PHYTOCHEMICAL AND ANTIMICROBIAL STUDIES ON THE STEM-BARK OF COMMIPHORA MOLLIS (Oliv. ) Engl. (BURSERACAEA)


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TABLE OF CONTENTS

Title page
Abstract
Table of Content
List of Abbreviation

CHAPTER ONE:
1.0       INTRODUCTION
1.1       Traditional Medicine
1.2       Anti microbial
1.3       Statement of Research Problem
1.4       Justification of the Study
1.5       Aim of the Research Work
1.5       Objective of the study
1.6       Research Hyphothesis

CHAPTER TWO:
2.0       LITERATURE REVIEW
2.1       Morphology of Commiphora mollis
2.2       Ethnomedicinal Uses of Commiphora mollis and other Species of the Genus
2.3       Flavonoids
2.3.1    Stucture diversity of flavonoids
2.3.2    Flavonoids- their therapeutic potential
2.3       Phytochemistry of Commiphora mollis and other Species of the Genus
2.4       Biology Action of Commiphora mollis and other Species of the Genus

CHAPTER THREE:
3.0       MATERIAL AND METHODS
3.1       Solvents/Reagents
3.2       Collection, Identification and Preparation of plant materials
3.3       Praparation of Extract
3.4       Preliminiry Phytochemical Screening
3.4.1    Test for sterols/terpenes
3.4.2    Test for flavonoids
3.4.3    Test for alkaloids
3.4.4    Test for tannins
3.4.5    Test for saponins
3.4.6    Test for antharquinone
3.5       Chromatography Procedure
3.5.1    Thin layer chromatography
3.5.2    Column chromatography
3.6       Chromatographic Separation
3.6.1 Thin layer chromatography of the methanol extract of Commiphora mollis
3.6.2 Column chromatography of ethylacetate fraction
3.6.3 Spectral analysis
3.7       Anti microbial Screening
3.7.1 Susceptibility tests
3.7.2 Minimum inhibition concentration
3.7.3 Minimum bacteriacidal / fungicidal concentration

CHAPTER FOUR:
4.0       RESULT
4.1       Preliminary Phytochemical Studies
4.2       Result of Column Chromatographic Separation
4.2.1 Gel- filteration chromatography of fraction D5
4.3       Thin Layer Chromatographic Profile of X1
4.4       Spectral Analysis of X1
4.4.1 1H NMR of X1 (δppm, 600 MHz, CD3OD)
4.4.2 13C NMR of X1 (δppm, 125 MHz, CD3OD)
4.4.3 DEPT experiment of X1
4.4.4 COSY experiment of X1
4.4.5 HSQC experiment of X1
4.4.6 HMBC experiment of X1

CHAPTER FIVE:
5.0       DISCUSSION
5.1       Preliminary Phytochemistry
5.2       NMR Spectroscopic Data of Compound X1
5.3       The Antimicrobial Profile

CHAPTER SIX:
6.0       CONCLUSSION AND RECOMMENDATIONS
6.1       Conclusion
6.2       Recommendation
REFREENCE




ABSTRACT

Commiphora mollis is traditionally used in the treatment of fever (malaria and typhoid) wound healing, cancer, ulcer and rheumatic condition. The plant grows in Nigeria and across Africa. Phytochemical analysis of methanol extract of the stem-bark of Commiphora mollis showed the presence of flavonoids, saponins, tannins, terpenoids and alkaloids. Extensive

Phytochemical studies of  ethylacetate   water soluble fraction of methanol extract resulted in

isolation (55 fractions, 100ml each were collected and pooled together based on similarities in their TLC profile to gave 8 major fractions and repeated gel filtration chromatography of Fraction 5 on Sephadex LH-20 packed column then preparative gave 7.5 mg of compound X1) and characterization using 1HNMR (9 proton signal 5 aromatic proton δ 7.0- 5.94 ppm, 2 oxymithine protons at 4.8 and 4.2 ppm and 2 methylene protons at 2.8 and 2.75 ppm),

13CNMR (15 carbon signals 12 in aromatic region, 2 oxymethine carbon and 1 methylene carbon.), DEPT (7 methine 1 methylene 1 quatenery ), 1H 1H COSY ( correlation between

protons at  7.0, H2’ //  6.83, H6’    7.0, H2’  //  4.8, H2      6.83, H6’ //  4.8, H2     4.8, H2 //  4.2,

H3  4.8, H2 //
2.8, H4   4.2 , H3  //  2.8, H4   ), HSQC  (correlation between proton and
carbon at
7.0, H2’ // 114, C2’
6.83, H6’ // 118.10, C6’  6.8, H5’ // 114.60, C5’  5.8, H6  //
94, C6
4.82,
H2  // 78.48, C2
4.21, H3 // 66.10, C3  2.8, H4 and 2.7, H4 // 27.84, C4 ),

HMBC and NOESY. The antimicrobial studies of the crude methanol extract, Chloroform and ethylacetate fractions were carried out using disc diffusion and broth agar dilution methods on clinical isolates of Corynebacterium ulcerans, Salmonella typhi, Proteus mirabilis, Bacillus sabtilis, Staphylococcus aureus, Streptococcus pyogens, Escherichia coli, Candida brusei, Shigella dysenteriae and Candida albicans. The crude methanol extract, chloroform and ethylacetate fractions showed strong inhibitory activity against all tested microoganisms with exception of Corynebacterium ulcerans, Salmonella typhi and Proteus mirabilis. Crude methanol extract was found to have MIC at 10 mg/ml for all organisms and

variable value of MBC/MFC and that of chloroform and ethylacetate fractions have variable value for both MIC and MBC. The stem-bark of Commiphora mollis was rich in bioactive phytochemicals which have antimicrobial activity and could serve as a potential source of compounds effective against disease causing micro- organisms.





CHAPTER ONE


1.0 INTRODUCTION


Natural products are the chemical compounds found in nature that usually have a pharmacological or biological activity for use in pharmaceutical drug discovery and drug design (Samuel, 1999). According to the World Health Organization (WHO), about 80 % of the world’s population relies on traditional medicine for their primary health care need (WHO, 2002). For thousands of years natural products have played a very important role in health care and prevention of diseases. The ancient civilizations of the Chinese, Indians and North Africans provide written evidence for the use of natural sources for curing various diseases (Phillipson, 2001).

Secondary metabolites are chemical compounds derived from living organisms. The study of natural products involves isolation in a pure form of these compounds and investigation of their structure. Secondary metabolites appear to function primarily in defense against predators and pathogens and in providing reproductive advantage as intraspecific and interspecific attractants. They may also act to create competitive advantage as poisons of rival species (Croteau, et al., 2000). Many plant terpenoids are toxins and feeding deterrents to herbivores or are attractants, and many possess pharmacological activity. Tannins, lignans, flavonoids, and some simple phenolic compounds serve as defenses against herbivores and pathogens, also many flavonoid pigments are important attractants for pollinators and seed dispersers.

The study of natural products has had a number of rewards. It has led to the discovery of a variety of useful drugs for the treatment of diverse ailments and contributed to the development of separation science and technology, spectroscopic methods of structure elucidation and synthetic methodologies that now make up the basics of analytical organic.....


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