PHYTOCHEMICAL ANALYSIS AND THE ANTI-INFLAMMATORY ACTIVITIES OF DICHLOROMETHANE FRACTION OF METHANOL EXTRACT OF CRATEVA ADANSONII

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PHYTOCHEMICAL ANALYSIS AND THE ANTI-INFLAMMATORY ACTIVITIES OF DICHLOROMETHANE FRACTION OF METHANOL EXTRACT OF CRATEVA ADANSONII

ABSTRACT

Inflammation is a complex biological response of vascular tissue to harmful stimuli such as pathogen, damage cells or irritants (Ryan & Majno, 1983). The urgency generated by increased rate of stroke, atherosclerosis attribute due to prolonged use of cyclooxygenase-1 and Cyclooxygenase- 2 inhibitors have accelerated anti-inflammatory drug research over the last decade while synthetic pharmaceutical agents continued to dominate research (Ryan & Majno, 1983). Attention increasingly has been directed to natural products. These are often more affordable and available and sometimes are perceived as more effective than conventional anti- inflammatory drugs.

Anti-inflammation was carried out using 12 rats which was divided into three groups of 4 rats each. Group 1 and 2 served as the negative and positive control respectively. Group 3 received 25mg/kg b.w. of the dichloromethane fraction of methanol extract of Crateva adansonii showed significance anti-inflammatory activity when compared with the standard difference used.

TABLE OF CONTENT

Title page
Certification

Dedication

Acknowledgement

Abstract

Table of Content

CHAPTER ONE

1.1 Introduction

1.2 Crateva adansonii as a plant

1.3 Research aim and objectives

CHAPTER TWO

LITERATURE REVIEW

2.0 Definition of Inflammation

2.1 Characteristics of Inflammation

2.2 Classes of Inflammation

2.3 Types of Inflammation

2.4 Chemical mediators of Inflammation

2.4.0 Properties of Mediators

2.4.1 Histamine

2.4.2 Serotonin (5- Hydroxy tryptamine)

2.4.3 Cytokines

2.4.4 Arachidonic acid

2.4.5 Platelet Activating Factor (PAF

2.4.6 Free Radicals

2.4.6.1 Nitric Oxide (NO

2.4.6.2 Oxygen-derived free radicals

2.4.7.0 Complement system

2.4.7.1 The clotting system/Coagulation system.

2.4.7.2 The Kinin System

2.4.7.3 The fibrinolytic system

2.5.0 Anti-Inflammatory agents

2.5.1 Non-steroidal Anti-inflammatory Drugs (NSAIDS)

2.5.1.0 Mechanism of Action of NSAIDS

2.5.2 Steroidal Anti-inflammatory drugs

CHAPTER THREE

MATERIAL AND METHODS

3.1 Materials

3.1.1 Equipment/Apparatus used

3.1.2 Chemical, Solvents and Reagents

3.1.3 Animals

3.2 Methodology

3.2.1 Collection and preparation of Plant Materials

3.2.2 Extraction

3.3 Fractionation

3.3.1 Gel Column Chromatography

3.4. Identification of Phytochemical groups in the extract

3.4.1 Preparation of Reagents for phytochemical analysis

3.4.2 Qualitative phytochemical analysis of the Extract

3.5 Thin Layer Chromatography

3.6 Anti-inflammatory activity test

CHAPTER FOUR

Result

CHAPTER FIVE

Discussion

Conclusion

References

Appendixes

CHAPTER ONE

INTRODUCTION AIM AND OBJECTIVES

Inflammation is one of body’s natural ways of protecting itself. It is also an essential reaction of the body to infection. Too little of an inflammatory response, and the body is unable to repel microbial invasions or heal injuries. Too much of an inflammatory response, and the immune system begins attacking the body’s own organs eventually leading to chronic disease (Sears, 2000).

Anti-Inflammatory refers to the property of a substance or treatment that reduces inflammation. There are obvious clinical markers of inflammation which include pain, redness, fever, swelling and loss of function (Pai et al. 2004). However if inflammation is allowed to continue unchecked it may results in neurodegenerative disease or cancer (Pai et al. 2004). A variety of safe and effective anti-inflammatory agents are available including non-steroidal anti-inflammatory drugs, corticosteroids.

Non-steroidal anti-inflammatory drugs (NSAID are usually indicated for the treatment of acute and chronic inflammation. NSAIDS work by reducing the production of prostaglandins (Bayness & Marek, 2005). Prostaglandins are chemicals that promote inflammation, pain and fever. The enzymes that produce prostaglandins are phospholipaseA2, PGH2synthase and Cyclooxygenase (Cox). There are two types of COX enzymes, cyclooxygenase-1 (Cox-1) and cyclooxygenase-2 (Cox-2). NSAIDS reduce the production of prostaglandin by inhibiting COX enzymes resultant effect is that, therefore, inflammation, pain and fever are reduced. Some herbs and plants possess anti-inflammatory and they include:

Crateva adansonii belonging to the family Capparaceae and phylum Magnoliophyta is small tree of forest and savanna woodland, often on river-banks, widely distributed in Nigeria and across Africa. The leaves are applied externally to relieve pain in joint, the fresh juice of leaves is used for the relief of earache, eye infection and anodyne in toothache. Powder of bark is used in rheumatism, itch, epilepsy and asthma (Sivarajan & Balachandran, 1994).

1.2 Crateva adansonii as a plant.

The flowering tree Crateva adansonii is called the sacred garlic pear and temple plant. The tree is sometimes called the spider tree because the showy flowers bear long, spidery stamens. It is native to Japan, Australia, much of south East Asia and several south pacific Islands. It grows in forest and savanna woodland, often on river-banks from Senegal to Nigeria, and across Africa to Zaire, Tanganyika and Madagascar. The plant’s common name is amakarode in Igbo, In Yoruba, it is egun-orun and in Hausa, it is ungududu. The tree attains approximately 15m height. The trunk is irregular, seldom straight, but is worthy of cultivation as an ornamental for its dense masses of white flowers borne at the ends of all the shoots. In the bush, owing to grass burning which it survives and repeated stripping of its leaves, the tree is often stunted. The wood is soft and yellow and strong-smelling when cut. The leaves are however eaten in soups or mixed with cereals. They are boiled and added to mixture called in Hausa “Kwadwo” containing a paste of locust beans, Parkia Spp. (Leguminosae Mimosoideae). The Yoruba consumes the leaves as a potherb.

In upper volta they are the ingredient of sauces. To some people the leaves are taken only in time of dearth and they are sold in northern markets. The bark is widely used for stomach-troubles in Nigeria. In Jebel Marra a bark-paste is used as a poultice on swellings.

Crateva adansonii is a deciduous plant with three-palmate leaves. The leaves are arranged opposite one another. They are elliptic with entire margins. The flowers are pale green. The plants bloom from March to May. The flowers are arranged in racemes. The fruits are berries. It can withstand temperatures above 1oc and 2oc (Burkill, 1985).

Taxonomy

Family: Capparaceae

Genus: Crateva

Species: Crateva adansonii DC

Phylum/Division: Magnoliophyta

Sub division: Magnoliophyta

Classes: Rosopsida

Subclass:  Dilleniidae

Superorder:  Violanae

Order:  Capparales

The medicinal effect of Crataeva adansonii plant bark is includes general healing, leprosy, stomach troubles. For the leaf, it is used in eye treatments, liver, for leaf; a pain-killer, for roots, dropsy, swellings, edema, gout, venereal diseases. Powder of bark is used in rheumatism, itch, epilepsy and asthma.

1.3 Research Aim and Objectives

Aim:

To study the anti-inflammatory effect of dichloromethane fraction extracts from Crataeva adansonii on rats.

Objectives:

  1. To determine the anti-inflammatory effect of Crataeva adansonii extracts in methanol on rats.
  2. To determine the phytochemicals present in Crateva adansonii extract in methanol on rats.

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