PHYTOCHEMICAL SCREENING, ANTIOXIDANT CAPACITY AND ANTIMALARIAL ACTIVITIES OF UNRIPE COCONUT FLUID
ABSTRACT
The phytochemical, antioxidant, antimicrobial and antimalarial activities of the fluid inside an unripe coconut fluid were investigated and at the end of the day, the phytochemicals were found to be terpenoids, flavonoids, alkaloids, tannins and saponins and the percentage composition of these classes of compounds were also determined. This component of an unripe coconut had fluid which is very effective in destroying malaria parasites in the same way the strength showed on the organism by the control drug which is Atesurnate. The antioxidant capacity of the fluid was also determined vis-à-vis and they were interestingly good results.
TABLE OF CONTENTS
Title page i
Certification iii
Dedication iv
Acknowledgement v
Abstract vi
Table of Contents vii
CHAPTER ONE
Introduction1
Natural products and medicinal chemistry1
Statement of problem2
Objectives of study2
Justification of study3
CHAPTER TWO
Review of Related Literature4
Phytochemical analysis of unripe coconut fluid4
Alkaloids4
Classification of alkaloids5
Properties6
Tannins6
Structure of tannins7
Classification of tannins8
Terpenoids10
Structure of terpenoids10
Flavanoids18
2.1.4 Saponins 25
Phytohormones present coconut fluid26
Auxin26
Cytokinins27
Gibberellins28
Nutritional benefits of unripe coconut fluid28
Natural products with antimalarial activities30
Structures of some established antimalarial drugs30
CHAPTER THREE
Experimental Section32
Qualitative and quantitative phytochemical screening of unripe coconut
Fluid 32
Qualitative phytochemical screening test32
Test for tepernoids32
Test for flavonoids32
Test for alkaloids33
Test for tannins33
Test for saponins34
Quantitative phytochemical screening test34
Terpenoids34
Flavonoids34
Alkaloids34
Tannins35
Saponins35
Antimicrobial screening test35
Minimum Inhibitory Concentration (MIC) testing36
Acute toxicity screening test37
Innoculation of the parasitaemia37
Determination of malaria parasite (MP+)38
Determination of antioxidant activities38
Determination of Superoxide Dismutase (SOD)38
Determination of vitamin C39
Determination of vitamin E40
Haematological test40
Determination of total white blood cell count40
Determination of total red blood cell count41
Determination of haemoglobin concentration41
Liver function test41
Determination of the aspartate aminotransferase activity42
Determination of the alanine transaminase activity42
Determination of total bilirubin43
The kidney function test activity43
Determination of urea44
Determination of uric acid44
Determination of acid phosphatase45
Determination of creatinine45
Determination of trace elements45
Serum chloride determination45
Serum sodium determination46
Serum potassium determination46
Serum zinc determination47
CHAPTER FOUR
Results and discussion48
Results of phytochemical screening of unripe coconut fluid49
Results of antimicrobial activity50
Result of acute toxicity test (LD50)51
Result of percentage parasitaemia52
Results of antioxidant activities53
Results of haematological test56
Results of liver function test activities59
Results of kidney function test activities61
Results of trace element determination63
CHAPTER FIVE 66
5.0 Conclusion 67
References
LIST OF TABLES
Table 1: Qualitative phytochemical screening of unripe coconut fluid 48
Table 2: Quantitative Phytochemical Screening of unripe coconut fluid 49
Table 3: Antimicrobial Sensitivity Testing of the unripe coconut fluid 50
Table 4: Inhibition Zones Diameter (mm) 50
Table 5: Acute toxicity result table 51
Table 6: 4days after inoculation of malaria parasite 52
Table 7: 7days treatment of malaria parasite 52
Table 8: Determination of Superoxide Dismutase (SOD) 54
Table 9: Determination of vitamin E 55
Table 10: Determination of vitamin C 55
Table 11: Determination of Total White Blood cell count 56
Table 12: Determination of Total Red Blood cell Count 57
Table 13: Determination of Haemoglobin Concentration 58
Table 14: Aspartate Aminotransferase (AST) Activity 59
Table 15: Alanine Aminotransferase (ALT) Activity 59
Table 16: Determination of total bilirubin 60
Table 17: Determination of Creatinine 61
Table 18: Determination of Urea 61
Table 19: Determination of Uric acid 62
Table 20: Determination of Acid phosphate 62
Table 21: Serum Potassium determination 63
Table 22: Serum Sodium determination 64
Table 23: Serum Chloride determination 64
Table 24: Serum Zinc determination 65
LIST OF FIGURES
Fig 1: UV-Visible spectral of unripe coconut fluid
Fig 2: FTIR spectral of unripe coconut fluid
ABBREVIATIONS
EDTA Ethylene diamine tetra acetic acid
MIC Minimum inhibitory concentration
UC Untreated control
SC Standard control
NC Normal control
NST No sign of toxicity
ND No death
GV Gential violet
HB Haemoglobin
ALT Alanine transaminase
AST Aspartate aminoferase
ACP Acid phosphatase
SOD Superoxide dismutase
SDA Saboround dextrose agar
CHAPTER ONE
INTRODUCTION
NATURAL PRODUCTS AND MEDICINAL CHEMISTRY
A natural product is a chemical compound or a substance produced by a living organism that is found in nature1.Within the field of organic chemistry, the definition of natural products is usually restricted to mean purified organic compounds isolated from natural sources.2 Natural products sometimes have pharmacological or biological activity that can be of therapeutic benefit in treating diseases. As such, natural products are active components not only for most traditional medicines but also for many modern medicines.3
A significant number of anti-infectives are based on natural products. The first antibiotic to be discovered is Penicillin [1].
R NH O
O
H
ImageS
CH3
N CH3
O
HO
1
A large number of currently prescribed drugs have been directly derived from natural products. For example the bark of the Willow tree has been known from antiquity to have pain relieving properties4; this is due to the presence of the natural product salicin [2] which in turn may be hydrolyzed to salicylic acid [3], a synthetic derivative acetylsalicylic acid [4] better known as aspirin widely usedas a pain reliever was obtained when salicylic acid is treated with acetic anhydride (scheme 1)
HO
O
O
O
HO
OH
HO O
OH OH
O CH3
hydrolysis
acetic anhydri
conc H SO O
2 4
OH
H3C
O
de
2 3 4
Scheme 1
Unripe coconut fluid is one of the world’s most versatile natural products with increasing scientific evidence that support its role in health and medicinal application. Unripe coconut fluid is the liquid endosperm of immature coconut. The drink is extracted from immature coconut, which implies it has not yet accumulated fat and has low sugar content5. The sugar and protein contents increase as the coconut matures. It is recommended for weight loss phases because it increases the rate of metabolism. Unripe coconut fluid is a hydrating drink, a natural isotonic drink, a universal donor which can be used in blood transfusion because it has the same salt concentration as cells and blood.6
STATEMENT OF THE PROBLEM
Despite the use of the various parts of coconut palm in herbal medicine, reports on antimalarial activities, antioxidant capacity and phytochemical screening of unripe coconut water are rather scanty. Hence it is imperative to examine its antimalarial activities, its antioxidant capacity in blood serum and the phytochemicals in unripe coconut water.
OBJECTIVES OF THE STUDY
The specific objectives of this study are:
i) to determine the antimalarial activities of the unripe coconut water
ii) qualitative and quantitative phytochemical screening of unripe coconut water.
iii) to determine its antimicrobial activities
iv) to determine itsacute toxicity (LD50)
v) to determine hematological assay.
vi) to determine the antioxidant capacity of the unripe coconut water in the blood serum and
vii) to determine the effect of the unripe coconut water in the body organs (Liver and kidney).
JUSTIFICATION OF THE STUDY
The wide applications of unripe coconut water especially in medical fields can be justified by its unique chemical composition of sugar, vitamins, minerals, amino acids, antioxidants, phytochemicals and phytohormones4,7. For example Vitamin E is an important antioxidant that protects unsaturated oil from being destroyed in the body by oxygen. It is also a potent water- soluble antioxidant in humans and most flavonoids have antioxidant properties8. This antioxidant blocks some of the damage caused by free radicals and substances that damage DNA9. Some alkaloids and terpenoids have antimalarial activities.10
.