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PHYTOCHEMICAL SCREENING AND ANTIMICROBIAL ACTIVITY OF FOUR NIGERIAN MEDICINAL PLANTS

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Product Code: 00009715

No of Pages: 60

No of Chapters: 1-5

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ABSTRACT


The antimicrobial activity and phytochemical screening of the leaves of Piper guineense Schumach, Psidium guajava Linn, Gongrenema latifolium Endl and Terminalia catappa Linn was carried out. The four plant species indicated the presence of alkaloid, phenol, tannin, spaonin and flavonoid which are bioactive component. The percentage of the phytochemcials are as follows alkaloid (0.46.139%), phenol (0.37-1.66%, saponin (0.31-0.67%), flavonoid (0.23-0.55%) and tannin (0.13-037%). The antimicrobial activity of the ethanolic and aqueous extracts of the leaves of the plant species were tested against four microorganisms, Candida albican, Klebsiella pneumonia, Escherchia coli and Bacillus subtilis. The sensitivity of the microorganisms was determined using disk diffusion method. Ethanolic plant extract and aqueous extract inhibited the growth of these pathogens. The zone of inhibition of the growth of these microorganisms by the plant extract depends on the concentration of the extract. Ethanolic plant extract had high sensitivity than the aqueous plant extract which shows less. Gongronema latifolium contained more alkaloids (1.39±0.03%) than other phytochemicals and Terminalia catappa had less tannin (0.13±0.00%) than other phytochemicals. The ethanolic extracts of Gongronema latifolium had the highest inhibitory activity (24.00±2.00mm) which shows more sensitivity against the test organisms while the aqueous extracts of Terminalia catappa has low inhibitory activity (8.33±0.58mm) which showed less sensitivity against the test organism. This result reveals that these plant species have potentials as source of raw materials in the pharmaceutical industry.  






TABLE OF CONTENTS

Cover page

Title page                                                                                                                                i

Certification                                                                                                                            ii

Dedication                                                                                                                               iii

Acknowledgement                                                                                                                  iv

Table of content                                                                                                                      v

List of tables                                                                                                                           vii

List of plates                                                                                                                           viii

Abstract                                                                                                                                   ix

CHAPTER ONE

1.0       Introduction                                                                                                                1

1.1.2    Aims and objective of the research                                                                            3

1.1.3    Justification of the study                                                                                             4

CHAPTER TWO

2.0       Literature review                                                                                                        5

2.1       Phytochemical screening of plants                                                                             5

2.1.2    Antimicrobial activity of plants                                                                                  7

2.3       Botany of plants                                                                                                          9

2.3.1    Botany of Terminalia catappa                                                                                   9

2.3.2    Botany of Gongronema latifolium                                                                             10

2.2.3    Botany of Piper guineenses                                                                                        10

2.2.4    Botany of Psidium guajava                                                                                        11

CHAPTER THREE

3.0       Source of materials                                                                                                     12

3.1       Sample preparation                                                                                                     12

3.2       Phytochemical screening                                                                                            12

3.2.1    Qualitative analysis of phytochemicals                                                                      13

3.2.1.1 Test of tannins                                                                                                            13

3.2.1.2 Test of saponin                                                                                                          13

3.2.1.3 Test of flavonoids                                                                                                      13

3.2.1.4 Test of alkaloids                                                                                                        14

3.2.1.5 Test of Phenols                                                                                                           14

3.3       Quantitative determination of the phytochemcials                                                    14

3.3.1    Determination of alkaloids                                         `                                               14

3.3.2    Determination of flavonoids                                                                                       15

3.3.3    Determination of tannins                                                                                            16

3.3.4    Determination of saponin                                                                                           17

3.3.5    Determination of Phenols                                                                                           18

3.4       Antimicrobial activity of the leaves of the plants Terminalia catappa,                     

Piper Guineenses, Gongronema Latifolium and Psidium Guajava.                                    19

3.4.1    Sample Preparation                                                                                                     19

3.4.2    Production of extracts                                                                                                 19

3.4.3    Sources of test organism                                                                                             19

3.4.4    Antimicrobial activity test                                                                                          20

3.4.5    Determination of Minimum Inhibition Concentration                                               20

3.4.6    Statistical Analysis                                                                                                     21

CHAPTER FOUR

4.0       Results                                                                                                                        25

4.1       Phytochemical components                                                                                        25

4.1.1    Qualitative analysis                                                                                                    25

4.1.2    Quantitative analysis                                                                                                  26

CHAPTER FIVE

5.0       Discussion and conclusion                                                                                         33

5.1       Discussion                                                                                                                   33

5.2       Conclusion                                                                                                                  36

References

 

 




 

 

LIST OF TABLES


Table 1:     The qualitative analysis of Piper guineenses, Gongronema latifolium,

Terminalia catappa and Psidum guajava                                                       25

Table 2:      The percentage yield of Alkaloid, Saponin, Tannin, Phenol and flavonoid

of Piper guineenses, Gongronema latifolium, Terminalia catappa and

Psidium guajava                                                                                          26

Table 3:     Antimicrobial activity of Aqueous and Ethanolic extract of Piper guineenses, Gongronema latifolium, Teminalia catappa and Psidium guajava                    27

Table 4:      Minimum inhibitory concentration of Candida albican                                    29

Table 5:      Minimum inhibitory concentration of Klebsiella pneumonia                                    30

Table 6:        Minimum Inhibitory concentration of E. coli                                                   31

Table 7:        Minimum Inhibitory concentration of Bacillus subtilis                                     32

 

 

 

 

 

 

 

LIST OF PLATES

Plate 1: The leaves of Piper guineenses                                                                                 22

Plate 2: The leaves of Gongronema latifolium                                                                      22

Plate 3: The leaves of Terminilia catappa                                                                             23

Plate 4: The leaves of Psidium guajava                                                                                 23

Plate 5: Solidified Muller Hinton agar in sterile petri dishes                                                 24

 

 

 

 

 

 


 

 

 

CHAPTER ONE

1.0                                                   INTRODUCTION

Medicinal plants are considerably useful and economically essential since they contain active constituents called phytochemicals. Phytochemicals are the non-nutritive plant chemicals that have protective or disease preventive properties. The most important of these bioactive constituents of plants are alkaloids, tannins, flavonoids, saponins and phenolic compounds (Okwu, 2001).

Though criticized, plant based medicines have survived through ages and it is still catering for the health needs of millions all over the world (Kapor, 2001). Herbs and natural products do not possess much of the toxicity that is present in synthetic chemical thus enhancing their appeal for long term preventive strategies (Zou, 2005). The use of plant extracts and phytochemicals, both with known antimicrobial properties can be of great significance in the treatments of diseases. Many plants have been used because of their antimicrobial traits, which are chiefly due to syntheses of secondary metabolism by the plant (Prusti, 2008). Discovery of new antimicrobial compounds with diverse chemical structures and novel mechanisms of action becomes is of urgent attention. Currently, the development of resistant strains of bacteria has increased the need for new antibiotics (Eloff, 1998). The antimicrobial compounds produced by plants are active against plants and human pathogenic microorganisms (Sarac and Ugur, 2007).

There are several reports in the literature regarding the antimicrobial activity of plant crude extracts and bioassay guided fractionations of  these extracts that yielded active principles (Rabe and Van Stadan et al., 2001; Srinivasan et al., 2001 and El-Seedi et al., 2002).

Isolation of bacteria less susceptible to regular antibiotics and recovery of resistant isolates during antibacterial therapy is now a global problem (Muhammad and Muhammad, 2005). In the developing world, the situation is even worse because of poor sanitation and ignorance of good hygienic practices thus exposing a large number of people to infectious agents.

Some of the bacteria implicated in causing enteric infections include but not limited to E. coli, Salmonella spp, Proteus spp, Shigella spp, Pseudomonas spp and the Staphylococci. These bacteria which are usually present as commensals have several virulent factors and colourize in a biofilm fashion, causing a variety of intestinal and extra intestinal diseases (Lino and Deogracious, 2006).

There is therefore the need to develop some newer, safer, effective and above all, cheaper antimicrobial agents to tackle this problem.


1.1          Aim and objectives

The aim and objectives of this work are to:

i)              determine the phytochemicals of Breynia nivosa, Manihot esculenta, Euphorbia hirta and Euphorbia heterophylla

ii)            determine antimicrobial activities of the aqueous and ethanolic  leaf extracts of Breynia nivosa, Manihot esculenta, Euphorbia hirta and Euphorbia heterophylla on Pseudomonas aeruginosa, Salmonella pullorum, Proteus vulgaris and Enterobacter aeruginese.

1.2          Justification

The use of medicinal plants in the treatment of most pathogenic diseases has been a public discourse, with people having different views on the use of herbal drugs, stating that it causes more harm than good because it does not have a prescribed dosage and may lead to serious life threatening diseases. The result of this study will therefore clarify these views which may lead to the discovery of an alternative form of treatment by the use of the MIC (Minimum inhibition concentration) which will take care of the problem of dosage compared to antibiotics being used at present, to which many of the bacteria are developing resistance.


 

1.3          Botany of the plants

1.3.1    Botany of Manihot esculenta Crantz(cassava)

Cassava is a woody shrub native to South America of the spurge family, Euphorbiaceae. It is extensively cultivated as an annual crop in tropical and subtropical regions for its edible starchy tuberous root, a major source of carbohydrate (Oriola and Raji, 2013). In addition to this, it tolerates even long prolonged drought, high temperature and other adverse environmental conditions following the initial establishment period after planting to marginal soil and reliable yields in areas where other crops such as maize, sorghum, beans and soybeans die or do not perform, well (El-Sharkawy, 2012). Its production worldwide has increased and is expected to increase further due to the tremendous demand for good quality food, animal feeds, raw industrial materials as well as its antioxidant potential. It also possess anticarcinogenic, antidiabetic, anti-tumor properties (El-Sharkawy, 2012; and Suresh et al., 2011).

Cassava leaves are largely consumed as African indigenous vegetables rich in nutrients (Lancaster and Brooks, 1983 and Achidi et al., 2005). Indeed, cassava leaves are excellent sources of proteins (21-39%), minerals, antioxidant compounds and vitamins for the human diet (Ek-Sharkawy, 2012; Balamurugan and Anbuselvi, 2013). Recent findings in plant biochemistry have proved that the proteins embedded in cassava leaves are equal in quality to those in eggs (Soudy, 2012).

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

Breynia nivosa is a rounded shrub in the family of Euphorbiaceae, about 2m high, that is primarily used for its attractive foliage and found in villages and towns. It has mall, mottled, multi-coloured variegated leaves with white green and red coloured leaves (Smith, 1981). In southern Nigeria, it is used as chewing stick (Amadi et al., 2007). Ethnomedically, Breynia nivosa is used in treating headaches, toothaches and tooth infections (Onyegbule et al., 2014) and in the treatment of fever and malaria by the Ibibios of Niger Delta region of Nigeria. The leaf extract of the plant has been reported to contain flavonoids, tannins, alkaloids, glycosides and starch, saponins and proteins (Onyegbule et al., 2007). Biological activities of the leaf include; antimicrobial (Amadi et al., 2014; Onyegbule et al., 2014), analgesic, anti-inflammatory and antioxidant activities (Onyegbule et al., 2014).

1.3.3    Botany of Euphorbia hirta Linn (Asthma plant)

Euphorbia hirta belongs to the family Euphorbiaceae which is frequently seen occupying open waste spaces and grasslands, roadsides and pathways. Though native to Central America, the herb is widely cultivated throughout the tropics especially Central and East Africa (Adedapo et al., 2005). It is usually erect slender-stemmed, spreading up to 45cm tall, though sometimes can be seen lying down (Burkill, 1994). The medicinal usefulness of the herb has been the subject of numerous chemical and microbial studies. Some of the reported phytochemicals of the herb included triterpenoids, sterols, alkaloids, glycosides, flavonoids, tannin, phenols, choline and shikimic acid while some of the reported scientific uses includes as antispasmodic, antiasthmatic, expectorant, anti-catarrhal and antisyphilic compounds (Burkill, 1994; Adedapo et al., 2005 and Falodun et al., 2006).

1.3.4    Botany of Euphorbia heterophylla Linn (Spurge weed)

Euphorbia heterophylla is a medicinal plant with the common name “Spurge weed” belonging to the family Euphorbiaceae. It is native plant to Mexico and its original distribution includes an area spreading from California to East Texas and Central America. It is a hardy species, growing between 30cm and 70cm in height. The medicinal usefulness of the plant has the subject of numerous chemical and pharmacological studies. It grows in semi-humid places especially in cassava, cowpea and soybean plantations. Report of previous chemical study on E. heterophylla is however scanty. Falodun et al. (2004) reported the isolation of a flavonoid quercetin from the leaves of this plant. The leaves are known to possess antibacterial activity (Falodun et al., 2003).

In Igbo community of Nigeria, E. heterophylla is used as a purgative (Erden et al., 1999). Extracts of the decoction of the leaves is also used in the treatment of respiratory tract infections and asthma by traditional medicine practitioners.



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