ABSTRACT
The phytochemical composition, antimicrobial and antioxidant activities of leaf of Croton gratissimus (lavender croton) was studied. Extraction of the leaf were obtained in different solvents (water, ethanol and ethyl acetate) and test at invitro level against, Staphylococcus aureus, Escherichia coli and Candida albicans. Results of preliminary phytochemical screening shows the presence of many phytochemicals including alkaloids, saponins, flavonoid, Taninnis, steroids and cyanogic glucosides at varying levels of detection. Quantative analysis of the leaf showed varied concentrations of the phytochemicals such as alkaloids (0.82%), Flavonoid (1.02%), saponin (0.37%), Tannin (0.58%), Phenols (0.82%), phytate (0.33%) and HCN (6.70mg/kg). The antimicrobial tests result indicated varying levels of inhibition of the three microbial pathogens, (Staphylococcus aureus, Escherichia coli and Candida albicans) by the plant extracts in different solvents. Water extract caused inhibition of diameters 12.33mm, 14.33mm, and 14.33mm, against Staphylococcus aureus, Escherichia coli, Candida albicans respectively while that of ethanol extract were 19.00mm, 24.33mm and 15.67mm and ethyl acetate extract had 16.33mm, 14.33mm and 18.67mm for the same test organisms respectively. The activity of the extracts were lower than that of standard antibiotic, which recorded 24.33mm, 23.33mm and 25.67mm against the same organisms. Further antimicrobial tests show minimum inhibition concentrations (MIC) of 150mg/ml for the water extract against each of the organisms. while, the ethanol extract was active at 100mg/ml 50mg/ml and 100mg/ml against Staphylococcus aureus Escherichia coli and Candida albicans respectively. Concentration of phytochemicals with antioxidant potency in the leaf was 21.38 mg /100g Vitamin C, 1.02% Flavnoid and 0.82 % Phenolics. Results of the antioxidant activity showed scavenging activity of 53.03%, chelating activity of 40.08% and reducing activity of standard antioxidant, Butylated Hydro toluene (BHT). Potential health benefits of Croton gratissimus leaves were recognized.
TABLE OF CONTENTS
Title page i
Certification ii
Dedication iii
Acknowledgements iv
Table of contents v
List of tables xi
Abstract xii
CHAPTER ONE
1.1 Introduction 1
1.2 Main Aim and Objective of the Research 3
CHAPTER TWO
2.0 LITERATURE REVIEW 4
2.1 Description and Habitat of Croton gratissimus 6
2.2 Nomenclature of Croton gratissimus 6
2.3 Growing Croton gratissimus 7
2.4 UsesVApplications of Croton Species 7
2.5 Phytochemical composition of Croton 8
2.5.1 Biological activities of the important phytochemicals found in plant 9
2.6 Mechanism of Antimicrobial Action of Bioactive Components of Plants 14
2.7 Some Phytochemical Groups And Mechanism of Anti-Infective Action 15
2.8 Plant as Alternative source of Antimicrobials
2.8.1 Plants having activity against multi drug resistance 17
2.8.2 Prospective of plants having activity against multi drug resistance 18
CHAPTER THREE
MATERIALS AND METHOD 20
3.1 Materials 20
3.2 Methods 20
3.2.1 Sample and media preparation 20
3.2.2 Media preparation 20
3.3 Preparation of Leave Extracts 21
3.4 Determination of Phytochemicals Content of the Plant Sample 21
3.4.1 Test for presence of alkaloids 21
3.4.2 Test for the presence of flavonoid 22
3.4.3 Test for the presence of phenols 22
3.4.4 Test for the presence of saponin 22
3.4.5 Test for the presence of tannin 23
3.4.6 Test for the presence of cyanogen glycosides 23
3.4.7 Test for steroid 23
3.4.8 Test for starch 24
3.5 Quantitative Determination of Phytochemical Constituents of the Plant Samples 24
3.5.1 Determination of alkaloid 24
3.5.2 Determination of flavonoid 25
3.5.3 Determination of phenols 25
3.5.4 Determination of saponins 26
3.5.5 Determination of tannin 27
3.5.6 Determination of cyanide 27
3.5.7 Determination of phytate 28
3.5.8 Determination of steroid 29
3.6. Preparation of Inoculant 30
3.6.1 Antimicrobial activity test 30
3.6.2 Minimum inhibitory concentration (mic) test 30
3.7. Determination of antioxidants 31
3.7.1 Determination of vitamin c 31
3.7.2 Chelating activity 31
3.7.3 Scavenging activity 32
3.7.4 Reducing activity 32
3.8 Statistical methods 33
CHAPTER FOUR
4.0 RESULTS 34
CHAPTER FIVE
DISCUSSION, CONCLUSION AND RECOMMENDATIONS 40
5.1 Discussion 40
5.2 Conclusion 42
5.3 Recommendations 43
References
Appendix
LIST OF TABLES
Table Title Page
1: Phytochemical Screening of Extract of Croton gratissimus 34
2: Qualitative phytochemical screening 35
3: Antimicrobial activity of Croton gratissimus extracts 36
4: Minimum Inhibitory Concentration, (mg/ml) of crude extracts of Croton gratissimus
against Staphylococcus aureus, Escherichia, coli and Candida albicans 37
5: Antioxidant components of leaf of Croton gratissimus 38
6: Antioxidant activity of Croton gratissimus 39
CHAPTER ONE
1.0 INTRODUCTION
Since ancient times plants have been used all over the world as unique sources of medicines and constitute the most common human use of biodiversity (Ribeiro et al, 2010). In Africa and in other developing countries many people depend on medicinal plant because of limited access to modern medicine (Runyoro et al 2006). The dependence on medicinal plants and traditional healers may be attributed to the low proportion of orthodox doctors to patients in Africa (Bekalo et al, 2009) reported the following ratios: South Africa 1:1639; Ethiopian 1: 33000; Kenya 1:7142; Tanzania 1:33000; Uganda 1:25000; Malawi 1:50000; Mozambique1:50000; Swaziland 1:10000. In these communities’ traditional healers operate closer to the people taking advantage of plant species in such areas to treat various disease and ailments (Kambizi and Afolayan 2011).
Phytochemicals (from the Greek word phyto, meaning plant) are biologically active, naturally occurring chemical compounds found in plants, which provide health benefits for humans further than those attributed to macronutrients and micronutrients (Mamta et al., 2013). They protect plants from disease and damage and contribute to the plant’s colour, aroma and flavor. In general, the plant chemicals that protect plant cells from environmental hazards such as pollution, stress, drought, UV exposure and pathogenic attack are called as phytochemicals [Mamta, et al, 2013). Recently, it is clearly known that they have roles in the protection of human health, when their dietary intake is significant. More than 4,000 phytochemicals have been catalogued and are classified by protective function, physical characteristics and chemical characteristics and About 150 phytochemicals have been studied in detail (Singh et al 2013)
In wide-ranging dietary phytochemicals are found in fruits, vegetables, legumes, whole grains, nuts, seeds, fungi, herbs and spices. Broccoli, cabbage, carrots, onions, garlic, whole wheat bread, tomatoes, grapes, cherries, strawberries, raspberries, beans, legumes, and soy foods are common sources (Mamta et al 2013) Phytochemicals accumulate in different parts of the plants, such as in the roots, stems, leaves, flowers, fruits or seeds. Many phytochemicals, particularly the pigment molecules, are often concentrated in the outer layers of the various plant tissues. Levels vary from plant to plant depending upon the variety, processing, cooking and growing conditions. Phytochemicals are also available in supplementary forms, but evidence is lacking that they provide the same health benefits as dietary phytochemicals (Mamta et al., 2013).
These compounds are known as secondary plant metabolites and have biological properties
such as antioxidant activity, antimicrobial effect, modulation of detoxification enzymes, stimulation of the immune system, decrease of platelet aggregation and modulation of hormone metabolism and anticancer property. There are more than thousand known and many unknown phytochemicals. It is well-known that plants produce these chemicals to protect themselves, but recent researches demonstrate that many phytochemicals can also protect human against diseases (Narasinga, 2003).
Historically, plant have provided a source of inspiration for novel drug compounds, as plant derived medicines have made large contribution to human health and wellbeing (Igbinosaoo and Aiyegoro 2009). Aromatic plants have been used in folk medicine as antimicrobial agents since ancient times. Although approximately 20% of the world plants have been submitted to pharmacology or biological test. It could be concluded that natural products from plant origin are an important source to discover new leads with economical and pharmaceutical importance and great possibilities to be developed as drugs, dyes fragrance and pesticides, among others. To obtain novel and promissory substance many plant extracts have to be assayed (Medes et al., 2011).
Furthermore, the screening of plant extracts as antimicrobial agent is necessary to go insight into medicinal flora and get molecules responsible for this activity and add value to natural resources from tropical areas (Rios et al., 2005) Many Euphobiaceae are well known in different parts of the world as toxic / or medicinal plants. The high diversity of described effects is a reflex of the high oxidation damage to a target molecule.
1.1 AIMS AND OBJECTIVES OF THE RESEARCH
· To produce extracts from the leaf of Croton gratissimus using water, ethanol and ethyl acetate
· To identify phytochemical components of Croton gratissimus
· To evaluate the Anti-oxidant properties of Croton gratissimus.
SPECIFIC OBJECTIVES
· To screen for the anti-microbial activity of leaf extracts of Croton gratissimus.
· To ascertain the Minimum Inhibitory Concentration of extracts of Croton gratissimus against Escherichia coli, Staphylococcus aerues and Candida albican.
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