TABLE OF CONTENTS
Title page
Abstract
Table of contents
List of abbreviations
CHAPTER ONE
1.0 Introduction
1.1 Definition of a drug
1.2 Medicinal plants
1.3 Medicinal plant research
1.4 Aim
1.4.1 Objectives
1.5 Scope and limitations of research
1.6 Justification of the research
CHAPTER TWO
2.0 Literature review
2.1 Botanical description of the genus Indigofera arrecta
2.1.1 Other botanical information
2.1.2 Origin and geographical distribution
2.2 Chemical constituents of indigofera arrecta
2.2.1 Traditional medicinal uses
2.2.2 Uses of some of the genus
2.3 Production and international trade
2.3.1 Pharmacological properties
2.3.2 Adulterations and substitutes
2.3.3 Growth and development
2.3.4 Ecology
2.3.5 Management
2.3.6 Propagation and planting
2.3.7 Diseases and pests
2.3.8 Harvesting
2.3.9 Yield
2.4 Handling after harvest
2.4.1 General description of Indigofera arrecta (HOCHST EX.A.RICH )
2.5 Review of some natural products from plants, tests and their uses
2.5.1Alkaloids
2.5.2Flavonoids
2.5.3 Saponins
2.5.4 Glycosides
2.5.5 Tannins
2.5.6 Steroids
2.5.7. Terpenoids
2.6 Factors which can affect the level or the composition of the active ingredients in medicinal plant
2.7 Some microorganisms and their effects on the human body
2.7.1 Staphylococci
2.7.2 Streptococci
2.7.3 Candida
2.7.4 Enterobacteriaceae
2.7.5 Klebsiella
2.7.6 Escherichia coli
2.7.7 Salmonellae
CHAPTER THREE
3.0 Materials and method
3.1 Materials/reagents/equipment and analytical procedure
3.1.1 Solvents
3.1.2 Equipment
3.1.3 Reagent
3.1.4 Microbial media, test organisms and equipment for antimicobial test
3.1.5 The identification and preparation of plant material
3.1.6 Extraction procedure for crude extract
3.2 Preliminary phytochemical screening
3.2.1 Test for steroids/terpenes
3.2.2 Test for flavonoids
3.2.3 Test for alkaloids
3.2.4 Test for tannins
3.2.5 Test for anthraquinones
3.2.6 Test for saponins
3.2.7 Test for glycoside (fecl3 test)
3.3 Antimicrobial screening
3.3.1 Preparation of bacterial test organisms
3.3.2 Preparation of fungal test organisms
3.3.3 The stock dilution of the plant extracts
3.3.4 Preparation of the nutrient agar
3.3.5 Preparation of the sabouraud dextrose agar media
3.3.6 The punched agar diffusion method [bryant, 1972]
3.3.7 Preparation of inoculums of test organisms
3.3.8 Sensitivity test of the extract using agar diffusion method
3.3.9 Determination of minimum inhibitory concentration using tube dilution method
3.4 Minimum bactericidal concentration (mbc)
3.4.1 Chromatographic purification of extracts
3.4.2 Thin layer chromatography (TLC)
3.4.3 Column chromatography
3.4.4 Solvents system/elution
3.4.5 Gel filtration chromatography
3.4.6 Thin layer chromatography of the n-hexane extract
3.4.7 Column chromatography of n-hexane fraction
CHAPTER FOUR
4.0 Results
4.1 Result of extraction
4.2 Result of phytochemical screening
4.3 Results of antimicrobial activity
4.4 Result of chromatographic separation
4.5 Column chromatography of n-hexane fraction
4.6 Isolation of EB
4.7 TLC analysis of EB
4.8 Result of antimicrobial activity of compound EB
CHAPTER FIVE
5.0 Discussion of result
5.1 Extraction
5.2 Phytochemical screening
5.3 Antimicrobial screening
5.4 Physical and chemical properties of EB
5.4.1 Spectral analysis
5.4.2 FTIR
5.4.3 1H NMR
5.4.4 13C NMR
5.4.5 DEPT
CHAPTER SIX
6.0 Summary, conclusion and recommendation
6.1. Summary
6.2 Conclusion
6.3 Recommendation
References
ABSTRACT
The Pulverized stem bark of Indigofera arrectawas exhaustively extracted with methanol and concentrated in vacuo using rotary evaporator at 40 0C.The extract was later subjected to solvent partitioning to yield soluble extracts of n-hexane, ethyl acetate, chloroform, and methanol. Genernal phytochemical screening of the fractions revealed the presence of secondary metabolites such as cardiac glycoside,steroid, terpenes flavonoids and tannins. The antimicrobial activity against S. aureus,S. pyogenes,S.feacalis, S.typhii, E.coli C. ulcerans,P. vulgaris and C.albicans was tested using the tube dilution and agar diffusion methods as outlined by the NCCLS. The results of the antimicrobial activity as indicated by the zonesof inhibition of growth of microorganism ranged from 20mm to 40mm for the n-hexane extract, 16mm to 21mm for ethyl acetate extract and 20mm to 27mm for the methanol extract. The MIC result for the n-hexane, ethyl acetate and methanol extracts ranged from 7.5mg/ml to 15mg/ml. The MIC of 15mg/ml exhibited by the n-hexane extract against both gram positive and gram negative bacteria indicates broad spectrum activity of Indigofera arrect. The n-hexane fractions was subjected to Column Chromatography using silica gel to yield 87 fractions, which were combined based on their thin layer chromatography analysis and recrystallized in methanol to give a pure white crystalline powder, which melts at 144oC. The structure of the isolated compound was established by spectroscopic analysis and by direct comparison of the data obtained with those reported in literature to be Stigmasterol (3β,22E-Sigmasta-5,22-dien-3-ol).
CHAPTER ONE
1.0 INTRODUCTION
1.1 Definition of a drug
A drug can be described as any chemical substance that has no nutritional value when
introduced into the body but causes some physiological effects within the system (Mbah, 2000). Drugs are classified under pharmaceuticals. Pharmaceutical drug, according to Dey (2006), also refers to as medicine or medicament, can be loosely defined as any substance intended for use in the diagnosis, cure, mitigation, treatment or prevention of diseases.
Some pharmaceuticals occur naturally in plants. These can be called phyto pharmaceuticals. By the strictest definition, they are drugs or chemicals that may have health related effects but are not considered essential nutrients. Protein, carbohydrates, fats, minerals and vitamins are regarded as essential nutrients. Some pharmaceuticals found in plants include gedunin and nimbolide from Azadirachta indica (Neem) (Khalid and Duddeck, 1993); santonin, a sesquiterpenoid lactone is found in species ofArtemisia which grows in Asia, quinine and alkaloid occurs in the bark ofcinchona tree; penincillin- a beta-lactam is produced by fungi in the genus Penicillium (Finar, 2003) and reserpine-an alkaloid is isolated fromRauwolfia plant.
1.2 Medicinal plants
According to biblical records, Prophet Ezekiel reported that the fruits will serve as food and their leaves for healing (Ezekiel 47:12). Thus, the use of plants for medicinal purposes dates back to thousands of years ago as the earliest humans used various plants to treat illness....
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