EXTRACTION, ISOLATION, CHARACTERIZATION AND ANTI–MICROBIAL ACTIVITIES OF BRIDELIA FERRUGINEA BARK EXTRACTS

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TABLE OF CONTENT

Title page……………………………………………………………………..i

Certification………………………………………………………………….ii                    

Dedication …………………………………………………………………..iii                   

Acknowledgment ……………………………………………………………iv

Abstract………………………………………………………………………v

CHAPTER ONE

1.0 Introduction……………………………………………………………..1

1.1 Micro-organisms………………………………………………………….1

1.2 Classification of micro-organism…………………………………………2

1.2.1 Bacteria…………………………………………………………………3

1.2.2 Virus……………………………………………………………………4

1.2.3 Algae…………………………………………………………………..4

1.2.4 Fungi…………………………………………………………………..5

1.2.5 Protozoa ………………………………………………………………6

 CHAPTER TWO

2.0 Literature review………………………………………………………..7

2.1 Bacteria…………………………………………………………………7

2.2 Bacteria and its cells……………………………………………………9

2.3 Shape of the bacteria……………………………………………………12                    

2.4 Environmental effect of bacteria……………………………………….14

2.5 Medicinal plants…………………………………………………………16

2.5.1 Characteristics of medicinal plants……………………………………17

2.6   Medicinal plants as antimicrobial………………………………………26

2.7 Why considering medicinal plants as antimicrobial…………………….28

2.8    Botanical profile of Bridelia ferruginea ………………………………30

2.8.1   Taxonomy of plant……………………………………………………31

2.8.2   Plant description…………………………………………………….31

2.8.3   Properties……………………………………………………………32

2.8.4   Cultivation details……………………………………………………32

2.8.5   Edible uses…………………………………………………………..32

2.8.6   Medicinal uses……………………………………………………….32

2.8.7   Propagation………………………………………………………….33

2.9 Bridelia ferruginea as antimicrobial…………………………………….33

CHAPTER THREE

3.0 Materials and methods………………………………………………….35

3.1 Aims and objectives…………………………………………………….35

3.2 Materials…………………………………………………………………35

3.2.1 Chemicals,solvents and chemical reagents……………………………35

3.2.2 Equipment and materials………………………………………………36

3.3    Methods………………………………………………………………. 36

3.3.1 Collection and preparation of plant materials…………………………36

3.3.2 Extraction of plant materials…………………………………………..36

3.3.3 Phytochemical screening of Methanol extract…………………………37

3.3.4 Phytochemical screening of Hexane extract………………………….40

3.3.5 Phytochemical screening of Acetone extract…………………………43

3.4    Thin-layer chromatography……………………………………………47

3.5    Isolation and characterization of the crude extract……………………49

3.5.1 Column chromatography………………………………………………49

3.5.2 Mass of fractions gotten from the column chromatography…………..49

3.6 Instrumentation techniques………………………………………………50

3.6.1 Sample analysis for GC-MS…………………………………………..50

3.6.2 Sample analysis for AAS………………………………………………51

3.7 Anti-microbial activity of Methanol extract of Bridelia ferruginea……………51

3.7.1 Method……………………………………………………………………….51

3.8 Flow diagram of the entire experiment carried out in this

      research work…………………………………………………………………..52

CHAPTER FOUR

4.0 Result and discussion ……………………………………………………………53

4.1 Result of phytochemical analysis of methanol, N-Hexane, and              

      Acetone extract of Bidelia ferruginea……………………………………………53      

4.2 Instrumentation techniques……………………………………………………..55

4.2.1 Result for GC-MS analysis……………………………………………………55

4.2.2 Result for AAS analysis            ………………………………………………………61

4.3 Anti-microbial activities result…………………………………………………62

CHAPTER FIVE

Conclusion………………………………………………………………………….64

Recommendation…………………………………………………………………..64

Reference

ABSTRACT

 Medicinal plants are those plants that are used in treating and preventing specific ailments and diseases that affect human beings. Hence the important role of medicinal plants in health care delivery cannot be over emphasized. Bridelia ferruginea is atropical medicinal plant which belongs to the family of euphorbiaceae commonly used in traditional Africa medicine for treating various diseases. My research in this study was concentrated on the bark of bridelia ferruginea to detect the active ingredient responsible for the antimicrobial activities extraction. Phytochemical screening, thin layer chromatography, column chromatography, GC–MS, AAS and biological activities was carried out on the crude methanol extract of  Bridelia ferruginea. The pytochemical screening for the methanol crude extract indicate the presence of tannins, alkaloids, saponins, steroids, resins and phlobatahin, while the phytochemical screening for acetone crude extract indicate the presence of tannins, alkaloids, flavonoids, phlobatanin and reducing sugar. And the photochemical tests on the hexane crude extract gave negative result all through. Isolation was done using Column chromatography, AAS was used for the determination of some certain heavy metals present in both the isolate and the crude which may be toxic to man health and the GC–MS was used to determine the presences of unknown substances in the sample. Result for GC-MS shown the presence of seven compounds which are medicinal namely, 2-[4-methyl-6-(2,6,6-trimethylcyclohex-1-enyl)hexa-1,3,5-trienyl]cyclohex-1-en-1-carboxaldehyde, limonene-6-ol,pivalate, 1-Heptatriacotanol, Benzene,1-4-dicloro, cyclobarbital, 2-nonadecanone 2,4-dinitrophenylhydrazine, 1-monolinoleylglycerol trimethylsilylether. While for AAS, two metals were detected namely iron (2.1ppm) and zinc (0.05ppm) which are non-toxic for human consumption. And also result from the antibacterial activities shows that Bridelia ferruginea was highly susceptible to Escherichia coliand highly resistant to Proteus vulgaris.

         CHAPTER ONE

1.0 INTRODUCTION

1.1 MICRO-ORGANISM

A microorganism or microbe is an organism (form of life) that is microscopic (too small to see by the unaided human eye) [Delong et al 2001]. Is a microscopic living organism, which may be single-celled or multicellular. The study of microorganisms is called microbiology, a subject that began with the discovery of microorganisms in 1674 by Antonie van Leeuwenhoek, using a microscope of his own design [Madigan M et al 2006].

Microorganisms are very diverse and include all bacteria, archaea and most protozoa. This group also contains some species of fungi, algae, and certain microscopic animals, such as rotifers. Many Macroscopic animals and plants have microscopic juvenile stages. Some microbiologists also classify viruses as microorganisms, but others consider these as nonliving. In July 2016, scientists reported identifying a set of 355 genes from the last universal common ancestor of all life, including microorganisms, living on Earth [Madigan M et al 2006].

Microorganisms are often described as single-celled, or unicellular, organisms; however, some unicellular protists are visible to human eye, and some multicellular species are microscopic [Delong et al 2001].

Single-celled microorganisms were the first forms of life to develop on Earth, approximately 3–4 billion years ago. Further evolution was slow, and for about 3 billion years in the Precambrian eon, all organisms were microscopic. So, for most of the history of life on Earth, the only forms of life were microorganisms. Bacteria, algae and fungi have been identified in amber that is 220 million years old, which shows that the morphology of microorganisms has changed little since the Triassic period. [Madigan M et al 2006].

 Merging the idea of the microscopic and the very small with the older idea of an organism as a living entity or cell, the concept of a microorganism enabled a real appreciation of the microbial world as one that is amenable to study using similar tools and approaches even though representing distinctly different types of reproductive units and cell organizations [John A 2014].

EXTRACTION, ISOLATION, CHARACTERIZATION AND ANTI–MICROBIAL ACTIVITIES OF BRIDELIA FERRUGINEA BARK EXTRACTS