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Daniellia Pharmacognostic and Biological Studies of the Root of Oliveri (Rolfe) Hutch. & Dalz. (Caesalpiniaceae).

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Daniellia Pharmacognostic and Biological Studies of the Root of Oliveri (Rolfe) Hutch. & Dalz. (Caesalpiniaceae). 

ABSTRACT 

Daniellia oliveri is one of the most extensively utilized medicinal plants; it is a tree all of whose parts are used in Nigeria and some West African countries for the treatment of various ailments example the root is used as chewing stick. In the bid to standardize this plant, pharmacognostic and biological (anti-Candida) studies were carried out on its root and root bark. The anatomical and powdered samples of the root and root bark of D. oliveri were investigated for their microscopical profiles.

The results indicated the presence of xylem and phloem tissues, parenchyma cells, cork cells, prisms of calcium oxalate crystals, abundant starch grains and secretory ducts. Phytochemical screening of the methanol extract and its fractions (ethyl acetate, n-butanol, and aqueous fractions) revealed the presence of carbohydrates, saponins, flavonoids, cardiac glycosides, steroids and tannins.

The solvent system chloroform: methanol (7:3v/v) and n – butanol: acetic acid: water (2:1:2v/v) were found to be the best solvent systems for the separation of the constituents of the crude methanol extract and the different fractions using thin layer chromatography.Physico-chemical constants of the powdered root bark were determined; as 21.25+ 1.0 (total ash value), 21.75 + 1.0 (acid insoluble ash value), 11 + 1.0 (water soluble extractive value), 15.33+ 2.4 (alcohol soluble extractive value) and 7 + 1.0 (moisture content).

The total phenolic content of the root bark extracts of D. oliveri were also determined; as 57±4.55mgg-l (Crude methanol extract), 59.33±6.65mgg-l (ethylacetate fraction), 60±5.72mgg-l (n-butanol fraction), 17±0.00mgg-l (aqueous fraction) and 12±2.94mgg-l (Crude water extract). The methanol extract showed an LD50 of 565.69mgkg-l i.p in rats. 

TABLE OF CONTENTS

Title page – – – – – – – i
Declaration – – – – – – – ii
Certification – – – – – – – iii
Dedication – – – – – – – – iv
Acknowledgment – – – – – – v
Abstract – – – – – – – vii
List of tables – – – – – – – xiv
List of figures – – – – – – – xv
List of plates – – – – – – – xvi

CHAPTER ONE
1.0 Introduction – – – – – – 1
1.1 Fungi – – – – – – – 2
1.1.1 Candida species – – – – – – 3
1.1.2 Forms of Candidiasis- – – – – – 5
1.1.3 Signs – – – – – – – – 7
1.1.4 Symptoms – – – – – – – 7
1.1.5 Causes – – – – – – – 8
1.1.6 Treatment – – – – – – – 9
1.1.7 Plants used as antifungal agents – – – – 14
1.2 Standardization of medicinal plants -` – – – 19
1.2.1 Pharmacognostic standardization – – – – 19
1.2.2 Biological Standardization – – – – 21
1.3 Statement of Research problem – – – – 21
1.4 Justification – – – – – – 22
1.5 Hypothesis – – – – – – – 23
1.6 Aim – `- – – – – – – 23
1.7 Objectives – – – – – – – 23

CHAPTER TWO
2.0 Literature review – – – – – – 24
2.1 Daniellia oliveri (Rolfe) Hutch. and Dalz. – – – 24
2.2 Botanical Description – – – – – 24
2.3 Ethnomedical uses of D. oliveri – – – – 25
2.4 Biological activities from extracts of D. oliveri – – 26
2.5 Phytochemical constituents of D. oliveri – – – 29
2.6 Microscopical investigation of D. oliveri – – – 33

CHAPTER THREE
3.0 Materials and Methods – – – – – 35
3.1 Collection, identification and preparation of
plant material – – – – – – – 35
3.2 Macroscopical examination of the powdered
root bark of D. oliveri – – – – – 35
3.3 Microscopical examination of the root and
powdered root bark D. oliveri – – – – 35
3.3.1 Anatomical sectioning of the root of D. oliveri – – 35
3.3.2 Microscopical examination of powdered root bark
of D. oliveri – – – – – – – 38
3.4 Chemomicroscopical examination of the root
bark of D. oliveri – – – – – – 39
3.4.1 Test for cellulose – – – – – – 39
3.4.2 Test for lignin – – – – – – 39
3.4.3 Test for tannins – – – – – – 40
3.4.4 Test for mucilage – – – – – – 40
3.4.5 Test for starch – – – – – – 40
3.4.6 Test for fats – – – – – – – 40
3.4.7 Test for proteins – – – – – – 40
3.4.8 Test for calcium oxalate crystals – – – – 41
3.4.9 Test for calcium carbonate – – – – – 41
3.5 Physico – chemical analytical methods – – – 41
3.5.1 Determination of total ash value – – – – 41
3.5.2 Determination of acid-insoluble ash value – – – 42
3.5.3 Determination of moisture content – – – – 42
3.5.4 Determination of extractive values – – – – 43
3.6 Extraction and fractionation of methanol extract – – 44
3.7 Phytochemical screening of methanol extract
and its fractions – – – – – – 46
3.7.1 Test for carbohydrates – – – – – 46
3.7.2 Test for saponins – – – – – – 47
3.7.3 Test for flavonoids – – – – – – 48
3.7.4 Test for cyanogenetic glycosides – – – – 48
3.7.5 Test for steroids / triterpenoids – – – – 49
3.7.6 Test for anthraquinones – – – – – 49
3.7.7 Test for tannins – – – – – – 50
3.7.8 Test for alkaloids – – – – – – 50
3.7.9 Test for cardiac glycosides – – – – – 51
3.8 Thin layer chromatographic studies on methanol
extract and fractions – – – – – – 52
3.9 Total phenolic content- – – – – – 53
3.10 Biological evaluation – – – – – – 53
3.10.1 Acute toxicity testing of the methanol extract – – 53
3.10.2 Anti Candida activity of water extract, methanol
extract and its fractions – – – – – 54
3.10.2.1 Preparation of media – – – – – 54
3.10.2.2 Preparation of solvents – – – – – 55
3.10.2.3 Study population – – – – – – 56
3.10.2.4 Preparation of the test organism – – – – 56
3.10.2.5 Sensitivity test – – – – – – 58

CHAPTER FOUR
4.0 Results – – – – – – – 61
4.1 Collection, identification and preparation of plant
materials – – – – – – – 61
4.2.1 Macroscopical analysis of the powdered root bark
of D. oliveri – – – – – – – 61
4.2.2 Microscopical examination of the powdered root
and root bark of D. oliveri – – – – – 64
4.2.3 Chemomicroscopical examination of the powdered
root bark of D. oliveri – – – – – 68
4.2.4 Physico-chemcial analytical method – – – 71
4.2.5 Phytochemcal screening of the methanol extract and
its fractions – – – – – – – 71
4.2.6 Thin layer chromatographic analysis of methanol extract and
the three different fractions – – – – – 75
4.2.7 Total phenolic content – – – – – 79
4.3.0 Biological evaluation – – – – – – 82
4.3.1 Acute toxicity study of the methanol extract in mice – – 82
4.3.2 Anti-Candida activity of the methanol extract and
its fractions – – – – – – – 84
4.3.2.1 Identification of isolates – – – – – 84
4.3.2.2 Sensitivity test – – – – – – 87

5.0 Discussion – – – – – – – 94

6.0 Summary, conclusion and recommendations – – – 99
References – – – – – – – – 101

INTRODUCTION  

Medicinal plants are not only important to the millions of people for whom traditional medicine is the only resource for health care and to those who use plants for various purposes in their daily lives, but also serve as a source of new structures leading to drugs in all major disease areas.

Traditional medicine practice in the treatment of diseases and infections have assumed a more scientific and wider dimension as the emphasis on ethnomedicine is on the increase, especially in the developing countries, where the primary health care needs of the populace are not easily met. Today there are at least 120 distinct chemical substances derived from plants that are considered as important drugs currently in use in one or more countries in the world. Several of the drugs sold today are simple synthetic modifications or copies of the naturally obtained substances.

For example, the drug Ipecac derived from Cephaelis ipecacuanha Brot. which was used for many years to induce vomiting mostly if someone accidently swallowed a poisonous or harmful substance; the plant chemical quinine which was discovered in a rainforest tree Cinchona ledgeriana Moens ex. Tremen 100 years ago, for many years the quinine chemical extracted from the bark of this tree was processed into pills for the treatment of malaria; cynarin drug derived from Cynara scolymus L. is still clinically in use today for liver problems and hypertension (Taylor, 2000). 

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Ahmadu, A.A.; Haruna, A.K.; Garba, M. and Yaro, A.H. (2003) Antispasmadic Actions
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Ahmadu, A.; Haruna, A.K.; Garba, M. Ehinmidu, J.O. and Sarker, S.D. (2004)
Phytochemical and antimicrobial activities of the Daniellia oliveri leaves.
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Ali, M. (2002). Textbook of pharmacognosy. CBS Publishers and Distributors. New
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