CHARLENE CASSANDRA CHATTERTON
Transcript of CHARLENE CASSANDRA CHATTERTON
EXTRACTING BIOACTIVE COMPOUNDS FROM MORINGA
OLEIFERA LEAVES FOR ANTICANCER PRODUCT
CHARLENE CASSANDRA CHATTERTON
BACHELOR OF CHEMICAL ENGINEERING
UNIVERSITI MALAYSIA PAHANG
EXTRACTING BIOACTIVE COMPOUNDS FROM MORINGA
OLEIFERA LEAVES FOR ANTICANCER PRODUCT
CHARLENE CASSANDRA CHATTERTON
Thesis submitted in partial fulfilment of the requirements
for the award of the degree of
Bachelor of Chemical Engineering
Faculty of Chemical & Natural Resources Engineering
UNIVERSITI MALAYSIA PAHANG
DECEMBER 2016
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SUPERVISOR’S DECLARATION
We hereby declare that we have checked this thesis and in our opinion, this thesis is
adequate in terms of scope and quality for the award of the degree of Bachelor of
Chemical Engineering.
Signature :
Name of main supervisor :
Position : SENIOR LECTURER
Date :
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STUDENT’S DECLARATION
I hereby declare that the work in this thesis is my own except for quotations and
summaries which have been duly acknowledged. The thesis has not been accepted for
any degree and is not concurrently submitted for award of other degree.
Signature :
Name : CHARLENE CASSANDRA CHATTERTON
ID Number : KE 13061
Date :
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Dedicated to my family
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ACKNOWLEDGEMENT
I would like to express my deepest gratitude and appreciation to my beloved
supervisor, Dr. Eman N. Ali for the endless motivation and guidance throughout my
undergraduate research project. I am thankful for the suggestions, honest comments and
advice on my research project as they served as a continuous encouragement in order
for me to work harder and improve my research work. You have been a wonderful
supervisor as well as a friend to me, always responsive to any question that I bring
forward to you regardless on weekends or even public holidays. Without your support,
my journey would have been a difficult one.
A special thanks to my family for showering me with love and support
throughout my entire research project as well as keeping me in your prayers. Apart from
that, I thank my friends and course mates who have helped me in one way or another in
my research project in terms of knowledge sharing, inspiration and support towards
achieving my goal.
I am also indebted to the Ministry of Higher Education and Universiti Malaysia
Pahang for funding my study.
Besides that, I am sincerely grateful to the staffs of Chemical Engineering and
Natural Resources Faculty who have been kind and resourceful in assisting me
whenever I had any enquiries.
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TABLE OF CONTENTS
Page
SUPERVISOR’S DECLARATION ii
STUDENT’S DECLARATION iii
ACKNOWLEDGEMENT v
ABSTRACT vi
ABSTRAK vii
TABLE OF CONTENTS viii
LIST OF TABLES x
LIST OF FIGURES xi
LIST OF ABBREVIATIONS 1
CHAPTER 1 INTRODUCTION 2
1.1 Background of the Study 2
1.1.1 Botanical Description of Moringa oleifera 5
1.1.2 Nutritional value of Moringa oleifera 7
1.1.3 Chemical composition of Moringa oleifera leaves 8
1.2 Motivation 1
1.3 Problem Statement 1
1.4 Objectives 1
1.5 Scopes of Study 1
CHAPTER 2 LITERATURE REVIEW 2
2.1 Introduction 2
2.2 Anticancer properties of Moringa oleifera leaves against various cancer cells 2
2.3 Bioactive compounds extracted from Moringa oleifera leaves 8
2.4 Summary of literature review 13
CHAPTER 3 METHODOLOGY 15
3.1 Introduction 15
3.2 Extraction 16
3.3 Total phenolic content and total flavonoid content 21
3.4 DPPH radical scavenging activity 23
3.5 Cytotoxic activity 25
CHAPTER 4 RESULTS AND DISCUSSION 27
4.1 Introduction 27
4.2 Total phenolic content and total flavonoid content 27
4.3 The DPPH radical scavenging activity 30
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4.4 Cytotoxic activity of Moringa oleifera leaves extracts 33
CHAPTER 5 CONCLUSION AND RECOMMENDATION 34
5.1 Conclusion 34
5.2 Recommendation 34
REFERENCES 35
Appendix 41
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LIST OF TABLES
Table No. Title Page
Table 1: Chemical composition of dried Moringa oleifera leaves. 8
Table 2: Amino acids composition of dried Moringa oleifera leaves. 9
Table 3: Mineral contents of dried Moringa oleifera leaves. 9
Table 4: Fatty acids composition of dried Moringa oleifera leaves. 10
Table 5: The nutritional value of Moringa oleifera. 11
Table 6: The A549 cell viability after exposure to Moringa oleifera leaves extract. 7
Table 7: Yields of crude extracts, contents of total phenolics and flavonoids. 8
Table 8: Phenolic acids content in Moringa oleifera leaves. 9
Table 9: Flavonoids content in Moringa oleifera leaves. 10
Table 10: Test conducted to identify the phytochemicals. 11
Table 11: The phytochemicals present from extraction using different solvents. 12
Table 12: Disadvantages of various extraction methods. 20
Table 13: The solvents and method used in the extraction of the samples. 26
Table 14: TPC and TFC values of the S1, S2, S3, S4 and S5 extracts. 28
Table 15: TPC and TFC values of the water extracts. 28
Table 16: DPPH IC50 values of the samples. 30
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LIST OF FIGURES
Figure No. Title Page
Figure 1: Moringa oleifera flowers. 6
Figure 2: Moringa oleifera leaves. 6
Figure 3: Moringa oleifera pods. 7
Figure 4: Moringa oleifera seeds. 7
Figure 5(A): Microscopic view of the cytotoxic effect, (B): Number of viable colonies. 5
Figure 6: The percentage survival of cancer cells and normal cells after treatment. 5
Figure 7: The set-up of Soxhlet extraction apparatus used in the actual experiment. 17
Figure 8: Rotary Evaporator. 19
Figure 9: Layout of 96-well plates for cytotoxicity study. 25
Figure 10: TPC test for S1, S2, S3, S4 and S5 extracts. 27
Figure 11: TFC test for S1, S2, S3, S4 and S5 extracts. 27
Figure 12: TPC test for water extracts by Soxhlet (left) and boiling (right). 27
Figure 13: TFC test for water extracts by Soxhlet (left) and boiling (right). 27
Figure 14: DPPH radical scavenging activity of S1-S5. 29
Figure 15: The DPPH test for water extracts by Soxhlet (left) and boiling (right). 30
Figure 16: The % inhibition of DPPH radicals of S1-S5. 31
Figure 17: The % inhibition of DPPH radicals of the water extracts. 31
Figure 18: The effect of the extracts on MCF-7 cells after 72 hours of treatment. 32
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LIST OF ABBREVIATIONS
EBV Epstein-Barr virus
HCT-8 ileocecal adenocarcinoma cancer cells of a 67 year old male
MDA-MB-231 breast mammary gland cancer cells
DPPH 2,2’diphenylpicrylhydrazyl assay
ALL acute lymphoblastic leukemia
AML acute myeloid leukemia
Panc-1 human pancreatic cancer cell lines
COLO-357 human pancreatic cancer cell lines
p34 human pancreatic cancer cell lines
A549 human lung cancer cell
MCF 7 epithelial breast cancer cell line
MDA MB 231 mesenchymal breast cancer cell line
CAE chlorogenic acid equivalents
1QE isoquercetin equivalents
SZ Squeezing
DF Decoction of fresh leaves
DD Decoction of dried leaves
MF70 Maceration of fresh leaves with 70% ethanol
MD50 Maceration of dried leaves with 50 % ethanol
MD70 Maceration of dried leaves with 70% ethanol
PD50 Percolation of dried leaves with 50 ethanol
PD70 Percolation of dried leaves with 70% ethanol
SD50 Soxhlet extraction of dried leaves with 50 % ethanol
SD70 Soxhlet extraction of dried leaves with 70% ethanol
ND Not detected
N/A Not available
LE Leucocyanidin equivalent
DE Diosgenin equivalent.
IC50 50% inhibitory concentration
HepG2 human hepatocellular carcinoma
Caco-2 colorectal adenocarcinoma
MCF-7 breast adenocarcinoma
COS-7 African green monkey normal cells
TPC Total phenolic content
TFC Total flavonoid content
GAE gallic acid equivalence
CE catechin equivalence
QE quercetin equivalence