TABLE OF CONTENTS
Title Page
Certification
Dedication
Acknowledgement
Abstract
Table of Contents
List of Figures
List of Tables
List of Abbreviations
CHAPTER ONE: INTRODUCTION
1.1 Sweeteners
1.1.1 Common Sweeteners and Their Production
1.1.1.2 Natural Sweeteners
1.1.1.2.1 Honey
1.1.1.2.2 Maple Syrup
1.1.1.2.3 Molasses
1.1.1.2.4 Stevia
1.1.1.2.5 Sucrose
1.1.1.3 Artificial Sweeteners
1.2 Synsepalum dulcificum
1.3 Nutrients
1.3.1 Carbohydrates
1.3.2 Proteins
1.3.3 Fats
1.4 Phytochemicals
1.5 Antinutrients
1.6 Vitamins
1.6.1 VitaminA
1.6.2 Vitamin C
1.6.3 Vitamin D
1.6.4 Vitamin E
1.6.5 Vitamin K
1.7 Antioxidant
1.8 Some Minerals and Their Biological Functions
1.8.1 Calcium (Ca)
1.8.1.1 Metabolic Functions and Deficiency Symptoms of Calcium
1.8.2 Magnesium (Mg)
1.8.2.1 Metabolic Functions and Deficiency Symptoms of Magnesium
1.8.3 Zinc (Zn)
1.8.3.1 Metabolic Functions and Deficiency Symptoms of Zinc
1.8.4 Iron (Fe)
1.8.4.1 Metabolic Functions and Deficiency Symptoms of Iron
1.8.5 Copper (Cu)
1.8.5.1 Metabolic Functions and Deficiency Symptoms of Copper
1.9 Blood Glucose
1.9.1 Blood Glucose Regulation
1.10 Lipids
1.10.1 Lipoproteins: Types and Functions
1.10.1.1 Chylomicrons
1.10.1.2 Very Low Density Lipoprotein (VLDL)
1.10.1.3 Low Density Lipoprotein (LDL)
1.10.1.3.1 Metabolism of Low Density Lipoprotein via LDL Receptor
1.10.1.3.2 Regulation of LDL Receptor
1.10.1.4 High Density Lipoprotein (HDL)
1.11 Total Cholesterol andCholesterol Balance in Tissues
1.11.1 Diet and Cholesterol Regulation
1.12 Liver Function Biomarkers
1.12.1 Alanine Aminotransferase
1.12.2 Aspartate Aminotransferase
1.12.3 Alkaline Phosphatase
1.12.4 Clinical and Diagnostic Significance of Liver Function Enzymes
1.12.5 Bilirubin
1.12.6 Serum Protein
1.12.7 Serum Albumin
1.13 Renal Function Biomarkers
1.13.1 Blood Urea Nitrogen (BUN)
1.13.2 Creatinine
1.14 Lipid Peroxidation
1.14.1 Initiation
1.14.2 Propagation
1.14.3 Termination
1.14.4 Types of Lipid Peroxidation
1.14.4.1 Non- Enzymatic Lipid Peroxidation
1.14.4.2 Enzymatic Lipid Peroxidation
1.15 Research Objectives
1.15.1 General Objectives
1.15.2 Specific Objectives
CHAPTER TWO : MATERIALS AND METHODS
2.1 Materials
2.1.1 Plant materials
2.1.2 Animals
2.1.3 Chemicals and Reagents
2.1.4 Equipment /Instruments
2.2 Methods
2.2.1 Experimental Design
2.2.2 Extraction of Plant Material
2.2.3 Determination of the Extract Yield
2.2.4 Toxicological studies
2.2.4.1 Acute Toxicity Studies and Lethal Dose (LD50) Test
2.2.5 Proximate Analysis
2.2.5.1 Moisture
2.2.5.2 Crude Protein
2.2.5.3 Crude Fat
2.2.5.4 Crude Fibre
2.2.5.5 Ash/Mineral Matter
2.2.5.6 Carbohydrate or Nitrogen Free Extract (NFE)
2.2.6 Estimation of Vitamins
2.2.6.1 Determination of Vitamin A
2.2.6.2 Determination of Vitamin C
2.2.6.3 Determination of Vitamin D
2.2.6.4 Determination of Vitamin E
2.2.6.5 Determination of Vitamin K
2.2.7 Determination of Mineral Content of S. dulcificum Pulp
2.2.7.1 Determination of Phosphorus
2.2.8 Determination of Amino Acid Profile
2.2.8.1 Defatting of the Pulp
2.2.8.2 Hydrolysis of the Pulp
2.2.8.3 Nitrogen Determination
2.2.8.4 Loading of the Hydrolysate into TSM Analyzer
2.2.8.5 Method of Calculating Amino Acid values using Chromatogram Peaks
2.2.9 Qualitative Phytochemical Studies on Synsepalum dulcificum Pulp
2.2.9.1 Test for Alkaloids
2.2.9.2 Test for Glycosides
2.2.9.3 Test for Cyanogenic Glycosides
2.2.9.4 Test for Tannins
2.2.9.5 Test for Saponins
2.2.9.6 Test for Flavonoids
2.2.9.7 Test for Resins
2.2.9.8 Test for Terpenoids and Steroids
2.2.10 Quantitative Phytochemical Analysis of S.dulcificum Pulp
2.2.10.1 Determination of Alkaloids
2.2.10.2 Determination of Cyanogenic Glycosides
2.2.10.3 Determination of Saponins
2.2.10.4 Determination of Flavonoids
2.2.10.5 Determination of Tannins
2.2.10.6 Determination of Steroids
2.2.10.7 Determination of Terpenoids
2.2.11 Antinutrient Analysis of S. dulcificum Pulp
2.2.11.1 Determination of Oxalates
2.2.11.2 Determination of Phytates
2.2.11.3 Determination of Haemagglutanins
2.2.12 Blood Sample Collection for Biochemical Analysis
2.2.13 Biochemical Assays
2.2.13.1 Assay of Alanine Aminotransferase (ALT) Activity
2.2.13.2 Assay of Aspartate Aminotransferase Activity
2.2.13.3 Assay of Alkaline Phosphatase (ALP) Activity
2.2.13.4 Determination of Bilirubin Concentration Using Colorimetric Method
2.2.13.4.1 Determination of Total Bilirubin (TB) Concentration
2.2.13.5 Total Serum Protein Assay
2.2.13.6 Serum Albumin Concentration
2.2.13.7 Creatinine
2.2.13.8 Urea
2.2.13.9 Blood glucose Assay
2.2.13.10 Estimation of Serum Lipid Concentrations
2.2.13.10.1 Estimation of Total Cholesterol Concentration
2.2.13.10.2 Estimation of Low Density Lipoprotein-Cholesterol Concentration
2.2.13.10.3 Estimation of High Density Lipoproteins (HDL)–Cholesterol Concentration
2.2.13.10.4 Estimation of Triacylglycerol
2.2.13.11 Estimation of Lipid Peroxidation
2.2.14 Histopathological Examination
2.2.15 Statistical Analysis
CHAPTER THREE: RESULTS
3.1 Proximate Composition of S. dulcificum Pulp
3.2 Mineral Composition of S. dulcificum Pulp
3.3 Vitamin Content of S.dulcificum Pulp
3.4 Amino Acid Profile of S. dulcificum Pulp
3.5 Phytochemical Composition of S. dulcificum Pulp
3.6 Antinutritional Composition of S.dulcificum Pulp
3.7 Acute toxicity (LD50) Studies
3.8 Mean Body Weights of Animals
3.9 Effect of S. dulcificumMethanolic Extract Administration on Alkaline Phosphatase (ALP) Activity in Rats
3.10 Effect of S. dulcificumMethanolic Extract Administration on Alanine Aminotransferase (ALT) Activity in Rats
3.11 Effect of S. dulcificumMethanolic Extract Administration on Aspartate Aminotransferase (AST) Activity in Rats
3.12 Effect of S. dulcificumMethanolic Extract Administration on Bilirubin levels in Rats
3.13 Effect of S. dulcificumMethanolic Extract Administration on Total Serum Protein concentration in rats
3.14 Effect of S. dulcificumMethanolic Extract Administration on Serum Albumin Concentration in Rats
3.15 Effect of S. dulcificumMethanolic Extract Administration on Serum Globulinin Rats
3.16 Effect of S. dulcificumMethanolic Extract Administration on Creatinine Level in Rats
3.17 Effect of S. dulcificumMethanolic Extract Administration on Urea Level in Rats
3.18 Effect of S. dulcificumMethanolic Extract Administration on Blood Glucose Concentration in Rats
3.19 Effect of S. dulcificumMethanolic Extract Administration on Cholesterol Concentration in Rats
3.20 Effect of S. dulcificumMethanolic Extract Administration on High Density Lipoprotein Cholesterol Concentration in Rats
3.21 Effect of S. dulcificumMethanolic Extract Administration on Low Density Lipoprotein Cholesterol Concentration in Rats
3.22 Effect of S. dulcificumMethanolic Extract Administration on Triacylglycerol Concentration in Rats
3.23 Effect of S. dulcificumMethanolic Extract Administration on Malondialdehyde Concentration in Rats
3.24 Effect of S. dulcificumMethanol Extract Administration on the Histopathology of Rat Liver [14 days duration]
3.25 Effect of S. dulcificumMethanol Extract Administration on the Histopathology of Rat Liver [28 days duration]
3.26 Effect of S. dulcificumMethanol Extract Administration on the Histopathology of Rat Kidney [14 days duration]
3.27 Effect of S. dulcificumMethanol Extract Administration on the Histopathology of Rat Kidney [28 days duration]
CHAPTER FOUR: DISCUSSION
4.1 Discussion
4.2 Conclusion
4.3 Suggestions For Further Studies
REFERENCES
APPENDICES
ABSTRACT
The nutritive and antinutritive compositions of S. dulcificum pulp were analysed to augment the available information on the anti-diabetic effect of the plant. Biochemical parameters like liver function enzymes (ALT, AST, ALP) and bilirubin concentrations,serum total protein, serum albumin and globulin, kidney function parameters (creatinine and urea concentrations), blood glucose, serum lipid profile and lipid peroxidation were determined in rats that were administered different concentrations of the methanolic extract to ascertain their effects. The internal organs (liver and kidney) were also removed and used for histopathological studies. From the result of the study, the proximate composition shows that S. dulcificum contains 7.75% protein, 59.55% moisture content, 4.36% ash, 6.24% crude fibre, 3.26% fat and 18.84% carbohydrate.The result of the mineral analysis shows that S.dulcificum pulp contains 100 mg/g calcium, 24.20 mg/g iron, 9.49 mg/g zinc, 6.22 mg/g copper, 0.01 mg/g chromium and 0.01 mg/g cobalt. Vitamin analyses shows that the S. dulcificum pulp contains 0.04% vitamin A, 22.69% vitamin C, 0.01% vitamin D and 0.02% vitamin K. Antinutrient analyses of the pulp show 5.67% oxalate, 0.03% phytates and 0.02% hemagglutanin. Amino acid profile shows that S.dulcificum pulp contains 8.055% tryptophan, 1.35% phenylalanine, 0.7% isoleucine, 0.5% tyrosine, 1.05% methionine, 0.4% proline, 0.69% valine, 1.1% threonine, 0.4% histidine, 0.5% alanine, 1.02% glutamine, 1.6% glutamic acid, 0.7% glycine, 0.3% serine, 1% arginine, 0.1% aspartic acid, 1.23% asparagine, 0.6% lysine and 0.6% leucine. Quantitative phytochemical analysis shows that the pulp contains 3.45% saponins, 57.01%`flavonoids, 7.12% tannins, 0.0001% alkaloids, 0.0001% glycosides, 0.0003% resins, 0.0002% terpenoids, 0.0001% steroids and 0.0003% cyanogenic glycosides.The results of the acute toxicity show that the methanol extract is not toxic to the mice at concentrations up to 5000mg/kg body weight. From the results obtained, the animals receiving 100mg/kg b.w of the methanolic extract showed significantly reduced (p<0 .05="" 14="" 28="" after="" alt="" and="" bilirubin="" cholesterol="" compared="" day="" density="" difference="" glucose="" however="" levels="" lipoprotein="" low="" no="" of="" p="" serum="" significant="" study.="" study="" the="" to="">0.05) was also observed across the groups in their serum ALP, AST, creatinine, urea, cholesterol, TAG, albumin and globulin levels on the 14th day compared with the 28th day. A significant difference (p
CHAPTER ONE
INTRODUCTION
The worsening food crisis and the consequent widespread prevalence of malnutrition in developing and under-developed countries have resulted in high mortality and morbidity rates, especially among infants and children in low-income groups (Enujiugba and Akanbi, 2005). Food has been defined as any substance containing primarily carbohydrates, fats, water, protein, vitamins and minerals that can be taken by an animal or human to meet its nutritional needs and sometimes for pleasure. Items considered as food may be sourced from plants, animals or fungus as well as fermented products like alcohol. Food is also anything solid or liquid that has a chemical composition which enables it provide the body with the material from which it can produce heat or any form of energy, provide material to allow for growth, maintenance, repair or reproduction to proceed and supply substances, which normally regulate the production of energy or the process of growth, repair or reproduction. Food is therefore, the most basic necessity of life (Turner, 2006).
Nutrition is the science that deals with all the various factors of which food is composed and the way in which proper nourishment is brought about. The average nutritional requirements of groups of people are fixed and depend on such measurable characteristics as age, sex, height, weight, degree of activity and rate of growth. Good nutrition requires a satisfactory diet which is capable of supporting the individual consuming it, in a state of good health by providing the desired nutrients in required amounts. It must provide the right amount of nutrients and fuel to execute normal physical activity. If the total amount of nutrients provided in the diet is insufficient, a state of under- nutrition develops.
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