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PHYTOCHEMICAL AND ANTIMICROBIAL STUDIES OF THE METHANOL EXTRACT OF THE ROOT OF NAPOLEONAEA HEUDELOTTI (A.JUSS)

Chemistry Project Topics & Materials

ABSTRACT

 

The extracts of the root part of Napoleonaea heudelotii were subjected to phytochemical and anti-microbial studies. Extraction was done by continuous Soxhlet extraction using methanol. The phytochemical screening of the crude methanol extract, chloroform and ethyl acetate fractions revealed the presence of carbohydrate, cardiac glycosides, saponins, steroids, triterpenes, flavanoids and tannins. The result of the antimicrobial screening of the crude methanol extract, ethyl acetate and chloroform fractions showed activity against Staphylococcus aureus, Streptococcus pyogenes, Bacillus subtilis, Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa, Proteus vulgaris, and Candida albicans. However, the chloroform fraction was the most active fraction against the test microoganisms. The zone of inhibition of the methanol extract ranged between 16 mm and 21 mm, the chloroform fraction ranged between 17 mm and 25 mm while the ethyl acetate fraction ranged between 15 mm and 21 mm. The MIC results of methanol extract, ranged between 12.5 mg/ml and 1.562 mg/ml, chloroform fraction ranged between 12.5 mg/ml and 1.562 mg/ml, while ethyl acetate ranged between 6.25 mg/ml and 1.625 mg/ml. The MBC of methanol extract and chloroform fraction ranged between 12.5 mg/ml and 1.562 mg/ml, while that of ethyl acetate fraction ranged between 6.2 mg/ml and 1.562 mg/ml. The chloroform fraction being the most active fraction was subjected to extensive chromatographic purification; white crystalline solid labelled NHPE were isolated. The structures of the isolated compounds were determined to be a mixture α-amyrin and β-amyrin using 1D and 2D NMR.

TABLE OF CONTENTS

 

Title Page

Abstract

Table of Contents

List of Abbreviations

CHAPTER ONE

1.0       INTRODUCTION

1.1       Secondary Metabolites

1.1.1    Alkaloids

1.1.2    Flavonoids

1.1.3    Terpenes

1.1.4    Steroids

1.1.5    Saponins

1.1.6    Tannins

1.1.7    Glycosides

1.3       Justification

1.4       Aim and Objectives

 

CHAPTER TWO

2.0       LITERATURE REVIEW

2.1       Botanical Description of Napoleonaea Species

2.1.1    Taxonomy of Napoleonaea Species

2.2       Origin and geographical distribution

2.3       Medicinal uses

2.4       Phytochemical Constituents of Napoleonaea species

2.5       Pharmacological Activities of Napoleonaea species

2.6       Scientific Classification of Napoleonaea heudelotti

 

CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Materials

3.1.1 Solvents

3.1.2 Apparatus

3.1.3 Reagents for phytochemical screening

3.1.4 Test microorganisms used

3.1.5 Materials used for chromatographic techniques

3.2       Collection of the Plant Material

3.3       Extraction of the Plant Material

3.4       Phytochemical Screening

3.4.1 Test for glycosides

3.4.2 Test for cardiac glycoside

3.4.3 Test for tannins

3.4.4 Test for saponins

3.4.5 Test for flavonoids

3.4.6 Test for carbohydrates

3.4.7 Test for combined reduced sugar

3.4.8 Test for steroids/terpenoids

3.4.9 Test for alkaloids

3.5       Antimicrobial Screening

3.6       Purification of chloroform fraction

3.6.1 Thin layer chromatography

3.6.2 Preparation of preparative TLC

3.6.3 Isolation of the pure compound

3.6.4 Melting point determination

3.6.5 Spectra Analysis

 

CHAPTER FOUR

4.0 RESULTS

4.1       Percentage ( %) Recovery on Extraction

4.2       Phytochemical Screening of extract and fractions

4.3       Antimicrobial Susceptibility test

4.4       Zone of Inhibition (mm)

4.5       Minimum inhibitory concentration (MIC) of extract and fractions

4.6       Minimum bactericidal concentration (MBC) of extract and fractions

4.7       Preparative thin layer chromatography

4.8       Chemical test of the isolated compound

4.9       Spectroscopic analysis of sample NHPE

 

CHAPTER FIVE

5.0 DISCUSSION

 

CHAPTER SIX

6.0 SUMMARY, CONCLUSION, AND RECOMMENDATION

6.1 Summary

6.2       Conclusion

6.3       Recommendation

REFERENCES

CHAPTER ONE

INTRODUCTION

Medicinal plants have been identified and used throughout human history. Plants have the ability to synthesize a wide variety of chemical compounds that are used to perform important biological functions, and to defend against attack from predators such as insects, fungi and herbivorous mammals (Babalola, 2009). Chemical compounds in plant mediate their effects on the human body through processes identical to those already well understood for the chemical compounds in conventional drugs; thus herbal medicines do not differ greatly from conventional drugs in terms of how they work. This enables herbal medicines to be as effective as conventional medicines, but also gives them the same potential to cause harmful side effects. Ethnobotany (the study of traditional human uses of plants) is recognized as an effective way to discover future medicines. In 2001, researchers identified 122 compounds used in modern medicine which were derived from ethnomedical plant sources (Babalola, 2009). Many of the pharmaceuticals currently available to physicians have a long history of use, as herbal remedies, including aspirin, digitalis, quinine, and opium. Treatment of diseases is almost universal among non-industrialized societies, and is often more affordable than purchasing expensive modern pharmaceuticals (Beltrame et al., 2002). The World Health Organization (WHO) estimates that 80 percent of the population of some Asian and African countries presently use herbal medicine for some aspect of primary health care (Beltrame et al., 2002). Studies in the United States and Europe have shown that the use of herbal madicine is less common in clinical settings, but has become increasingly more in recent years as scientific evidence about it effectiveness has become more widely available. The annual global export value of pharmaceutical plants in 2011 accounted for over US$ 2.2 billion. Plants have continued to be major source of medicine either in the form of traditional medicine preparations or as pure active principles (Hill, 2011). This has made it important to identify plants with useful therapeutic actions for possible isolation and characterization of their active constituents. About 80 % of the world population relies on the use of traditional medicine which is predominantly based on plant materials (Brunton et al., 2006). Plant have been part of our lives since beginning of time, we get numerous products from plants, most of them, not only good and beneficial but also crucial to our existence. The use of plant to heal or combats illness is probably as old as human kind. Out of these simple beginning came the pharmaceutical industry. Yet the current view of plant is very different from how it all started. The acceptance of traditional medicine as an alternate form of health care and the development of microbial resistance to the available antibiotics has led researchers to investigate the antimicrobial herbal extract (WHO, 1993). In Africa, particularly Nigeria is rich in plants which are used in herbal medicine to cure diseases and to heal injuries. Some of these plants exhibit a wide range of biological and pharmacological activities such as antihelmenthics, oxytoxic laxative (Hostettmann et al., 2012). The secondary metabolites of plant provide human with numerous biological active components which have been used extensively as drugs, foods, additives, flavours, insecticides and chemicals. They exhibited remarkable biological activities, which include inhibitory effects on enzymes, modulatory effects on some cell types, protect against allergies antioxidants (Dongmo et al., 2001).

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