PHYTOCHEMICAL AND ANTI-INFLAMMATORY STUDIES ON THE HEXANE EXTRACT OF THE STEM BARK OF STEGANOTAENIA ARALIACEA HOSCHTS (APIACEAE)

TABLE OF CONTENTS
TITLE PAGE
ABSTRACT
TABLE OF CONTENTS

CHAPTER ONE
1.0       INTRODUCTION
1.1       Definition and Scope of Pharmacognosy
1.2       Taxonomy of medicinal plants
1.3       Chemistry and Biological Activity of Plant Extracts
1.4       Extraction, Isolation and Identification of Bioactive Constituents from Plants
1.5       Statement of the Research Problem
1.6       Justification of this Study
1.7       Hypothesis
1.8       General Aim
1.8       Specific Aims and Scope of the Study

CHAPTER TWO
2.0       LITERATURE REVIEW
2.1       Taxonomic Classification of the Family Apiaceae
2.2       Botanical Description of the Plant Family Apiaceae (Umbeliferae)
2.3       Reported Phytochemical Constituents of the Apiaceae Family
2.4       Description of the Subfamily Saniculoidae
2.5       Botanical Description of the Genus Steganotaenia
2.5.1    Ethanobotanical Uses of Species S. araliacea
2.5.2    Phytochemistry and Biological activity of the Species S. araliacea
2.6       Inflammation
2.6.1    Acute Inflammation
2.6.2    Chronic Inflammation
2.7       Plant-Derived Compounds with Anti-Inflammatory Activity

CHAPTER THREE
3.0       MATERIALS AND METHODS
3.1       List of Chemicals and Reagents
3.1.1    List of Chemicals
3.1.2    List of Reagents for Chromatographic Studies
3.2       List of Materials and Equipments
3.2.1    Chromatographic Materials
3.2.2    Materials for Anti-inflammatory Studies
3.2.3    Source of Laboratory Animals Used
3.3       Plant Collection, Preparation and Identification
3.4       Pharmacognostic Evaluation of the Stem Bark of S. araliacea
3.4.1 Macroscopical evaluation of whole and powdered stem bark of S. araliacea
3.4.2  Physico-Chemical Evaluation of Powdered Stem Bark of S. araliacea
3.5       Extraction of Plant Material
3.6       Phytochemical Studies on the Hexane Extract of the Stem Bark of S. araliacea
3.6.1    Thin Layer Chromatographic (TLC) Profiling
3.6.2    Column Chromatography
3.6.3 Identification of Compound UF
3.7Acute Toxicity Testing
3.7.1   Determination of Median Lethal Dose (LD50)
3.8Anti-Inflammatory Studies
3.9Statistical Analysis

CHAPTER FOUR
4.0       RESULT
4.1       Pharmacognostic Evaluation of S. araliacea Stem Bark
4.1.1    Macroscopical Evaluation of S. araliacea Stem Bark
4.1.2    Physico-Chemical Evaluation of the Powdered Stem Bark of S. araliacea
4.2       Extraction Yield of Powdered S. araliacea Stem Bark with Hexane
4.3       Phytochemistry of the Hexane Extract of Stem Bark of S.araliacea
4.3.1    Thin Layer Chromatographic Profile
4.3.2    Result of the Colum Chromatography of the hexane extract of the stem bark
4.3.3    Isolation of Compound UF
4.5       Identification of Compound UF
4.5.1    Physical Properties of Compound UF
4.5.2    Proton NMR of Compound UF
4.5.3    Proposed structure of Compound UF
4.6       Acute Toxicity Testing (Median Lethal Dose LD50)
4.7       Anti-Inflammatory Studies

CHAPTER FIVE
5.0       DISCUSSION

CHAPTER SIX
6.0 SUMMARY CONCLUSIONS AND RECOMMENDATIONS
6.1       Summary
6.2       Conclusions
6.3       Recommendations
REFERENCES
APPENDECIES

ABSTRACT
Inflammation has been implicated in virtually all human and animal diseases. It has become the focus of global scientific research, more so, since the currently used anti-inflammatory agents both steroidal and non-steroidal are prone to evoking serious adverse reactions.

Steganataeniaaraliacea (Apiaceae)stem bark has been reported to be used in traditional medicine for the treatment of asthma and rheumatism in East Africa. In this study, thehexane extract of the stem bark of the plant was evaluated for its anti-inflammatory property and with a view to isolating bioactive compounds from it.Methods for evaluating macroscopical features and physico-chemical properties were used for the pharmacognostic studies on the stem bark, chromatographic techniques including the thin layer chromatography (TLC) and column chromatography using silica gel were used for the phytochemical studies on the hexane extract of the plant while the anti-inflammatory study was also carried out on the hexane extractusing formalin induced paw oedema in rats. The macroscopical features of the whole stem bark was described as yellowish-green in colour, waxy and short fractured with an aromatic odour while the powdered stem bark was described as brown in colour, aromatic in odour with a sweet bitter taste. The physico-chemical properties of the bark determined include total ash value 10.67%, acid-insoluble ash value 4.00%, water-soluble ash value 1.25%, moisture content 8%, water-soluble extractive value 2.87%, alcohol-soluble extractive value 6.67% and petroleum ether-soluble extractive value 1.27%. The TLC profiling of the hexane extract of the bark showed the presence of steroids/terpenes. The column chromatography of the hexane extract of the stem bark lead to isolation of Compound UF which was characterized using the 1H-NMR spectroscopy and its physical properties. Compound UF was identified as Hentriacotane (C31H64) based on the available spectroscopic data and physical properties which agreed well with those reported in the literature. The acute toxicity studies of the hexane extract revealed (LD50) of 282.84mg/kg suggesting the extract is moderately toxic. The effect of the hexane extract on formalin induced paw oedema in rats revealed a dose dependant inhibition of the paw oedema when compared to the reference. The concentrations of the extract at 35, 70 and 140 mg/kg of the extract showed 41%, 42% and 50% inhibition respectively at the peak of the paw oedema which were statistically significant at P<0.05. This indicates a dose dependent anti-inflammatory activity of the extract.

CHAPTER ONE
1.0 INTRODUCTION
Medicinal preparations derived from natural sources, especially from plants, have been in widespread use since time immemorial. Ancient texts of India and China contain exhaustive depictions of the use of a variety of plant-derived medications (Samuelsson, 2004). In fact, plants remain the main source of medicines for a large proportion of the world‟s population, particularly in the developing world, despite the advent of the pharmaceutical chemistry during the early twentieth century, which brought with it the ability to synthesize an enormous variety of medicinal drug molecules and allowed the treatment of previously incurable and/or life-threatening diseases(Ahmad et al., 2006).

Over the years, however, synthetic drugs have been plagued by unwanted side-effects, toxicity, and inefficiency, among other problems. In addition, the search for new drugs against a variety of illnesses through chemical synthesis and other modern approaches has not been encouraging. These factors, as well as the emergence of new infectious diseases, the proliferation of disorders such as cancer, and growing multidrug resistance in pathogenic microorganisms, have prompted renewed interest in the discovery of potential drug molecules from medicinal plants. (Ahmad et al., 2006)

1.1 Definition and Scope of Pharmacognosy

The term pharmacognosy was first used between 1811 and 1815, and originally referred to

“materia medica”. The knowledge of drug materials or pharmacology. It is derived from two words , pharmakon (a drug) and gignosko (to acquire a knowledge of ) (Evans, 2002). Later on, pharmacognosy became restricted to that branch of pharmacy investigating “medicinal....

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