 < 4 < 3.5 < 3 < 2.5 < 2 < 1.5
Yield = Distance Weighted Least Squares
> 4 < 4 < 3.5 < 3 < 2.5 < 2 < 1.5 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5
pH
0
10
20
30
40
50
60
70
80
90
100
Tim
e
154
Figure – 35: Response surface graph and contour plot of effect of temperature and
time on yield of pectin at constant pH
Yield = -0.3684+0.0773*x-0.0038*y-0.0005*x*x+3.75E-5*x*y-1.0216E-5*y*y
> 2.2 < 2.2 < 2 < 1.8 < 1.6
Yield = Distance Weighted Least Squares
> 2.5 < 2.5 < 2 < 1.5 < 1 < 0.5 40 50 60 70 80 90 100 110
Temperature
0
10
20
30
40
50
60
70
80
90
100
Tim
e
155
Table – 87: Flow properties of granules made for paracetamol tablets
Test
Formulatio
n
Mass
(g)
Bulk
Volume
(ml)
Tapped
Volume
(ml)
Bulk
Density
(g/ml)
Tapped
Density
(g/ml)
Angle of
Repose
(θ⁻¹)
Compressibilit
y Index
(%)
Hausner
Ratio
-
Standard 2.004±0.00
1
4.5068±0.11
5
4.123±0.12
5
0.446±0.05
1
0.487±0.05
7
17.955±1.15
8 8.41±0.507
1.091±0.00
6
F1 2.010±0.00
5 3.867±0.115
3.067±0.11
5
0.520±0.01
4
0.656±0.02
3
18.747±0.54
3 20.702±0.608
1.261±0.01
0
F2 2.003±0.00
1 3.867±0.115
3.067±0.11
5
0.518±0.01
5
0.654±0.02
4
16.494±1.68
0 20.702±0.608
1.261±0.01
0
F3 2.008±0.00
1 4.067±0.115
3.067±0.11
5
0.494±0.01
4
0.655±0.02
4
16.558±1.79
2 24.603±0.687
1.326±0.01
2
F4 2.008±0.00
1 4.333±0.115
4.133±0.11
5
0.464±0.01
2
0.486±0.01
4
17.545±1.54
0 4.618±0.125
1.048±0.00
1
F5 2.004±0.00
1 3.533±0.306
3.333±0.30
6
0.570±0.05
1
0.605±0.05
7
17.955±1.15
8 5.690±0.507
1.060±0.00
6
F6 2.005±0.00
1 4.133±0.115
3.667±0.46
2
0.485±0.01
4
0.552±0.06
5
17.453±0.17
1 11.032±13.884
1.141±0.16
3
F7 2.016±0.02
1 4.000±0.200
3.333±0.30
6
0.505±0.02
1
0.608±0.05
6
17.987±0.61
8 16.700±5.888
1.204±0.08
2
F8 2.020±0.01 4.067±0.115 3.067±0.11 0.497±0.01 0.659±0.02 21.546±0.44 24.603±0.687 1.326±0.01
156
0 5 4 5 1 2
F9 2.017±0.00
6 4.133±0.115
3.333±0.11
5
0.488±0.01
2
0.605±0.02
0
16.911±0.38
3 19.365±0.550
1.240±0.00
8
F1-F9 = test formulations 1 till 9 Each value is a Mean±SD of three determination
Table - 88: Pharmaceutical characteristics of compressed formulation of paracetamol tablet
Test Formulation
Pharmacopoeial Limits
(USP 32/NF 27)
Wt. Variation
(Mean ± S.D)
(mg)
±5%
Thickness
(Mean ±
S.D)
(mm)
±5%
Diameter
(Mean ± S.D)
(mm)
₋
Hardness
(Mean ± S.D)
(kg)
At least 5 kg
Loss on drying
%
Not more than 1.5%
Standard 702.67± 2.52 5.60±0.20 9.47±0.05 5.22±0.01 4.0%
F1 705.00±5.00 5.22±0.03 9.42±0.03 2.54±0.06 4.0%
F2 672.33±2.52 5.24±0.05 9.39±0.01 1.37±0.07 3.0%
F3 701.33±3.21 5.32±0.05 9.45±0.04 4.16±0.08 4.3%
F4 695±5.00 5.35±0.06 9.44±0.05 5.06±0.21 4.2%
F5 705±4.51 5.21±0.03 9.41±0.03 5.15±0.07 4.6%
F6 701.67±3.97 5.19±0.04 9.42±0.03 5.21±0.07 4.3%
F7 708.3±2.89 5.29±0.04 9.31±0.03 6.07±0.12 4.2%
F8 694.67±4.51 5.27±0.13 9.27±0.06 6.67±0.58 4.5%
157
F9 704.00±3.61 5.27±0.10 9.42±0.04 7.40±0.33 4.5%
F1-F9 = test formulations 1 till 9 Each value is a Mean±SD of three determination.
Table – 89: Dissolution studies of paracetamol tablet
FORMULATION NUMBER
F1 F2 F3 F4 F5 F6 F7 F8 F9
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
0.304 43.43 0.339 50.64 0.401 58.77 0.269 71.52 0.564 84.17 0.078 23.00 0.084 26.77 0.123 36.73 0.111 32.73
0.291 41.67 0.338 50.49 0.412 60.39 0.283 70.28 0.557 83.12 0.087 25.65 0.080 23.59 0.111 32.73 0.094 27.97
0.362 51.83 0.564 84.25 0.382 55.99 0.331 83.13 0.418 91.93 0.112 33.03 0.093 27.42 0.108 31.85 0.108 31.85
0.372 53.26 0.557 83.20 0.389 57.02 0.333 87.28 0.620 92.53 0.105 30.96 0.096 28.31 0.120 35.39 0.101 29.78
0.313 44.82 0.418 62.44 0.577 84.58 0.285 85.20 0.406 93.13 0.096 28.313 0.076 22.41 0.120 35.39 0.099 29.19
0.323 46.24 0.406 60.65 0.590 86.48 0.312 85.20 0.632 94.32 0.116 34.21 0.091 26.83 0.107 31.55 0.088 25.95
STANDARD FOR FORMULATION
0.692 99.10 0.664 99.10 0.676 99.10 0.478 99.10 0.664 99.10 0.336 99.10 0.336 99.10 0.336 99.10 0.336 99.10
F1-F9 are test formulation while Abs = absorbance and %release = percent release
158
159
F1 F2
F3
Figure -36: Formulated paracetamol tablest ( F1, F2, F3)
160
F4 F5
F6
Figure 37: Formulated paracetamol tablest ( F4, F5, F6)
161
F7 F8
F9
Figure 38: Formulated paracetamol tablest (F7, F8, F9)
162
Table - 90: Flow properties ofgranules for ibuprofen tablet
Test
Formulation
Mass
Bulk
Volume
Tapped
Volume
Bulk
Density
Tapped
Density
Angle of
Repose
Compressibility
Index
Hausner
Ratio
(g) (ml) (ml) (g/ml) (g/ml) (θ⁻¹) (%) ₋
Std 2.02±0.013 4.25±0.015 3.75±0.092 0.475±0.002 0.538±0.020 11.71±0.210 11.71±0.116 1.132±0.061
R1 2.02±0.013 4.20±0.015 3.76±0.093 0.49±0.003 0.55±0.019 15.23±0.208 4.82±0.115 1.13±0.061
R2 2.01±0.016 4.35±0.042 3.61±0.010 0.46±0.005 0.56±0.004 13.83±0.153 9.74±0.344 1.24±0.059
R3 2.01±0.007 4.60±0.080 3.93±0.064 0.42±0.021 0.52±0.010 13.73±0.306 7.40±0.352 1.17±0.028
R4 2.01±0.004 4.51±0.090 4.15±0.050 0.44±0.002 0.47±0.003 11.13±0.153 3.77±0.252 1.09±0.009
R1= Test formulation one, R2= Test formulation two, R3= Test formulation three, R4= Test formulation four. Each value is a Mean±SD of three determination
Table - 91: Pharmaceutical characteristics of compressed formulation of ibuprofen tablet
Test Formulation
Pharmacopoeial Limits
(USP 32/NF 27)
Wt. Variation
(Mean ± S.D)
(mg) ±5%
Thickness
(Mean ± S.D)
(mm) ±5%
Length x width
(Mean ± S.D)
(mm) ₋
Hardness
(Mean ± S.D)
(kg) At least 5 kg
Loss on
Drying %
Standard 833.33±2.08 6.33± 0.06 20 x 9.5 6.13± 0.14 4.5%
R1 843.33±2.08 6.23± 0.06 20 x 9.5 6.23± 0.14 4.9%
R2 848.33±2.89 6.37±0.06 20 x 9.5 8.40±0.40 4.8%
R3 847.67±2.52 6.22±0.03 20 x 9.5 9.23±0.14 4.9%
R4 842.67±2.52 6.27±0.06 20 x 9.5 10.43±0.18 4.7%
163
Table- 92: dissolution studies of ibuprufen tablets
FORMULATION NUMBER
R1 R2 R3 R4
243nm (Abs) % release 243nm (Abs) % release 243nm (Abs) % release 243nm (Abs) % release
0.405 89.6 0.166 36.72 0.092 33.45 -0.018 -6.54
0.415 91.81 0.141 31.190 0.090 32.72 -0.017 -6.182
0.420 92.91 0.147 32.51 0.112 40.72 -0.020 -7.27
0.425 94.02 0.154 34.06 0.113 41.92 0.008 -2.90
0.429 94.95 0.143 31.62 0.030 10.90 -0.008 -2.90
0.430 95.13 0.161 35.60 0.036 13.091 -0.008 -2.90
STANDARD FOR FORMULATION
0.503 111.33 0.503 111.33 0.306 111.33 0.306 111.33
R1-R4 are test formulation while Abs = absorbance and %release = percent release
164
R1 R2
R3 R4
Figure – 39: Formulated ipubrufen tablets ( R1, R2, R3, R4)
91
Table -93: Basic evaluation test of antidiarrheal formulation prepared from
sapodilla pectin
Parameters Suspension made from
sapodilla pectin Comparative suspension *
Color Pinkish white White
Odor vanilla Vanilla
Taste sweet sweet
pH 6.1 5.56
Viscosity 14.14 13.13
Sedimentation rate 0.3 0.1
Redispersity +++ +++
WHC 32.69 33.56
+ denotes the number of times the cylinder was moved. * Keptin antidiarrheal preparation
92
Figure- 40 Formulated antidiarrheal preparation
93
Table - 94: Effect of different concentration of sapodilla pectin on the chemical
properties of the jam samples
Sample PH
TSS
(mg)
TTA
(ml)
MC
(%)
Vit C
(mg)
Ash
(%)
TS
(mg)
Viscosity
(Cp)
S 3.06 61.9 1.14 35.9 18.3 1.78 64.1 71
F1 3.16 56.7 1.08 41 17.6 1.69 59 56
F2 3.25 62.6 1.01 35.7 18.1 1.73 64.3 63
F3 3.25 62.4 1.03 35.1 17.9 1.71 64.9 67
WhereS = Jam with standard pectin, F1 = Jam with 2g pectin, F2 = Jam with pectin in same concentration
as in standard, F3 = Jam with 10g pectin while MC is moisture content . TSS total soluble solids, TTA
total titratable acid and TS is total solid.
Table - 95: Scores for sensory parameters as judged by twenty (20) panelists.
Sample Appearance Taste Aroma Spreadability Texture Mouth
feel
Overall
acceptability
S 7.7 7.9 7.9 8.2 7.8 7.5 7.8
F1 7.5 7.3 7.2 7.3 7.2 7 7.2
F2 7.5 7.6 7.7 7.8 7.8 7.5 7.6
F3 6.2 6.4 6.2 5.6 6.3 5.5 6
WhereS = Jam with standard pectin, F1 = Jam with 2g pectin, F2 = Jam with pectin in same concentration
as in standard, F3 = Jam with 10g pectin
(S)=Jam made from 5g standard pectin (F1) Jam made from 5g pectin
94
(F2) Jam made from 7 g pectin (F3) Jam made from 10g pectin
Figure -41 formulation of Jam made from extracted pectin
Figure -42: formulation of pudding made from extracted pectin.
95
Table - 9: List of fruits used in the screening study for the presence of pectin
(a) is Munarin et al., 2012; (b) is Bhat and Singh, 2014; (c) is Baker, 1997; (d) is Baissise
et al., 2010. (e) is Rasheed, 2008; (f) is Aina et al., 2012.
S no
Fruit
%Yield Reported
amount Common name Parts
used
1 Orange peel 20 6-26% a
2 Apple exocarp 6 2-19%a
3 Grape Fruit peel 3.5 13-32%a
4 Sapodilla peel 5.5 Notfound
5 Muskmelon peel 2 Not found
6 Guava (unripe) exocarp 0 Not found
7 Guava(ripe) exocarp 4 16.8% b
8 Chinese apple (Jungle Fruit) exocarp 0 Notfound
9 Strawberry exocarp 0 0.35–0.44%.C
10 Sweet lime peel 3 6-26%a
11 Peach exocarp 0 4-18%a
12 Apricot exocarp 0 4.97%d
13 Canteloup peel 3 Not found
14 Plum exocarp 0 Not found
15 Watermelon peel 2.35 15.70% e
16 Mango(Unripe) peel 0 Not found
17 Mango (ripe) peel 3.5 9-29%a
18 Banana peel 3.5 2-15%a
19 Lemon peel 3.5 2.7% f
96
Table - 10: Showing percent yield of pectin from five selected fruits
Mechanical
procedure
p
H
Sapodilla Banana Muskmelo
n Apple Orange
B M B M B M B M B M
Homogenizin
g
1 0.5 1.0
5
3.2
5 5.6 0.55 0.55 0.5 1
16.1
5
15.2
5
3 0.7
5 1.6
6.8
5
4.3
5 2 1
1.9
5
2.7
5 18.2 19.8
5 1.2 1.5 8.8
5 8.4 1.45 2.3
2.3
5 3.7 19.4 22.7
6 1.5 2 5.8
5
9.7
5 1.15 1 1.5 1.4
13.9
5 2.85
7 0.5 0.5 5.4
5 3.1 0.55 1.8
0.0
5
0.3
5 21.7 7.5
Grinding
1 0.5
5 0.8
2.4
5 3.2 2.25 2.65
2.4
5 2.5 2.6 3.95
3 4 3.5 4.2 6.9 1.4 1.8 0.4
5
1.1
5 7.8 5.3
5 2.4
5 2.9 3.5 4.5 1.95 2.4 1 1.1 2.3 7.1
6 2 2 3.6
5
2.6
5 0.95 1.6 2.4
2.8
5 6.45 6.6
7 1.9
5 2.3
1.2
5
0.2
5 0.55 1
0.0
5
0.5
5 0.6 0.2
Cutting
1 0.8 0.9 0 2.5 0.5 0.5 2.8
5 3.9 8.1 7
3 1.1 1.5 1.9
5 1.2 2.45 2.55
2.9
5 4.5 8.5 12
5 0.4 1 5 5.5 1 1 2.7 3.7 11.8
5
20.6
5
6 1 1.2 1 5.4 0.7 0.95 1.1
5 2.9 6.95 10.2
7 0.7 1 3 6.5 1.3 2.2 0.0
5 0.7 4 7.4
Chopping
1 2.7 4.7 1.6
5 2.1 0.05 0.7
2.8
5
3.3
5 2.6 3.35
3 3.9 3.4
5 4 3.3 0.05 0.5
1.2
5
2.7
5 7.95
10.0
5
5 3.9
5
4.0
5
5.9
5 6.6 1.7 2.1
0.0
5
1.7
5 10.4 15.8
6 2.3 2.4
5
5.8
5
6.3
5 2.25 1.75 0.1 0.9
15.2
5
17.1
5
7 3.9 2.4 1.8 1.6 0.5 1.05 0.2 0.6 5.35 9.35
97
5 5 5
Hammering
1 1.5 2.0
5 4.5 7 0.65 1.25 1.5 1.7
13.5
5
16.6
5
3 0.6 1.5
5 5.5 5.5 0.5 0.8 2.7 3.5 7.2 8.35
5 0.8 1.9
5 8.5
10.
5 0.65 0.3
3.0
5
4.8
5 7.4
12.1
5
6 0.8 1.4 5 5.6 0.95 0.5 1.2
5
2.0
5 5.55 10.7
7 0.5 0 0.5 0.5 0.45 0.05 1.7 2.0
5 5.5 8.85
B= Boiling on burner, M= heating in microwave
98
Table - 11: Analysis of variance (mean squares) of yield for five selected fruits
Source of variation Degrees of
freedom
Mean squares
Sapodilla Banana Muskmelon Apple Orange
Mech. procedure (MP)
pH level
Boiling method (BM)
MP pH
MP BM
pH BM
MP pH x BM
Error
Total
4
4
1
16
4
4
16
100
149
32.6575**
3.3917**
3.2413**
2.5462**
0.3392**
1.0112**
0.6258**
0.0266
51.245**
84.818**
22.349**
16.774**
4.640**
6.332**
4.698**
0.191
4.3869**
1.5751**
2.0184**
2.7688**
0.2814**
0.4112**
0.4141**
0.0112
9.5920**
17.3200**
22.6981**
5.6967**
0.8120**
0.8975**
0.2720**
0.0314
493.729**
142.586**
59.914**
88.941**
81.797**
44.377**
21.750**
1.0100
NS = Non-significant (P>0.05); * = Significant (P<0.05); ** = Highly significant (P<0.01) , Mech = mechanical
99
Table – 12: Means comparison of yield for sapodilla fruit peel (Mechanical procedure pH interaction mean±SE )
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 0.78 ± 0.13ij 0.68 ± 0.06j 0.85 ± 0.03hij 3.70 ± 0.47a 1.78 ± 0.14ef 1.56 ± 0.23C
3 1.18 ± 0.19gh 3.75 ± 0.15a 1.30 ± 0.09g 3.68 ± 0.13a 1.08 ± 0.21ghi 2.20 ± 0.24A
5 1.35 ± 0.08g 2.68 ± 0.11c 0.70 ± 0.14j 4.00 ± 0.12a 1.38 ± 0.26g 2.02 ± 0.23B
6 1.75 ± 0.12f 2.00 ± 0.06ef 1.10 ± 0.05ghi 2.38 ± 0.10cd 1.10 ± 0.14ghi 1.67 ± 0.10C
7 0.50 ± 0.01jk 2.13 ± 0.10de 0.85 ± 0.07hij 3.18 ± 0.35b 0.25 ± 0.11k 1.38 ± 0.22D
Mean 1.11 ± 0.10C 2.25 ± 0.19B 0.96 ± 0.05D 3.39 ± 0.16A 1.12 ± 0.12C
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 13: Means comparison of yield for sapodilla fruit peel (Mechanical procedure boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 0.89 ± 0.11e 2.19 ± 0.30b 0.80 ± 0.07e 3.36 ± 0.20a 0.84 ± 0.09e 1.62 ± 0.14B
M 1.33 ± 0.14c 2.30 ± 0.25b 1.12 ± 0.06d 3.41 ± 0.25a 1.39 ± 0.20c 1.91 ± 0.13A
B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean.
Table - 14: Means comparison of yield for sapodilla fruit peel (Mechanical procedure pH boiling method
interaction mean ± SE)
BM pH Mechanical procedure BM pH
100
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 0.50 ± 0.03qrs 0.55 ± 0.02pqr 0.80 ± 0.04n-r 2.70 ± 0.14ef 1.50 ± 0.06i-l 1.21 ± 0.22E
B 3 0.75 ± 0.03n-r 4.00 ± 0.15bc 1.10 ± 0.03j-o 3.90 ± 0.14bcd 0.60 ± 0.03o-r 2.07 ± 0.41B
B 5 1.20 ± 0.07j-n 2.45 ± 0.10efg 0.40 ± 0.02rs 3.95 ± 0.22bcd 0.80 ± 0.03n-r 1.76 ± 0.35C
B 6 1.50 ± 0.07i-l 2.00 ± 0.10ghi 1.00 ± 0.04l-q 2.30 ± 0.13fg 0.80 ± 0.03n-r 1.52 ± 0.16D
B 7 0.50 ± 0.02qrs 1.95 ± 0.08ghi 0.70 ± 0.03n-r 3.95 ± 0.14bcd 0.50 ± 0.01qrs 1.52 ± 0.36D
M 1 1.05 ± 0.06k-p 0.80 ± 0.03n-r 0.90 ± 0.03m-r 4.70 ± 0.26a 2.05 ± 0.14gh 1.90 ± 0.40BC
M 3 1.60 ± 0.08hij 3.50 ± 0.16cd 1.50 ± 0.06i-l 3.45 ± 0.14d 1.55 ± 0.05h-k 2.32 ± 0.26A
M 5 1.50 ± 0.04i-l 2.90 ± 0.06e 1.00 ± 0.03l-q 4.05 ± 0.14b 1.95 ± 0.11ghi 2.28 ± 0.29A
M 6 2.00 ± 0.09ghi 2.00 ± 0.08ghi 1.20 ± 0.04j-n 2.45 ± 0.16efg 1.40 ± 0.02j-m 1.81 ± 0.13C
M 7 0.50 ± 0.02qrs 2.30 ± 0.12fg 1.00 ± 0.04l-q 2.40 ± 0.08efg 0.00 ± 0.00s 1.24 ± 0.26E
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
.
101
Table – 15: Means comparison of yield for banana fruit peel (Mechanical procedure pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 4.43 ± 0.55fgh 2.83 ± 0.19j 1.25 ± 0.56klm 1.88 ± 0.12k 5.75 ± 0.60cd 3.23 ± 0.36D
3 5.60 ± 0.57cde 5.55 ± 0.64cde 1.58 ± 0.17kl 3.65 ± 0.19hij 5.50 ± 0.18cde 4.38 ± 0.34C
5 8.63 ± 0.31ab 4.00 ± 0.25ghi 5.25 ± 0.21def 6.28 ± 0.27c 9.50 ± 0.53a 6.73 ± 0.41A
6 7.80 ± 0.90b 3.15 ± 0.26ij 3.20 ± 0.99ij 6.10 ± 0.22cd 5.30 ± 0.19def 5.11 ± 0.42B
7 4.28 ± 0.54gh 0.75 ± 0.23lm 4.75 ± 0.80efg 1.73 ± 0.07k 0.50 ± 0.01m 2.40 ± 0.38E
Mean 6.15 ± 0.41A 3.26 ± 0.33D 3.21 ± 0.40D 3.93 ± 0.37C 5.31 ± 0.55B
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 16: Means comparison of yield for banana fruit peel (Mechanical procedure boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 6.05 ± 0.50a 3.01 ± 0.29e 2.19 ± 0.46f 3.86 ± 0.50cd 4.80 ± 0.69b 3.98 ± 0.27B
M 6.24 ± 0.67a 3.50 ± 0.59de 4.22 ± 0.55c 3.99 ± 0.57cd 5.82 ± 0.87a 4.75 ± 0.31A
B= Boiling on burner , M= heating in microwave In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
102
Table - 17: Means comparison of yield for banana fruit peel (Mechanical procedure pH boiling method interaction mean ±
SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 3.25 ± 0.19l-q 2.45 ± 0.11o-u 0.00 ± 0.00x 1.65 ± 0.12r-w 4.50 ± 0.16h-l 2.37 ± 0.41D
B 3 6.85 ± 0.24def 4.20 ± 0.19i-m 1.95 ± 0.06q-v 4.00 ± 0.19j-n 5.50 ± 0.27e-i 4.50 ± 0.44C
B 5 8.85 ± 0.51b 3.50 ± 0.09l-p 5.00 ± 0.31g-k 5.95 ± 0.22d-h 8.50 ± 0.29b 6.36 ± 0.56B
B 6 5.85 ± 0.26d-h 3.65 ± 0.24k-o 1.00 ± 0.03u-x 5.85 ± 0.31d-h 5.00 ± 0.21g-k 4.27 ± 0.49C
B 7 5.45 ± 0.30e-j 1.25 ± 0.07t-x 3.00 ± 0.12m-s 1.85 ± 0.08q-v 0.50 ± 0.03vwx 2.41 ± 0.46D
M 1 5.60 ± 0.30d-i 3.20 ± 0.16l-q 2.50 ± 0.14o-t 2.10 ± 0.09p-u 7.00 ± 0.43cd 4.08 ± 0.52C
M 3 4.35 ± 0.12i-m 6.90 ± 0.40de 1.20 ± 0.07t-x 3.30 ± 0.15l-q 5.50 ± 0.31e-i 4.25 ± 0.53C
M 5 8.40 ± 0.43bc 4.50 ± 0.23h-l 5.50 ± 0.26e-i 6.60 ± 0.46def 10.50 ± 0.57a 7.10 ± 0.59A
M 6 9.75 ± 0.46ab 2.65 ± 0.14n-t 5.40 ± 0.27f-j 6.35 ± 0.29d-g 5.60 ± 0.21d-i 5.95 ± 0.62B
M 7 3.10 ± 0.09l-r 0.25 ± 0.01wx 6.50 ± 0.32def 1.60 ± 0.07s-w 0.50 ± 0.02vwx 2.39 ± 0.61D
B= Boiling on burner , M= heating in microwave; BM= boiling method
. In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table -18: Means comparison of yield for muskmelon fruit peel (Mechanical procedure pH interaction mean±SE)
pH level Mechanical procedure Mean
103
Homogenizing Grinding Cutting Chopping Hammering
1 0.55 ± 0.02m-p 2.45 ± 0.12a 0.50 ± 0.01n-q 0.38 ± 0.15pqr 0.95 ± 0.14ijk 0.97 ± 0.15D
3 1.50 ± 0.23fg 1.60 ± 0.10ef 2.50 ± 0.05a 0.28 ± 0.10qr 0.65 ± 0.07l-o 1.31 ± 0.15B
5 1.88 ± 0.20cd 2.18 ± 0.13b 1.00 ± 0.02ij 1.90 ± 0.10cd 0.48 ± 0.08o-r 1.49 ± 0.13A
6 1.08 ± 0.05hi 1.28 ± 0.15gh 0.83 ± 0.06jkl 2.00 ± 0.12bc 0.73 ± 0.10k-n 1.18 ± 0.09C
7 1.18 ± 0.28hi 0.78 ± 0.10j-m 1.75 ± 0.21de 0.78 ± 0.13j-m 0.25 ± 0.09r 0.95 ± 0.12D
Mean 1.24 ± 0.11C 1.66 ± 0.12A 1.32 ± 0.14B 1.07 ± 0.15D 0.61 ± 0.06E
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 19: Means comparison of yield for muskmelon fruit peel (Mechanical procedure boiling method interaction
mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 1.14 ± 0.15d 1.42 ± 0.17b 1.19 ± 0.18d 0.91 ± 0.24e 0.64 ± 0.05f 1.06 ± 0.08B
M 1.33 ± 0.17bc 1.89 ± 0.16a 1.44 ± 0.21b 1.22 ± 0.16cd 0.58 ± 0.11f 1.29 ± 0.09A
B= Boiling on burner , M= heating in microwave;
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table -20: Means comparison of yield for muskmelon fruit peel (Mechanical procedure pH boiling method interaction
mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 0.55 ± 0.03qr 2.25 ± 0.13bcd 0.50 ± 0.03qr 0.05 ± 0.00s 0.65 ± 0.03pqr 0.80 ± 0.20F
104
B 3 2.00 ± 0.04def 1.40 ± 0.05i-l 2.45 ± 0.06abc 0.05 ± 0.00s 0.50 ± 0.02qr 1.28 ± 0.24BCD
B 5 1.45 ± 0.05h-k 1.95 ± 0.10d-g 1.00 ± 0.04m-p 1.70 ± 0.05f-i 0.65 ± 0.03pqr 1.35 ± 0.13B
B 6 1.15 ± 0.06k-n 0.95 ± 0.03m-p 0.70 ± 0.05opq 2.25 ± 0.10bcd 0.95 ± 0.04m-p 1.20 ± 0.15DE
B 7 0.55 ± 0.02qr 0.55 ± 0.01qr 1.30 ± 0.07j-m 0.50 ± 0.03qr 0.45 ± 0.02qr 0.67 ± 0.09G
M 1 0.55 ± 0.03qr 2.65 ± 0.12a 0.50 ± 0.02qr 0.70 ± 0.04opq 1.25 ± 0.05j-m 1.13 ± 0.22E
M 3 1.00 ± 0.03m-p 1.80 ± 0.09e-h 2.55 ± 0.09ab 0.50 ± 0.02qr 0.80 ± 0.06n-q 1.33 ± 0.20BC
M 5 2.30 ± 0.09a-d 2.40 ± 0.14abc 1.00 ± 0.03m-p 2.10 ± 0.08cde 0.30 ± 0.01rs 1.62 ± 0.22A
M 6 1.00 ± 0.05m-p 1.60 ± 0.08g-j 0.95 ± 0.05m-p 1.75 ± 0.04e-i 0.50 ± 0.02qr 1.16 ± 0.12DE
M 7 1.80 ± 0.11e-h 1.00 ± 0.04m-p 2.20 ± 0.09bcd 1.05 ± 0.07l-o 0.05 ± 0.00s 1.22 ± 0.20CDE
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
105
Table -21: Means comparison of yield for apple fruit peel (Mechanical procedure pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 0.75 ± 0.12ijk 2.48 ± 0.06d 3.38 ± 0.26bc 3.10 ± 0.15c 1.60 ± 0.05gh 2.26 ± 0.19B
3 2.35 ± 0.19de 0.80 ± 0.16ij 3.73 ± 0.36ab 2.00 ± 0.34ef 3.10 ± 0.20c 2.40 ± 0.22A
5 3.03 ± 0.31c 1.05 ± 0.04i 3.20 ± 0.24c 0.90 ± 0.38i 3.95 ± 0.42a 2.43 ± 0.26A
6 1.45 ± 0.05h 2.63 ± 0.12d 2.03 ± 0.39ef 0.50 ± 0.18jkl 1.65 ± 0.18fgh 1.65 ± 0.16C
7 0.20 ± 0.07l 0.30 ± 0.11l 0.38 ± 0.15kl 0.43 ± 0.08jkl 1.88 ± 0.10fg 0.64 ± 0.12D
Mean 1.56 ± 0.20B 1.45 ± 0.18BC 2.54 ± 0.26A 1.39 ± 0.22C 2.44 ± 0.20A
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 22: Means comparison of yield for apple fruit peel (Mechanical procedure boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 1.27 ± 0.23e 1.27 ± 0.27e 1.94 ± 0.31c 0.90 ± 0.29f 2.04 ± 0.19c 1.48 ± 0.12B
M 1.84 ± 0.33c 1.63 ± 0.24d 3.14 ± 0.36a 1.87 ± 0.28c 2.83 ± 0.32b 2.26 ± 0.15A
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 23: Means comparison of yield for apple fruit peel (Mechanical procedure pH boiling method interaction
mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
106
B 1 0.50 ± 0.02r-u 2.45 ± 0.09fgh 2.85 ± 0.16d-g 2.85 ± 0.13d-g 1.50 ± 0.05i-m 2.03 ± 0.25C
B 3 1.95 ± 0.06hij 0.45 ± 0.02r-u 2.95 ± 0.14c-f 1.25 ± 0.08k-o 2.70 ± 0.15efg 1.86 ± 0.25C
B 5 2.35 ± 0.10gh 1.00 ± 0.05m-r 2.70 ± 0.05efg 0.05 ± 0.00u 3.05 ± 0.15cde 1.83 ± 0.30C
B 6 1.50 ± 0.08i-m 2.40 ± 0.07fgh 1.15 ± 0.06l-p 0.10 ± 0.01u 1.25 ± 0.09k-o 1.28 ± 0.20D
B 7 0.05 ± 0.00u 0.05 ± 0.00u 0.05 ± 0.00u 0.25 ± 0.00tu 1.70 ± 0.09i-l 0.42 ± 0.17F
M 1 1.00 ± 0.06m-r 2.50 ± 0.10e-h 3.90 ± 0.19b 3.35 ± 0.20bcd 1.70 ± 0.04i-l 2.49 ± 0.29B
M 3 2.75 ± 0.16efg 1.15 ± 0.04l-p 4.50 ± 0.21a 2.75 ± 0.14efg 3.50 ± 0.10bc 2.93 ± 0.30A
M 5 3.70 ± 0.15b 1.10 ± 0.04m-q 3.70 ± 0.18b 1.75 ± 0.09ijk 4.85 ± 0.23a 3.02 ± 0.37A
M 6 1.40 ± 0.07j-n 2.85 ± 0.11d-g 2.90 ± 0.06d-g 0.90 ± 0.04n-s 2.05 ± 0.05hi 2.02 ± 0.21C
M 7 0.35 ± 0.02stu 0.55 ± 0.02q-u 0.70 ± 0.04o-t 0.60 ± 0.02p-u 2.05 ± 0.12hi 0.85 ± 0.16E
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
107
Table -24: Means comparison of yield for orange fruit peel (Mechanical procedure pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 15.70 ± 0.45b 3.28 ± 0.32j 7.55 ± 0.30fgh 2.98 ± 0.19j 15.10 ± 0.88bc 8.92 ± 1.05C
3 19.00 ± 0.91a 6.55 ± 0.58ghi 10.25 ± 0.81d 9.00 ± 0.54def 7.78 ± 0.33e-h 10.52 ± 0.87B
5 21.05 ± 1.02a 4.70 ± 1.09ij 16.25 ± 2.07b 13.10 ± 1.27c 9.78 ± 1.11de 12.98 ± 1.18A
6 8.40 ± 2.51d-g 6.53 ± 0.21ghi 8.58 ± 0.81d-g 16.20 ± 0.61b 8.13 ± 1.17d-g 9.57 ± 0.84C
7 14.60 ± 3.20bc 0.40 ± 0.09k 5.70 ± 0.78hi 7.35 ± 0.92fgh 7.18 ± 0.78fgh 7.05 ± 1.07D
Mean 15.75 ± 1.14A 4.29 ± 0.49C 9.67 ± 0.82B 9.73 ± 0.91B 9.59 ± 0.65B
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table -25: Means comparison of yield for orange fruit peel (Mechanical procedure boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 17.88 ± 0.78a 3.95 ± 0.73e 7.88 ± 0.69d 8.31 ± 1.17d 7.84 ± 0.80d 9.17 ± 0.65B
M 13.62 ± 2.02b 4.63 ± 0.67e 11.45 ± 1.35c 11.14 ± 1.34c 11.34 ± 0.83c 10.44 ± 0.68A
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
108
Table -26: Means comparison of yield for orange fruit peel (Mechanical procedure pH boiling method interaction
mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 16.15 ± 0.44d-g 2.60 ± 0.14uvw 8.10 ± 0.33m-r 2.60 ± 0.11uvw 13.55 ± 0.48g-j 8.60 ± 1.49DE
B 3 18.20 ± 1.36b-e 7.80 ± 0.21m-r 8.50 ± 0.20l-r 7.95 ± 0.33m-r 7.20 ± 0.41n-s 9.93 ± 1.14CD
B 5 19.40 ± 0.80a-d 2.30 ± 0.10uvw 11.85 ± 0.64i-l 10.40 ± 0.66j-n 7.40 ± 0.20m-r 10.27 ± 1.52BC
B 6 13.95 ± 0.76f-i 6.45 ± 0.40p-t 6.95 ± 0.41o-s 15.25 ± 0.72e-h 5.55 ± 0.24q-u 9.63 ± 1.12B
B 7 21.70 ± 0.72a 0.60 ± 0.03vw 4.00 ± 0.21stu 5.35 ± 0.20r-u 5.50 ± 0.18q-u 7.43 ± 1.97BC
M 1 15.25 ± 0.80e-h 3.95 ± 0.16s-v 7.00 ± 0.17o-s 3.35 ± 0.14t-w 16.65 ± 1.12c-g 9.24 ± 1.52A
M 3 19.80 ± 1.29abc 5.30 ± 0.28r-u 12.00 ± 0.46h-k 10.05 ± 0.51k-o 8.35 ± 0.22m-r 11.10 ± 1.33CD
M 5 22.70 ± 1.36a 7.10 ± 0.39n-s 20.65 ± 1.25ab 15.80 ± 0.54efg 12.15 ± 0.66h-k 15.68 ± 1.55CD
M 6 2.85 ± 0.19uvw 6.60 ± 0.22p-t 10.20 ± 0.66j-o 17.15 ± 0.67c-f 10.70 ± 0.46i-m 9.50 ± 1.28EF
M 7 7.50 ± 0.35m-r 0.20 ± 0.01w 7.40 ± 0.35m-r 9.35 ± 0.43k-p 8.85 ± 0.48k-q 6.66 ± 0.90F
Means sharing similar letter in a row or in a column are statistically non-significant (P>0.05). Small letters represent comparison among interaction means and
capital letters are used for overall mean
109
Table - 27: Percentage yield of pectin from sapodilla fruit peel after using different
physicomechanical procedures ( pH, mechanical procedure, boiling method and time of
boiling) with 0.1N HCl
Mechanical procedure
pH %yield 10 Min %yield 20min %yield 40 min %yield 60min
B M B M B M B M
Ham
mer
ing
1 1.5 2.05 0.5 1.1 0.05 0.05 1.2 1.8
3 0.6 1.55 2.2 4.85 0.05 0.1 4.1 3.15
5 0.8 1.95 5.5 6.35 0.05 0.05 4.5 4.6
6 0.8 1.4 3.5 5.2 1.65 1.65 3 3.45
7 0.5 0 2.6 4.35 0.05 0.95 1.75 3.85
Gri
nd
ing
1 0.55 0.8 0.75 1.6 0.1 1.15 0.25 0.35
3 4 3.5 3.55 4 1 1.35 0.05 0.05
5 2.45 2.9 2.05 3.8 2.4 2.75 0.05 2.1
6 2 2 3.4 2.7 2.1 3.1 0.05 0.05
7 1.95 2.3 2.85 3.3 0.95 1.75 0.05 2.3
Cu
ttin
g
1 0.8 0.9 1.45 1.5 0.05 0.05 0.05 0.1
3 1.1 1.5 1.95 3.15 0.45 0.05 0.4 1.4
5 0.4 1 1.5 2.2 1.2 3.95 0.05 0.5
6 1 1.2 1.45 1.85 0.65 1 0.1 0.9
7 0.7 1 1.35 1.8 0.1 1.5 0.05 1.05
Ho
mo
gen
izin
g
1 0.5 1.05 0.05 0.15 0.05 0.05 0.05 0.05
3 0.75 1.6 0.05 1 2.5 1.3 2.75 2.8
5 1.2 1.5 0.05 0.1 3 3.2 2.7 6
6 1.5 2 0.05 0.05 2.25 2.45 2.7 4.5
7 0.5 0.5 0.1 0.05 2.45 2.8 0.15 2.65
Ch
op
pin
g
(mort
ar/
pes
tle
1 2.7 4.7 0.05 0.05 0.05 0.05 1.85 2.75
3 3.9 3.45 4.75 6.5 1.6 1.35 1.25 3
5 3.95 4.05 2.25 3.05 2.05 2.35 1.95 5.6
6 2.3 2.45 2.5 2.8 2.7 3.6 3.25 5
7 3.95 2.4 0.1 0.7 1 2 3.6 3.65
B= Boiling on burner , M= heating in microwave
110
Table -28: Analysis of variance (mean squares) of yield of pectin from sapodilla fruit peel after using different
physicomechanical procedure different fruits with 0.1N HCl
Source of
variation
Degrees of
freedom
Mean squares
Yield 10 min Yield 20 min Yield 40 min Yield 60 min
MP
pH
BM
MP x pH
MP x BM
pH x BM
MP x pH x BM
Error
Total
4
4
1
16
4
4
16
100
149
32.6575**
3.3917**
3.2414**
2.5462**
0.3392**
1.0112**
0.6258**
0.0081
49.7412**
27.4284**
18.6772**
7.5601**
1.7538**
1.4724**
0.4729**
0.0129
12.1446**
20.1921**
6.1206**
2.3355**
1.0438**
1.6176**
0.5655**
0.0070
55.6906**
15.5938**
39.7837**
4.3065**
2.0295**
3.7639**
1.5634**
0.0122
NS = Non-significant (P>0.05); * = Significant (P<0.05); ** = Highly significant (P<0.01). Mech = Mechanical
Table – 29: Means comparison of yield for sapodilla fruit peel after 10 min of boiling using 0.1N HCl (Mechanical procedure
pH interaction mean±SE)
111
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 1.78 ± 0.13g 0.68 ± 0.06kl 0.85 ± 0.02k 3.70 ± 0.45b 0.78 ± 0.12k 1.56 ± 0.23D
3 1.08 ± 0.21j 3.75 ± 0.14b 1.30 ± 0.09hi 3.68 ± 0.11b 1.18 ± 0.19ij 2.20 ± 0.24A
5 1.38 ± 0.26h 2.68 ± 0.10d 0.70 ± 0.13k 4.00 ± 0.07a 1.35 ± 0.07hi 2.02 ± 0.23B
6 1.10 ± 0.13j 2.00 ± 0.04f 1.10 ± 0.05j 2.38 ± 0.04e 1.75 ± 0.12g 1.67 ± 0.10C
7 0.25 ± 0.11m 2.13 ± 0.08f 0.85 ± 0.07k 3.18 ± 0.35c 0.50 ± 0.01l 1.38 ± 0.22E
Mean 1.12 ± 0.12C 2.25 ± 0.19B 0.96 ± 0.05D 3.39 ± 0.15A 1.11 ± 0.09C
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 30: Means comparison of yield for sapodilla fruit peel after 10 min of boiling using 0.1N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 0.84 ± 0.09f 2.19 ± 0.30c 0.80 ± 0.07f 3.36 ± 0.19a 0.89 ± 0.11f 1.62 ± 0.14B
M 1.39 ± 0.20d 2.30 ± 0.24b 1.12 ± 0.06e 3.41 ± 0.24a 1.33 ± 0.14d 1.91 ± 0.13A B= Boiling on burner , M= heating in microwave; BM= boiling method In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table -31: Means comparison of yield for sapodilla fruit peel after 10 min of boiling using 0.1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 1.50 ± 0.02ij 0.55 ± 0.01qr 0.80 ± 0.02m-q 2.70 ± 0.05de 0.50 ± 0.01qr 1.21 ± 0.22F
B 3 0.60 ± 0.02pqr 4.00 ± 0.10b 1.10 ± 0.02klm 3.90 ± 0.09b 0.75 ± 0.02n-q 2.07 ± 0.41B
B 5 0.80 ± 0.01m-q 2.45 ± 0.03ef 0.40 ± 0.01r 3.95 ± 0.12b 1.20 ± 0.03jkl 1.76 ± 0.35D
112
B 6 0.80 ± 0.01m-q 2.00 ± 0.05gh 1.00 ± 0.02l-o 2.30 ± 0.05fg 1.50 ± 0.04ij 1.52 ± 0.15E
B 7 0.50 ± 0.02qr 1.95 ± 0.06h 0.70 ± 0.02o-r 3.95 ± 0.11b 0.50 ± 0.01qr 1.52 ± 0.36E
M 1 2.05 ± 0.05gh 0.80 ± 0.02m-q 0.90 ± 0.02l-p 4.70 ± 0.10a 1.05 ± 0.01lmn 1.90 ± 0.39C
M 3 1.55 ± 0.05i 3.50 ± 0.14c 1.50 ± 0.03ij 3.45 ± 0.02c 1.60 ± 0.04i 2.32 ± 0.25A
M 5 1.95 ± 0.05h 2.90 ± 0.05d 1.00 ± 0.01l-o 4.05 ± 0.10b 1.50 ± 0.05ij 2.28 ± 0.29A
M 6 1.40 ± 0.03ijk 2.00 ± 0.08gh 1.20 ± 0.03jkl 2.45 ± 0.04ef 2.00 ± 0.05gh 1.81 ± 0.12CD
M 7 0.00 ± 0.00s 2.30 ± 0.02fg 1.00 ± 0.02l-o 2.40 ± 0.06ef 0.50 ± 0.01qr 1.24 ± 0.26F
B= Boiling on burner , M= heating in microwave; BM= boiling method In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table -32: Means comparison of yield for sapodilla fruit peel after 20 min of boiling using 0.1N HCl (Mechanical procedure
pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 0.80 ± 0.13k 1.18 ± 0.19j 1.48 ± 0.03i 0.05 ± 0.00m 0.10 ± 0.02m 0.72 ± 0.11E
3 3.53 ± 0.60e 3.78 ± 0.12d 2.55 ± 0.27g 5.63 ± 0.40b 0.53 ± 0.21l 3.20 ± 0.34A
5 5.93 ± 0.19a 2.93 ± 0.40f 1.85 ± 0.16h 2.65 ± 0.19g 0.08 ± 0.01m 2.69 ± 0.37B
6 4.35 ± 0.38c 3.05 ± 0.17f 1.65 ± 0.09hi 2.65 ± 0.08g 0.05 ± 0.00m 2.35 ± 0.28C
7 3.48 ± 0.40e 3.08 ± 0.12f 1.58 ± 0.10i 0.40 ± 0.13l 0.08 ± 0.01m 1.72 ± 0.27D
Mean 3.62 ± 0.35A 2.80 ± 0.19B 1.82 ± 0.10D 2.27 ± 0.38C 0.17 ± 0.05E In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 33: Means comparison of yield for sapodilla fruit peel after 20 min of boiling using 0.1N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
113
B 2.86 ± 0.44c 2.52 ± 0.28d 1.54 ± 0.06g 1.93 ± 0.47f 0.06 ± 0.01i 1.78 ± 0.18B
M 4.37 ± 0.47a 3.08 ± 0.23b 2.10 ± 0.15e 2.62 ± 0.61d 0.27 ± 0.10h 2.49 ± 0.22A B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 34: Means comparison of yield for sapodilla fruit peel after 20 min of boiling using 0.1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 0.50 ± 0.01ab 0.75 ± 0.01YZa 1.45 ± 0.03UVW 0.05 ± 0.00c 0.05 ± 0.00c 0.56 ± 0.14H
B 3 2.20 ± 0.04OPQ 3.55 ± 0.10GH 1.95 ± 0.01P-S 4.75 ± 0.09D 0.05 ± 0.00c 2.50 ± 0.42C
B 5 5.50 ± 0.07B 2.05 ± 0.03PQR 1.50 ± 0.05TUV 2.25 ± 0.06NOP 0.05 ± 0.00c 2.27 ± 0.48D
B 6 3.50 ± 0.05GHI 3.40 ± 0.13HIJ 1.45 ± 0.03UVW 2.50 ± 0.06MNO 0.05 ± 0.00c 2.18 ± 0.35D
B 7 2.60 ± 0.02MN 2.85 ± 0.09KLM 1.35 ± 0.02VWX 0.10 ± 0.01c 0.10 ± 0.01c 1.40 ± 0.31F
M 1 1.10 ± 0.01WXY 1.60 ± 0.03S-V 1.50 ± 0.05TUV 0.05 ± 0.00c 0.15 ± 0.01bc 0.88 ± 0.18G
M 3 4.85 ± 0.13CD 4.00 ± 0.12EF 3.15 ± 0.06IJK 6.50 ± 0.17A 1.00 ± 0.03XYZ 3.90 ± 0.49A
M 5 6.35 ± 0.05A 3.80 ± 0.12FG 2.20 ± 0.03OPQ 3.05 ± 0.10JKL 0.10 ± 0.01c 3.10 ± 0.55B
M 6 5.20 ± 0.04BC 2.70 ± 0.08LM 1.85 ± 0.05Q-T 2.79 ± 0.10KLM 0.05 ± 0.01c 2.52 ± 0.45C
M 7 4.35 ± 0.14E 3.30 ± 0.10HIJ 1.80 ± 0.05R-U 0.70 ± 0.02Za 0.05 ± 0.00c 2.04 ± 0.43E
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 35: Means comparison of yield for sapodilla fruit peel after 40 min of boiling using 0.1N HCl (Mechanical procedure
pH interaction mean±SE)
114
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 0.05 ± 0.00l 0.63 ± 0.23ij 0.05 ± 0.00l 0.05 ± 0.00l 0.05 ± 0.00l 0.17 ± 0.06D
3 0.08 ± 0.01kl 1.18 ± 0.08g 0.25 ± 0.09k 1.48 ± 0.06ef 1.90 ± 0.27d 0.98 ± 0.14C
5 0.05 ± 0.00l 2.58 ± 0.08b 2.58 ± 0.62b 2.20 ± 0.08c 3.10 ± 0.09a 2.10 ± 0.23A
6 1.65 ± 0.02e 2.60 ± 0.23b 0.83 ± 0.08h 3.15 ± 0.20a 2.35 ± 0.07c 2.12 ± 0.16A
7 0.50 ± 0.20j 1.35 ± 0.18fg 0.80 ± 0.31hi 1.50 ± 0.23ef 2.63 ± 0.09b 1.36 ± 0.16B
Mean 0.47 ± 0.12D 1.67 ± 0.16B 0.90 ± 0.21C 1.68 ± 0.20B 2.01 ± 0.20A
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 36: Means comparison of yield for sapodilla fruit peel after 40 min of boiling using 0.1N HCl ( Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 0.37 ± 0.17f 1.31 ± 0.22d 0.49 ± 0.11e 1.48 ± 0.24c 2.05 ± 0.28a 1.14 ± 0.12B
M 0.56 ± 0.17e 2.02 ± 0.21a 1.31 ± 0.38d 1.87 ± 0.31b 1.96 ± 0.31ab 1.54 ± 0.14A
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 37: Means comparison of yield for sapodilla fruit peel after 40 min of boiling using 0.1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 0.05 ± 0.00r 0.10 ± 0.00r 0.05 ± 0.00r 0.05 ± 0.00r 0.05 ± 0.01r 0.06 ± 0.01H
B 3 0.05 ± 0.00r 1.00 ± 0.01p 0.45 ± 0.01q 1.60 ± 0.05lm 2.50 ± 0.09fgh 1.12 ± 0.23E
B 5 0.05 ± 0.00r 2.40 ± 0.05h 1.20 ± 0.01op 2.05 ± 0.05j 3.00 ± 0.11cd 1.74 ± 0.28D
115
B 6 1.65 ± 0.02l 2.10 ± 0.02ij 0.65 ± 0.01q 2.70 ± 0.03efg 2.25 ± 0.08hij 1.87 ± 0.19C
B 7 0.05 ± 0.00r 0.95 ± 0.01p 0.10 ± 0.01r 1.00 ± 0.06p 2.45 ± 0.05gh 0.91 ± 0.23F
M 1 0.05 ± 0.00r 1.15 ± 0.01op 0.05 ± 0.00r 0.05 ± 0.01r 0.05 ± 0.00r 0.27 ± 0.12G
M 3 0.10 ± 0.00r 1.35 ± 0.02mno 0.05 ± 0.00r 1.35 ± 0.05mno 1.30 ± 0.04no 0.83 ± 0.17F
M 5 0.05 ± 0.00r 2.75 ± 0.06def 3.95 ± 0.10a 2.35 ± 0.09hi 3.20 ± 0.13c 2.46 ± 0.35A
M 6 1.65 ± 0.03l 3.10 ± 0.09c 1.00 ± 0.01p 3.60 ± 0.08b 2.45 ± 0.07gh 2.36 ± 0.25B
M 7 0.95 ± 0.01p 1.75 ± 0.03kl 1.50 ± 0.03lmn 2.00 ± 0.06jk 2.80 ± 0.07de 1.80 ± 0.16CD
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
116
Table - 38: Means comparison of yield for sapodilla fruit peel after 60 min of boiling using 0.1N HCl Mechanical procedure
pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 1.50 ± 0.13g 0.30 ± 0.02lm 0.08 ± 0.01mn 2.30 ± 0.21f 0.05 ± 0.00n 0.85 ± 0.17D
3 3.63 ± 0.21c 0.05 ± 0.00n 0.90 ± 0.22j 2.13 ± 0.40f 2.78 ± 0.04e 1.90 ± 0.26C
5 4.55 ± 0.06a 1.08 ± 0.46ij 0.28 ± 0.10lmn 3.78 ± 0.82c 4.35 ± 0.74ab 2.81 ± 0.40A
6 3.23 ± 0.10d 0.05 ± 0.00n 0.50 ± 0.18kl 4.13 ± 0.40b 3.60 ± 0.41c 2.30 ± 0.33B
7 2.80 ± 0.47e 1.18 ± 0.50hi 0.55 ± 0.22k 3.63 ± 0.04c 1.40 ± 0.56gh 1.91 ± 0.27C
Mean 3.14 ± 0.21A 0.53 ± 0.16C 0.46 ± 0.09C 3.19 ± 0.24A 2.44 ± 0.34B In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 39: Means comparison of yield for sapodilla fruit peel after 60 min of boiling using 0.1N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 2.91 ± 0.34d 0.09 ± 0.02i 0.13 ± 0.04i 2.38 ± 0.24e 1.67 ± 0.34f 1.44 ± 0.17B
M 3.37 ± 0.25b 0.97 ± 0.27g 0.79 ± 0.12h 4.00 ± 0.30a 3.20 ± 0.54c 2.47 ± 0.21A B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 40: Means comparison of yield for sapodilla fruit peel after 60 min of boiling using 0.1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 1.20 ± 0.01qr 0.25 ± 0.01st 0.05 ± 0.00t 1.85 ± 0.05o 0.05 ± 0.00t 0.68 ± 0.19G
B 3 4.10 ± 0.03e 0.05 ± 0.00t 0.40 ± 0.01st 1.25 ± 0.03qr 2.75 ± 0.05l 1.71 ± 0.41E
117
B 5 4.50 ± 0.04d 0.05 ± 0.00t 0.05 ± 0.00t 1.95 ± 0.03no 2.70 ± 0.02l 1.85 ± 0.45D
B 6 3.00 ± 0.06jkl 0.05 ± 0.00t 0.10 ± 0.01t 3.25 ± 0.09hij 2.70 ± 0.10l 1.82 ± 0.38DE
B 7 1.75 ± 0.03op 0.05 ± 0.01t 0.05 ± 0.01t 3.60 ± 0.04fgh 0.15 ± 0.01st 1.12 ± 0.37F
M 1 1.80 ± 0.02o 0.35 ± 0.00st 0.10 ± 0.01t 2.75 ± 0.09l 0.05 ± 0.01t 1.01 ± 0.29F
M 3 3.15 ± 0.05ijk 0.05 ± 0.00t 1.40 ± 0.03pq 3.00 ± 0.12jkl 2.80 ± 0.08kl 2.08 ± 0.32C
M 5 4.60 ± 0.11d 2.10 ± 0.04no 0.50 ± 0.02s 5.60 ± 0.07b 6.00 ± 0.21a 3.76 ± 0.57A
M 6 3.45 ± 0.03ghi 0.05 ± 0.00t 0.90 ± 0.02r 5.00 ± 0.17c 4.50 ± 0.14d 2.78 ± 0.53B
M 7 3.85 ± 0.06ef 2.30 ± 0.08mn 1.05 ± 0.02qr 3.65 ± 0.07fg 2.65 ± 0.11lm 2.70 ± 0.27B
B= Boiling on burner , M= heating in microwave; BM= boiling method In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
118
Table - 41: Percentage yield of pectin from sapodilla fruit peel after using different
physicomechanical procedures (pH, mechanical procedure, boiling method and time of
boling) with IN HCl.
Mechanical
procedure pH
% yield 10 min %Yield 20 min %yield 40 min % yield 60 min
B M B M B M B M
Hammering
1 2.65 3.15 1.5 2.2 3.5 3 0.3 2
3 5.65 6.55 3.45 5.45 4.5 4.5 0.4 2.85
5 2.3 4.6 4.4 4.5 4.2 5.25 0.15 2.3
6 2.15 2.95 2.45 4.3 2 3.4 3.45 2.6
7 2.5 3.1 2.4 2.45 2 2.55 3.5 0.05
Grinding
1 3.1 4.15 1.55 1.65 1.4 3.7 1.4 3
3 5.6 4.8 2.55 5.4 3.25 5.15 3.15 5
5 1.05 6.1 4.2 5.1 3.8 5.5 1 3.3
6 1.45 3.55 4.3 3.65 3.5 3.3 0.8 1.2
7 0.35 2.75 1.3 2.5 2.15 2.6 1.65 1.6
Cutting
1 0.05 0.05 0.5 1.4 3.65 4 0.65 0.05
3 0.05 1.6 1 2.35 2.5 3.5 0.75 0.1
5 2.95 5.3 2.05 2.7 4.1 5.5 0.1 0.05
6 0.15 3.5 3.35 3.7 1.5 3 0 0.6
7 0.05 1.2 2.6 1.8 2 3.25 0.1 0.05
Homogenizing
1 1.55 4.95 2.1 3.6 3.4 4.6 0.6 0
3 2.4 5.35 3.55 5.25 3.65 4.55 0.05 0.15
5 1.7 4.85 3.55 5.9 2.7 3 0.15 3.4
6 0.6 1.5 3.8 3.15 3.9 3.5 0.05 0.25
7 1.2 2.55 1.25 2.2 3.35 3.3 0.1 0.6
Chopping 1 3 5.5 2.5 5.9 1.95 3.2 0.05 3.55
(mortor/pastle 3 3.7 5.4 1.4 6 1.15 4.85 0.2 0.1
5 4.1 6.7 1.25 6.1 4.5 6.5 0.45 0.5
6 3.75 3.8 1.2 4.9 2.5 3.1 0 1
7 0.05 0.25 1.8 2.5 1 3.95 0.1 0.6
B= Boiling on burner , M= heating in microwave
119
Table - 42: Analysis of variance (mean squares) of yield of pectin from sapodilla fruit peel after using different
physicomechanical procedure different fruits with 1N HCl
Source of
variation
Degrees of
freedom
Mean squares
Yield 10 min_NHCl Yield 20 min_NHCl Yield 40 min_NHCl Yield 60 min_NHCl
MP
pH
BM
MP x pH
MP x BM
pH x BM
MP x pH x BM
Error
Total
4
4
1
16
4
4
16
100
149
23.648**
38.736**
106.361**
8.396**
1.944**
4.779**
2.360**
0.022
8.5850**
21.8147**
71.7604**
3.9668**
10.4560**
4.7719**
1.5394**
0.0173
0.5439**
16.3404**
42.5601**
3.5228**
3.5189**
0.9204**
1.2315**
0.0168
21.8563**
0.8645**
14.8838**
4.0399**
2.1424**
4.7014**
3.3080**
0.0054
NS = Non-significant (P>0.05); * = Significant (P<0.05); ** = Highly significant (P<0.01); Mech= mechanical
120
Table - 43: Means comparison of yield for sapodilla fruit peel after 10 min of boiling using 1N HCl ( Mechanical procedure
pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 2.90 ± 0.12j 3.63 ± 0.24fg 0.05 ± 0.00p 4.25 ± 0.56cd 3.25 ± 0.76i 2.82 ± 0.33C
3 6.10 ± 0.21a 5.20 ± 0.19b 0.83 ± 0.35no 4.55 ± 0.39c 3.88 ± 0.66ef 4.11 ± 0.37A
5 3.45 ± 0.51ghi 3.58 ± 1.13fgh 4.13 ± 0.53de 5.40 ± 0.59b 3.28 ± 0.71hi 3.97 ± 0.34B
6 2.55 ± 0.18k 2.50 ± 0.47k 1.83 ± 0.75lm 3.78 ± 0.07f 1.05 ± 0.20n 2.34 ± 0.24D
7 2.80 ± 0.14jk 1.55 ± 0.54m 0.63 ± 0.26o 0.15 ± 0.05p 1.88 ± 0.30l 1.40 ± 0.22E
Mean 3.56 ± 0.27A 3.29 ± 0.34B 1.49 ± 0.33D 3.63 ± 0.38A 2.67 ± 0.31C In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 44: Means comparison of yield for sapodilla fruit peel after 10 min of boiling using 1N HCl ( Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 3.05 ± 0.35d 2.31 ± 0.50e 0.65 ± 0.31g 2.92 ± 0.40d 1.49 ± 0.16f 2.08 ± 0.19B
M 4.07 ± 0.37b 4.27 ± 0.31a 2.33 ± 0.50e 4.33 ± 0.60a 3.84 ± 0.41c 3.77 ± 0.21A
B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 45: Means comparison of yield for sapodilla fruit peel after 10 min of boiling using 1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 2.65 ± 0.03n-q 3.10 ± 0.05lmn 0.05 ± 0.01w 3.00 ± 0.06mno 1.55 ± 0.03st 2.07 ± 0.31F
121
B 3 5.65 ± 0.12cd 5.60 ± 0.07cd 0.05 ± 0.01w 3.70 ± 0.10jk 2.40 ± 0.08pq 3.48 ± 0.56C
B 5 2.30 ± 0.05pq 1.05 ± 0.03tu 2.95 ± 0.11no 4.10 ± 0.15ij 1.70 ± 0.04rs 2.42 ± 0.28E
B 6 2.15 ± 0.04qr 1.45 ± 0.01st 0.15 ± 0.01vw 3.75 ± 0.08jk 0.60 ± 0.01uv 1.62 ± 0.34G
B 7 2.50 ± 0.05opq 0.35 ± 0.01vw 0.05 ± 0.01w 0.05 ± 0.01w 1.20 ± 0.03st 0.83 ± 0.25H
M 1 3.15 ± 0.09lmn 4.15 ± 0.05ij 0.05 ± 0.00w 5.50 ± 0.08d 4.95 ± 0.12e-h 3.56 ± 0.52C
M 3 6.55 ± 0.08ab 4.80 ± 0.13gh 1.60 ± 0.05s 5.40 ± 0.20de 5.35 ± 0.10def 4.74 ± 0.45B
M 5 4.60 ± 0.02hi 6.10 ± 0.20bc 5.30 ± 0.07d-g 6.70 ± 0.24a 4.85 ± 0.16fgh 5.51 ± 0.22A
M 6 2.95 ± 0.09no 3.55 ± 0.05kl 3.50 ± 0.05klm 3.80 ± 0.13jk 1.50 ± 0.05st 3.06 ± 0.22D
M 7 3.10 ± 0.01lmn 2.75 ± 0.09nop 1.20 ± 0.03st 0.25 ± 0.02vw 2.55 ± 0.05opq 1.97 ± 0.29F
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 46: Means comparison of yield for sapodilla fruit peel after 20 min of boiling using 1N HCl Mechanical procedure
pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 1.82 ± 0.15kl 1.60 ± 0.03l 0.95 ± 0.20m 4.20 ± 0.77cd 2.85 ± 0.34h 2.28 ± 0.27D
3 4.45 ± 0.45abc 3.98 ± 0.64de 1.68 ± 0.30kl 3.70 ± 1.03ef 4.40 ± 0.39bc 3.64 ± 0.32B
5 4.45 ± 0.06abc 4.65 ± 0.21ab 2.38 ± 0.15i 3.68 ± 1.09f 4.73 ± 0.53a 3.98 ± 0.28A
6 3.38 ± 0.41g 3.98 ± 0.15de 3.53 ± 0.10fg 3.05 ± 0.83h 3.48 ± 0.16fg 3.48 ± 0.19C
7 2.43 ± 0.03i 1.90 ± 0.27jk 2.20 ± 0.18i 2.15 ± 0.16ij 1.73 ± 0.21kl 2.08 ± 0.09E
Mean 3.30 ± 0.23BC 3.22 ± 0.27C 2.15 ± 0.18D 3.36 ± 0.37AB 3.44 ± 0.25A
122
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
123
Table - 47: Means comparison of yield for sapodilla fruit peel after 20 min of boiling using 1N HCl ( Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 2.84 ± 0.27d 2.78 ± 0.34d 1.90 ± 0.28f 1.63 ± 0.13g 2.85 ± 0.27d 2.40 ± 0.13B
M 3.77 ± 0.34c 3.66 ± 0.39c 2.39 ± 0.21e 5.08 ± 0.37a 4.02 ± 0.37b 3.78 ± 0.18A
B= Boiling on burner , M= heating in microwave In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
124
Table - 48: Means comparison of yield for sapodilla fruit peel after 20 min of boiling using 1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 1.50 ± 0.02qrs 1.55 ± 0.03qrs 0.50 ± 0.01u 2.50 ± 0.06j-m 2.10 ± 0.05mno 1.63 ± 0.18F
B 3 3.45 ± 0.02ghi 2.55 ± 0.05jkl 1.00 ± 0.02t 1.40 ± 0.03q-t 3.55 ± 0.06ghi 2.39 ± 0.28D
B 5 4.40 ± 0.06e 4.20 ± 0.07ef 2.05 ± 0.03nop 1.25 ± 0.02rst 3.55 ± 0.05ghi 3.09 ± 0.33C
B 6 2.45 ± 0.04j-n 4.30 ± 0.04e 3.35 ± 0.05hi 1.20 ± 0.03st 3.80 ± 0.10fg 3.02 ± 0.29C
B 7 2.40 ± 0.05j-n 1.30 ± 0.01rst 2.60 ± 0.09jk 1.80 ± 0.05opq 1.25 ± 0.01rst 1.87 ± 0.15E
M 1 2.13 ± 0.07l-o 1.65 ± 0.01pqr 1.40 ± 0.01q-t 5.90 ± 0.19a 3.60 ± 0.08gh 2.94 ± 0.45C
M 3 5.45 ± 0.08b 5.40 ± 0.13b 2.35 ± 0.02j-n 6.00 ± 0.15a 5.25 ± 0.14bc 4.89 ± 0.35A
M 5 4.50 ± 0.10de 5.10 ± 0.10bc 2.70 ± 0.06j 6.10 ± 0.08a 5.90 ± 0.17a 4.86 ± 0.33A
M 6 4.30 ± 0.03e 3.65 ± 0.06gh 3.70 ± 0.14gh 4.90 ± 0.11cd 3.15 ± 0.07i 3.94 ± 0.16B
M 7 2.45 ± 0.04j-n 2.50 ± 0.08j-m 1.80 ± 0.04opq 2.50 ± 0.05j-m 2.20 ± 0.06k-o 2.29 ± 0.08D
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
125
Table - 49: Means comparison of yield for sapodilla fruit peel after 40 min of boiling using 1N HCl (Mechanical procedure
pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 3.25 ± 0.12gh 2.55 ± 0.51k-n 3.83 ± 0.09ef 2.58 ± 0.28j-m 4.00 ± 0.28de 3.24 ± 0.17C
3 4.50 ± 0.05c 4.20 ± 0.43d 3.00 ± 0.23hi 3.00 ± 0.83hi 4.10 ± 0.21de 3.76 ± 0.22B
5 4.73 ± 0.24bc 4.65 ± 0.38bc 4.80 ± 0.32b 5.50 ± 0.46a 2.85 ± 0.08ij 4.51 ± 0.21A
6 2.70 ± 0.32jkl 3.38 ± 0.04g 2.25 ± 0.34o 2.80 ± 0.14ijk 3.70 ± 0.10f 2.97 ± 0.13D
7 2.28 ± 0.13no 2.38 ± 0.10mno 2.63 ± 0.28j-m 2.48 ± 0.66l-o 3.33 ± 0.04g 2.62 ± 0.15E
Mean 3.49 ± 0.20B 3.43 ± 0.22B 3.30 ± 0.20C 3.27 ± 0.31C 3.60 ± 0.11A
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 50: Means comparison of yield for sapodilla fruit peel after 40 min of boiling using 1N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 3.24 ± 0.29e 2.81 ± 0.24f 2.75 ± 0.26f 2.22 ± 0.34g 3.40 ± 0.11d 2.88 ± 0.12B
M 3.74 ± 0.27c 4.05 ± 0.30b 3.85 ± 0.24c 4.32 ± 0.34a 3.79 ± 0.18c 3.95 ± 0.12A
B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 51: Means comparison of yield for sapodilla fruit peel after 40 min of boiling using 1N HCl ( Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 3.50 ± 0.05j-n 1.40 ± 0.02uv 3.65 ± 0.05i-m 1.95 ± 0.04t 3.40 ± 0.10k-o 2.78 ± 0.25G
126
B 3 4.50 ± 0.11def 3.25 ± 0.04mno 2.50 ± 0.05rs 1.15 ± 0.02uv 3.65 ± 0.12i-m 3.01 ± 0.30F
B 5 4.20 ± 0.04efg 3.80 ± 0.11g-k 4.10 ± 0.09fgh 4.50 ± 0.04def 2.70 ± 0.10pqr 3.86 ± 0.17C
B 6 2.00 ± 0.04t 3.47 ± 0.03j-n 1.50 ± 0.01u 2.50 ± 0.03rs 3.90 ± 0.05g-j 2.67 ± 0.24G
B 7 2.00 ± 0.04t 2.15 ± 0.03st 2.00 ± 0.03t 1.00 ± 0.03v 3.35 ± 0.08l-o 2.10 ± 0.20H
M 1 3.00 ± 0.11opq 3.70 ± 0.05h-l 4.00 ± 0.10ghi 3.20 ± 0.05no 4.60 ± 0.09de 3.70 ± 0.16D
M 3 4.50 ± 0.04def 5.15 ± 0.14bc 3.50 ± 0.03j-n 4.85 ± 0.09cd 4.55 ± 0.09de 4.51 ± 0.15B
M 5 5.25 ± 0.09bc 5.50 ± 0.07b 5.50 ± 0.08b 6.50 ± 0.19a 3.00 ± 0.05opq 5.15 ± 0.31A
M 6 3.40 ± 0.10k-o 3.30 ± 0.03l-o 3.00 ± 0.04opq 3.10 ± 0.06nop 3.50 ± 0.08j-n 3.26 ± 0.06E
M 7 2.55 ± 0.07rs 2.60 ± 0.05qr 3.25 ± 0.07mno 3.95 ± 0.14ghi 3.30 ± 0.05l-o 3.13 ± 0.14EF
B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 52: Means comparison of yield for sapodilla fruit peel after 60 min of boiling using 1N HCl (Mechanical procedure
pH interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 1.15 ± 0.38fg 2.20 ± 0.36c 0.35 ± 0.13hi 1.80 ± 0.78d 0.30 ± 0.13ij 1.16 ± 0.23B
3 1.63 ± 0.55e 4.08 ± 0.42a 0.43 ± 0.15hi 0.15 ± 0.02jk 0.10 ± 0.02k 1.28 ± 0.31A
5 1.23 ± 0.48f 2.15 ± 0.52c 0.08 ± 0.01k 0.48 ± 0.02h 1.78 ± 0.73de 1.14 ± 0.24B
6 3.03 ± 0.20b 1.00 ± 0.09g 0.30 ± 0.13ij 0.50 ± 0.22h 0.15 ± 0.05jk 1.00 ± 0.21C
7 1.78 ± 0.77de 1.63 ± 0.03e 0.08 ± 0.01k 0.35 ± 0.11hi 0.35 ± 0.11hi 0.84 ± 0.20D
Mean 1.76 ± 0.25B 2.21 ± 0.24A 0.25 ± 0.05E 0.66 ± 0.19C 0.54 ± 0.18D In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 53: Means comparison of yield for sapodilla fruit peel after 60 min of boiling using 1N HCl (Mechanical procedure
boiling method interaction mean±SE)
127
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 1.56 ± 0.42c 1.60 ± 0.22c 0.32 ± 0.08f 0.16 ± 0.04g 0.19 ± 0.06g 0.77 ± 0.12B
M 1.96 ± 0.27b 2.82 ± 0.36a 0.17 ± 0.06g 1.15 ± 0.33d 0.88 ± 0.34e 1.40 ± 0.17A
B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 54: Means comparison of yield for sapodilla fruit peel after 60 min of boiling using 1N HCl (Mechanical procedure
pH boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 0.30 ± 0.01r-u 1.40 ± 0.05kl 0.65 ± 0.03op 0.05 ± 0.01vw 0.60 ± 0.02opq 0.60 ± 0.12E
B 3 0.40 ± 0.02q-t 3.15 ± 0.07de 0.75 ± 0.03o 0.20 ± 0.01t-w 0.05 ± 0.00vw 0.91 ± 0.31D
B 5 0.15 ± 0.01uvw 1.00 ± 0.03mn 0.10 ± 0.01uvw 0.45 ± 0.01p-s 0.15 ± 0.01uvw 0.37 ± 0.09F
B 6 3.45 ± 0.09bc 0.80 ± 0.03no 0.00 ± 0.00w 0.00 ± 0.00w 0.05 ± 0.01vw 0.86 ± 0.36D
B 7 3.50 ± 0.08bc 1.65 ± 0.03j 0.10 ± 0.01uvw 0.10 ± 0.01uvw 0.10 ± 0.01uvw 1.09 ± 0.36C
M 1 2.00 ± 0.05i 3.00 ± 0.08ef 0.05 ± 0.00vw 3.55 ± 0.07b 0.00 ± 0.00w 1.72 ± 0.39B
M 3 2.85 ± 0.05f 5.00 ± 0.10a 0.10 ± 0.01uvw 0.10 ± 0.01uvw 0.15 ± 0.01uvw 1.64 ± 0.53B
M 5 2.30 ± 0.09h 3.30 ± 0.12cd 0.05 ± 0.00vw 0.50 ± 0.02pqr 3.40 ± 0.05bc 1.91 ± 0.37A
M 6 2.60 ± 0.07g 1.20 ± 0.04lm 0.60 ± 0.03opq 1.00 ± 0.05mn 0.25 ± 0.02s-v 1.13 ± 0.22C
M 7 0.05 ± 0.01vw 1.60 ± 0.05jk 0.05 ± 0.00vw 0.60 ± 0.02opq 0.60 ± 0.02opq 0.58 ± 0.15E B= Boiling on burner , M= heating in microwave; BM= boiling method
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
128
Table - 55: Percentage yield of pectin from sapodilla fruit peel after using different
organic acid
Mechanical
procedure Inorganic acid
% of inorganic
acid pH
% yield
B M h
om
og
en
izin
g
Citric acid 1%
3 to 5
2.55 3.25
10% 2.55 4.24
Oxalic acid 1% 3.95 4.95
10% 0.5 3.7
Tartaric acid 1% 1.8 3.25
10% 2.5 2.55
ch
op
pin
g
Citric acid 1% 2.4 4.3
10% 0.05 0.1
Oxalic acid 1% 0.05 1.5
10% 0.15 1.85
Tartaric acid 1% 0.75 1.25
10% 0.35 3.2
Grin
din
g
Citric acid 1% 1.35 2.15
10% 0.05 1.2
Oxalic acid 1% 0.1 0.3
10% 1.25 1.05
Tartaric acid 1% 1.8 2.43
10% 1.65 3.15
Cu
ttin
g
Citric acid 1% 1.4 2.64
10% 0.1 1.1
Oxalic acid 1% 0.05 1.2
10% 0.13 1.32
Tartaric acid 1% 0.2 1.87
10% 0.32 1.45
Ha
mm
erin
g
Citric acid 1% 1.6 1.8
10% 0.54 1.67
Oxalic acid 1% 0.1 1.3
10% 0.43 2
Tartaric acid 1% 0.33 1.4
10% 0.44 2.1
B= Boiling on burner , M= heating in microwave
129
Table - 56 : Analysis of variance (mean squares) of yield of pectin from sapodilla
fruit peel after using different organic acid
Source of
variation
Degrees of
freedom
Mean squares
Yield (citric acid) Yield (oxalic
acid)
Yield (tartaric
acid)
BM
MP
Acid%
BM x MP
BM x Acid%
MP x Acid%
BM x MP x Acid%
Error
Total
1
4
1
4
1
4
4
40
59
14.5829**
7.7555**
21.0278**
0.1237**
0.0049NS
5.6981**
1.0202**
0.0126
23.2877**
14.9075**
0.1882**
1.8114**
0.9077**
5.0161**
0.7483**
0.0090
23.4750**
6.0553**
1.0375**
0.3754**
0.5245**
0.3919**
1.5337**
0.0101
NS = Non-significant (P>0.05); * = Significant (P<0.05); ** = Highly significant (P<0.01) BM= Boiling method. MP= Mechanical procedure
Table - 57: Means comparison of yield for sapodilla fruit peel after using citric acid
(Boiling method x mechanical procedure interaction mean±SE)
Mechanical
procedure
Boiling Method Mean
B M
Homogenizing 2.55 ± 0.05b 3.75 ± 0.23a 3.15 ± 0.21A
Chopping 1.23 ± 0.53e 2.20 ± 0.94c 1.71 ± 0.54B
Grinding 0.70 ± 0.29f 1.68 ± 0.21d 1.19 ± 0.23D
Cutting 0.75 ± 0.29f 1.87 ± 0.34d 1.31 ± 0.27CD
Hammering 1.07 ± 0.24e 1.74 ± 0.04d 1.40 ± 0.15C
Mean 1.26 ± 0.18B 2.25 ± 0.24A
B= Boiling on burner , M= heating in microwave.In a row and column statistically non-significant (P>0.05)
represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 58: Means comparison of yield for sapodilla fruit peel after using citric acid
Acid (% x boiling method interaction mean±SE)
Inorganic Boiling Method Mean
130
acid B M
1% 1.86 ± 0.14b 2.83 ± 0.24a 2.34 ± 0.16A
10% 0.66 ± 0.26d 1.66 ± 0.37c 1.16 ± 0.24B B= Boiling on burner , M= heating in microwave Among the two concentrations of acid 1% has found more effective with microwave as heating procedure
Table - 59: Means comparison of yield for sapodilla fruit peel after using citric acid
(Boiling method x Acid% x mechanical procedure interaction mean±SE)
Acid% x MP Boiling Method
Mean B M
1% x MP1 2.55 ± 0.08c 3.25 ± 0.10b 2.90 ± 0.17B
1% x MP2 2.40 ± 0.09cd 4.30 ± 0.16a 3.35 ± 0.43A
1% x MP3 1.35 ± 0.04fg 2.15 ± 0.04d 1.75 ± 0.18D
1% x MP4 1.40 ± 0.02fg 2.64 ± 0.04c 2.02 ± 0.28C
1% x MP5 1.60 ± 0.04ef 1.80 ± 0.05e 1.70 ± 0.05D
10% x MP1 2.55 ± 0.08c 4.24 ± 0.12a 3.40 ± 0.38A
10% x MP2 0.05 ± 0.00i 0.10 ± 0.00i 0.08 ± 0.01G
10% x MP3 0.05 ± 0.00i 1.20 ± 0.02g 0.63 ± 0.26F
10% x MP4 0.10 ± 0.00i 1.10 ± 0.01g 0.60 ± 0.22F
10% x MP5 0.54 ± 0.01h 1.67 ± 0.05ef 1.11 ± 0.25E
BM= Boiling method. MP= Mechanical procedure; B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by
similar letters. Comparison among interaction means are represented by lower case alphabets while capital
alphabets are used for overall mean
Table - 60: Means comparison of yield for sapodilla fruit peel after using oxalic acid
(Boiling method x mechanical procedure interaction mean±SE)
Mechanical
procedure
Boiling Method Mean
B M
Homogenizing 2.23 ± 0.77b 4.33 ± 0.29a 3.28 ± 0.51A
Chopping 0.10 ± 0.02f 1.68 ± 0.08c 0.89 ± 0.24B
Grinding 0.68 ± 0.26e 0.68 ± 0.17e 0.68 ± 0.15C
Cutting 0.09 ± 0.02f 1.26 ± 0.03d 0.68 ± 0.18C
Hammering 0.27 ± 0.07f 1.65 ± 0.16c 0.96 ± 0.22B
Mean 0.67 ± 0.21B 1.92 ± 0.24A
131
B= Boiling on burner , M= heating in microwave; B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by
similar letters. Comparison among interaction means are represented by lower case alphabets while capital
alphabets are used for overall mean
Table - 61: Means comparison of yield for sapodilla fruit peel after using oxalic acid
Acid% x boiling method interaction mean±SE
Inorganic
acid
Boiling Method Mean
B M
1% 0.85 ± 0.42 1.85 ± 0.43 1.35 ± 0.31A
10% 0.49 ± 0.11 1.98 ± 0.25 1.24 ± 0.19B
B= Boiling on burner , M= heating in microwave
Among the two concentrations of acid 1% has found more effective with microwave as
heating procedure
Table - 62: Means comparison of yield for sapodilla fruit peel after using oxalic acid
(Boiling method x Acid% x mechanical procedure interaction mean±SE)
Acid% x MP Boiling Method
Mean B M
1% x MP1 3.95 ± 0.15b 4.95 ± 0.14a 4.45 ± 0.24A
1% x MP2 0.05 ± 0.00h 1.50 ± 0.02d 0.78 ± 0.32E
1% x MP3 0.10 ± 0.01h 0.30 ± 0.01fgh 0.20 ± 0.05F
1% x MP4 0.05 ± 0.00h 1.20 ± 0.01e 0.63 ± 0.26E
1% x MP5 0.10 ± 0.01h 1.30 ± 0.02de 0.70 ± 0.27E
10% x MP1 0.50 ± 0.02f 3.70 ± 0.10b 2.10 ± 0.72B
10% x MP2 0.15 ± 0.01gh 1.85 ± 0.03c 1.00 ± 0.38D
10% x MP3 1.25 ± 0.04de 1.05 ± 0.02e 1.15 ± 0.05CD
10% x MP4 0.13 ± 0.01h 1.32 ± 0.02de 0.73 ± 0.27E
10% x MP5 0.43 ± 0.02fg 2.00 ± 0.05c 1.22 ± 0.35C
BM= Boiling method. MP= Mechanical procedure ; B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by
similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
132
Table - 63: Means comparison of yield for sapodilla fruit peel after using tartaric
acid (Boiling method x mechanical procedure interaction mean±SE)
Mechanical
procedure
Boiling Method Mean
B M
homogenizing 2.15 ± 0.16b 2.90 ± 0.17a 2.53 ± 0.16A
Chopping 0.55 ± 0.09d 2.23 ± 0.44b 1.39 ± 0.33C
Grinding 1.73 ± 0.04c 2.79 ± 0.17a 2.26 ± 0.18B
Cutting 0.26 ± 0.03e 1.66 ± 0.10c 0.96 ± 0.22D
Hammering 0.39 ± 0.03de 1.75 ± 0.16c 1.07 ± 0.22D
Mean 1.01 ± 0.15B 2.27 ± 0.14A
B= Boiling on burner , M= heating in microwave
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by
similar letters. Comparison among interaction means are represented by lower case alphabets while capital
alphabets are used for overall mean
Table - 64: Means comparison of yield for sapodilla fruit peel after using tartaric
acid (Acid% x boiling method interaction mean±SE)
Inorganic
acid
Boiling Method Mean
B M
1% 0.98 ± 0.19c 2.04 ± 0.20b 1.51 ± 0.17B
10% 1.05 ± 0.24c 2.49 ± 0.18a 1.77 ± 0.20A
B= Boiling on burner , M= heating in microwave
Among the two concentrations of acid 1% has found more effective with microwave as heating procedure
133
Table - 65: Means comparison of yield for sapodilla fruit peel after using tartaric
acid (Boiling method x Acid% x mechanical procedure interaction mean±SE)
Acid% x MP Boiling Method
Mean B M
1% x PM1 1.80 ± 0.03cd 3.25 ± 0.08a 2.53 ± 0.33A
1% x PM2 0.75 ± 0.02g 1.25 ± 0.02f 1.00 ± 0.11E
1% x PM3 1.80 ± 0.05cd 2.43 ± 0.09b 2.12 ± 0.15B
1% x PM4 0.20 ± 0.02h 1.87 ± 0.08cd 1.04 ± 0.38E
1% x PM5 0.33 ± 0.02h 1.40 ± 0.02ef 0.87 ± 0.24E
10% x PM1 2.50 ± 0.07b 2.55 ± 0.12b 2.53 ± 0.06A
10% x PM2 0.35 ± 0.02h 3.20 ± 0.08a 1.78 ± 0.64C
10% x PM3 1.65 ± 0.03de 3.15 ± 0.10a 2.40 ± 0.34A
10% x PM4 0.32 ± 0.02h 1.45 ± 0.02ef 0.89 ± 0.25E
10% x PM5 0.44 ± 0.03h 2.10 ± 0.05c 1.27 ± 0.37D
B= Boiling on burner, M= heating in microwave; BM= Boiling method. MP= Mechanical procedure
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by
similar letters. Comparison among interaction means are represented by lower case alphabets while capital
alphabets are used for overall mean
134
Table - 66: percentage yield of pectin from sapodilla fruit peel using different
strengths of same inorganic acid.
Mechanical
procedure pH
0.1N HCl 0.5N HCl 1N HCl
B M B M B M
Hammering
1 1.5 2.05 0.65 1 2.65 3.15
3 0.6 1.55 0.05 3.3 5.65 6.55
5 0.8 1.95 0.3 2.2 2.3 4.6
6 0.8 1.4 0.45 1.65 2.15 2.95
7 0.5 0 0.25 0.05 2.5 3.1
Grinding
1 0.55 0.8 0.3 4.6 3.1 4.15
3 4 3.5 0.1 2.85 5.6 4.8
5 2.45 2.9 0.1 1 1.05 6.1
6 2 2 3.15 4.05 1.45 3.55
7 1.95 2.3 2.5 2.8 0.35 2.75
Cutting
1 0.8 0.9 3.65 4.2 0.05 0.05
3 1.1 1.5 4 2.25 0.05 1.6
5 0.4 1 4.35 3.75 2.95 5.3
6 1 1.2 0.55 4.75 0.15 3.5
7 0.7 1 1.25 0.5 0.05 1.2
Homogenizing
1 0.5 1.05 4.15 4.95 1.55 4.95
3 0.75 1.6 3.2 4.95 2.4 5.35
5 1.2 1.5 0.15 4.55 1.7 4.85
6 1.5 2 1.8 0.3 0.6 1.5
7 0.5 0.5 3.6 0.6 1.2 2.55
Chopping
(mortor/pastle
1 2.7 4.7 4.4 4.65 3 5.5
3 3.9 3.45 4.8 4.95 3.7 5.4
5 3.95 4.05 2.25 2.65 4.1 6.7
6 2.3 2.45 1.5 0.65 3.75 3.8
7 3.95 2.4 1 0.1 0.05 0.25
BM= Boiling method. MP= Mechanical procedure B= Boiling on burner , M= heating in microwave
133
Table - 67: Analysis of variance (mean squares) of yield of pectin from sapodilla fruit peel after using different strength of
same inorganic acid
Source of
variation
Degrees of
freedom
Mean squares
Yield_0.1N HCl Yield_0.5N HCl Yield_1N HCl
MP
pH
BM
MP x pH
MP x BM
pH x BM
MP x pH x BM
Error
Total
4
4
1
16
4
4
16
100
149
32.6575**
3.3917**
3.2413**
2.5462**
0.3392**
1.0112**
0.6258**
0.0093
19.2084**
20.5359**
21.2064**
8.7942**
4.8684**
6.8626**
5.2626**
0.0104
24.375**
37.244**
109.739**
7.934**
1.832**
4.535**
2.303**
0.047
NS = Non-significant (P>0.05); * = Significant (P<0.05); ** = Highly significant (P<0.01) B= Boiling on burner , M= heating in
microwave; BM= Boiling method. MP= Mechanical procedure
134
Table - 68: Means comparison for yield of pectin from sapodilla fruit peel after using 0.1N HCl (Mechanical procedure pH
interaction mean±SE )
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 0.78 ± 0.12k 0.68 ± 0.06kl 0.85 ± 0.03k 3.70 ± 0.45b 1.78 ± 0.12g 1.56 ± 0.23D
3 1.18 ± 0.19hij 3.75 ± 0.13b 1.30 ± 0.09hi 3.68 ± 0.12b 1.08 ± 0.21j 2.20 ± 0.24A
5 1.35 ± 0.07h 2.68 ± 0.10d 0.70 ± 0.13kl 4.00 ± 0.09a 1.38 ± 0.26h 2.02 ± 0.23B
6 1.75 ± 0.11g 2.00 ± 0.04f 1.10 ± 0.05ij 2.38 ± 0.05e 1.10 ± 0.14ij 1.67 ± 0.10C
7 0.50 ± 0.01l 2.13 ± 0.09f 0.85 ± 0.07k 3.18 ± 0.35c 0.25 ± 0.11m 1.38 ± 0.22E
Mean 1.11 ± 0.09C 2.25 ± 0.19B 0.96 ± 0.05D 3.39 ± 0.15A 1.12 ± 0.12C
B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 69: Means comparison for yield of pectin from sapodilla fruit peel after using 0.1N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 0.89 ± 0.11e 2.19 ± 0.30b 0.80 ± 0.07e 3.36 ± 0.19a 0.84 ± 0.09e 1.62 ± 0.14B
M 1.33 ± 0.14c 2.30 ± 0.24b 1.12 ± 0.06d 3.41 ± 0.24a 1.39 ± 0.20c 1.91 ± 0.13A
B= Boiling on burner, M= heating in microwave; BM= Boiling method. MP= Mechanical procedure In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are
represented by lower case alphabets while capital alphabets are used for overall mean
Table - 70: Means comparison for yield of pectin from sapodilla fruit peel after using 0.1N HCl (Mechanical procedure pH
boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 0.50 ± 0.02qr 0.55 ± 0.01qr 0.80 ± 0.02m-q 2.70 ± 0.05de 1.50 ± 0.04ij 1.21 ± 0.22F
135
B 3 0.75 ± 0.02n-q 4.00 ± 0.10b 1.10 ± 0.03klm 3.90 ± 0.10b 0.60 ± 0.03pqr 2.07 ± 0.41B
B 5 1.20 ± 0.03jkl 2.45 ± 0.04ef 0.40 ± 0.01r 3.95 ± 0.11b 0.80 ± 0.04m-q 1.76 ± 0.35D
B 6 1.50 ± 0.02ij 2.00 ± 0.05gh 1.00 ± 0.03l-o 2.30 ± 0.05fg 0.80 ± 0.03m-q 1.52 ± 0.15E
B 7 0.50 ± 0.01qr 1.95 ± 0.08h 0.70 ± 0.01o-r 3.95 ± 0.10b 0.50 ± 0.01qr 1.52 ± 0.36E
M 1 1.05 ± 0.03lmn 0.80 ± 0.01m-q 0.90 ± 0.02l-p 4.70 ± 0.10a 2.05 ± 0.03gh 1.90 ± 0.39C
M 3 1.60 ± 0.02i 3.50 ± 0.10c 1.50 ± 0.02ij 3.45 ± 0.08c 1.55 ± 0.05i 2.32 ± 0.25A
M 5 1.50 ± 0.03ij 2.90 ± 0.05d 1.00 ± 0.03l-o 4.05 ± 0.15b 1.95 ± 0.03h 2.28 ± 0.29A
M 6 2.00 ± 0.03gh 2.00 ± 0.08gh 1.20 ± 0.02jkl 2.45 ± 0.08ef 1.40 ± 0.05ijk 1.81 ± 0.12CD
M 7 0.50 ± 0.01qr 2.30 ± 0.08fg 1.00 ± 0.01l-o 2.40 ± 0.05ef 0.00 ± 0.00s 1.24 ± 0.26F
B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
136
Table - 71:Means comparison for yield of pectin from sapodilla fruit peel after using 0.5N HCl (Mechanical procedure pH
interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 4.55 ± 0.19b 2.45 ± 0.96fg 3.93 ± 0.13c 4.53 ± 0.08b 0.83 ± 0.08l 3.26 ± 0.32A
3 4.08 ± 0.39c 1.48 ± 0.62i 3.13 ± 0.39e 4.88 ± 0.08a 1.68 ± 0.73i 3.05 ± 0.32B
5 2.35 ± 0.99g 0.55 ± 0.20m 4.05 ± 0.14c 2.45 ± 0.10fg 1.25 ± 0.43j 2.13 ± 0.30C
6 1.05 ± 0.34jk 3.60 ± 0.21d 2.65 ± 0.94f 1.08 ± 0.19jk 1.05 ± 0.27jk 1.89 ± 0.28D
7 2.10 ± 0.67h 2.65 ± 0.07f 0.88 ± 0.17kl 0.55 ± 0.20m 0.15 ± 0.05n 1.27 ± 0.22E
Mean 2.83 ± 0.34B 2.15 ± 0.29D 2.93 ± 0.29A 2.70 ± 0.33C 0.99 ± 0.19E
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 72: Means comparison for yield of pectin from sapodilla fruit peel after using 0.5N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 2.58 ± 0.39c 1.23 ± 0.35e 2.76 ± 0.41b 2.79 ± 0.41b 0.34 ± 0.05f 1.94 ± 0.19B
M 3.07 ± 0.57a 3.06 ± 0.33a 3.09 ± 0.41a 2.60 ± 0.53c 1.64 ± 0.29d 2.69 ± 0.20A B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 73: Means comparison for yield of pectin from sapodilla fruit peel after using 0.5N HCl (Mechanical procedure pH
boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 4.15 ± 0.09def 0.30 ± 0.02s-v 3.65 ± 0.07h 4.40 ± 0.05cd 0.65 ± 0.03r 2.63 ± 0.48D
B 3 3.20 ± 0.02j 0.10 ± 0.01v 4.00 ± 0.03fg 4.80 ± 0.10ab 0.05 ± 0.01v 2.43 ± 0.53E
B 5 0.15 ± 0.00uv 0.10 ± 0.01v 4.35 ± 0.10cde 2.25 ± 0.05n 0.30 ± 0.01s-v 1.43 ± 0.45H
B 6 1.80 ± 0.02o 3.15 ± 0.07jk 0.55 ± 0.00rst 1.50 ± 0.04op 0.45 ± 0.02r-u 1.49 ± 0.26H
137
B 7 3.60 ± 0.08hi 2.50 ± 0.05mn 1.25 ± 0.03pq 1.00 ± 0.02q 0.25 ± 0.02tuv 1.72 ± 0.32G
M 1 4.95 ± 0.12a 4.60 ± 0.05bc 4.20 ± 0.05def 4.65 ± 0.11abc 1.00 ± 0.03q 3.88 ± 0.39A
M 3 4.95 ± 0.05a 2.85 ± 0.06kl 2.25 ± 0.03n 4.95 ± 0.13a 3.30 ± 0.11ij 3.66 ± 0.30B
M 5 4.55 ± 0.12bc 1.00 ± 0.02q 3.75 ± 0.05gh 2.65 ± 0.08lm 2.20 ± 0.08n 2.83 ± 0.33C
M 6 0.30 ± 0.01s-v 4.05 ± 0.11efg 4.75 ± 0.05ab 0.65 ± 0.03r 1.65 ± 0.03o 2.28 ± 0.48F
M 7 0.60 ± 0.02rs 2.80 ± 0.05lm 0.50 ± 0.01rst 0.10 ± 0.01v 0.05 ± 0.01v 0.81 ± 0.27I B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure
In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison
among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
138
Table - 74: Means comparison for yield of pectin from sapodilla fruit peel after using 1N HCl (Mechanical procedure pH
interaction mean±SE)
pH level Mechanical procedure Mean
Homogenizing Grinding Cutting Chopping Hammering
1 3.25 ± 0.76ghi 3.63 ± 0.24fg 0.05 ± 0.00n 4.25 ± 0.56cd 2.90 ± 0.12hij 2.82 ± 0.33B
3 3.88 ± 0.66def 5.20 ± 0.19b 0.83 ± 0.35lm 4.55 ± 0.39c 6.10 ± 0.23a 4.11 ± 0.37A
5 3.28 ± 0.71gh 3.58 ± 1.13fg 4.13 ± 0.53cde 5.40 ± 0.59b 3.45 ± 0.52fg 3.97 ± 0.34A
6 1.05 ± 0.20l 2.50 ± 0.47j 1.83 ± 0.75k 3.78 ± 0.04ef 2.55 ± 0.19j 2.34 ± 0.24C
7 1.88 ± 0.30k 1.55 ± 0.54k 0.63 ± 0.26lm 0.48 ± 0.35mn 2.80 ± 0.14ij 1.47 ± 0.21D
Mean 2.67 ± 0.31C 3.29 ± 0.34B 1.49 ± 0.33D 3.69 ± 0.36A 3.56 ± 0.27A In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 75: Means comparison for yield of pectin from sapodilla fruit peel after using 1N HCl (Mechanical procedure
boiling method interaction mean±SE)
Boiling Mechanical procedure Mean
Method Homogenizing Grinding Cutting Chopping Hammering
B 1.49 ± 0.16f 2.31 ± 0.50e 0.65 ± 0.31g 2.92 ± 0.40d 3.05 ± 0.35d 2.08 ± 0.19B
M 3.84 ± 0.41c 4.27 ± 0.31ab 2.33 ± 0.50e 4.46 ± 0.55a 4.07 ± 0.37bc 3.79 ± 0.21A
B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
Table - 76: Means comparison for yield of pectin from sapodilla fruit peel after using 1N HCl (Mechanical procedure pH
boiling method interaction mean±SE)
BM pH Mechanical procedure BM pH
Homogenizing Grinding Cutting Chopping Hammering interaction mean
B 1 1.55 ± 0.05rst 3.10 ± 0.03i-m 0.05 ± 0.01w 3.00 ± 0.08j-n 2.65 ± 0.05l-o 2.07 ± 0.31F
139
B 3 2.40 ± 0.02m-p 5.60 ± 0.13bc 0.05 ± 0.01w 3.70 ± 0.10hij 5.65 ± 0.15bc 3.48 ± 0.56C
B 5 1.70 ± 0.03p-s 1.05 ± 0.02s-v 2.95 ± 0.04k-n 4.10 ± 0.09gh 2.30 ± 0.03n-q 2.42 ± 0.28E
B 6 0.60 ± 0.01uvw 1.45 ± 0.03rst 0.15 ± 0.01w 3.75 ± 0.05hi 2.15 ± 0.07o-r 1.62 ± 0.34G
B 7 1.20 ± 0.01stu 0.35 ± 0.01vw 0.05 ± 0.00w 0.05 ± 0.01w 2.50 ± 0.10l-o 0.83 ± 0.25H
M 1 4.95 ± 0.12cde 4.15 ± 0.10fgh 0.05 ± 0.00w 5.50 ± 0.10bcd 3.15 ± 0.08i-l 3.56 ± 0.52C
M 3 5.35 ± 0.12cd 4.80 ± 0.08d-g 1.60 ± 0.05q-t 5.40 ± 0.10bcd 6.55 ± 0.20a 4.74 ± 0.45B
M 5 4.85 ± 0.13def 6.10 ± 0.12ab 5.30 ± 0.10cde 6.70 ± 0.18a 4.60 ± 0.12efg 5.51 ± 0.22A
M 6 1.50 ± 0.03rst 3.55 ± 0.09h-k 3.50 ± 0.09h-k 3.80 ± 0.08hi 2.95 ± 0.10k-n 3.06 ± 0.22D
M 7 2.55 ± 0.09l-o 2.75 ± 0.05l-o 1.20 ± 0.03stu 0.92 ± 0.66tuv 3.10 ± 0.06i-m 2.10 ± 0.26F
B= Boiling on burner , M= heating in microwave; BM= Boiling method. MP= Mechanical procedure In a row and column statistically non-significant (P>0.05) represents mean sharing and are denoted by similar letters. Comparison among interaction means are represented by lower case alphabets while capital alphabets are used for overall mean
140
Table - 77: Identification tests for the presence of pectin from three selected fruits
Test Name Banana Sapodilla Muskmelon
Stiff gel test - + -
Test with 95% Ethanol + + Not clear
Test with Potassium Hydroxide (KOH) + + +
Iodine test + + +
Table - 78: Biochemical Characterization of the purified pectin extracted from
sapodilla pectin at different pH
Test Name Sapodilla
( pH5)
Sapodilla
(pH3)
Sapodilla
(pH1) FGP*
Quantitative test for ammonia. No
ammonia
No
ammonia
No
ammonia No ammonia
Moisture (%) 5.29 6.02 5.52 7.02
Ash (%) 5.11 4.3 4.89 1.16
Equivalent weight 1700 2680 2290 1271
Methoxyl Content (%) 5.1 4.4 4.9 8.16
Anhydrouronic Acid. (%) 39.33 31.78 34.56 70.50
Jelly Grade 100 99 98 150
Galacturonic Acid Content (%) 77.7 65.8 60.5 76.19
Protien 1.71 3.36 4.04
Degree of esterificatoion (%) 73.63 72.1 70.2 76.41
*FGP = Food grade pectin
141
Table -79: Water holding, water binding and fat binding capacity of sapodilla pectin
Test Name Sapodilla peel pectin Apple
pectin
Orange
pectin
Fibers
Apple Orange Grape
Fruit
WBC g/g 7.6 - - 1.62 1.65 2.09
WHC g/g 6.99 16.51 28.07 - - -
FBC g/g 2.2 - - 0.95 1.81 1.52
Table – 80: Showing FTIR spectral values of sample and standard pectin along with
associated functional groups.
Functional groups FOOD
GRADE
Sapodilla
(pH5)
Sapodilla
(pH3)
Sapodilla
(pH1)
O-H stretching 3319.2 3287.3 3271.1 3289.2
C-H stretching , symmetric,
asymmetric 2932.4 2923.6 2929.6 2930.2
C=O esterified 1733.3 1735.6 1738.6 1737.8
COO- asymmetric stretching
1622.2 1611.2 1612.2
COO- symmetric stretching 1426.4 1424.3 1423.3 1425.8
C-H bending 1335.4 1362.0 1363.0
C=O stretching 1234.9 1234.1 1234.5 1228.6
142
Figure – 23: FTIR spectra of food grade pectin
Figure – 24: FTIR spectra of sapodilla peel pectin extracted at pH 5
403.
3
467.
1
523.
2
679.
9
858.
2
909.
2
995.
2
1052
.6
1234
.9
1273
.913
35.4
1426
.4
1733
.3
1996
.0
2068
.9
2349
.0
2932
.4
3319
.2
3554
.1
*Pectin f ood grade
62
64
66
68
70
72
74
76
78
80
82
84
86
88
90
92
94
96
98
100
102
%T
500 1000 1500 2000 2500 3000 3500 4000
Wavenumbers (cm-1)
143
Figure – 25: FTIR spectra of sapodilla peel pectin extracted at pH 3
Figure – 26: FTIR spectra of Sapodilla peel pectin extracted at pH 1
DLS studies
Table – 81: Summary the physical characterization of standard pectin and the
different pectin extracted from various source and at different pH by DLS
144
Sample Extraction at
pH
Time of
extraction RH (nm)
PD (%)
(Đ)
Standard -- -- 73.7-1979.92 8.6-37.9
Apple 5.0 10 94.37-1357 18.4-47.8
Orange 5.0 10 473.07-482.92 36.8-36.9
Sapodilla 3.0 10 464.37-548.53 29.3-30.1
Sapodilla 5.0 10 390.21-421.17 28.1-29.3
RH= Radius of hydration ; PD = polydispersity
145
Standard
Figure – 27: Physical characterization of standard pectin by DLS. (A) DLS results of
standard pectin illustrating the experimental conditions i.e., the mean autocorrelation
function (a), monodispersity and radius plot (b-c), respectively. (B)Comparative
corresponding radius distribution of pectin standard (a) and standard pectin (b). All
experiments were performed with an auto–piloted run of 50 measurements at every 20 s,
with a wait time 1 s (at 25 °C).
Figure- 28: Physical characterization of apple pectin extracted at pH5.0. by DLS. (A)
DLS results of apple pectin extracted at pH5.0. Illustrating the experimental conditions
i.e., the mean autocorrelation function (a), monodispersity and radius plot (b-c),
respectively. (B) Comparative corresponding radius distribution of pectin standard (a)
and apple pectin extracted at pH5.0 (b). All experiments were performed with an auto–
piloted run of 50 measurements at every 20 s, with a wait time 1 s (at 25 °C).
146
Figure – 29: Physical characterization of Orange pectin extracted at pH5.0. by DLS. (A)
DLS results of Orange pectin extracted at pH5.0. Illustrating the experimental conditions
i.e., the mean autocorrelation function (a), monodispersity and radius plot (b-c),
respectively. (B) Comparative corresponding radius distribution of pectin standard (a)
and Orange pectin extracted at pH5.0 (b). All experiments were performed with an auto–
piloted run of 50 measurements at every 20 s, with a wait time 1 s (at 25 °C).
147
Figure – 30: Physical characterization of sapodilla pectin extracted at pH3.0. by DLS.
(A) DLS results of sapodilla pectin extracted at pH3.0. illustrating the experimental
conditions i.e., the mean autocorrelation function (a), monodispersity and radius plot (b-
c), respectively. (B) Comparative corresponding radius distribution of pectin standard (a)
and sapodilla pectin extracted at pH3.0 (b). All experiments were performed with an
auto–piloted run of 50 measurements at every 20 s, with a wait time 1 s (at 25 °C).
Figure – 31: Physical characterization of sapodilla pectin extracted at pH5.0. by DLS.
(A) DLS results of sapodilla pectin extracted at pH5.0. illustrating the experimental
conditions i.e., the mean autocorrelation function (a), monodispersity and radius plot (b-
c), respectively. (B) Comparative corresponding radius distribution of pectin standard (a)
and sapodilla pectin extracted at pH5.0 (b). All experiments were performed with an
148
auto–piloted run of 50 measurements at every 20 s, with a wait time 1 s (at 25 °C). Also
see Table 5 for details.
Response surface methodology
Table - 82: Box-Bechen experimental design and levels of factors used for
optimization of pectin yield
Variables Symbol Low High
pH X1 1 5
Temperature(oC) X2 50 100
Time ( minutes) X3 10 90
Table - 83: Box-Behnken experimental design and corresponding results for
responses
Std Order Run Order Pt Type Blocks pH Temperature Time Yield
10 1 2 1 3 100 10 1.6
3 2 2 1 1 100 50 1.7
14 3 0 1 3 75 50 1.5
1 4 2 1 1 50 50 1.6
2 5 2 1 5 50 50 3.5
6 6 2 1 5 75 10 2.6
9 7 2 1 3 50 10 1.5
12 8 2 1 3 100 90 1.65
4 9 2 1 5 100 50 2.5
8 10 2 1 5 75 90 3.5
11 11 2 1 3 50 90 1.4
13 12 0 1 3 75 50 2
5 13 2 1 1 75 10 3
15 14 0 1 3 75 50 1.8
149
7 15 2 1 1 75 90 1.5
Table - 84: Analysis of Variance for Yield (Response surface methodology)
Source DF Adj SS Adj MS F-value Prob.
Model
Linear
pH
Temperature
Time
Square
pH*pH
Temperature*Temperature
Time*Time
2-Way Interaction
pH*Temperature
pH*Time
Temperature*Time
Error
Lack-of-Fit
Pure Error
Total
9
3
1
1
1
3
1
1
1
3
1
1
1
5
3
2
14
7.15996
2.40188
2.31125
0.03781
0.05281
3.00996
2.57694
0.28348
0.00848
1.74813
0.30250
1.44000
0.00563
0.51104
0.38438
0.12667
7.67100
0.79555
0.80063
2.31125
0.03781
0.05281
1.00332
2.57694
0.28348
0.00848
0.58271
0.30250
1.44000
0.00563
0.10221
0.12813
0.06333
7.78*
7.83*
22.61**
0.37
0.52
9.82*
25.21**
2.77
0.08
5.70*
2.96
14.09*
0.06
2.02
0.018
0.025
0.005
0.570
0.504
0.015
0.004
0.157
0.785
0.045
0.146
0.013
0.824
0.348
* = Significant (P<0.05); ** = Highly significant (P<0.01)
S = 0.3197
R² = 93.34%
R²(adj) = 81.35%
R²(pred) = 16.11%
150
Table - 85: Estimated Regression Coefficients for Yield (Box-Bechen experimental
design, response surface methodology)
Term Effect Coef. SE(Coef.) t-value Prob. VIF
Constant
pH
Temperature
Time
pH*pH
Temperature*Temperature
Time*Time
pH*Temperature
pH*Time
Temperature*Time
1.075
-0.137
-0.163
1.671
-0.554
0.096
-0.550
1.200
0.075
1.767
0.538
-0.069
-0.081
0.835
-0.277
0.048
-0.275
0.600
0.038
0.185
0.113
0.113
0.113
0.166
0.166
0.166
0.160
0.160
0.160
9.57**
4.76**
-0.61
-0.72
5.02**
-1.67
0.29
-1.72
3.75*
0.23
0.000
0.005
0.570
0.504
0.004
0.157
0.785
0.146
0.013
0.824
1.00
1.00
1.00
1.01
1.01
1.01
1.00
1.00
1.00
* = Significant (P<0.05); ** = Highly significant (P<0.01)
Yield = 0.76 - 0.947 pH + 0.0784 Temperature - 0.0303 Time + 0.2089 pH² -
0.000443 Temperature² + 0.000030 Time² - 0.00550 pH*Temperature
+ 0.00750 pH*Time + 0.000037 Temperature*Time
Table - 86: Predicted values of yield. (Box-Bechen experimental design, response
surface methodology)
Yield Composite
Solution pH Temperature Time Fit Desirability
1 5 61.1111 90 3.79087 1.00000
2 5 85.2577 86.8718 3.48497 0.99284
3 5 67.7177 17.6639 2.83699 0.68428
151
4 5 67.7177 17.6639 2.83699 0.68428
5 1 90.6592 10 2.79074 0.66226
Figure – 32: Showing the optimal conditions for the extraction of pectin from
sapodilla fruit peel (Box-Bechen experimental design, response surface
methodology)
152
Figure – 33: Response surface graph and contour plot of effect of pH and
temperature on yield of pectin at constant time
Yield = -0.6969-0.5663*x+0.0811*y+0.2079*x*x-0.0055*x*y-0.0004*y*y
> 3.5 < 3.5 < 3 < 2.5 < 2 < 1.5
Yield = Distance Weighted Least Squares
> 4 < 4 < 3.5 < 3 < 2.5 < 2 < 1.5 < 1 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5
pH
40
50
60
70
80
90
100
110
Tem
pe
ratu
re
153
Figure- 34: Response surface graph and contour plot of effect of pH and time on
yield of pectin at constant temperature
Yield = 4.0523-1.3913*x-0.0289*y+0.2142*x*x+0.0075*x*y+4.3269E-5*y*y
> 4.5 < 4.5 < 4 < 3.5 < 3 < 2.5 < 2 < 1.5
Yield = Distance Weighted Least Squares
> 4 < 4 < 3.5 < 3 < 2.5 < 2 < 1.5 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5
pH
0
10
20
30
40
50
60
70
80
90
100
Tim
e
154
Figure – 35: Response surface graph and contour plot of effect of temperature and
time on yield of pectin at constant pH
Yield = -0.3684+0.0773*x-0.0038*y-0.0005*x*x+3.75E-5*x*y-1.0216E-5*y*y
> 2.2 < 2.2 < 2 < 1.8 < 1.6
Yield = Distance Weighted Least Squares
> 2.5 < 2.5 < 2 < 1.5 < 1 < 0.5 40 50 60 70 80 90 100 110
Temperature
0
10
20
30
40
50
60
70
80
90
100
Tim
e
155
Table – 87: Flow properties of granules made for paracetamol tablets
Test
Formulatio
n
Mass
(g)
Bulk
Volume
(ml)
Tapped
Volume
(ml)
Bulk
Density
(g/ml)
Tapped
Density
(g/ml)
Angle of
Repose
(θ⁻¹)
Compressibilit
y Index
(%)
Hausner
Ratio
-
Standard 2.004±0.00
1
4.5068±0.11
5
4.123±0.12
5
0.446±0.05
1
0.487±0.05
7
17.955±1.15
8 8.41±0.507
1.091±0.00
6
F1 2.010±0.00
5 3.867±0.115
3.067±0.11
5
0.520±0.01
4
0.656±0.02
3
18.747±0.54
3 20.702±0.608
1.261±0.01
0
F2 2.003±0.00
1 3.867±0.115
3.067±0.11
5
0.518±0.01
5
0.654±0.02
4
16.494±1.68
0 20.702±0.608
1.261±0.01
0
F3 2.008±0.00
1 4.067±0.115
3.067±0.11
5
0.494±0.01
4
0.655±0.02
4
16.558±1.79
2 24.603±0.687
1.326±0.01
2
F4 2.008±0.00
1 4.333±0.115
4.133±0.11
5
0.464±0.01
2
0.486±0.01
4
17.545±1.54
0 4.618±0.125
1.048±0.00
1
F5 2.004±0.00
1 3.533±0.306
3.333±0.30
6
0.570±0.05
1
0.605±0.05
7
17.955±1.15
8 5.690±0.507
1.060±0.00
6
F6 2.005±0.00
1 4.133±0.115
3.667±0.46
2
0.485±0.01
4
0.552±0.06
5
17.453±0.17
1 11.032±13.884
1.141±0.16
3
F7 2.016±0.02
1 4.000±0.200
3.333±0.30
6
0.505±0.02
1
0.608±0.05
6
17.987±0.61
8 16.700±5.888
1.204±0.08
2
F8 2.020±0.01 4.067±0.115 3.067±0.11 0.497±0.01 0.659±0.02 21.546±0.44 24.603±0.687 1.326±0.01
156
0 5 4 5 1 2
F9 2.017±0.00
6 4.133±0.115
3.333±0.11
5
0.488±0.01
2
0.605±0.02
0
16.911±0.38
3 19.365±0.550
1.240±0.00
8
F1-F9 = test formulations 1 till 9 Each value is a Mean±SD of three determination
Table - 88: Pharmaceutical characteristics of compressed formulation of paracetamol tablet
Test Formulation
Pharmacopoeial Limits
(USP 32/NF 27)
Wt. Variation
(Mean ± S.D)
(mg)
±5%
Thickness
(Mean ±
S.D)
(mm)
±5%
Diameter
(Mean ± S.D)
(mm)
₋
Hardness
(Mean ± S.D)
(kg)
At least 5 kg
Loss on drying
%
Not more than 1.5%
Standard 702.67± 2.52 5.60±0.20 9.47±0.05 5.22±0.01 4.0%
F1 705.00±5.00 5.22±0.03 9.42±0.03 2.54±0.06 4.0%
F2 672.33±2.52 5.24±0.05 9.39±0.01 1.37±0.07 3.0%
F3 701.33±3.21 5.32±0.05 9.45±0.04 4.16±0.08 4.3%
F4 695±5.00 5.35±0.06 9.44±0.05 5.06±0.21 4.2%
F5 705±4.51 5.21±0.03 9.41±0.03 5.15±0.07 4.6%
F6 701.67±3.97 5.19±0.04 9.42±0.03 5.21±0.07 4.3%
F7 708.3±2.89 5.29±0.04 9.31±0.03 6.07±0.12 4.2%
F8 694.67±4.51 5.27±0.13 9.27±0.06 6.67±0.58 4.5%
157
F9 704.00±3.61 5.27±0.10 9.42±0.04 7.40±0.33 4.5%
F1-F9 = test formulations 1 till 9 Each value is a Mean±SD of three determination.
Table – 89: Dissolution studies of paracetamol tablet
FORMULATION NUMBER
F1 F2 F3 F4 F5 F6 F7 F8 F9
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
243nm
(Abs)
%
release
0.304 43.43 0.339 50.64 0.401 58.77 0.269 71.52 0.564 84.17 0.078 23.00 0.084 26.77 0.123 36.73 0.111 32.73
0.291 41.67 0.338 50.49 0.412 60.39 0.283 70.28 0.557 83.12 0.087 25.65 0.080 23.59 0.111 32.73 0.094 27.97
0.362 51.83 0.564 84.25 0.382 55.99 0.331 83.13 0.418 91.93 0.112 33.03 0.093 27.42 0.108 31.85 0.108 31.85
0.372 53.26 0.557 83.20 0.389 57.02 0.333 87.28 0.620 92.53 0.105 30.96 0.096 28.31 0.120 35.39 0.101 29.78
0.313 44.82 0.418 62.44 0.577 84.58 0.285 85.20 0.406 93.13 0.096 28.313 0.076 22.41 0.120 35.39 0.099 29.19
0.323 46.24 0.406 60.65 0.590 86.48 0.312 85.20 0.632 94.32 0.116 34.21 0.091 26.83 0.107 31.55 0.088 25.95
STANDARD FOR FORMULATION
0.692 99.10 0.664 99.10 0.676 99.10 0.478 99.10 0.664 99.10 0.336 99.10 0.336 99.10 0.336 99.10 0.336 99.10
F1-F9 are test formulation while Abs = absorbance and %release = percent release
158
159
F1 F2
F3
Figure -36: Formulated paracetamol tablest ( F1, F2, F3)
160
F4 F5
F6
Figure 37: Formulated paracetamol tablest ( F4, F5, F6)
161
F7 F8
F9
Figure 38: Formulated paracetamol tablest (F7, F8, F9)
162
Table - 90: Flow properties ofgranules for ibuprofen tablet
Test Formulation
Mass Bulk Volume Tapped Volume Bulk Density Tapped Density Angle of Repose Compressibility Index Hausner Ratio
(g) (ml) (ml) (g/ml) (g/ml) (θ⁻¹) (%) ₋
Std 2.02±0.013 4.25±0.015 3.75±0.092 0.475±0.002 0.538±0.020 11.71±0.210 11.71±0.116 1.132±0.061
R1 2.02±0.013 4.20±0.015 3.76±0.093 0.49±0.003 0.55±0.019 15.23±0.208 4.82±0.115 1.13±0.061
R2 2.01±0.016 4.35±0.042 3.61±0.010 0.46±0.005 0.56±0.004 13.83±0.153 9.74±0.344 1.24±0.059
R3 2.01±0.007 4.60±0.080 3.93±0.064 0.42±0.021 0.52±0.010 13.73±0.306 7.40±0.352 1.17±0.028
R4 2.01±0.004 4.51±0.090 4.15±0.050 0.44±0.002 0.47±0.003 11.13±0.153 3.77±0.252 1.09±0.009
R1= Test formulation one, R2= Test formulation two, R3= Test formulation three, R4= Test formulation four. Each value is a Mean±SD of three determination
Table - 91: Pharmaceutical characteristics of compressed formulation of ibuprofen tablet
Test Formulation
Pharmacopoeial Limits
(USP 32/NF 27)
Wt. Variation
(Mean ± S.D)
(mg) ±5%
Thickness
(Mean ± S.D)
(mm) ±5%
Length x width
(Mean ± S.D)
(mm) ₋
Hardness
(Mean ± S.D)
(kg) At least 5 kg
Loss on
Drying %
Standard 833.33±2.08 6.33± 0.06 20 x 9.5 6.13± 0.14 4.5%
R1 843.33±2.08 6.23± 0.06 20 x 9.5 6.23± 0.14 4.9%
R2 848.33±2.89 6.37±0.06 20 x 9.5 8.40±0.40 4.8%
R3 847.67±2.52 6.22±0.03 20 x 9.5 9.23±0.14 4.9%
R4 842.67±2.52 6.27±0.06 20 x 9.5 10.43±0.18 4.7%
Table- 92: dissolution studies of ibuprufen tablets
163
FORMULATION NUMBER
R1 R2 R3 R4
243nm (Abs) % release 243nm (Abs) % release 243nm (Abs) % release 243nm (Abs) % release
0.405 89.6 0.166 36.72 0.092 33.45 -0.018 -6.54
0.415 91.81 0.141 31.190 0.090 32.72 -0.017 -6.182
0.420 92.91 0.147 32.51 0.112 40.72 -0.020 -7.27
0.425 94.02 0.154 34.06 0.113 41.92 0.008 -2.90
0.429 94.95 0.143 31.62 0.030 10.90 -0.008 -2.90
0.430 95.13 0.161 35.60 0.036 13.091 -0.008 -2.90
STANDARD FOR FORMULATION
0.503 0.503 111.33 0.306 111.33 0.306 111.33
R1-R4 are test formulation while Abs = absorbance and %release = percent release
164
R1 R2
R3 R4
Figure – 39: Formulated ipubrufen tablets ( R1, R2, R3, R4)
165
Table -93: Basic evaluation test of antidiarrheal formulation prepared from
sapodilla pectin
Parameters Suspension made from
sapodilla pectin Comparative suspension *
Color Pinkish white White
Odor vanilla Vanilla
Taste sweet sweet
pH 6.1 5.56
Viscosity 14.14 13.13
Sedimentation rate 0.3 0.1
Redispersity +++ +++
WHC 32.69 33.56
+ denotes the number of times the cylinder was moved. * Keptin antidiarrheal preparation
166
Figure- 40 Formulated antidiarrheal preparation
167
Table - 94: Effect of different concentration of sapodilla pectin on the chemical
properties of the jam samples
Sample PH
TSS
(mg)
TTA
(ml)
MC
(%)
Vit C
(mg)
Ash
(%)
TS
(mg)
Viscosity
(Cp)
S 3.06 61.9 1.14 35.9 18.3 1.78 64.1 71
F1 3.16 56.7 1.08 41 17.6 1.69 59 56
F2 3.25 62.6 1.01 35.7 18.1 1.73 64.3 63
F3 3.25 62.4 1.03 35.1 17.9 1.71 64.9 67
WhereS = Jam with standard pectin, F1 = Jam with 2g pectin, F2 = Jam with pectin in same concentration
as in standard, F3 = Jam with 10g pectin while MC is moisture content . TSS total soluble solids, TTA
total titratable acid and TS is total solid.
Table - 95: Scores for sensory parameters as judged by twenty (20) panelists.
Sampl
e
Appearanc
e
Tast
e
Arom
a
Spreadabilit
y
Textur
e
Mout
h feel
Overall
acceptabilit
y
S 7.7 7.9 7.9 8.2 7.8 7.5 7.8
F1 7.5 7.3 7.2 7.3 7.2 7 7.2
F2 7.5 7.6 7.7 7.8 7.8 7.5 7.6
F3 6.2 6.4 6.2 5.6 6.3 5.5 6
WhereS = Jam with standard pectin, F1 = Jam with 2g pectin, F2 = Jam with pectin in same concentration as in standard, F3 = Jam with 10g pectin
168
(S)=Jam made from 5g standard pectin (F1) Jam made from 5g
pectin
(F2) Jam made from 7 g pectin (F3) Jam made from 10g
pectin
Figure -41 formulation of Jam made from extracted pectin
169
Figure -42: formulation of pudding made from extracted pectin.
170
DISCUSSION
Appropriate utilization of waste products after fruit processing is one of the most
propitious field which needs comprehensive research, evaluation and interpretation by the
scientists of related discipline. Surprisingly, wastes from food processing industries are
among the largest mass produced all over the world as a result of pre-consumer
procedures. It is sometimes around 50% of the total processing waste and are very
difficult to manage for further developments because of improper utilization techniques,
available resources, cost and regulatory issues. Making useful functional compounds
from fruit wastes and their proper utilization can be beneficial economically and will also
help in reducing risk to the changing global environment. Initial studies on pectin has
been reported in some articles published in 1750 with the utilization to develop food
products specially apple jelly (Kertesz, 1951). Although extraction and identification of
crude pectin has been reported after this study, however the first commercial extraction
process started in Germany in 1908 and subsequently the process was granted a patent
(U.S. Patent no. 1,082.682) in 1913 (Canteri, 2012). This has triggered various
researchers as well as food industries to develop new and improved method of extractions
of pectin to obtain considerable quantities of pectin to utilize in developing various food
and pharmaceutical products. The commercial production of pectin and its utilization
needs proper handling selection of fruits and processing of waste products to achieve the
desired results. This demands a very comprehensive and dedicated research, evaluation
and interpretation of the results by the scientist working in the particular field. In the
initial phase most studies on pectin production and its utilization have been reported in
western world because of the resources and techniques available with them. However
171
with the passage of time this has moved to under developed or developing countries and
as a result a large number of people have been noted to be engaged in such studies to
develop an economical extraction procedure for pectin. The research in this field is still
continued and despite huge commercial production by some of the well reputed
companies (such as Copenhagen pectin, Denisco pectin Mexico, Sanofi bio- industries
and Cargill and CP Kelco ) researchers are still trying to find new methods modified
resources and optimization of extraction conditions to increase pectin yield.
Pakistan is primarily classified under agricultural based country where huge quantity of
fruits are produced and sold in the market. However the processing industries are lacking
which results in the commercial production of various food products with the utilization
of pectin. Every year considerable quantity of fruits are spoiled and discarded due to
improper handling and non-availability of basic manufacturing units which can utilize
these fruits and their waste to convert into valuable products. Ultimately Pakistan is
mostly dependent upon the imports of such basic raw materials including pectin to cater
its food and pharmaceutical industries to utilize and develop products for commercial and
therapeutic applications consequently a huge quantity of pectin is imported in Pakistan
against the expenditure of valuable foreign exchange currency. The June 2015 to October
2016 import figures were 1,120,029.15US $ import of pectin and most important
exporting countries include Brazil, China, Czech Republic, Germany, Denmark
and Spain. With this aim and objective present studies were organized and conducted to
evaluate an economical and easily available source of fruit for developing process to
extract pectin and its utilization in some food and pharmaceutical formulations. The
integral part in the production of high quality pectin is based on the method of extraction
172
used to recover pectin from the desired source. The selection of fruits as well as the
nature of waste and the experimental design used for the extraction of pectin greatly
affect the physico-mechanical properties and yield. Recent studies indicated successful
recovery of pectin from the lime biomass using enzyme cellulose obtained from
Penicillium funiculosum and designated as Laminex CK2 (Dominiak et al., 2014). The
current study focus on a number of methods which may affect pectin yield, were studied
and repeated as a new system in order to acquire an efficient extraction procedure from
fruits wastes. Method for extraction and evaluation also moves around various physico-
mechanical factors for optimizing the yield of pectin from a new source which were not
reported earlier from this part of the world and also its utilization in pharmaceutical
preparations and as a thickening agent in food products.
The different sources investigated in this study are mentioned in Table - 9 with their
respective percent yield of pectin. During the preliminary study acid extraction was
mainly used because it generally results to acquire a good yield of pectin. Hydrochloric
acid for pH 3-5, ammonium hydroxide for 6 and above with conventional boiling
methods (Bunsen burner) and 10 min of boiling was applied for the extraction process.
The pH of the medium varied with the origin of the material used for pectin extraction
(May, 1990). The study conducted exibited a variable response in terms of yield and were
compared with reported values. Orange, apple, banana and lemon showed yield similar to
the range presented in previous studies ( Aina et al., 2012 ; Munarin et al., 2012 ) while
grapefruit (Munarin et al., 2012), guava (Bhat and Singh, 2014), sweet lime (Munarin et
al., 2012), watermelon (Rasheed, 2008) and mango (Munarin et al., 2012) presented
lesser yield, while there were few fruits which failed to produce any results, although a
173
considerable amount of pectin was reported from them in earlier studies. The fruits were
peach (Munarin et al., 2012) and apricot (Baissise et al., 2010). Few fruits showed a
good quantity of pectin, although there was no previous study found on them, the fruits
were sapodilla, muskmelon and cantaloupe. Moreover, there were fruits which were
unable to present any resulted pectin and no previous study was found on them such
as guava (unripe), jungle fruit and mango (unripe). The difference in yield of pectin from
the screened fruits from their past studies was due to the fact that that the
physicochemical properties of pectin depend upon the breed, characteristic features and
atmospheric conditions during growth of the fruit and fruit sources (El-Nawawi and
Shehata, 1987; Chan and Choo., 2013).
Following the preliminary screening, three fruits were selected for further evaluation
while two already explored fruits; apple and orange were studied for their comparing
effects. The extraction conditions used during the isolation of pectin was designed in a
way that all the preliminary yield-affecting variables of pectin were studied very
carefully. pH, boiling method and mechanical methods are factors which can affect the
yield from fruit as described earlier. The extraction regime selected for the study was pH
(1 to 7), boiling method (Bunsen burner and microwave), and mechanical procedures
(homogenising, grinding, chopping, cutting and hammering). The overall objective was
to explore the fruits for better yields of pectin after applying different phyicomechanical
conditions which later can identify a single source for further evaluation to enhance yield
of pectin from it. The data was also analyzed statistically to study the significant impact
of all the variables (pH, boiling method and mechanical method) used during the study.
Table -10 shows the yield of pectin achieved from five selected fruits (sapodilla, banana,
174
muskmelon, apple and orange) and indicates difference in yield after applying different
combination of variables. Statistically analyzed by Analysis of variance (ANOVA),
results indicated that different variables like mechanical procedure (MP), pH and boiling
method (BM) have significant impact on the yield of all five fruits at 1% level of
significance (Table - 11 ). The difference among the means regarding different factors
during evaluation of all five fruits was also determined by Tukey’s test and are shown
under respective tables of investigated fruit.
The yields recorded from different fruits proved to be dependent upon their natural pectin
content and efficiency of the process used and are showed in Table- 10. Previous study
showed that sapodilla fruit was investigated for total dietary fibers and pectin contents
(0.35g ± 0.01/100g fruit) in its edible portion (Mahattanatawee et al., 2006). However, in
the present study, pectin was extracted from peel of sapodilla fruit and was best extracted
(4% yield) through grinding at pH 3, using conventional heating method. A further
increase in yield was observed (4.7%) at lower pH (pH 1) when heating mode was
changed to microwave (Table-10), with the indication that intensity of heating has more
impact on pectin yield from sapodilla on that specific pH. The difference among the
means regarding different factors using Tukey’s test are given in Table – 12-14 for
sapodilla peel. The overall mean for mechanical procedure, pH and boiling method
recommended chopping, pH 3 (Table - 12) and microwave boiling (Table -13) has a
significant impact (P<0.05) on the yield of pectin from sapodilla fruit. Table 12 and 13
also show the status of remaining mechanical procedure, pH and boiling method and their
order of influence is given from most to least effective. Alphabet A was considered the
most and subsequent alphabets lesser in effectiveness than the preceding alphabet.
175
Overall interaction effect of boiling method with pH (BM x pH) on sapodilla peel showed
(Table - 14) that M x 3 and M x 5 (Heating using microwave at pH 3 and 5) were
significantly non-significant with each other, giving significantly (P<0.05) highest yield
while yield at B x 1 ( Heating using Bunsen burner at pH1) was the lowest.
Table- 10 also showed the comparison of yields form banana fruit peel. The microwave
heating was also noted as a better option to obtain good yield using all mechanical
methods. The banana peels gave good yield of pectin and was observed to be very much
close to the yield of pectin as reported in an earlier study (Christy et al., 2014).
Homogenizing was the best mechanical procedure, as it gave the maximum yield (8.85%
yield) at pH 5, using conventional method. For microwave heating extraction, hammering
was noted as the best method (10.5% yield) for banana peel at the same pH (pH 5). The
difference among the means regarding different factors using Tukey’s test are given in
Table – 15-17 for banana peel. The overall mean for mechanical procedure, pH and
boiling method recommended homogenising, pH 5 (Table - 15) and microwave boiling
(Table - 16) has a significant impact (P<0.05) on the yield of pectin from banana fruit.
Table 15 and 16 also show the status of remaining mechanical procedure, pH and boiling
method and their order of influence is given from most to least effective. Alphabet A was
considered the most and subsequent alphabets lesser in effectiveness than the preceding
alphabet. The Overall interaction effect of boiling method and pH (BM X pH) on the
yield of banana showed that M x 5 (Heating using microwave at pH 5) gave highest yield
and significantly (P<0.05) different from other levels of BM x pH. Yield at B x 1
(Heating using Bunsen burner) was lowest which was statistically significantly different
from all other levels (Table -16).
176
Muskmelon was also studied previously for total dietary fibers (Mahattanatawee et
al., 2006). For muskmelon peels, pH 3 using conventional heating and cutting as
mechanical process was noted as the best option to achieve high yield (2.45%) of pectin.
However, using microwave technique, extraction after grinding as mechanical process at
pH 1 provided best yield (2.65%) (Table-10). The difference among the means regarding
different factors using Tukey’s test are given in Table – 18 - 20 for muskmelon peel. The
overall mean for mechanical procedure, pH and boiling method recommended grinding,
pH 5 (Table 18) and microwave boiling (Table 19) has a significant impact (P<0.05) on
the yield of pectin from muskmelon fruit. Table 18 and 19 also show the status of
remaining mechanical procedure, pH and boiling method and their order of influence is
given from most to least effective. Alphabet A was considered the most and subsequent
alphabets lesser in effectiveness than the preceding alphabet. Overall interaction effect of
BM x pH showed that M x 5 (Heating using microwave at pH 5) gave highest yield and
significantly (P<0.05) different from other levels of BM x pH while yield at B x 7
(Heating using Bunsen burner at pH1) was lowest (Table-20).
Although literature contains the amount of pectin that can be extracted from apple
pomace (10 to 15%) (Ziari et al., 2010), it was not comparable with the present study,
since to synchronize the study, apple peels, instead of pomace was used. For apple peels
the best mechanical method was found hammering for both the heating methods at pH 5
(3.05% to 4.85% yield) Table – 10. The difference among the means regarding different
factors using Tukey’s test are given in Table – 21 -23 for apple peel. The overall mean
for mechanical procedure, pH and boiling method recommended cutting and hammering,
pH 3 and 5 (Table - 21) and microwave boiling (Table - 22) has a significant impact
177
(P<0.05) on the yield of pectin from apple fruit. Table - 21 and 22 also show the status of
remaining mechanical procedure, pH and boiling method and their order of influence is
given from most to least effective. Alphabet A was considered the most and subsequent
alphabets lesser in effectiveness than the preceding alphabet. Overall interaction effect of
boiling method and pH (BM x pH) showed that M x 3 and M x 5 (Heating using
microwave at pH 3 and 5) gave highest yield and significantly (P<0.05) different from
other levels of BM x pH while yield at B x 7 ( Heating using Bunsen burner at pH7) was
lowest Table – 23.
The extracted pectin yield from orange peels was also comparable with previously
extracted pectin (Sayah et al., 2014). For orange peels grinding was found producing
highest yield at pH=7 in conventional method (21.7 %) and slightly higher yield with
microwave method (22.7%) at lower pH 5 (Table -10). The difference among the means
regarding different factors using Tukey’s test are given in Table – 24-26 for orange peel.
The overall mean for mechanical procedure, pH and boiling method recommended
homogenising, pH 1 (Table - 24) and microwave boiling (Table - 25) has a significant
impact (P<0.05) on the yield of pectin from orange fruit. Table 24 and 25 also show the
status of remaining mechanical procedure, pH and boiling method and their order of
influence is given from most to least effective. Alphabet A was considered the most and
subsequent alphabets lesser in effectiveness than the preceding alphabet The overall
interaction effect of boiling method and pH (BM x pH) showed that M x 1 (Heating using
microwave at pH 1) gave highest yield and significantly (P<0.05) different from other
levels of BM x pH while yield at M x 7 (Heating using Bunsen burner at pH7) was lowest
Table – 26.
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In earlier studies, the effect of particle size reduction was also reported to have significant
impact, based on mechanical procedure for yield of pectin. This is because reduction in
particle size enhances protopectin release which ultimately increases the yield of pectin
when precipitated with ethanol (Canteri-Schemin et al., 2005). Likewise, pH of the
extracting medium also affects significantly on the yield of pectin (Ziari et al., 2010). In
the present study the effect of interaction between variable combination such as
mechanical procedure with pH, mechanical procedure with boiling methods, pH with
boiling methods and also the interaction of all mechanical procedures, pH and boiling
method on yield of all five fruits were also investigated. Results highlighted in Table -8
showed that interactions of all have positive impact on the yield of pectin from all five
fruits.
Preliminary investigation pointed towards a new source sapodilla fruit peel from which
extraction of pectin has not been investigated to date. Sapodilla ranked top among the
high pulp fruit with about 85% as edible portion. But the fruit has been neglected and did
not achieve the position it deserved. The current research can highlight one of the many
advantages of this special fruit grown in large areas of Pakistan and rest of the world. The
extraction of pectin from sapodilla fruit peel were conducted on the aforementioned
variables used for the study of five selected fruits (Table-8) with the exception that time
of boiling was added ( 10, 20, 40 and 60 min). The objective of adding time of boiling
was to evaluate the best condition for maximum yield that could be employed in the
commercialization of sapodilla peel pectin. The added extraction technique was applied
after using two strengths of inorganic acid (0.1N and 1N HCl).
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Time of boiling has a profound effect on pectin yield which was studied among the other
affecting variable on yield by different scientists (Kliemann et al 2009, Khan et al
2015), while the mode of boiling also has a positive effect (Guo et al 2012). The highest
yield (6.5%) obtained at pH 3, chopping as mechanical procedure after 20 min of boiling
in Microwave (Table- 27). As reported earlier, the extraction results obtained after
3 hrs of conventional heating were the same with yield obtained by 15 min of extraction
in microwave (Salam et al., 2012). The maximum yield obtained by conventional heating
was 5.5% but at pH 5 and hammering as mechanical procedure (Table – 27). To
understand the overall effects of each variable, the data was also analyzed statistically by
the same software and procedure as used in aforestated five fruit analysis. Analysis of
variance (ANOVA) in the Table - 28 shows that the mechanical procedures have a
significant (P<0.01) impact on yield on all time of boiling selected for the study.
Similarly, pH level and Boiling method also significantly (P<0.01) impact on yield when
using different times of boiling. All the two way interactions i.e. mechanical procedure
and pH, (MPxpH), mechanical procedure and boiling method (MPxBM), pH and boiling
method (pHxBM) and three factor interaction of mechanical procedure, pH and boiling
method MPxpHxBM showed significant (P<0.01) impact on yield. Table 29- 31 shows
the overall mean comparisons and their effect after 10min of boiling.
The difference among the means regarding different factors using Tukey’s test are given
in Table – 29- 31 for 10 min of boiling. The overall mean for mechanical procedure, pH
and boiling method showed chopping, pH 3 (Table - 29) and microwave boiling (Table -
30) has a significant impact (P<0.05) on the yield of pectin after 10 min of boiling of
peels. Table 29 and 30 also show the status of remaining mechanical procedure, pH and
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boiling method and their order of response is given from most to least effective. Alphabet
A was considered the most and subsequent alphabets lesser in effectiveness than the
preceding alphabet. Overall interaction effect in Table – 31 shows interaction of BM x
pH showed that M x 3 and M x 5 (Heating in microwave at pH 3and 5) gave highest yield
and significantly (P<0.05) different from other levels of BM x pH while yield at B x 1
and Bx7 (Heating on Bunsen burner) was lowest.
For 20 min of boiling the difference among the means regarding different factors using
Tukey’s test are given in Table – 32-34. The overall mean for mechanical procedure, pH
and boiling method showed homogenising pH 3 (Table - 32) and microwave boiling
(Table - 33) has a significant impact (P<0.05) on the yield of pectin after 20 min boiling
of peels. Table-32 and 33 also show the status of remaining mechanical procedure, pH
and boiling method and their order of response is given from most to least effective.
Alphabet A was considered the most and subsequent alphabets lesser in effectiveness
than the preceding alphabet. Overall interaction effect in Table – 34 shows interaction of
BM x pH showed that M x 3 and M x 5 (Heating in microwave at pH 3and 5) gave
highest yield and significantly (P<0.05) different from other levels of BM x pH while
yield at B x 1 and Bx7 (Boiling on Bunsen burner at pH 1 and 7) was lowest.
The difference among the means regarding different factors using Tukey’s test are given
in Table – 35-37 for 40 min of boiling. The overall mean for mechanical procedure, pH
and boiling method showed hammering pH 5 and 6 (Table 35) and microwave boiling
(Table 36) has a significant impact (P<0.05) on the yield of pectin after 40 min of boiling
of peels. Table-35 and 36 also show the status of remaining mechanical procedure, pH
and boiling method and their order of response is given from most to least effective.
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Alphabet A was considered the most and subsequent alphabets lesser in effectiveness
than the preceding alphabet. Overall interaction effect in Table – 37 shows interaction of
BM x pH showed M x 5 (Heating on microwave at pH 5) gave highest yield and
significantly (P<0.05) different from other levels of BM x pH while yield at B x 1
(Heating on Bunsen burner at pH1) was lowest.
The difference among the means regarding different factors using Tukey’s test are given
in Table – 38- 40 for 60 min of boiling. The overall mean for mechanical procedure, pH
and boiling method showed chopping and homogenising, pH 5 (Table-38) and
microwave boiling (Table-39) has a significant impact (P<0.05) on the yield of pectin
after 60 min of boiling of peels. Table-38 and 39 also show the status of remaining
mechanical procedure, pH and boiling method and their order of response is given from
most to least effective. Alphabet A was considered the most and subsequent alphabets
lesser in effectiveness than the preceding alphabet. Overall interaction effect of BM x pH
level showed that M x 5 (Heating on microwave at pH 5) gave highest yield and
significantly (P<0.05) different from other levels of BM x pH and yield at B x 1(Heating
on Bunsen burner at pH1) was the lowest (Table-40).
In the subsequent part extraction was carried out by same procedure as followed in
foregoing study with the exception of increasing the strength of acid from 0.1N to 1N
HCl. Strength of acid showed to influence positively on the yield of pectin in numerous
earlier studies (Yapo 2009, Kalapathy and Proctor 2001) and was tested in current study
for extraction of pectin from sapodilla fruit peel. Table - 41 presented the yield acquired
by the use of increasing strength of acid. It was observed that maximum yield was
achieved from chopped peels only after 10 min of boiling in microwave (6.7%) at pH 5.
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Whereas 0.1 N HCl showed lesser highest yield 6.5% after 20 min of boiling at pH3.
Table - 42 shows analysis of variance (mean squares) of yield for different fruits. The
Table showed that the mechanical procedures has a significant (P<0.01) effect on yield
on all time of boiling selected for the study. Similarly, pH and boiling method have
significantly (P<0.01) affected the yield when using different times of boiling. All the
two way interactions i.e. mechanical procedure and pH (MPxpH), mechanical procedure
and boiling method (MPxBM), Ph and boiling method (pHxBM) and three factor
interaction mechanical procedure, pH and boiling method (MPxpHxBM) showed
significant (P<0.01) effect on yield. The difference among the means regarding different
factors using Tukey’s test are given in Table – 43-45 for 10 min of boiling using 1NHCl.
The overall mean for mechanical procedure, pH and boiling method showed chopping
and homogenising, pH 3 (Table - 43) and microwave boiling (Table - 44) has a
significant impact (P<0.05) on the yield of pectin after 10 min of boiling of peels using
1N HCl. Table- 43 and 44 also show the status of remaining mechanical procedure, pH
and boiling method and their order of response is given from most to least effective.
Alphabet A was considered the most and subsequent alphabets lesser in effectiveness
than the preceding alphabet. Overall interaction effect of boiling method and pH (BM x
pH) showed that M x 5 (Heating on microwave at pH 5) gave highest yield and
significantly (P<0.05) different from other levels of BM x pH while yield at B x 7
(Heating on Bunsen burner at pH 7) was lowest Table - 45.
The difference among the means regarding different factors using Tukey’s test are given
in Table – 46-48 for 20 min of boiling using 1NHCl. The overall mean for mechanical
procedure, pH and boiling method showed hammering, pH 5 (Table 46) and microwave
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boiling (Table 47) has a significant impact (P<0.05) on the yield of pectin after 20 min of
boiling of peels using 1N HCl. Table- 46 and 47 also show the status of remaining
mechanical procedure, pH and boiling method and their order of response is given from
most to least effective. Alphabet A was considered the most and subsequent alphabets
lesser in effectiveness than the preceding alphabet. Overall interaction effect of boiling
method and pH (BM x pH) showed that M x 3 and M x 5 (Heating on microwave at pH 3
and pH 5) gave highest yield and significantly (P<0.05) different from other levels of BM
x pH while yield at B x 1 (Heating on Bunsen burner at pH1) was lowest (Table – 48).
The difference among the means regarding different factors using Tukey’s test are given
in Table – 49-51 for 40 min of boiling using 1NHCl. The overall mean for mechanical
procedure, pH and boiling method showed hammering, pH 5 (Table - 49) and microwave
boiling (Table - 50) has a significant impact (P<0.05) on the yield of pectin after 40 min
of boiling of peels using 1N HCl. Table- 49 and 50 also show the status of remaining
mechanical procedure, pH and boiling method and their order of response is given from
most to least effective. Alphabet A was considered the most and subsequent alphabets
lesser in effectiveness than the preceding alphabet. Overall interaction effect of boiling
method and pH (BM x pH) showed that M x 5 (Heating in microwave at pH 5) gave
highest yield and significantly (P<0.05) different from other levels of BM x pH while
yield at B x 7 (Heating on Bunsen burner at pH at 7) was lowest Table – 51.
The difference among the means regarding different factors using Tukey’s test are given
in Table – 52 -54 for 60 min of boiling using 1NHCl. The overall mean for mechanical
procedure, pH and boiling method showed grinding, pH 3 (Table - 52) and microwave
boiling (Table - 53) has a significant impact (P<0.05) on the yield of pectin after 60 min
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of boiling of peels using 1N HCl. Table- 50 and 51 also show the status of remaining
mechanical procedure, pH and boiling method and their order of response is given from
most to least effective. Alphabet A was considered the most and subsequent alphabets
lesser in effectiveness than the preceding alphabet. Overall interaction effect of boiling
method and pH (BM x pH) showed that M x 5 (Heating in microwave at pH 5) gave
highest yield and significantly (P<0.05) different from other levels of BM x pH and yield
at B x 5 (Heating on Bunsen burner at pH5) was lowest (Table - 54).
Organic acids are also widely used for optimizing the yield of pectin from various fruits.
The use of organic acid instead of inorganic was also studies in the current study. These
include citric, oxalic and tartaric acid. Oxalic and citric acid gave comparable high yield
of pectin when used in the study. Organic acids like oxalic and citric acid have been used
to extract pectin and gave similar results 4.95% and 4.24% yield respectively after 10 min
of extraction time (Table - 55). Chan and Choo, 2013 studied effect on yield of pectin
after using citric acid and extraction time from 1.5 to 3.0 h and reported increase in yield
with increase extraction time. While Vriesmann et al. (2012) and Canteri-Schemin et al.
(2005) at the end of their study drawn conclusion that extraction time can increase yield
of pectin from coca husk and apple pomace. However the most effective model for
extraction of pectin using organic acid was after using 1% oxalic acid with homogenizing
as a mechanical tool in the present study. Two different concentration of organic acid 1%
and 10% were used in the present study. Low concentration of all the organic acids (1%)
has been found more effective with microwave as heating procedure. Table – 56 shows
Analysis of variance (mean squares) of yield for pectin from sapodilla fruit using organic
acid. The table shows that the mechanical procedures has a significant (P<0.01) effect on
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yield using different types of acid (citric acid, oxalic acid and tartaric acid) selected for
the study. Similarly, the concentration of organic acids (% age) and boiling method was
also been noted to have significant effect (P<0.01) on the yield of pectin. The two way
interactions i.e. boiling method (BM x MP) and mechanical procedure and acid
concentration (MP x Acid %) and three factor interaction boiling method, mechanical
procedure and acid concentration (BM x MP x Acid %) showed significant (P<0.01)
effect on yield. Only boiling method and acid concentration (BM x Acid %) has no
significant (P>0.05) effect on yield.
The difference among the mean values related to various factors (e.g. mechanical
procedure and method of boiling using Tukey’s test are given in Table – 57 -59 for yield
of pectin using citric acid from sapodilla fruit. The overall mean for mechanical
procedure, indicated homogenising and 1% strength of organic acid (Table – 57- 58) has
a significant impact (P<0.05) on the yield of pectin. Table- 57 and 58 also show the
status of remaining mechanical procedure,% strength and boiling method and their order
of response is given from most to least effective. Alphabet A was considered the most
and subsequent alphabets lesser in effectiveness than the preceding alphabet. While
overall interaction for 1% citric acid and mechanical procedure 2 (Chopping) and for
10% mechanical procedure 1 (Homogenizing) gave most effective results as compare to
others (Table – 59).
The difference among the mean values related to various factors (e.g. mechanical
procedure and method of boiling using Tukey’s test are given in Table –60 -62 for yield
of pectin using oxalic acid from sapodilla fruit. The overall mean for mechanical
procedure, indicated homogenising and 1% strength of organic acid (Table – 60 -61) has
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a significant impact (P<0.05) on the yield of pectin. Table- 60 and 61 also show the status
of remaining mechanical procedure, % strength and boiling method and their order of
response is given from most to least effective. Alphabet A was considered the most and
subsequent alphabets lesser in effectiveness than the preceding alphabet. While overall
interaction for 1% oxalic acid with mechanical procedure, mechanical procedure
1(Homogenizing) gave most effective results as compare to others Table – 62.
The difference among the mean values related to various factors (e.g. mechanical
procedure and method of boiling using Tukey’s test are given in Table – 63 - 65 for yield
of pectin using tartaric acid from sapodilla fruit. The overall mean for mechanical
procedure, indicated homogenising (Table- 63 ) and 10% strength of organic acid (Table
- 64 ) has a significant impact (P<0.05) on the yield of pectin Table- 63 and 64 also show
the status of remaining mechanical procedure,% strength and boiling method and their
order of response is given from most to least effective. Alphabet A was considered the
most and subsequent alphabets lesser in effectiveness than the preceding alphabet. While
overall interaction of mechanical procedure strength of organic acid and boiling method,
1% tartaric acid with mechanical procedure 1 ( Homogenizing) , 10% tartaric acid with
mechanical procedure 1 and 10% tartaric acid with mechanical procedure 3 (Grinding) is
equally effective for better yield from sapodilla fruit as compare to others (Table – 65).
Finally the effect of strength of inorganic acid on the yield of pectin is investigated using
three strength of inorganic acid 0.1, 0.5 and 1N HCl (Table - 66). The yield under 0.1N
HCL and 1N HCl (Table – 27 and 41 respectively) has already been discussed in
previous part of this discussion hence a statistical analysis was performed. Table - 67
shows Analysis of variance (mean squares) of yield for pectin from sapodilla fruit using
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different strength inorganic acid (HCl). The table shows that the mechanical procedures
has a significant (P<0.01) effect on yield using different strength of acid selected for the
study. Similarly pH and Boiling method have significantly (P<0.01) affected the yield
when using different strength of acids. The two way interactions i.e. boiling method and
pH (BM x MP), mechanical procedure and pH (MP x pH) and three factor interaction
boiling method, mechanical procedure and pH (BM x MP x pH) and boiling method and
pH (BM x pH) showed significant (P<0.01) effect on yield.
The difference among the mean values related to various factors (e.g. mechanical
procedure and method of boiling using Tukey’s test are given in Table – 68 - 70 for
pectin yield from sapodilla fruit using 0.1NHCl. The overall mean for mechanical
procedure, pH and boiling method showed chopping, pH 3 (Table - 68) and microwave
boiling (Table - 69) has a significant impact (P<0.05) on the yield of pectin after using
0.1N HCl. Table- 68 and 69 also show the status of remaining mechanical procedure, pH
and boiling method and their order of response is given from most to least effective.
Alphabet A was considered the most and subsequent alphabets lesser in effectiveness
than the preceding alphabet. Overall interaction effect of boiling method and pH (BM x
pH) showed that M x 5 (heating in microwave at pH 5) gave highest yield and
significantly (P<0.05) different from other levels of BM x pH (Table - 70)
The difference among the mean values related to various factors (e.g. mechanical
procedure and method of boiling using Tukey’s test are given in Table – 71 -73 for pectin
yield from sapodilla fruit using 0.5 NHCl. The overall mean for mechanical procedure,
pH and boiling method showed cutting, pH 1 (Table - 71) and microwave boiling (Table -
72) has a significant impact (P<0.05) on the yield of pectin after using0.5N HCl. Table-
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71 and 72 also show the status of remaining mechanical procedure, pH and boiling
method and their order of response is given from most to least effective. Alphabet A was
considered the most and subsequent alphabets lesser in effectiveness than the preceding
alphabet. Overall interaction effect of boiling method and pH (BM x pH) showed that M
x 1 (Heating in microwave at pH 1) showed highest yield and significantly (P<0.05)
different from other levels of BM x pH (Table - 73)
The difference among the mean values related to various factors (e.g. mechanical
procedure and method of boiling using Tukey’s test are given in Table – 74 -76 for pectin
yield from sapodilla fruit using 1N HCl. The overall mean for mechanical procedure, pH
and boiling method showed chopping, pH 3 and 5 (Table - 74) and microwave boiling
(Table - 75) has a significant impact (P<0.05) on the yield of pectin after using 1N HCl.
Table- 74 and 75 also show the status of remaining mechanical procedure, pH and boiling
method and their order of response is given from most to least effective. Alphabet A was
considered the most and subsequent alphabets lesser in effectiveness than the preceding
alphabet. Overall interaction effect of boiling method and pH (BM x pH) showed that M
x 5 (heating in microwave at pH 51) showed highest yield and significantly (P<0.05)
different from other levels of BM x pH (Table - 76).
In the conducted study, maximum yield of pectin was obtained in acidic medium,
indicating it as an important factor to be considered while extracting pectin. Earlier
studies also suggested positive effect of acidic medium (low pH) upon yield of pectin and
a drastic effect on pectin yield with the increase in pH (Canteri-Schemin et al., 2005
and Yapo et al., 2007). The lower value of pH supports the theory of the disruption of the
cell wall, release of protopectin containing cellulose pectin in the medium and its
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hydrolysis, thus releasing high quantity of pectin in the liquid phase which can thus be
recovered by precipitation. It was also observed that among the five fruit peels
investigated in the study, sapodilla peels gave considerably good yield in basic medium
too that is pH 6.5 and 7, which is quite interesting but in accordance with an earlier
published report (Kirtchev et al., 1989) in which authors suggested that both acidic and
alkaline medium with elevated temperatures can help the rupture of cell
walls to release protopectin and its hydrolysis to release pectin.
The data showed that mechanical method does have effect on yield as shown in Table -
10. Different kinds of techniques have been used for their specific advantages by various
workers while working on pectin extraction process optimization. Cutting of peels into
smaller pieces, help to increase surface area of the material for better extraction
(Rudolph and Petersen, 2012). Likewise, mortar and pestle facilitated the size reduction
of the raw material selected for the extraction of pectin (Poovaiah and Nukaya, 1979).
Hammer mill is extensively used in studies to ground cell wall material to extract pectin
( Besson et al., 2013), while grinding in mechanical blender and in mills procedure has
also been termed a successful procedure to extract pectin (Loyola et al., 2011).
Homogenizing is also an effective way of forming loose slurry used in extraction of
pectin (Slavov et al., 2013). Different fruits indicated different yield when subjected to
various mechanical methods adopted in the study. The effectiveness of a mechanical
process in increasing pectin yield depends upon its capability of disrupting the cell wall
of the peels to release pectic substance. One mechanical process may not be suitable for
different fruit peels because of the structure and composition of the cell wall. Chopping
with mortar and pestle has been used for size reduction of raw materials with or without
190
adding solvents since long in the extraction processes (Lamotte et al., 1969; Poovaiah and
Nukaya, 1979).
With the conventional or direct heating procedure there is a chance of pectin
degradation. Therefore, microwave heating was studies in the present study to learn the
effect of extraction time and to improve the quality of pectin. Among the two boiling
methods, the overall yield was recorded better with microwave. The mineral acid used in
the extraction of pectin was used at low strengths (0.1 till 1N HCl), in order to keep the
process environmental friendly because of corrosive nature of HCl (Vriesmanna and
Petkowicz, 2013). The methods used in the present study were kept simple, in order to
get the required preliminary knowledge about the fruits like sapodilla, muskmelon and
banana, while comparing it at the same time with known high content pectin fruits, in
order to get an authentic and reproducible results for more effective and economical way
for the better extraction of pectin.
Apparently it is recognized that physicochemical properties of pectin influence their
functional properties consequently affecting its application in food and pharmaceutical
systems. In order to study the basic character of sapodilla pectin basic identification and
bio characterization tests were performed. Table - 77 describes the basic identification
test based on USP for isolated sapodilla, banana and muskmelon pectin. The basic
identification tests of the current study showed positive response from sapodilla and
banana and affirms the isolated pectin has all the basic characteristics of being a
carbohydrate material (Yadav and Agarwala 2011). However, banana pectin failed to
form stiff gel which is the basic and most important functional character of pectin.
Whereas muskmelon also failed to form gel and tests performed for identification. Hence
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the two fruits (banana and muskmelon) were omitted for further analysis and sapodilla
peel pectin was given full consideration. The preceding extraction analysis revealed the
impact of pH on the yield of pectin from sapodilla fruit peel. Recent studies on isolation
of pectin from different fruits have also indicated that pH has a key role both on the yield
and the functionality of pectin (Ziari et al., 2011; Chan and Choo 2013; Kaya et al.,
2014). Hence further studies were also involved the sapodilla peel pectin extracted at
different pH (Table-78).
As mentioned earlier the basic functional property of pectin is to form gel with a
specified system. The gel formation capacity of pectin depends upon many factors which
were also included in the studies. Sapodilla being a new source was studied for its basic
bio-character and are shown in Table - 78. The dried pectin was stored in a cool dry
place. Before analysis it was necessary to confirm that the dried pectin is free from
ammonia. Presence of ammonia may obstruct with the other tests used for the
characterization of pectin. If ammonium is detected, the sample was washed with
acidified 60% alcohol, followed by neutral alcohol to remove the acid. The presence of
ammonia can be identified by the distinctive smell of ammonia (Ranganna, 1986). In the
present study all the samples were observed free rom ammonia.
The moisture content of pectin appeared in the range of 5.52% to 6.02% .The high
moisture content is likely to increase the susceptibility of dried pectin for the growth of
microorganisms in it and may also lead to the formation of pectinase enzyme which also
has a supplementary role in affecting the quality of pectin (Muhamadzadeh et al.,
2010).In the present study the moisture in the extracted pectin was in accordance with the
192
former studies as done by Acikgoz, 2011 and it is also under the required standard limit.(
Brejnholt , 2010).
The standard ash content of the extracted pectin reported to be less than 10% on free
moisture basis. Low values of ash content is among the judge for the purity of pectin
along with other important chemical parameters like galacturonic acid content which
should be 65% (Acikgoz, 2011).The ash content in the current study exhibited in the
range of 4.3% till 5.11% for extracted sapodilla pectin .The equivalent weight of the
extracted pectin showed to be little higher than the food grade pectin for sapodilla. The
pectin extracted at pH3 and pH1 indicated higher equivalent weight (1700 at pH3 and
1975 at pH1) as compared to the pH5 which is 1325. The equivalent weight at pH5 was
comparable with the apple (833.33 to 1666.30, Kumar & Chauhan, 2010) and
commercial pectin (1271±18.4) equivalent weights. Equivalent weight also plays a key
role in the jelling property of pectin, the good jelling ability pectin have higher molecular
weight. (Vaclavik and Christian, 2008). This is due to the fact that equivalent weight can
be influenced by number of free acid and maturity of fruit and the low values of
equivalent weight might be due to some degradation of pectin (Ramli and Asmawati,
2011). It was learned further that increased temperature may results in greater yield of
pectin but it affects negatively on the methoxy content and equivalent weigh of pectin
(Kulkarni and Vijayanand, 2010).
Methoxyl content of the extracted pectin appeared to be 4.4% - 5.1%. Methoxyl content
is an important factor in controlling the setting time of pectins and the ability of the
pectin to form gels (Constenla and Lozano, 2003). Table - 78 shows that the sapodilla
pectin at pH 5 gave higher methoxyl content (10.25%) followed by pH3 (4.4%) and then
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at pH1 (4.9%). These values were approximately similar to earlier reported values as
found for peel of mango (7.33%), pomelo peel (8.57%), Lime (9.92%), (Madhav and
Pushpalatha, 2002), passion (8.81%-9.61%) but higher than dragon fruit pectin (2.98% to
4.34%) (Ismail et al., 2012). Methoxyl content of extracted pectin vary from 0.2-12%
depending on the source and mode of extraction, the percentage methoxyl content
obtained falls within the range. Since all the values obtained experimentally were below
or almost 7%, for sapodilla means that the pectin were of low ester characterization
indicating that the pectin is good in terms of quality .Aina, 2012.
Estimation of anhydrouronic acid content is essential to determine the purity and degree
of esterification, and to evaluate the physical properties. It was found in a range from
61.78% to 69.33% in case of sapodilla which is the desired value in an extracted pectin as
indicated by the food chemical codex 1996 which states that AUA should not be less than
65%. Similar values were observed in apple pomace pectin, commercial apple pectin and
dragon fruit pectin which were 59.52% to 70.50%, (Kumar & Chauhan, 2010), 61.72%
and 45.25% to 52.45% (Ismail et al., 2012) respectively. Low value of AUA means that
the extracted pectin might have a high amount of protein (Ismail et al., 2012).
The gelling capacity of extracted pectin was also evaluated and observed to be in range of
99 to 100 for the extracted pectin. The jelling capacity of fruit pectin was calculated low
which can be due to numerous gelling affecting property for a pectin. One of the reason is
the presence of acetyl group and presence of neutral side chains of sugars which in case
of sugar beet pectin hinders in making gel (Fishman et al 2010). The other factor such as
ash cannot be the cause here as it was already in required limits and it is learned that
lower ash values aids in forming better gels (Ismail et al., 2011).
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The GA contents of sapodilla pectin was observed in the range of 60.5% to 77.7%. The
difference in the galacturonic acid is also due to many factors as Garna et al (2007)
reported that pH and temperature at which the extraction is done influenced the
galacturonic acid content of apple pectin. Also it may be a result of the side sugar chain
hydrolysis of pectin which in turn is a reaction of high temperature extraction. (Fraeye et
al., 2007). However, this could also be the attribute of the type of acid used, as when
citric acid was added for extraction opposite phenomenon was observed and low
temperature resulted in high contents of uronic acid. (Garna et al., 2004). The
hydrochloric acid used in extraction can also have an effect on the galacturonic acid
content as it was also learned that apple when only hot water used during extraction gave
better GA contents compared to the extraction using hydrochloric acid (Hwang, Kim, and
Kim (1998). Moreover increased extraction time also influenced uronic acid value as
studied by Vriesmann et al. (2011). Protein contents are necessary to determine in order
to assess the purity of extracted pectin ( Alba et al 2015) the gelling property of pectin is
also influenced by protein content as in case of sugar beet pectin ( Yapo et al., 2007). The
pectin content for the sapodilla pectin at different pH showed smaller values (1.71- 4.04),
and within limit which also has an effect on its gelling property.
Water holding capacity is also an important parameter in order to understand the physical
characteristic of a compound. The affinity of a particular compound to binds with water
determine its water holding capacity which consequently exerts great influence in
determining the physiological property the compound may carry. The water binding
capacity was greater than the reported values of apple and orange fiber (7.6 for sapodilla)
(Table- 79) while the water holding capacity was less than the reported values of apple
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and orange pectin (6.99 for sapodilla) (Lecumberri et al., 2007) while the fat binding
capacity was quite comparable with the apple, orange and grape fruit fibers (Figuerola et
al., 2005). Physicochemical properties such as water holding, water binding and fat
binding capacities are important to understand as structural information alone cannot
predict the functional properties of dietary fibers and their health benefits. Hence the
physiological effects of fibers rely on inter related combination of physical, chemical and
structural properties (Blackwood at al., 2000). Due to the many reported advantages of
dietary fibers present day researchers and industries are giving it great attention. Dietary
fibers, including pectin have proved affective against the control of cardiovascular, blood
cholesterol, diabetes and colon cancer. With the comprehensive knowledge about the
nutritional as well as functional properties it is assumed that new value added foods and
convenience food products can be developed which are high in demand along with
functional properties of the foods.
The Fourier Transform Infra-Red (FT-IR) study was also carried out to understand the
structural similarities of extracted and food grade pectin available in market. The
absorbance (400-4000 cm-1) of sapodilla peel pectin extracted at different pH were
presented in Figure- 23 to 26.The absorbance region of important peaks are tabulated in
Table - 80. The spectra revealed no prominent structural differences in commercial and
pectin extracted from peel of sapodilla at different pH. The IR absorption peaks achieved
in between 1000-2000 cm-1 comprised of the main functional groups in pectin as these
field of spectra are commonly employed in describing different types of pectin
(Kalapathy and Proctor, 2001). Similar spectral peaks are seen in the absorption range of
1300-1000 cm-1. The spectra also revealed C=O stretching in a range of 1640-1750 cm-1.
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These peaks are of particular interest when degree of esterification (DE) values of pectin
are calculated the reason being the esterified carboxylic group shows an increase in
intensities and band regions as the DE value increases. DE was more than the standard
pectin in all the samples and appeared similar as in C limon which exhibited strong C=O
stretch at 1710-1665 cm-1 Kanmani, P 2014.The C-H stretching of the CH3 group was
also observed in all samples of pectin including the food grade . These O-H stretching
vibrations appeared within a broad range of frequencies (Singthong et al. 2004), followed
by the absorption band of C-H stretching of CH, CH2 or CH3 roughly at 2900 cm-1
(Singthong et al. 2004; Tamaki et al. 2007). The moisture in the samples typically
broadens the band from 2400-3600 which mainly form due to the stretching of the
hydroxyl group. Among the samples least moisture was found in food grade while pectin
at pH 5 has the most. Similar sharp and strong peaks were observed by Kanmanim, P.,
2014 when studying C.limon at 3595.31
The presence of peaks at 1728.22 cm_1 and 1242.16 cm_1 shows the existence of α, β-
unsaturated esters and aliphatic amine functional groups. These absorption peaks are
usually the result of inter and intra molecular hydrogen bonding of galacturonic acid
backbone. The finger print region (1300e800 cm_1) can also identify variations in
monosaccharide composition of pectin (Kamnev et al., 1998; Monsoor et al., 2001),
however, some researchers believe that it is challenging to interpret because of spectrum
overlap (Gnanasambandam& Proctor, 2000), this was also found in the present study and
it was difficult to assign the peaks of fingerprint region to corresponding chemical
analysis results. However, some symbolic characteristic peaks of pectin, for example, 882
cm_1 (pyranose ring), 1273 cm_1 (CeO dilatation vibration), 1453 cm_1 (eCH3
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antisymmetric deformation or the eCH2e symmetric deformation), were observed. The
weak peaks of amide bands (amide I: 1670 cm_1; amide II: 1588 cm_1) in samples
indicated that small amount of protein existed, which also corresponds with chemical
analysis (Table - 80).
Particle size determination was carried though dynamic light scattering studies (DLS)
which is one of most common methods use for particle size. It is used to explore
polydispersity index and size of different extracted and standard pectin. For the detailed
study of particle size apple and orange pectin was also extracted and used in the study.
The apple and orange pectin particle size study gave an insight of other extracted pectin
extracted in the same conditions (Ahmed and others 2016). Table - 81 summarizes the
results obtained after DLS studies, while the illustration of standard and other extracted
pectin are shown from Figure - 28 till Figure - 31. In the figure the autocorrelation
function (ACF) is shown in the top panel denoted by letter A (a). The ACF are usually in
the form of a curve which shows the size of the particle and which is formed after
plotting values of ACF against time. The curve formed specifically after the movement of
particles according to their size, greater the particle size, slower will be its movement and
vice versa. The decay of ACF, with time, is due to the faster agitation of the smaller
particles that leads to the faster decorrelation of scattered light intensity. The central
panel depicts the intensity of radius distribution of the particles denoted by A(b) and are
in the range of 1 nm to 10 μm. The solid line peak indicated the result of 50 experiments
mean calculated by the help of software, while the dotted lines is the basic data received
after the 50 measurements. The bottom panel A(c)) shows the radius plot of all the 50
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measurements which were taken over a period of 1000 s with measurement of every 20 s
and a delay time of 1 s in between.
It represents the differences in the particle size and monodispersity of pectin in each
individual results in 50 measurements. While in B it illustrate the final results of standard
(a) and sapodilla fruit pectin (b). Table -81 demonstrated the radius of hydration (RH)
which is the average particle size and percent polydispersity (% PD) characterized by the
DLS, of standard and the pectin extracted from various other fruits during the study.
Table - 81, presents the average particle size which is expressed in radius of hydration
(RH) and the polydispersity (% PD) which were characterized by DLS studies. It is
observed that most of the extracted pectin (orange, sapodilla extracted at pH 3 and 5)
acquired in relatively smaller and same sizes (~400 - 550 nm), while apple and standard
pectin has quite comparable large sizes (~70 - 1980 nm). While the percent polydispersity
were approximately in identical range for all samples (~20-45%). However it is
suggested that particle size decreases when the pH increases when pectin was
investigated in an aqueous medium (Gbassi and others 2013). It also determined that size
is an essential criteria in assessing the stability and chemical behavior of a particle in a
particular solution or formulation (Gruy 2011). The solubility enhances with the lowering
of particle size in a solution. The dispersion of smaller particle sizes in suspension
dominates the gravitational force which results in keeping the homogeneousness of
suspension intact (Gbassi and others 2013). Observations collected are quite comparable
with a previous study which reported identical particle size data of pectin, while a
contrasting estimation of polydispersity was achieved (Nguyen and others 2011).
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During the studies conducted under extraction of pectin from sapodilla fruit was noted
that a number of factors can influence the yield of pectin from the peel. When there are
many factors influencing a certain response, RSM or response surface methodology is an
effective tool to determine the optimal response (Sun et al., 2011). Box-Behnken design
was used in the RSM study to produce the experimental design that is shown in Tables 82
and 83.
The analysis depicting the variance for yield (ANOVA) is shown in Table- 84. This table
shows that the fitted model is in full quadratic model where the p-value of lack of fit is
0.348 which proposes that the data is adequately fitting for the model. Examination and
evaluation of the variance table shows the linear, quadratic and two-way interactions
between variables. Overall, the model is statistically significant, as indicated by the p-
value of 0.018 for the model. In addition, the small p-values for linear terms (i.e.
p=0.025), quadratic terms (i.e. p=0.015) and two-way interactions (i.e. p=0.045) indicates
a curvature in the response surface. These effects are statistically significant as indicated
by p-values for the pH by time interaction (p = 0.017) and pH squared (p = 0.004). The
value of R-square shows that the fitted model explain 81.35% variation due to pH, time
and temperature that is good but the predicted R-value explain 16.11% variation which is
not proper in the new dataset. R-square (adjusted) refers to the total variation in yield due
to pH, time and temperature. The error term, s = 0.3197. The F-value (22.61) indicates
that pH has a highly significant impact on the yield of pectin and when pH is changed
slightly it has a greater effect on the yield of pectin. Thus pH is an important factor
influencing yield of pectin from sapodilla fruit peel. Similar studies are carried which
also showed that pH has a major influence on yield of pectin (Oliveira and others 2016).
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In addition, the interaction of the pH with the time of heating or boiling is another
important factor that has a significant impact on the yield of pectin.
The coefficients with their standard error, t value and probability for all the terms in the
model are all illustrated in Table -84. In this case, an orthogonal design model was
exercised which led to each effect being assessed independently from each other. Value
for VIFs (variance influence factor) can all be rounded-off to 1, which shows that the
predictors are not interrelated, thus there is no multicollinearity present in the model. The
coefficients in Table-85 indicated the direction of individual variable impact on yield.
According to findings, pH, interaction of pH x pH and pH x time have significant (p <
0.05) impact on yield. All the three terms (i.e. the pH, pH2 and pH x time) denotes the
positive impact of independent variables on yield since they have the positive coefficient
value.
Using approximated coefficients for yield, the following predicted equation or regression
equation was derived which gave the yield of pectin from sapodilla fruit peel.
Yield = 0.76 – 0.947 pH + 0.0784 Temperature – 0.0303 Time + 0.2089 pH2 – 0.000443
Temperature2 + 0.000030 Time2 – 0.00550 pH*Temperature + 0.00750 pH* Time +
0.000037 Temperature*Time
The three dimensional response surface is plotted and illustrated in figure 33-35, where
the vertical axis represents the yield and the two horizontal axes represents any two of the
three affecting variables (i.e. pH, temperature and time). The response surface aids to
understand the optimal level of every variable which affects the pectin yield and also
indicates the interaction of variables that affect the response. The effect on pectin yield
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by the interaction of temperature and pH, the fact that pH has a substantial influence on
the pectin yield and that maximum yield of pectin can be reached at a pH greater than 5
are all shown in Figure-33. Figure -34 represents the effect on pectin yield influenced by
the interaction of pH and time at constant temperature. The result concludes that the
highest yield can be attained when the extraction is done after a heating span of 100
minutes at a pH greater than 5. Figure - 35 demonstrates the effect of the interaction of
temperature and time on the yield of pectin at constant pH. However, the graph shows
that no linear relationship exists between the two quantities and demonstrates that the
impact of the interaction of time and temperature is unresponsive and has no significant
effect on the yield.
In order to determine the optimized response lowest yield (1.4) and target yield (3.5) was
given to generate the best solution to optimize yield. The predicted values are given in
Table – 86 According to response optimization predicted model, the best extraction
variables to extract pectin is to keep a constant pH of 5 at 61.11oC for a heating span of
90 minutes will contribute a 3.7% yield. Pectin was extracted from sapodilla fruit peel
through the predicted model and it was verified, since a value close to the approximated
value of yield (i.e. 3.5%) was extracted when the predicted values of variables (pH,
temperature and time) were followed.
Sapodilla crude pectin extracted was used in the preparation of two common
pharmaceutical solid oral formulations, paracetamol and ibuprofen tablets .The extracted
sapodilla pectin was used as a binding agent in both formulations and were evaluated for
different physico-mechanical and micromeritics properties. As sapodilla pectin is a new
source of pectin which has not been reported in earlier studies in tablet formulation
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therefore, it is important to determine the micromeritics of granules .The nature and
concentration of a binder can effect on the compression, mechanical strength,
consolidation, flow and mechanical strength of a tablet (Rao et al., 2012). Table - 87 and
90 represents the micrometrics properties of granules formulated with different
concentrations of binder which is pectin in this case, in paracetamol and ibuprofen tablets
respectively. The compressibility index of paracetamol came out to be in range from
4.618 to 24.603 %. Among the different formulations F4 and F5 has excellent
compressibility index (4.618% and 5.690 % respectively) and Hausner ratio (F4=1.048
and F5=1.060), F6 was good for both compressibility index and Hausner ratio and F1,
F2, F7 and F9 were fair formulations while F3 and F8 were passable .The angle of repose
came out to be under 21.546. While for ibuprofen carr’s index was excellent for all four
formulations (Table - 90) and excellent to fair Hausner ratio for the four formulations. R1
showed excellent Hausner ratio (1.09) and compressibility index (3.77) while angle of
repose was also better for all four formulations which came between 11.13 to 15.25.
The weight variation of tablet is a credible sign of the uniformity of constituents present
in the formulation which is a basic requirement of good manufacturing practice as well as
important for keeping a constant size of the tablets( Nasrin et al., 2011). Uniformity in
weight of tablet is also required as the table contains the active compound or drug which
should be present in a specific ratio, weight is an important indicator that the drug in the
tablet is in required limit or not. The weight of both the types (paracetamol and ibuprofen
) remained under the limit of ± 5%. The diameter and thickness of all the tablets also
didn’t exceeded from the required level (Table - 88 and 91). Hardness is also an
important parameter which is useful to control chipping, abrasion or breakdown of tablets
203
during storage and transportation of tablets (Bano, 2011). Therefore, the method to check
hardness is the breaking of tablet by applying reasonable pressure on it. Hardness is also
an important parameter to ascertain the quality of a drug as it also effects disintegration
and dissolution of a tablet (Nasrin et al., 2011)
In paracetamol tablets,10 and 20 mg of pectin were found unsuitable to achieve desired
hardness granules when compressed into tablets were soft and thus further concentration
of pectin was increased. When the concentration increased from 30mg/tab desired
hardness was achieved (Table-88) however interestingly as the concentration of pectin
was increased upto a certain limit, dissolution was noted to decreased. The best hardness
and dissolution was achieved with the formulation F4 and F5 when the concentration of
40mg /tab and 50 mg/tab were used respectively. Dissolution was recorded as 80.43%
and 86.35% respectively. Further increased in pectin from 60 to 120mg not only
increased the hardness of tablet but also noted to decrease the dissolution significantly.
While the ibuprofen tablets showed a similar type of results. It is also noted that the
higher concentrations of pectin had detrimental effect on dissolution properties of tablets.
Among ibubrufen tablets 50mg of pectin gave the best dissolution results as compared to
the rest of the formulations containing 75mg, 100mg and 125 mg of pectin as a binder. It
was also observed that the hardness of the tablet was successfully increased by increasing
the binder but it significantly affected the tablets dissolution property of ibuprofen tablet.
The bulk and tap density results selected formulation shows that granules have good flow
and compressibility property and same observation supported with the Carr’s index and
Hausner ration results.Tablet hardness usually required between 6 - 14 KP and is
primarily based on the friability less than 0.1% and DT not more than 5 minutes results
204
which a drug formulation targets to achieve. The particle distribution result shows good
combination of cores and fine granules for weight variation control during compression.
The thickness of the tablets are also found consistent, indicating reasonable justification
to use blister packaging very common for these ( active pharmaceutical ingredients
(APIs) in Pakistan. Uniformity in blend shows good distribution of API while dissolution
result also support selection of compression parameters.
An antidiarrheal preparation was also formulated with the extracted sapodilla pectin .The
basic evaluation tests (Table- 93) of suspension indicated no major difference between
the physical attributes of suspension made from sapodilla pectin and marketed products
(Figure – 40). Pectin is an effective antidiarrheal agent and together with kaolin it acts as
adsorbent resulting in more solid form stool and it also has affinity to attach the digestive
mucus and toxins hence aids in reducing water loss (Abdullah and Firmansyah, 2013;
Smith, 2013). The use of kaolin-pectin antidiarrheal preparation is still common in
Middle east and Gulf (MOH Saudi Arabia ; HAAD) South Africa (Westhuizen, A. V.
(2010), Indonesia (Abdullah and Firmansyah, 2013) and Australia (Bis pectin,2016)
Although FDA has discontinued the use of kaolin /pectin suspension early 2000, the use
of kaolin pectin as adsorbent has proved effective in the pharmacological treatment of for
noninfectious HIV-associated diarrhea (Dikman et al., 2015). The primary function of
pectin based antidiarrheal is to provide water soluble fibers capable of stimulating
epithelial growth in the colon helping in the reduction of diarrheal frequency. Apart from
this, pectin possess strong water holding capacity, water and fat binding properties which
ultimately helps to change the consistency of stool from watery to soft and also helps in
eliminating excess mucus formed in GI tract. In the present study the water holding
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capacity of extracted pectin was noted as 6.99 g/g while the formulated suspension
indicated 32.69 g/g which was quite comparable with the reference sample (Kepect)
showing value of 33.96 g/g.
For the food preparation one of the formulation was sapodilla jam. The jam was prepared
with three different concentrations (5, 7 and 10 g) of extracted sapodilla pectin. A
standard jam preparation was also made with equal quantity of pectin as F1 of the sample
preparations. It was clearly seen through the results that the concentration of pectin
affected the physical and chemical attributes of jam. A slightly increased amount of
extracted sapodilla pectin was needed to prepare jam which was comparable with the
standard pectin jam.( Figure - 41). The pH of jam should be under limit specified by the
Food and Agriculture Organization of the United Nations to minimize any bacterial
growth which might contaminate the food product. The pH of all the samples prepared
showed with in the specified range (3.00 to 3.30) as shown is Table -94. The pH has also
some influence on the viscosity of finally prepared jam. The gel firmness also depends
upon pH of jam, its range should be between 3 to 3.5 for best results. However the
viscosity depends upon other factors too which makes it difficult to predict for the final
end product. The other factors which influence viscosity of jam are temperature, degree
of methylation and concentration of pectin or any alkaline earth salts which may be
present in it (Panda, 2011). Pectin concentration should increase the viscosity of jam and
was proved during the current study when the viscosity of jam increased with the
increasing concentration of pectin. This is also in accordance with some previous
studies.(Nwson et al.,2014 ; Kar and Arsalan, 1999). Moisture content is another
important determinant in assessing the shelf life or life span of a food product. It also
206
gives basic information for how long a product can be stored safely and effectively
(Ashaye and Adeleke, 2009). The higher the amount of moisture content greater will be
affinity for microbial growth specially fungus and molds. Pectin has a key role in
stabilizing the moisture content present in a finished product (Nwson et al., 2014). The
moisture content also came into the specified range which was reported in earlier studies
(Ajenifujah-Solebo and Aina, 2011).The moisture content was also observed six month
after the formulation of the test samples and no bacterial growth was found in it. Table –
94 also shows the value of ash present in the test as well as standard formulations which
is also an indicator of stability criteria of the formulated product. The increased value of
ash enhances the vulnerability of product in terms of stability. The test samples exhibits
ash values ranging from 1.69 to 1.73% which is same in value of the standard product
and hence showed to be a stable product.
Another important in jam processing is the level of acidity in fruit pulp which also affects
the gel formation. The total titratable acidity (TTA) of the jam under investigation also
observed to be within the range of standard values (1.01 to 1.14, Table - 94) which also
predicts that the jam prepared was of good quality. Total soluble sugar (TSS) was also
measured which is an important observation during the storage of jam for assessing its
quality. It was reported in earlier studies that with a 5% level of pectin and pH ranging
from 3.0 to 3.5 the TSS should be 65%. (Ndabikunze et al., 2011) Among all the test
formulation only F3 showed the values of TSS to be in desirable range. It was also
learned that TSS decreases if the product is not refrigerated and kept at room temperature
after formulation ( Sindumathi and Amutha, 2014). Vitamin c (ascorbic acid) contents
were also determined during the study which is also shown in Table 94. The values were
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in the range from 17.6 to 18.3. Vitamin C is a natural antioxidant which determines the
quality of a product. As pectin is used as a gelling agent and helps the jam in setting
faster at lower temperature also aids in preserving the heat sensitive nutrients present in
the jam (Nwson et al., 2014).
Sensory evaluation of sample as well as standard jam was performed on a nine point
hedonic scale as shown in Table - 95. The data collected from the test showed that the
amount of pectin used in each formulation affects positively the sensory attributes of jam.
Among the three formulations F1 showed good scores for appearance, aroma and
spreadibility (7.5, 7.2 and 7.3 respectively) but a moderate score for taste and mouth feel
( 7.3 and 7.0). The results can be linked to the amount of pectin present in the
formulation. The formulation F2 exhibited highest score and was comparable with the
scores achieved for standard formulation. Score for appearance, taste, mouth feel and
aroma were almost same as that of standard jam (7.5, 7.6 7.5, and 7.7 respectively).
While spreadibility of F2 jam was slightly lesser (7.8 for F2 and 8.2 for S). The least
acceptable formulation was F3 in which the highest amount of pectin was used. It was
observed that the high amount of pectin affected the formulation negatively and got the
least scores. The jam also became harder and hence the overall acceptability score was
observed as 6.03. The cooking of the jam usually caramelize the sugar and hydrolyzes
pectin in it which can be one reason of low scores of mouth feel and taste in formulation
F1 and F3 (Fishman and Jen, 1986). Pectin is used as a thickening agent in formulations
hence the increased amount in formulation F3 resulted in the thickening of jam more than
it is desired which also affects the texture, spreadibility and mouth feel of jam (Broomes
and Badrie 2010).
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Another preparation which is used to investigate the functional property of pectin was the
formulation of pudding with the extracted and standard commercial pectin. Textural
property of pudding was the main attribute which is studied here as it is the main desired
quality a good pudding should possess. The failure of the participant to establish any
difference in the pudding texture made from two sample concentration of sapodilla
pectin points towards the similar physical attributes which all the pudding possessed. The
study also gives a slight insight of the role of pectin in milk products. It was studied
earlier that pectin in milk preparations aids in minimizing the agglomeration of protein
which helps to keep the phase in the milk product intact and prevents separation. Pectin
due to its thickening property also has a role in the specific type of mouthfeel which is
required for a particular product (www.herbstreith-fox 2016) According to a some
details published on a company website (Cargill foods 2016). HM pectins are good
stabilizers in acid milk drinks and probably the two puddings pass the tests made from
sapodilla pectin due to this property. It was also understood from the site that HM pectins
creates a coat on the casein particles of milk products. The reason for making this food
formulation was to determine the effect of extracted sapodilla pectin in a formulation
other than the conventional use that is gel formulation.
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Conclusion
Comprehensive knowledge on carbohydrate chemistry and skills in hydrocolloids are the
basic and prerequisite for developing new pectin source as well as to ascertain the
quantity and quality of pectin along with its applications. On the basis of sound
background on both the disciplines, one can rely on the selection of raw material as a
starting source of pectin, establishment of extraction protocol and the bio-characterization
of the end product to find its proper utilization. In view of this, current study was
designed to investigate possible new indigenous source (fruits) of pectin, utilization of
different physico-mechanical procedures for the extraction of pectin from fruit wastes
and to explore one best fruit for in-depth studies. The best identified fruit in this study is
observed as Sapodilla (Manilkara zapota) and thus a comprehensive plan of work was
designed and attention was driven towards the pectin extracted from sapodilla for
physicochemical characterization and its application in food and pharmaceutical products
development. In conclusion, the whole study and suggestions for future studies have been
summarized below.
(i) Mechanical procedure noted to be a potential tool for the extraction of pectin.
Out of five mechanical procedures and five different pH, Chopping was
observed the best mechanical procedure at pH 3 for sapodilla fruit peel when
microwave was used for extraction of pectin. For future studies, suggest to
develop a pilot scale setup to optimize the selected procedure for possible
commercial exploitation.
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(ii) The bio-characterization along with FTIR and DLS were found excellent
methodologies to study type, structure and particle size of extracted pectin.
However, suggest to utilize more sophisticated spectroscopic analysis such as
GCMS to detect the monosaccharide composition for more knowledge of the
fine structure of pectin and for its specific application.
(iii) Since process optimization has significant effect on the yield of pectin,
therefore various parameters were utilized for high yield of pectin and results
were authenticated through RSM studies. For future studies, suggest to use
RSM technique to authenticate the mechanical procedure at different pH as
well as in the bio-characterization of pectin such as degree of methylation and
degree of acetylation.
(iv) The formulation of solid oral pharmaceutical (tablets) is a very vast field and
needs good and cost effective binders in developmental work. The extracted
Sapodilla pectin was observed a good binder to be utilized in some tablet
formulation. Although dissolution studies were conducted, however it is
suggested to perform kinetic studies as well in future for more authentic data
and effect of Sapodilla pectin as a binder.
(v) Antidiarrheal preparation formulated in the present studies were compared
with the marketed product showed similar results, especially when tested for
its water holding capacity. In addition, the water binding and fat binding
capacities of extracted pectin indicated a promising results. This confidently
pointed towards the potential use of sapodilla pectin in antidiarrheal
211
preparation. However, in vivo procedures can add more data if performed in
future studies.
(vi) Pectin has great food applications. Present study demonstrated potential use of
extended sapodilla pectin in the preparation of Jam and pudding. However,
the present formulation needs further optimization for reproducible results
despite the fact that both formulation showed similar results when compared
with the formulation prepared from standard pectin. Yogurt and milk
preparations should also be formulated and tested to find more specific role of
sapodilla pectin in future.
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REFERENCES
Abbaszadeh, A.H. (2008). Pectin and galacturonic acid from citrus wastes (Master of
Science dissertation). Available from www.academia.edu/1272315
Abdullah, M., and Firmansyah, M. A. (2013). Clinical Approach and Management of
Chronic Diarrhea. ActaMedicaIndonesiana-The Indonesian Journal of Internal
Medicine, 45(2), 157-165.
Acikgoz, C. (2011). Extraction and Characterization of Pectin Obtained from Quince
Fruits (Cydonia vulgaris pers) Grown in Turkey. Asian Journal of
Chemistry, 23(1), 149-152.
Acton, Q. A. (Ed.). (2013). Lactobacillus—Advances in Research and Application.
Atlanta, Georgia: Scholarly Editions.
Ahmad, I., Arsalan, A., Ali, S. A., Bano, R., Munir, I., and Sabah, A. (2016). Formulation
and stabilization of norfloxacin in liposomal preparations.European Journal of
Pharmaceutical Sciences, 91, 208-215.
Ahmed, T., Burhanuddin, M., Haque, M., & Hossain, M. (2012). Preparation of Jam from
Sapota (Achraszapota). The Agriculturists Agriculturists, 9(1-2).
Aina, V., Barau, M., Mamman, O., Zakari, A., Haruna, H., Hauwa Umar, M. and Abba,
Y. (2012). Extraction and Characterization of Pectin from Peels of Lemon (Citrus
limon), Grape Fruit (Citrus paradisi) and Sweet Orange (Citrus sinensis). British
Journal of Pharmacology and Toxicology, 3(6), 259-262.
Ajenifujah-Solebo, S., and Aina, J. (2011). Physico-chemical properties and sensory
evaluation of jam made from black-plum fruit (vitexdoniana). African Journal of
Food, Agriculture, Nutrition and Development, 11(3).
213
Alba, K., Laws, A., and Kontogiorgos, V. (2015). Isolation and characterization of
acetylated LM-pectins extracted from okra pods. Food Hydrocolloids, 43, 726-
735.
AOAC International. (2006). Official Methods of Analysis AOAC International
.Washington DC. USA: AOAC International.
AOAC International. (2012). Official Methods of Analysis AOAC International.
Washington DC. USA: AOAC International.
Archimedes Pharma US (2011) PecSys® is a registered trademark of Archimedes
Development, Ltd. © 2011 Archimedes Pharma US Inc. July 2011
Ashaye, O. A., and Adeleke, T. O. (2009). Quality attributes of stored Roselle jam.
International Food Research Journal, 16, 363-371.
Ashford, M., Fell, J., Attwood, D., Sharma, H., and Woodhead, P. (1993). An evaluation
of pectin as a carrier for drug targeting to the colon. Journal of Controlled
Release, 26(3), 213-220.
Aydin, Z., and Akbugˇa, J. (1996). Preparation and evaluation of pectin
beads. International Journal of Pharmaceutics, 137(1), 133-136.
Azad, A., Ali, M., Akter, M., Jiaur Rahman, M, and Ahmed, M. (2014). Isolation and
Characterization of Pectin Extracted from Lemon Pomace during Ripening. JFNS
Journal of Food and Nutrition Sciences, 2(2), 30-35.
Baissise, S., Ghannem, H., and Fahloul, D. (2010). Comparison of Structure and
Emulsifying Activity of Pectin Extracted from Apple Pomace and Apricot Pulp.
World Journal of Dairy and Food Sciences5 (1), 79-84.
Baker, R. (1997). Reassessment of Some Fruit and Vegetable Pectin Levels. Journal of
Food Science, 62(2), 225-229.
214
Banerjee, N. and Singh, S. (2013). Formulation, Evaluation and Optimization of
Effervescent Granules to be reconstituted into Suspension of Levetiracetam for
Sustained Release. International Journal of Pharmaceutical Sciences Review and
Research, 20(2), 181-186
Bennett, M. E. (1958). U.S. Patent No. 2828242. Washington, DC: U.S. Patent and
Trademark Office.
Besson, V., Yapo, B. and Koffi, K. (2013). Cinnamon Apple Pomace Pectins:
Physicochemical Characteristics and Gel-Forming Properties. Journal of Human
Nutrition & Food Science, 1(3), 1019.
Bhat, S. and Singh, E. (2014). Extraction And Characterization of Pectin From Guava
Fruit Peel. International Journal of Research in Engineering & Advanced
Technology, 2(3).
Bis-Pectin Suspension. (2016). Retrieved July 21, 2016, from http://www.nps.org.au/
medicines/digestive-system/antidiarrhoea-medicines/codeine-kaolin-aluminium-
hydroxide-pectin/bis-pectin-suspension
Bonnaillie, L., Zhang, H., Akkurt, S., Yam, K. and Tomasula, P. (2014). Casein Films:
The Effects of Formulation, Environmental Conditions and the Addition of Citric
Pectin on the Structure and Mechanical Properties. Polymers, 6(7), 2018-2036.
Boonrod, D., Reanma, K., andNiamsup, H. (2006). Extraction and Physicochemical
Characteristics of Acid-Soluble Pectin from Raw Papaya (Carica papaya) Peel.
Chiang Mai Journal of Science, 33(1), 129-135.
Brito, E. S., and Narain, N. (2002). Physical and chemical characteristics of sapota fruit at
different stages of maturation. Pesquisa AgropecuáriaBrasileira, 37(4), 567-572
Broomes, J., and Badrie, N. (2010). Effects of Low-Methoxyl Pectin on Physicochemical
and Sensory Properties of Reduced- Calorie Sorrel/ Roselle (Hibiscus sabdariffa
L.) Jams. The Open Food Science Journal, 4(1), 48-55
215
Brouns, F., Theuwissen, E., Adam, A., Bell, M., Berger, A. and Mensink, R. (2011).
Cholesterol-lowering properties of different pectin types in mildly hyper-
cholesterolemic men and women. European Journal of Clinical Nutrition, 66,
591-599.
Buchholt, H. C., and Larsen, P. F. (2005). U.S. Patent No. US 6,855,363 B1. Washington,
DC: U.S. Patent and Trademark Office.
Caffall, K., and Mohnen, D. (2009). The structure, function, and biosynthesis of plant cell
wall pectic polysaccharides. Carbohydrate Research, 344, 1879-1900.
Calderon, L. (2012). Characterization of Apple Pectin –A Chromatographic Approach.
InChromatography - the most versatile method of chemical analysis (p. 329).
Rijeka: InTech.
Canteri, M. H., Nogueira, A., Petkowicz, C. D., &Wosiacki, G. (2012). Characterization
of Apple Pectin – A Chromatographic Approach (L. Calderon, Ed.).
In "Chromatography - The Most Versatile Method of Chemical Analysis",
(pp.325-342). Retrieved from http://www.intechopen.com
Canteri-Schemin M., Fertonani H., Waszczynskyj N. and Wosiacki G. (2005).
Extraction of pectin from apple pomace. Brazilian Archives of Biology and
Technology, 48(2). 259-266.
Cargill foods (2016). Commercial production of pectin. Retrieved from
https://www.cargillfoods.com/lat/en/products/hydrocolloids/pectins/manufacturin
g-process/index.jsp.
Carr, J.M., Sufferling, K., and Poppe, (1995). Hydrocolloids and their use in the
confectionery industry. Food Technol. 49(7), 41-44.
Castillo-Israel, K. A., Baguio, S. F., Diasanta, M. D., Lizardo, R. C., Dizon, E. I., and
Mejico, M. I. (2015). Extraction and characterization of pectin from Saba banana
[Musa ‘saba’(Musa acuminata x Musa balbisiana)] peel wastes: A preliminary
study. International Food Research Journal, 22(1), 202-207.
216
Castillo-Israel, K. A., Baguio, S. F., Diasanta, M. D., Lizardo, R. C., Dizon, E. I., and
Mejico, M. I. (2015). Extraction and characterization of pectin from Saba banana
[Musa ‘saba’(Musa acuminata x Musa balbisiana)] peel wastes: A preliminary
study. International Food Research Journal, 22(1), 202-207.
Chan, S. and Choo, W. (2013). Effect of extraction conditions on the yield and chemical
properties of pectin from cocoa husks. Food Chemistry, 141, 3752-3758.
Chandran, S., Praveen, G., Snima, K., Nair, S., Pavithran, K., Chennazhi, K. and
Lakshmanan, V. (2013). Potential Use of Drug Loaded Nano Composite Pectin
Scaffolds for the Treatment of Ovarian Cancer. Current Drug Delivery, 10(3),
326-335.
Chen, H., and Chen, W. (2011). Orchid biotechnology. Singapore: World Scientific.
Christy E., Suganya K., Kiruba J., Madhumitha S., Suja S. and Kalaivani G.(2014).
Extraction of Pectin from Fruit wastes– an effective method of municipal solid
waste management.International Journal of Advanced Research. 2(2). 936-944.
Constenla, D., and Lozano, J. E. (2003). Kinetic model of pectin demethylation. Latin
American Applied Research, 33, 91-96.
Cosgrove, D. and Jarvis, M. (2012). Comparative structure and biomechanics of plant
primary and secondary cell walls. Frontiers in Plant Science, 3, 1-6.
Cui, S., and Chang, Y. (2014). Emulsifying and structural properties of pectin
enzymatically extracted from pumpkin. LWT - Food Science and Technology,
58(2), 396–403.
Cybulska, J., Zdunek, A., and Kozioł, A. (2015). The self-assembled network and
physiological degradation of pectins in carrot cell walls. Food Hydrocolloids, 43,
41-50.
217
Daou, C. and Zhang, H. (2012). Study on Functional Properties of Physically Modified
Dietary Fibres Derived from Defatted Rice Bran.Journal of Agricultural Science
JAS, 4(9).
Darvill, J., Mcneil, M., Darvill, A., and Albersheim, P. (1980). Structure of Plant Cell
Walls: XI. Glucuronoarabinoxylan, a second hemicellulose in the primary cell
walls of suspension-cultured sycamore cells. Plant Physiology, 66, 1135-1139.
Dhingra, D., Michael, M., Rajput, H. and Patil, R. (2011). Dietary fibre in foods: A
review. J Food SciTechnol Journal of Food Science and Technology, 49(3), 255-
266.
Di Lorenzo, C., Williams, C., Hajnal, F. and Valenzuela, J. (1988). Pectin delays gastric
emptying and increases satiety in obese subject.Gastroenterology, 95(5), 1211-5.
Dikman, A. E., Schonfeld, E., Srisarajivakul, N. C., and Poles, M. A. (2015). Human
Immunodeficiency Virus-Associated Diarrhea: Still an Issue in the Era of
Antiretroviral Therapy. Digestive Diseases and Sciences, 60(8), 2236-2245.
Dominiak, M., Søndergaard, K., Wichmann, J., Vidal-Melgosa, S., Willats, W., Meyer,
A. and Mikkelsen, J. (2014). Application of enzymes for efficient extraction,
modification, and development of functional properties of lime pectin. Food
Hydrocolloids, 40, 273-282.
Dorta, E., Lobo, M. and González, M. (2013). Improving the Efficiency of Antioxidant
Extraction from Mango Peel by Using Microwave-assisted Extraction. Plant
Foods for Human Nutrition,68,190-199.
Du, F., Xiao, X. and Li, G. (2007). Application of ionic liquids in the microwave-assisted
extraction of trans-resveratrol from RhizmaPolygoniCuspidati. Journal of
Chromatography A, 1140(1-2), 56-62
El-Nawawi, S. and Shehata, F. (1987). Extraction of Pectin from Egyptian Orange Peel.
Factors Affecting the Extraction. ChemieIngenieurTechnik, 20(4), 699-699.
218
Endreb H, Christensen S. Pectins. In G. Phillips and P. Williams (2009), Handbook of
hydrocolloids (2nd ed., pp. 274-297). Boca Raton, Fla; CRC Press.
Englyst, H., Hay, S., and Macfarlane*, G. (1987). Polysaccharide breakdown by mixed
populations of human faecal bacteria. FEMS Microbiology Letters, 45(3), 163-
171.
Ensymm( 2015 ). Showing commercial production of pectin (ensymm) . Retrieved from
http://ensymm.com/wp-content/uploads/2016/01/ensymm_pectin_producti
on_abstract.pdf
Ershoff, B. H., and Wells, A. F. (1962). Effects of Gum Guar, Locust Bean Gum and
Carrageenan on Liver Cholesterol of Cholesterol-Fed Rats. Experimental Biology
and Medicine, 110(3), 580-582.
Fertonani, H. C., Scabio, A., Carneiro, E. B., Schemim, M. H., Nogueira, A., and
Wosiacki, G. (2009). Extraction model of low methoxyl pectin from apple
pomace effects of acid concentration and time on the process and the
product. Brazilian Archives of Biology and Technology Braz. Arch. Biol.
Technol., 52(1), 177-185.
Figuerola, F., Hurtado, M. L., Estévez, A. M., Chiffelle, I., and Asenjo, F. (2005). Fibre
concentrates from apple pomace and citrus peel as potential fibre sources for food
enrichment. Food Chemistry, 91(3), 395-401.
Fishman G. and Jen F. Thickening and gelling Agents for food. 1st ed. Publ. Blackies
Academic and professional, London: 1986, p- 175.
Fishman, M. L., Chau, H. K., Hoagland, P., and Ayyad, K. (1999). Characterization of
pectin, flash-extracted from orange albedo by microwave heating, under
pressure. Carbohydrate Research, 323(1-4), 126-138.
Fishman, M., Chau, H., Cooke, P. and Jr., A. (2008). Global Structure of Microwave-
Assisted Flash-Extracted Sugar Beet Pectin. Journal of Agricultural and Food
Chemistry, 56, 1471-1478.
219
Food and Agriculture Organization of the United Nations. (2009). Pectin. Retrieved from
the Food and Agriculture Organization of the United Nations: http://www.fao.org/
ag/agn/jecfa-additives/specs/monograph7/additive-306-m7.pdf.
Food and Agriculture Organization of the United Nations. (2015). Jam.Retrieved from
the Food and Agriculture Organization of the United Nations: www.fao.org/3/a-
au115e.pdf.
Fraeye, I., Duvetter, T., Verlent, I., Sila, D. N., Hendrickx, M., and Loey, A. V. (2007).
Comparison of enzymatic de-esterification of strawberry and apple pectin at
elevated pressure by fungal pectinmethylesterase.Innovative Food Science &
Emerging Technologies, 8(1), 93-101.
Franchi, M., Marzialetti, M., Pose, G. and Cavalitto, S. (2014). Evaluation of Enzymatic
Pectin Extraction by a Recombinant Polygalacturonase (PGI) From Apples and Pears
Pomace of Argentinean Production and Characterization of the Extracted Pectin. J
Food Process Technol Journal of Food Processing & Technology, 5(8), 1-4.
Funami, T., Nakauma, M., Ishihara, S., Tanaka, R., Inoue, T. and Phillips, G. (2011).
Structural modifications of sugar beet pectin and the relationship of structure to
functionality. Food Hydrocolloids, 25(2), 221-229.
Gama, B. V., Silva, C. D., Silva, L. O., and Abud, A. D. (2015). Extraction and
Characterization of Pectin from Citric Waste. Chemical Engineering
Transactions, 44, 259-264.
Garna, H., Mabon, N., Nott, K., Wathelet, B., and Paquot, M. (2006). Kinetic of the
hydrolysis of pectin galacturonic acid chains and quantification by ionic
chromatography. Food Chemistry, 96(3), 477-484.
Garna, H., Mabon, N., Robert, C., Cornet, C., Nott, K., Legros, H., Wathelet B, and
Paquot, M. (2007). Effect of Extraction Conditions on the Yield and Purity of
Apple Pomace Pectin Precipitated but Not Washed by Alcohol.Journal of Food
Science J Food Science, 72(1), 1-9.
220
Garna, H., Mabon, N., Wathelet, B., and Paquot, M. (2004). New Method for a Two-
Step Hydrolysis and Chromatographic Analysis of Pectin Neutral Sugar
Chains. Journal of Agricultural and Food Chemistry, 52(15), 4652-4659.
Gbassi, G. K., Atheba, P., Yolou, F. S., and Vandamme, T. (2013). Macrobeads Based-
Polysaccharides: Development and Morphological Analysis. World Applied
Sciences Journal, 22(5), 732-737.
Genu (2016). Commecial production of pectin. Retrieved from http://www.cfs.purdue
.edu/fn/fn453/GENU%20Pectin%20Book%202005-10.pdf.
Georgiev. Yordan.,Ognyanov. Manol.,Yanakieva. Irina., Kussovski. Veselin, and
Kratchanova Maria. (2012). Isolation, characterization and modification of citrus
pectinsJournal of BioScience and Biotechnology, 1(3).223-233.
Ginter, E., Kubec, F.J., Vozar, J and Bobek, P. (1979). Natural hypocholesterolemic
agent: pectin plus ascorbic acid. International Journal of Viticulture and Natural
Resource,49, 406-408.
Gnanasambandam, R., and Proctor, A. (1999). Preparation of soy hull pectin. Food
Chemistry, 65(4), 461-467.
Gnanasambandam, R., and Proctor, A. (2000). Determination of pectin degree of
esterification by diffuse reflectance Fourier transform infrared spectroscopy. Food
Chemistry, 68(3), 327-332.
Gruy, F. (2011). Relationship between the morphology and the light scattering cross
section of optically soft aggregates. Journal of Quantitative Spectroscopy and
Radiative Transfer, 112(16), 2609-2618.
Gullón, B., Gómez, B., Martínez-Sabajanes, M., Yáñez, R., Parajó, J. and Alonso, J.
(2013). Pectic oligosaccharides: Manufacture and functional properties. Trends in
Food Science & Technology, 30(1), 153-161.
221
Guo, X., Han, D., Xi, H., Rao, L., Liao, X., Hu, X. and Wu, J. (2012). Extraction of
pectin from navel orange peel assisted by ultra-high pressure, microwave or
traditional heating: A comparison. Carbohydrate Polymers, 88(2), 441-448.
Guolin, H., Jeffrey, S., Kai, Z. and Xiaolan, H. (2012). Application of Ionic Liquids in
the Microwave-Assisted Extraction of Pectin from Lemon Peels. Journal of
Analytical Methods in Chemistry, 1-8.
Hagesaether, E. and Sande, S. (2007). In Vitro Measurements of Mucoadhesive
Properties of Six Types of Pectin. Drug Development and Industrial
Pharmacy, 33(4), 417-425.
Hagesaether, E., Hiorth, M. and Sande, S. (2009). Mucoadhesion and drug permeability
of free mixed films of pectin and chitosan: An in vitro and ex vivo
study. European Journal of Pharmaceutics and Biopharmaceutics, 71(2), 325-
331.
Hamza, S., Naseem, S., Erum, B., and Hina, B. (2013). Trace element geochemistry of
Manilkarazapota (L.) P. Royen, fruit from winder, Balochistan, Pakistan in
perspective of medical geology. Pakistan Journal of Pharmaceutical
Sciences,26(4), 805-811.
Harholt, J., Suttangkakul, A., and Scheller, H. V. (2010). Biosynthesis of Pectin. Plant
Physiology, 153(2), 384-395.
Hester, R. and Harrison, R. (2013). Waste as a resource. Cambridge: Royal Society of
Chemistry.
Holt, P., Dominguez, A., and Kwartler, J. (1979). Effect of sucrose feeding upon
intestinal and hepatic lipid synthesis. The American Journal of Clinical
Nutrition,32, 1792-1798.
Huang, J. (1973). Improved method for the extraction of pectin. Florida State
Horticultural Society.
222
Hussain, A., Yasmin, A. and Ali, J. (2010). Comparative study of chemical composition
of some dried apricot varieties grown in northern areas of Pakistan. Pakistan
Journal of Botany, 42(4), 2497-2502.
Hwang, J., Kim, C., and Kim, C. (1998). Extrusion of Apple Pomace Facilitates Pectin
Extraction. Journal of Food Science, 63(5), 841-844.
Imeson, A. (Ed.). (2010). Food stabilisers, thickeners and gelling agents. Chichester,
U.K.: Wiley-Blackwell Pub.
IMR International. (2016). Pie chart showing major sources of pectin used arround the
world. Retrieved from http://www.hydrocolloid.com/.
IMR International. (2016). Pie chart Showing major pectin producers and their market
share. Retrieved from http://www.hydrocolloid.com/
IMR International. (2016). Pie chart shwing the overall application and uses of pectin in
pharmaceutical, food, cosmetics and chemical industries. Retrieved from
http://www.hydrocolloid.com/
Ink, S. L., and Hurt, H. D. (1987). Nutritional implications of gums. Food Tech, 41, 77-82.
Islam, M., Monalisa, K, and Hoque, M. (2012). Effect of Pectin on the processing and
preservation of Strawberry (Fragariaananassa) jam and jelly. International
Journal of Natural Sciences, 2(1), 8-14.
Ismail, N., Ramli, N., Hani, N., and Meon, Z. (2012). Extraction and Characterization of
Pectin from Dragon Fruit (Hylocereuspolyrhizus) using Various Extraction
Conditions. SainsMalaysiana, 41(1), 41-45.
J.F.Hydrocolloids (2013). Showing different uses and properties of pectin. Retrieved
from www.jfhydrocolloids.com/applications.html# pectin.
Jain, J. L., Jain, S., and Jain, N. (2005). Fundamentals of biochemistry: For university
and college students in India and abroad. New Delhi: S. Chand &.& Co Ltd
223
Jameson, E., TFrancis N, T. N., and Clarence, W. P. (1924). U.S. Patent No. US 1497884
A. Washington, DC: U.S. Patent and Trademark Office.
Jenkins, D., Leeds, A., Wolever, T., Goff, D., George, K., Alberti, Gassull, M., Derek,
T.,Hockaday, R. (1976). Unabsorbable Carbohydrates And Diabetes: Decreased
Post-Prandial Hyperglycæmia. The Lancet, 308(7978), 172-174.
Jones, O., Lesmes, U., Dubin, P. and McClements, D. (2010). Effect of polysaccharide
charge on formation and properties of biopolymer nanoparticles created by heat
treatment of b-lactoglobulin–pectin complexes. Food Hydrocolloids, 24, 374–
383-374–383.
Kalapathy, U., and Proctor, A. (2001). Effect of acid extraction and alcohol precipitation
conditions on the yield and purity of soy hull pectin. Food Chemistry, 73(4), 393-
396.
Kamnev, A. A., Colina, M., Rodriguez, J., Ptitchkina, N. M., and Ignatov, V. V. (1998).
Comparative spectroscopic characterization of different pectins and their
sources. Food Hydrocolloids, 12(3), 263-271.
Kanmani, P., Dhivya, E., Aravind, J. and Kumaresan, K. (2014). Extraction and Analysis
of Pectin from Citrus Peels: Augmenting the Yield from Citrus limon Using
Statistical Experimental Design. Iranica Journal of Energy & Environment, 5(3),
303-312.
Kar, F., and Arslan, N. (1999). Characterization of orange peel pectin and effect of
sugars, l-ascorbic acid, ammonium persulfate, salts on viscosity of orange peel
pectin solutions. Carbohydrate Polymers, 40(4), 285-291.
Kaya, M., Sousa, A., Crepeau, M., Sorensen, S. and Ralet, M. (2014). Characterization of
citrus pectin samples extracted under different conditions: Influence of acid type
and pH of extraction. Annals of Botany, 1319-1326.
224
Kermani, Z., Shpigelman, A., Kyomugasho, C., Buggenhout, S., Ramezani, M., Loey, A.,
and Hendrickx, M. (2014). The impact of extraction with a chelating agent under
acidic conditions on the cell wall polymers of mango peel. Food Chemistry
Volume, 161, 199–207.
Kertesz, Z. I. (1951). The pectic substances. New York: Interscience.
Khan, M., Bibi, N., and Zeb, A. (2015). Optimization of Process Conditions for Pectin
Extraction from Citrus Peel. Science, Technology and Development, 34(1), 9-15.
Kim, H., Venkatesh, G., &Fassihi, R. (1998). Compactibility characterization of granular
pectin for tableting operation using a compaction. International Journal of
Pharmaceutics, 161, 149-159.
Kirtchev N., Panchev I. and Kratchanov C. (1989). Kinetics of acid-catalysed de–
esterification of pectin in a heterogeneous medium. International Journal of Food
Science and Technology, 24, 479-486.
Kliemann, E., De Simas, K., Amante, E., Prudêncio, E., Teófilo, E., Ferreira, M. and
Amboni, R. (2009). Optimisation of pectin acid extraction from passion fruit peel
(Passiflora edulis flavicarpa) using response surface methodology. International
Journal of Food Science & Technology, 44(3), 476–483.
Koffi, K., Yapo, B. and Besson, V. (2013). Extraction and characterization of gelling
pectin from the peel of Poncirustrifoliatafruit. Agricultural Sciences AS, 4(11),
614-619.
Koh, P., Leong,, C. and Noranizan, M. (2014). Microwave-assisted extraction of pectin
from jackfruit rinds using different power levels. International Food Research
Journal, 21(5), 2091-2097.
Kohn, R. (1982). Binding of toxic cations to pectin, its oligomeric fragment and plant
tissues. Carbohydrate Polymers, 2, 273-275.
225
Kopjar, M., Piližota, V., Nedićtiban, N., Šubarić, D., Babić, J., Ačkar, Đ. and Sajdl, M.
(2009). Strawberry Jams: Influence of Different Pectins on Colour and Textural
Properties. Czech Journal of Food Sciences, 27(1), 20-28.
Koubala, B., Kansci, G., Mbome, L., Crépeau, M., Thibault, J., and Ralet, M. (2008).
Effect of extraction conditions on some physicochemical characteristics of pectins
from “Améliorée” and “Mango” mango peels.Food Hydrocolloids, 22(7), 1345-
1351.
Kratchanova, M., Panchev, I., Pavlova, E., and Shtereva, L. (1994). Extraction of pectin
from fruit materials pretreated in an electromagnetic field of super-high
frequency. Carbohydrate Polymers, 25(3), 141-144.
Kratchanova, M., Pavlova, E., and Panchev, I. (2004). The effect of microwave heating
of fresh orange peels on the fruit tissue and quality of extracted
pectin.Carbohydrate Polymers, 56(2), 181-185.
Kratchanova, M., Pavlova, E., Panchev, I., and Kratchanov, C. (1996). Influence of
microwave pretreatment of fresh orange peels on pectin extraction. Progress in
Biotechnology Pectins and Pectinases, Proceedings of an International
Symposium, 941-946.
Krishnamurti, C. and Giri, K. (1949). Preparation, purification and composition of
pectins from Indian fruits and vegetables. Proceedings of the Indian Academy of
Sciences - Section B, 29(4), 155-16.
Krusteva, S., Lambov, N., and Velinov, G. (1990). Pharmaceutical investigation of a
bioerodible nystatin system. Die Pharmazie, 45(3), 197.
Kulkarni, S. and Vijayanand, P. (2010). Effect of extraction conditions on the quality
characteristics of pectin from passion fruit peel (Passiflora edulis f. flavicarpa
L.). LWT - Food Science and Technology, 43(7), 1026-1031.
226
Kumar, A. and Chauhan, G. (2010). Extraction and characterization of pectin from apple
pomace and its evaluation as lipase (steapsin) inhibitor. Carbohydrate
Polymers, 82(2), 454-459.
Kushwaha, P., Fareed, S., Nanda, S. and Mishra, A. (2011). Design & Fabrication of
Tramadol HCl loaded Multiparticulate Colon Targeted Drug Delivery
System. Journal of Chemical and Pharmaceutical Research, 3(5), 584-595.
Lamotte C., Gochnauer C., Lamotte L., Mathur J. and Davies L. (1969). Pectin Esterase
in Relation to Leaf Abscission in Coleus and Phaseolus. Plant Physiology,44, 21-
26
Leclere, L., Cutsem, P. and Michiels, C. (2013). Anti-cancer activities of pH- or heat-
modified pectin. Frontiers in Pharmacology, 4, 128-128.
Lecumberri, E., Mateos, R., Izquierdo-Pulido, M., Rupérez, P., Goya, L., and Bravo, L.
(2007). Dietary fibre composition, antioxidant capacity and physico-chemical
properties of a fibre-rich product from cocoa (Theobroma cacao L.). Food
Chemistry, 104(3), 948-954.
Li, D., Jia, X., Wei, Z. and Liu, Z. (2012). Box–Behnken experimental design for
investigation of microwave-assisted extracted sugar beet pulp pectin.
Carbohydrate Polymers, 88(1), 342-346.
Liu, L., Cao, J., Huang, J., Cai, Y. and Yao, J. (2010). Extraction of pectins with
different degrees of esterification from mulberry branch bark. Bioresource
Technology, 101(9), 3268-3273.
Liu, Y., Shi, J. and Langrish, T. (2006). Water-based extraction of pectin from flavedo
and albedo of orange peels. Chemical Engineering Journal, 120(3), 203-209.
Lochridge, E. H. (1951). U.S. Patent No. 2568752. Washington, DC: U.S. Patent and
Trademark Office.
227
Loyola N., Pavéz P and Lillo S. (2011). Pectin extraction from cv. Pink Lady (Malus
pumila) apples. Ciencia e investigación agrarian, 38(3). 425-434.
Ma, S., Yu, S., Zheng, X., Wang, X., Bao, Q. and Guo, X. (2013). Extraction,
characterization and spontaneous emulsifying properties of pectin from sugar beet
pulp. Carbohydrate Polymers, 98(1), 750-753.
Madhav, A., and Pushpalatha, P. B. (2002). Characterization of pectin extracted from
different fruit wastes. Journal of Tropical Agriculture, 40, 53-55.
Mahattanatawee, K., Manthey J., Luzio G., Talcott S., Goodner K. and Baldwin E.
(2006). Total Antioxidant Activity and Fiber Content of Select Florida-Grown
Tropical Fruits. Journal of Agricultural and Food Chemistry, 54, 7355-7363.
Maran, J., Sivakumar, V., Thirugnanasambandham, K. and Sridhar, R. (2014).
Microwave assisted extraction of pectin from waste Citrulluslanatus fruit
rinds. Carbohydrate Polymers, 101, 786-791.
Markets and Markets (2015). Graph showing current and growing market size of
hydrocolloids in the different regions of the world. Retrieved from http://www.
marketsandmarkets.com/PressReleases/hydrocolloid.asp
Maxwell, E., Belshaw, N., Waldron, K. and Morris, V. (2012). Pectin – An emerging
new bioactive food polysaccharide. Trends in Food Science & Technology, 24,
64-73.
May, C. (1990). Industrial pectins: Sources, production and applications. Carbohydrate
Polymers, 12(1), 79-99.
Menon, S., Basavaraj, B., Bharath, S., Deveswaran, R, and Madhavan, V. (2011).
Formulation and evaluation of ibuprofen tablets using orange peel pectin as
binding agent. Der Pharmacia Lettre, 3(4), 241-247.
228
Miettinen, T.A and Tarpila, S (1977): Effect of pectin on serum cholesterol fecal-bile
acid and biliary lipids in nonemolipidemic and hyperlipidemic individuals.
ClinicaChimicaActa, 60, 1429-1431.
Mikami, S., Iwano, K., Shiinoki, S. and Shimada, T. (1987). Purification and some
properties of acid-stable .ALPHA.-amylases from shochukoji (Aspergillus
kawachii). Agricultural and Biological Chemistry, 51(9), 2495-2501.
Milind, P and P. (2015). Chickoo: A Wonderful Gift from Nature. International Journal
of Research in Ayurveda and Pharmacy, 6(4), 544-550.
Ministry of health portal Kingdom of Saudi Arabia (2016). Ministry of Health Formulary
Drug list. Retrieved from Ministry of health portal Kingdom of Saudi Arabia:
http://www.moh.gov.sa.
Minjares-Fuentes, R., Femenia, A., Garau, M., Meza-Velázquez, J., Simal, S. and
Rosselló, C. (2014). Ultrasound-assisted extraction of pectins from grape pomace
using citric acid: A response surface methodology approach. Carbohydrate
Polymers, 106, 179-189.
Mittal, N. and Kaur, G. (2014). In situ gelling ophthalmic drug delivery system:
Formulation and evaluation. Journal of Applied Polymer Science, 131(2).
Mohnen, D. (2008). Pectin structure and biosynthesis. Current Opinion in Plant
Biology, 11, 266-277.
Molecular expressions. (2016). Artist's impression of Structure of plant cell showing
location of pectin in the cell wall. Retrieved from http://micro.magnet.
fsu.edu/cells/plants/cellwall.html
Monsoor, M. A., and Proctor, A. (2001). Preparation and functional properties of soy hull
pectin. Journal of the American Oil Chemists' Society, 78(7), 709-713.
229
Monsoor, M. A., Kalapathy, U., and Proctor, A. (2001). Improved Method for
Determination of Pectin Degree of Esterification by Diffuse Reflectance Fourier
Transform Infrared Spectroscopy. Journal of Agricultural and Food
Chemistry, 49(6), 2756-2760.
Morris, G., Kök, S., Harding, S. and Adams, G. (2010). Polysaccharide drug delivery
systems based on pectin and chitosan. Biotechnology and Genetic Engineering
Reviews, 27, 257-284.
Morris, V., Gromer, A., Kirby, A., Bongaerts, R. and Gunning, A. (2011). Using AFM
and force spectroscopy to determine pectin structure and (bio) functionality. Food
Hydrocolloids,25(2), 230-237.
Muhamadzadeh, J., Sadghi-Mahoonak, A. R., Yaghbani, M., and Aalam, M. (2010).
Extraction of Pectin from Sunflower Head Residues of Selected Iranian
Caltivers. World Applied Science Journal, 8, 21-24.
Muhammad, K., Zahari, N., Gannasin, S., Adzahan, N. and Bakar, J. (2014). High
methoxyl pectin from dragon fruit (Hylocereuspolyrhizus) peel. Food
Hydrocolloids, 2(Special Issue: International Conference on Halal Gums 2012),
289–297
Munarin, F., Tanzi, M. and Petrini, P. (2012). Advances in biomedical applications of
pectin gels. International Journal of Biological Macromolecules, 51, 681-689.
Naggar, V. F., El-Khawas, M., Ismail, F. A., and Boraie, N. A. (1992.). Pectin, a
possible matrix for oral sustained-release preparations of water-soluble
drugs. STP Pharma Sciences, 2, 227-234.
Naghshineh, M., Olsen, K. and Georgiou, C. (2013). Sustainable production of pectin
from lime peel by high hydrostatic pressure treatment. Food Chemistry, 136(2),
472-478.
230
Nanji, D. R. (1927). U.S. Patent No. 634 879. Washington, DC: U.S. Patent and
Trademark Office.
Nanji, H. and Chinoy, J. (1934). A simple method for the purification of Citrus
pectin.Biochemical Journal, 28(2), 456–462.
Nasrin, N. Asaduzzaman,M Mowla R, Rizwan F, Alam, A. (2011). A comparative study
of physical parameters of selected ketorolac tromethamine tablets available in the
pharma market of Bangladesh. Journal of Applied Pharmaceutical Science, 1(8),
101-103
Ndabikunze, B. K., Masambu, B. N., Tiisekwa, B. P., and Issa-Zacharia, A. (2011). The
production of jam from indigenous fruits using baobab (Adansoniadigitata L.)
powder as a substitute for commercial pectin. African Journal of Food
Science,5(3), 168-175.
Nguyen, S., Alund, S. J., Hiorth, M., Kjøniksen, A., and Smistad, G. (2011). Studies on
pectin coating of liposomes for drug delivery. Colloids and Surfaces B:
Biointerfaces, 88(2), 664-673.
Nurdjanah, S., Hook, J., Paton, J, and Paterson, J. (2013). Galacturonic Acid Content and
Degree of Esterification of Pectin from Sweet Potato Starch Residue Detected
Using 13C CP/MAS Solid State NMR. European Journal of Food Research &
Review, 3(1), 16-37.
Nwosu, J. N., Udeozor, L. O., Ogueke, C. C., Onuegbu, N., Omeire, G. C., and Egbueri,
I. S. (2014). Extraction and Utilization of Pectin from Purple Star-Apple
(Chrysophyllumcainito) and African Star-Apple (Chrysophyllumdelevoyi) in Jam
Production. Austin Journal of Nutrition and Food Sciences, 1(1), 1003.
Nwosu, J., Udeozor, L., Ogueke, C., Onuegbu, N., Omeire, G, and Egbueri, I. (2014).
Extraction and Utilization of Pectin from Purple Star- Apple (Chrysophyl
lumcainito) and African Star-Apple (Chrysophyllumdelevoyi) in Jam Production.
Austin Journal of Nutrition and Food Sciences, 1(1), 1003-1003.
231
Oliveira, T. Í, Rosa, M. F., Cavalcante, F. L., Pereira, P. H., Moates, G. K., Wellner N,
Mazetto SE, Waldron, KW., Azeredo H M C .(2016). Optimization of pectin
extraction from banana peels with citric acid by using response surface
methodology. Food Chemistry,198, 113-118.
Ovodov, Y. (2009). Current views on pectin substances. Russian Journal of Bioorganic
Chemistry, 35(3), 269-284.
Pagarra, H., Rahman, R. A., &Jusoh, M. (2014). Isolation of Pectin from
NephrolepisBiserrata Leaves at Different Extraction Time. JurnalTeknologi,
69(5).
Panda, H. (2011). The complete book on gums and stabilizers for food industry. Delhi:
Asia Pacific Business Press.
Pandharipande, S. and Makode, H. (2012). Separation of oil and pectin from orange peel
and study of effect of pH of extracting medium on the yield of pectin. Journal of
Engineering Research and Studies, 3(2).
Park, S., Venditti, R., Jameel, H. and Pawlak, J. (2006). Changes in pore size distribution
during the drying of cellulose fibers as measured by differential scanning
calorimetry. Carbohydrate Polymers, 66(1), 97-103.
Paskins-Hurlburt, A. J., Tanaka, Y., Skoryna, S., Moore, W., and Stara, J. (1977). The
binding of lead by a pectic polyelectrolyte. Environmental Research, 14(1), 128-
140.
Pinheiro, E., Silva, I., Gonzaga, L., Amante, E., Teófilo, R., Ferreira, M. and Amboni, R.
(2008). Optimization of extraction of high-ester pectin from passion fruit peel
(Passiflora edulis flavicarpa) with citric acid by using response surface
methodology. Bioresource Technology, 99(13), 5561-5566.
232
Poovaiah, B., Nukaya A. (1979). Polygalacturonase and Celiulase Enzymes in the
Normal Rutgers and Mutant rin Tomato Fruits and Their Relationship 'to the
Respiratory Climacteric'. Plant Physiology.64,534-537.
Ptichkina, N., Markina, O. and Rumyantseva, G. (2008). Pectin extraction from pumpkin
with the aid of microbial enzymes. Food Hydrocolloids, 22(1), 192-195.
Puri, M., Sharma, D. and Barrow, C. (2012). Enzyme-assisted extraction of bioactives
from plants. Trends in Biotechnology, 30(1), 37-44.
Rababah, T., Al-U'datt, M., Al-Mahasneh, M., Yang, W., Feng, H., Ereifej, K., Kilani, I.
and Ishmais, M. (2014). Effect of Jam Processing and Storage on Phytochemicals
and Physiochemical Properties of Cherry at Different Temperatures. Journal of
Food Processing and Preservation, 38(1), 247-254
Ramli, N., and Asmawati. (2011). Effect of ammonium oxalate and acetic acid at several
extraction time and pH on some physicochemical properties of pectin from cocoa
husks (Theobroma cacao). African Journal of Food Science, 5(15), 790-798.
Ranganna, S. (1986). Handbook of analysis and quality control for fruit and vegetable
products (2nd ed.). New Delhi: Tata McGraw-Hill.
Rao, M. R., Sadaphule, P., Khembete, M., Lunawat, H., Thanki, K., and Gabhe, N.
(2013). Characterization of Psyllium (Plantago ovata) Polysaccharide and its Use
as a Binder in Tablets. Indian Journal of Pharmaceutical Education and
Research, 47(2), 154-159.
Rasheed, A. (2008). Effect of Different Acids, Heating Time and Particle Size on Pectin
Extraction from Watermelon Rinds. Journal of Kerbala University, 6(4).
Raudnitz, J. (1979). U.S. Patent No. 4139612. Washington, DC: U.S. Patent and
Trademark Office.
Ridley, B. L., O'neill, M. A., and Mohnen, D. (2001). Pectins: Structure, biosynthesis,
and oligogalacturonide-related signaling. Phytochemistry, 57(6), 929-967.
233
Ridley, B., O'neill, M. and Mohnen, D. (2001). Pectins: Structure, biosynthesis, and
oligogalacturonide-related signaling. Phytochemistry, 57, 929-967.
Rinaldi, L., Rioux, L., Britten, M. and Turgeon, S. (2015). In vitro bioaccessibility of
peptides and amino acids from yogurt made with starch, pectin, or β-
glucan. International Dairy Journal, 46, 39-45.
Rouse, A. H., and Crandall, P. G. (1978). Pectin Content Of Lime And Lemon Peel As
Extracted By Nitric Acid. Journal of Food Science J Food Science, 43(1), 72-73.
Rubinstein, A., and Ezra, R. M. (1992). Adhesion of bacteria on pectin casted
films. Microbios., 70, 284-285.
Rubinstein, A., Radai, R., Ezra, M., Pathak, S., and Rokem, J. S. (1993). In vitro
evaluation of calcium pectinate: A potential colon-specific drug delivery
carrier. Pharmaceutical Research, 10(2), 258-263.
Rudolph B. and Petersen S.A. (2012). U.S. Patent No.U.S. 20120190831A1. Washington,
DC: U.S. Patent and Trademark Office.
Rungrodnimitchai, S. (2011). Novel source of pectin from young sugar palm by
microwave assisted extraction. Procedia Food Science, 1, 1553-1559.
Sakamoto, T., Hours, R. A., and Sakai, T. (1995). Enzymic pectin extraction from
protopectins using microbial protopectinases. Process Biochemistry, 30(5), 403-
409.
Salam, M., Jahan, N., Islam, M., &Hoque, M. (2014). Extraction of Pectin from lemon
peel: Technology development. Journal of Chemical Engineering, 27(2), 25-30.
Sandberg, A. S., Ahderinne, R., Andersson, H., Hallgren, B., and Hultén, L. (1983). The
effect of citrus pectin on the absorption of nutrients in the small intestine. Human
Nutrition. Clinical Nutrition, 37(3), 171-183.
234
Santos, J., Espeleta, A., Branco, A. and Assis, S. (2013). Aqueous extraction of pectin
from sisal waste. Carbohydrate Polymers, 92, 1997-2001.
Sayah M., Chabir R., El Madani N., Rodi El Kandri Y., OuazzaniChahdi F., Touzani H.
and Errachidi F.( 2014). Comparative Study on Pectin Yield According to the
State of the Orange Peels and Acids Used International journal of innovative
research in science, engineering and technology, 3(8).15658-15665.
Schols, H., Voragen, A., andVisser, R. (2009). Pectins and pectinases. Wageningen
Academic.
Seixas, F., Fukuda, D., Turbiani, F., Garcia, P., Petkowicz, C., Jagadevan, S. and
Gimenes, M. (2014). Extraction of pectin from passion fruit peel (Passiflora
edulis f. flavicarpa) by microwave-induced heating. Food Hydrocolloids, 38, 186-
192.
Seymour, G. B., and Knox, J. P. (2002). Pectins and their manipulation. Oxford:
Blackwell.
Shaha, R., Punichelvana, Y. and Afandi, A. (2013). Optimized Extraction Condition and
Characterization of Pectin from Kaffir Lime (Citrus hystrix). Research Journal of
Agriculture and Forestry Sciences, 1(2), 1-11.
Shahnawaz, M., Sheikh, S, and Nizamani, S. (2009). Determination of Nutritive Values
of Jamun Fruit (Eugenia jambolana) Products. Pakistan J. of Nutrition Pakistan
Journal of Nutrition, 8(8), 1275-1280.
Shailendra, P., Shikha, A, and Singh, L. (2012). Natural Binding Agents in Tablet
Formulation. International Journal of Pharmaceutical & Biological
Archives, 3(3), 466-473.
Sharma, H., Lahkar, S, and Nath, L. (2013). Extraction, CharacterisationAnd
Compatibility Study Of Polysaccharides FromDilleniaindica and Abelmoschu
235
sesculentus With Metformin Hydrochloride For Development of Drug Delivery
System. International Journal of PharmTech Research, 5(1), 275-283.
Shi, J., Mazza, G., and Maguer, M. (Eds.). (2002). Biochemical & processing
aspects (Vol. 2). Florida: CRC press.
Shi, J., Mazza, G., andMaguer, M. L. (1998). Functional foods: Biochemical and
processing aspects. Lancaster, PA: Technomic Pub.
Sila, D., Buggenhout, S., Duvetter, T., Fraeye, I., Roeck, A., Loey, A.andHendrickx, M.
(2009). Pectins in Processed Fruits and Vegetables: Part II-Structure-Function
Relationships. Comprehensive Reviews in Food Science and Food Safety, 8, 86-
104
Sindumathi, G., and Amutha, S. (2014). Processing and quality evaluation of coconut
based jam. IOSR Journal of Environmental Science, Toxicology and Food
Technology, 8(1), 10-14.
Singthong, J., Cui, S., Ningsanond, S., and Douglasgoff, H. (2004). Structural
characterization, degree of esterification and some gelling properties of Krueo Ma
Noy (Cissampelospareira) pectin. Carbohydrate Polymers, 58(4), 391-400.
Slany, J et al. (1981a): Evaluation of tablets with pectin as a binding agent.
FarmaceutickyObzor, 50, 491-498.
Slany, J et al. (1981b): Study of functional action of citrus pectins in tablets. Ceska a
SlovenskaFarmacie, 30,195-200.
Slavov A., Karagyozov V., Denev P., Kratchanova M. and Kratchanov C. (2013).
Antioxidant Activity of Red Beet Juices Obtained after Microwave and Thermal
Pretreatments. Czech Journal of Food Sciences,31(2).139-147.
Smith, B. C. (1999). Infrared spectral interpretation: A systematic approach. Boca
Raton: CRC Press
236
Smith, H. (2013). Gastroenteritis. Retrieved from South African Pharmacist's Assistant:
http://www.sapajournal.co.za/index.php/SAPA/article/download/510/751
Sotanaphun, U., Chaidedgumjorn, A., Kitcharoen, N., Satiraphan, M., Asavapichayont, P.
and Sriamornsak, P. (2011). Preparation of Pectin from Fruit Peel of Citrus
maxima. Silpakorn University Science and Technology Journal, 6(1), 42-48.
Sriamornsak, P. (1996). The utilization of pectin for the development of sustained release
theophylline pellets. (Master of Science dissertation).
Sriamornsak, P. (2001). Pectin, The role in health. Journal of Silpakorn University, 21-
22, 60-77.
Sriamornsak, P.(2002). Chemistry of pectin and its pharmaceutical uses: A review.
Journal of Pharmaceutical Sciences and Pharmacology,44, 207-228.
Sriamornsak, P., Prakongpan, S., Puttipipatkhachorn, S., and Kennedy, R. A. (1997).
Development of sustained release theophylline pellets coated with calcium
pectinate. Journal of Controlled Release, 47(3), 221-232.
Sriamornsak, P., Puttipipatkhachorn, S., and Prakongpan, S. (1997). Calcium pectinate
gel coated pellets as an alternative carrier to calcium pectinate
beads. International Journal of Pharmaceutics, 156(2), 189-194.
Sriamornsak, P., Wattanakorn, N. and Takeuchi, H. (2010). Study on the mucoadhesion
mechanism of pectin by atomic force microscopy and mucin-particle
method. Carbohydrate Polymers, 79, 54-59.
Srivastava, P and Malviya, R .(2011). Sources of pectin, extraction, and its application in
pharmaceutical industry – An overview, Indian Journal of Natural Products and
Resources,2(1), 10-18.
Steel, R. G., and Torrie, J. H. (1997). Principles and procedures of statistics: A
biometrical approach. New York: McGraw-Hill.
237
Steel, R.G.D., Torrie, J.H. and Dickey, D.A. 1997. Principles and procedures of statistics,
a biometrical approach. 3rd edition. McGraw-Hill Co. Inc., New York NY.
Stone, H., and Sidel, J. L. (2004). Sensory evaluation practices. Amsterdam: Elsevier
Academic Press.
Study blue, (2016) . Artist's impression of Showing glycoside
linkage g lycoside linkage. Retrieved from https://www.studyblue.com/note
s/note/n/ lecture-15-carbohydrates/deck/16767090.
Suh, D., Kim, Y., Kim, H., Ro, J., Cho, S., Yun, G., Choi, S and Lee, J. (2014).
Enhanced In Vitro Skin Deposition Properties of Retinyl Palmitate through Its
Stabilization by Pectin. Biomolecules Therapeutics, 22(1), 73–77.
Sun, J., Yin, G., Du, P. and Chen, L. (2011). Optimization of extraction technique of
polysaccharides from pumpkin by response surface method. Journal of Medicinal
Plants Research, 5(11). 2218-2222.
Sundar Raj, A.A., Rubila, S., Jayabalan, R and Ranganathan, T.V .(2012). A Review on
pectin: Chemistry due to general properties of pectin and its pharmaceutical uses.
Open Access Scientific Reports,1(12), 1-4.
Sungthongjeen, S., Pitaksuteepong, T., Somsiri, A., and Sriamornsak, P. (1999). Studies
on Pectins as Potential Hydrogel Matrices for Controlled-Release Drug
Delivery. Drug Development and Industrial Pharmacy,25(12), 1271-1276.
Surh, J., Decker, E. and Mcclements, D. (2006). Influence of pH and pectin type on
properties and stability of sodium-caseinate stabilized oil-in-water
emulsions. Food Hydrocolloids,20(5), 607-618.
Tamaki, Y., Konishi, T., Fukuta, M. and Tako, M. (2008). Isolation and structural
characterisation of pectin from endocarp of Citrus depressa. Food
Chemistry, 107(1), 352-361.
238
Tamaki, Y., Konishi, T., Fukuta, M., and Tako, M. (2008). Isolation and structural
characterisation of pectin from endocarp of Citrus depressa.Food
Chemistry, 107(1), 352-361.
Tan, Q., Lan, A., Kieu, T. and Thi, T. (2014). Optimisation of the pectin extraction from
pomelo peels by oxalic acid and microwave. Banat's Journal of Biotechnology,
V(9).
Thakur, B.R., Singh, R.K and Handa, A. K .(1997). Chemistry and uses of pectin--a
review. Critical Reviews in Food Science and Nutrition, 37, 47-73.
Thirawong, N., Nunthanid, J., Puttipipatkhachorn, S. and Sriamornsak, P. (2007).
Mucoadhesive properties of various pectins on gastrointestinal mucosa: An in
vitro evaluation using texture analyzer. European Journal of Pharmaceutics and
Biopharmaceutics, 67(1), 132-140.
Thirawong, N., Nunthanid, J., Puttipipatkhachorn, S., and Sriamornsak, P. (2007).
Mucoadhesive properties of various pectins on gastrointestinal mucosa: An in
vitro evaluation using texture analyzer. European Journal of Pharmaceutics and
Biopharmaceutics, 67(1), 132-140.
Tokusoglu, O. (2010). Fruit and cereal bioactives: Sources, chemistry, and applications.
Boca Raton, Fla.: CRC.
Tompkins, C. A. (1936). U.S. Patent No. 2 139 139. Washington, DC: U.S. Patent and
Trademark Office.
Trudsoe, J. E., and Olsen, H. B. (2014). U.S. Patent No. US 8,685,420 B2. Washington,
DC: U.S. Patent and Trademark Office.
Turakhozhaev, M. and Khodzhaev, M. (1993). Plant pectin substances. Methods of
isolating pectin substances. Chemistry of Natural Compounds, 29, 558-565
239
Turakhozhaev, M. T., &Khodzhaev, M. A. (1993). Plant pectin substances. Methods of
isolating pectin substances. Chemistry of Natural Compounds, 29(5), 558-565.
U S Food and Drug Administration Home Page. (2105, April 1). Code of Federal
regulations, Title 21, Volume 3. Retrieved January 05, 2016, from http://www.f
da.gov.
United Nations Industrial Development Organization (2016) Food Wastes. Retrieved
from United Nations Industrial Development Organization: http://www.unido.
org/fileadmin/import/32068_35FoodWastes.
USP 36-NF 31; United States Pharmacopeia: National Formulary. (2013). Rockville,
MD: United States Pharmacopeial Convention.
Vision (2015). Showing the global pectin market. Retrieved from https://www.sec.gov/
Archives/edgar/data/37785/000003778512000033/investorday2012.htm
Voragen, A., Schols, H., and Pilnik, W. (1986). Determination of the degree of
methylation and acetylation of pectins by h.p.l.c. Food Hydrocolloids, 1(1), 65-
70.
Vriesmann, L. C., Teófilo, R. F., and Petkowicz, C. L. (2011). Optimization of nitric
acid-mediated extraction of pectin from cacao pod husks (Theobroma cacao L.)
using response surface methodology. Carbohydrate Polymers, 84(4), 1230-1236.
Vriesmann, L., Teófilo, R. and Petkowicz, C. (2011). Optimization of nitric acid-
mediated extraction of pectin from cacao pod husks (Theobroma cacao L.) using
response surface methodology. Carbohydrate Polymers, 84(4), 1230-1236.
VriesmannaL.andPetkowicz C. (2013). Highly acetylated pectin from cacao pod husks
(Theobroma cacao L.) forms gel. Food Hydrocolloids, 33(1). 58-65.
240
Walsh, H., Cheng, J. and Guo, M. (2014). Effects of Carbonation on Probiotic
Survivability, Physicochemical, and Sensory Properties of Milk-Based Symbiotic
Beverages. Journal of Food Science , 79(4), M604–M613.
Walter, R. H. (1998). Polysaccharide association structures in food. New York: M.
Dekker
Wang, S., Chen, F., Wu, J., Wang, Z., Liao, X., and Hu, X. (2007). Optimization of
pectin extraction assisted by microwave from apple pomace using response
surface methodology. Journal of Food Engineering, 78(2), 693-700.
Wang, X., Chen, Q. and Lü, X. (2014). Pectin extracted from apple pomace and citrus
peel by subcritical water. Food Hydrocolloids, 38, 129-137.
Ware, P., Patel, D., Patel, J. and Krishnamurthy, R. (2013). Synthesis and
Characterization of Biofuel from Non-edible Seed Oil. Journal of Energy,
Environment & Carbon Credits, 3(2).
Watts, P. and Smith, A. (2009). PecSys: In situ gelling system for optimised nasal drug
delivery.ExpertOpin Drug Deliv, 6(5), 543-52.
Westhuizen, A. V. (2010). Evidence-based pharmacy practice:Gastroenteritis in
children. SA Pharmaceutical Journal, 10-19.
Wikiera, A., Mika, M. and Maja, M. (2015). Multicatalytic enzyme preparations as
effective alternative to acid in pectin extraction. Food Hydrocolloids, 44,156–161.
Wikipedia. ( 2016). Artist's impression of colour and texture of pectin. Retrieved from
https://en.wikipedia.org/wiki/Pectin
Willats, W. G., Knox, J. P., &Mikkelsen, J. D. (2006). Pectin: New insights into an old
polymer are starting to gel. Trends in Food Science & Technology, 17(3), 97-104.
Willats, W., McCartney, L., Mackie, W., & Knox, J. (2001). Pectin: Cell biology and
prospects for functional analysis. Plant Molecular Biology, 47(1-2), 9-27.
241
Willats, W., McCartney, L., Mackie, W., & Knox, J. (2001). Pectin: Cell biology and
prospects for functional analysis. Plant Molecular Biology, 47(1-2), 9-27.
Woo, K., Chong, Y., Li Hiong, S. and Tang, P. (2010). Pectin Extraction and
Characterization form Red Dragon Fruit ( Hylocereusployrhizus): A preliminary
Study. Journal of Biological Sciences, 10(7), 631-636
Wu, B., Degner, B. and Mcclements, D. (2014). Soft matter strategies for controlling
food texture: Formation of hydrogel particles by biopolymer complex
coacervation. Journal of Physics: Condensed Matter, 26(46), 464104.
Wuestenberg, T. (2014). Cellulose and Cellulose Derivatives in the Food Industry
Fundamentals and Applications (First ed.). Weinheim, Bergstr: Wiley-VCH
Xu, Y., Zhang, L., Bailina, Y., Ge, Z., Ding, T., Ye, X. and Liu, D. (2014). Effects of
ultrasound and/or heating on the extraction of pectin from grapefruit peel. Journal
of Food Engineering, 126, 72-81.
Yadav, R. S., and Agarwala, M. (2011). Phytochemical analysis of some medicinal
plants. Journal of Phytology, 3(12), 10-14.
Yapo, B. (2009). Pineapple and Banana Pectins Comprise Fewer Homogalacturonan
Building Blocks with a Smaller Degree of Polymerization as Compared with
Yellow Passion Fruit and Lemon Pectins: Implication for Gelling
Properties. BioMacroMolecules, 10(4), 717-721.
Yapo, B. and Koffi, K. (2014). Extraction and Characterization of Highly Gelling Low
Methoxy Pectin from Cashew Apple Pomace. Foods, 3, 1-12.
Yapo, B., Robert, C., Etienne, I., Wathelet, B. and Paquot, M. (2007). Effect of extraction
conditions on the yield, purity and surface properties of sugar beet pulp pectin
extracts. Food Chemistry, 100, 1356-1364.
242
Yeoh, S., Shi, J. and Langrish, T. (2008). Comparisons between different techniques for
water-based extraction of pectin from orange peels.Desalination, 218, 229-237.
Yuliarti, O., Matia-Merino, L., Goh, K., Mawson, J. and Brennan, C. (2011). Effect of
Celluclast 1.5L on the Physicochemical Characterization of Gold Kiwifruit
Pectin. International Journal of Molecular Sciences, 12, 6407-6417.
Zhang, C. and Mu, T. (2011). Optimisation of pectin extraction from sweet potato
(Ipomoea batatas, Convolvulaceae) residues with disodium phosphate solution by
response surface method. International Journal of Food Science &
Technology, 46(11), 2274-2280.
Zhongdong, L., Guohua, W., Yunchang, G. and Kennedy, J. (2006). Image study of
pectin extraction from orange skin assisted by microwave. Carbohydrate
Polymers, 64(4), 548-552.
Ziari, H., Ashtiani, F. and Mohtashamy, M. (2010). Comparing the effectiveness of
processing parameters in pectin extraction from apple pomace. Afinidad, 549,
374-379.