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PHYTOCHEMICAL SCREENING AND ANTIMICROBIAL ACTIVITY OF EUPORBIA HIRTA LINN, BREYNIA NIVOSA W. BULL, EUPHORBIA HETEROPHYLLA LINN AND MANIHOT ESCULENTA CRANTZ.

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Product Code: 00009708

No of Pages: 52

No of Chapters: 1-5

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ABSTRACT

Euporbia hirta, Breynia nivosa, Euphorbia heterophylla and Manihot esculenta powdered plant material were extracted using two solvents, ethanol and water. The ethanol extracts provided the higher yield and also more antimicrobial effectiveness than aqueous extracts. Phytochemical screening of the crude extracts revealed the presence of alkaloids, saponin, tannins, flavonoids, phenols and hydrogen cyanide (HCN). The plant extracts had a high percentage of HCN ranging from 13.48±0.03 to 39.15±0.04%. The presence of these bioactive constituents have been linked to the antimicrobial activity of the plant materials. The disc diffusion method was used to determine the antimicrobial activity against Pseudomonas aeruginosa, Enterobacter aerogenes, Salmonella pullorum and Proteus vulgaris.Minimum inhibitory concentrations was analysed using agar well diffusion method with values ranging from 50 to 200mg/ml. The growth of all the bacteria were inhibited through to varying degrees, thus justifying the use of the herbs in traditional medicine in treating enteric infections. P. vulgaris and P. aeruginosa were more susceptible with diameters ranging from 19mm and 16.33mm respectively. The antimicrobial activity of the plant materials is enhanced under acidic conditions and at elevated temperatures. The herbs E. hirta, B. nivosa, E. heterophylla and M. esculenta can be used as sources of oral drugs to fight infections caused by susceptible bacteria.

 





TABLE OF CONTENTS

Cover page

Title page                                                                                                                                i

Declaration                                                                                                                             ii

Certification                                                                                                                            iii

Dedication                                                                                                                               iv

Acknowledgement                                                                                                                  v

Table of content                                                                                                                      vi

List of tables                                                                                                                           viii

List of plates                                                                                                                           ix

Abstract                                                                                                                                   x


CHAPTER ONE

INTRODUCTION                                                                                            1       

1.1          Aim and Objectives                                                                                 2

1.2          Justification                                                                                              3

1.3          Botany of the plants                                                                                 3

1.3.1    Botany of Manihot esculenta Crantz (Cassava)                                       3

1.3.2    Botany of Breynia nivosa W. Bull(Small-Snow bush)                               4

1.3.3    Botany of Euphorbia hirta Linn (Asthma plant)                                     5

1.3.4    Botany of Euphorbia heterophylla Linn (Purge weed)                               6


CHAPTER TWO

2.1     Literature review                                                                                      7

2.1.1  Phytochemical screening of plants                                                          7


CHAPTER THREE

3.0     Materials and methods                                                                             10

3.1     Collection of plant materials and identification                                       10

3.1.1  Preparation of the samples for analysis                                                  10     

3.2     Phytochemical analysis                                                                            11

3.2.1  Qualitative analysis on leaf extract                                                          11

3.1.1. Test for the presence of tannins                                                               11

3.2.1.2 Test for the presence of saponins                                                           11

3.2.1.3 Test for flavonoids                                                                                 12

3.2.1.4 Test for the presence of Alkaloids                                                         12

3.2.1.5Test for the presence of phenols                                                             13

3.3     Quantitative analysis of phytochemicals                                                 13

3.3.1  Determination of tannins                                                                         13

3.3.2  Determination of alkaloids                                                                      14

3.3.3  Determination of phenols                                                                        15

3.3.4  Determination of flavonoids                                                                    16

3.3.5  Determination of saponin                                                                        17

3.3.6  Determination of hydrogen cyanide (HCN)                                            18

3.4     Determination of antimicrobial activity                                                   19

3.4.1  Production of plant extracts                                                                     19

3.4.2  Source of test organisms                                                                          20

3.4.4  Determination of minimum inhibitory concentration                               21

3.4.5  Statistical Analysis                                                                                   22


CHAPTER FOUR

4.0     Results                                                                                                     26

4.1     Phytochemical properties                                                                         26

4.1.1  Quantitative analysis                                                                                26

4.1.2  Qualitative analysis                                                                                  27

4.1.3  Antimicrobial activity                                                                              29

4.1.4  Minimum inhibition concentration (MIC)                                               30


CHAPTER FIVE

5.0     Discussion and conclusion                                                                       37

5.1     Discussion                                                                                                38

5.2     Conclusion                                                                                               43

References

 

  

 

 

 

LIST OF TABLES

Table 1:       Qualitative analysis of phytochemicals present in

E. hirta, B. nivosa, E. heterophylla and M. esculenta.               27

Table 2:       Quantitative analysis of phytochemicals present in E. hirta,

B. nivosa, E. heterophylla and M. esculenta.                                28

Table3:        Antimicrobial activity of the aqueous and ethanolic

extracts of  Euphorbia hirta, Breynia nivosa,                                30

Euphorbia heterophylla and Manihot esculenta against the

test organisms (P.aeruginosa, E.aerogenes, S.pullorum            

and P.vulgaris)

Table 4:       The minimum inhibitory concentrations (MIC – mm) of

the plant extracts against the test organism (P. aeruginosa) 31

Table 5:       The minimum inhibitory concentrations (MIC – mm) of the

 plant extract against the test organism (E. aeruginese)            32

Table 6:       The minimum inhibitory concentrations (MIC – mm) of the

plant extract against the test organism (S.pullorum)                               33

Table 7:       The minimum inhibitory concentration (MIC- mm) of the

plant extracts against P. vulgaris                                                   34

 

 

 

 

LIST OF PLATES

Plate 1: The leaves of Euphorbia hirta                                                              23

Plate 2: The leaves of Breynia nivosa                                                                23

Plate 3: The leaves of Euphorbia heterophylla                                                  24

Plate 4: The leaves of Manihot esculenta                                                          24

Plate 5: Solidified Muller Hinton agar in sterile petri dishes                           25

Plate 6: Zone inhibition for aqueous and ethanolic leaf extract of

Euphorbia heterophylla on Salmonella pullorum                                   35

Plate 7 Zone of inhibition for aqueous and ethanolic leaf extract of                

Euphorbia hirta on Salmonella pullorum                                                36

 


 

 

 

 

CHAPTER ONE

1.0         INTRODUCTION

Antimicrobial agents are substances that interfere with the growth and metabolism of microbes. In common usage the term denotes inhibition of growth and with reference to specific groups of organisms; such terms as antibacterial, antifungal, antiviral and antiprotozoal are frequently employed. Antimicrobial agents may either kill microorganism or inhibit their growth. Antimicrobial agents are particularly useful in situations which the normal host defenses cannot be relied on to remove or destroy pathogen (Nester et al., 2004).

Plants have provided a source of inspiration for novel drug compounds, as plant derived health and wellbeing. Plant extract have been used for a wide variety of purposes for many thousands of years (Jonnes, 1996).

Medicinal plants have continued to attract attention in the global search for effective antimicrobial agents that can combat resistant pathogens that have been rendering many conventional drug obsolete in the treatment of infections (Cox, 1990).

Many important drugs used in medicine today are directly or indirectly derived from plants. A number of plants have been used in traditional medicine for many years due to their antimicrobial properties (Sofowora, 1993). Specifically, the medicinal value of these plants lies in some chemical substance that produce a definite physiological action on the human or animal body (Edeoga et al.,2005). Since ancient times, plants with therapeutic properties have secured an important place in the healing practices and treatment of diseases (Mohammad, 2011).

Medicinal plants are of great importance to the health of individuals and communities. Modern scientists have made phenomenal step in developing this heritage handed over by our forefathers (Sofowara, 1986).

In Nigeria, application of medicinal plants especially in traditional medicine is currently well acknowledged and established as a visible profession (Kafaru, 1994).

Extraction of bioactive compounds from medicinal plants permits the demonstrations of their studies leading to synthesis of a more portend drugs with reduced toxicity (Ebana et al., 1991).

Presently in the developing countries, synthetic drugs are inadequate for the treatment of diseases but are also often with adulterations and side effects (Shariff, 2001). Therefore, there is need to search for suitable plants of medicinal value to be effective in the treatment of diseases, which must be harmless to human tissues. Medicinal plants still remains the primary source of supply of many important drugs used in orthodox medicine today. Investigation into the chemical and biological activities of plants in the past years have yielded compounds for the development of modern synthetic organic compounds as a major route for the discovery of novel and more effective therapeutic agents. The African continent is one which is endowed with one of the richest biodiversity in the world with abundance of many plants used as herbs, foods and for therapeutic purposes. Great need arises for the evaluation of the pharmacological properties and detailed screening of bioactive substances for chemotherapeutic purposes. Furthermore, investigation into the antimicrobial activities of these plants will show that plants are potential source of synthetic drugs (Clark, 1996; Kubmarawa et al., 2009).

The active principles for many drugs found in plants are secondary metabolites (Ghani, 1990). Phytochemicals are chemical compounds that occur naturally in plants. Medicinal properties of the plants are generally dependent on the presence of certain phytochemicals such as alkaloids, tannins, saponins, flavonoids, reducing sugar and anthraquinone with bioactive bases thought to be responsible for antimicrobial property (Ebana et al., 1993).

Alkaloids are a group of mildly alkaline compound because it is astringent. Saponins occurs widely in varieties of plant, they are used as cleaning agent. Some of the saponins are useful as raw materials for synthesis of steroid hormones. In recent years, secondary metabolites previously with unknown pharmacological activities have been extensively investigated as a source of medicinal agents (Krishnaraju et al., 2005).

Phytochemicals with adequate antimicrobial efficacy will be used for the treatment of the bacterial infections (Balandrin et al., 1985). Natural products are therefore gaining attention as an alternative for antimicrobial agents (Helal et al., 2015).

 

1.1.2    AIMS AND OBJECTIVE OF THE RESEARCH

(i) To determine the phytochemicals of the leaf extracts of Terminalia catappa, Gongronema latifolium, Piper guineenses and Psidium guajava.

(ii) To determine the plant with the best antimicrobial properties


1.1.3    JUSTIFICATION OF THE STUDY

Many important drugs used in medicine today are directly or indirectly derived from plants. A number of plants have been used in traditional medicine for many years due to their antimicrobial properties. The use of medicinal plant reduces side effect.

The result of this study will demonstrate the antimicrobial properties of the plant extracts that will kill or inhibit the growth of microorganisms and in the concentration in which the plant extracts will be effective in treating microbial infections. The minimum inhibitory concentration (MIC) will help in resolving the problem of dosage.

 


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