ABSTRACT
The hexane, chloroform, ethylacetate and methanol extracts of C. schweinfurthii leaves showed
high degree of ovicidal activity with 80 % inhibition at 10 μg/ml with chloroform crude extract
being the most potent with a zone of inhibition of 24-26 mm against Gram-positive bacteria with
a MIC of 12.5 mg/ml and MBC and MFC of 12.5 mg/ml. The chloroform and ethylacetate
extracts of C. schweinfurthii stem bark showed high degree of larvicidal activity with 74 %
inhibition at 10 μg/ml with ethylacetate crude extract showing a zone of inhibition of 22-33 mm
against Gram-negative bacteria with a MIC of 12.5 mg/ml and, MBC and MFC of 25 mg/ml.
The chloroform, ethylacetate and methanol extracts of M. pruriens showed potential ovicidal and
larvicidal activities with 58 % inhibition at 10 μg/ml with chloroform crude extract having a
zone of inhibition of 22-27 mm against Gram-negative bacteria with a MIC of 12.5 mg/ml and,
MBC and MFC of 25 mg/ml. Four compounds were isolated – 3β-hydroxyllup-20(29)-en-28-oic
acid (Betulinic acid) from the ethylacetate extract of C. schweinfurthii stem bark, a novel
derivative of α-amyrin identified as 3β-hydroxyl olean-12,18-diene from the chloroform extract
of C. schweinfurthii leaves, while stigmat-5-en-3β-ol (β-sitosterol) and naurol A were obtained
from the chloroform extract of M. pruriens. 3β-hydroxyllup-20(29)-en-28-oic acid has the
highest zone of inhibitions of 24 mm, 26 mm and 24 mm against Staphylococcus aureus,
Bacillus subtlis and Candida albicans with MIC of 6.25 μg/ml against all tested organisms and
MBC of 12.5 μg/ml and MFC of 25 μg/ml. 3β-hydroxyl olean-12,18-diene showed a
comparative potency with zone of inhibitions of 29 mm, 32 mm and 26 mm against Methicillin
Resistance Staphylococcus aureus , Bacillus subtlis and Candida albicans with MIC of 6.25
μg/ml and 12.5 μg/ml against the bacteria and candidas respectively, and MBC of 25 μg/ml and
MFC of 25 μg/ml. The finding proved that 3β-hydroxyllup-20(29)-en-28-oic acid and 3β-
hydroxyl olean-12,18-diene are the most potent isolated antimicrobial compounds.
The most ovicidal compounds were 3β-hydroxyl olean-12,18-diene and stigmat-5-en-3β-ol with
percentage inhibition of 62-65 % and 55-62 % respectively while the most larvicidal compounds
viii
were 3β-hydroxyllup-20(29)-en-28-oic acid ,3β-hydroxyl olean-12,18-diene and stigmat-5-en-
3β-ol at 78-83 %, 76-81% and 64–70 % at a concentration of 10-20 μg/ml respectively compared
to albendazole.
TABLE OF CONTENTS
Title Page
i
Cover Page ………………………………………………………………………………………………………………….
…………………………………………………………………………………………………………………. iii
Declaration ………………………………………………………………………………………………………………….
…………………………………………………………………………………………………………………. iv
Certification ………………………………………………………………………………………………………………..
……………………………………………………………………………………………………………………v
Acknowledgment …………………………………………………………………………………………………….. vi
Abstract vii
Table of Contents …………………………………………………………………………………………………….. ix
List of Tables …………………………………………………………………………………………………………..xv
List of Figures ……………………………………………………………………………………………………….. xvi
List of Scheme …………………………………………………………………………………………………….. xviii
List of Plates ……………………………………………………………………………………………………………….
……………………………………………………………………………………………………………….. xix
Abbreviations …………………………………………………………………………………………………………..xx
CHAPTER ONE ………………………………………………………………………………………………………1
1.0 INTRODUCTION …………………………………………………………………………………………….1
1.1 Natural Products and the Earliest Known Medicines to Man ………………………………1
1.2 Medicinal Plants in Folklore ………………………………………………………………………………2
1.3 Historically Important Natural Products ……………………………………………………………3
1.4 Natural Products from Plants …………………………………………………………………………….4
1.5 Statement of the Problem …………………………………………………………………………………..7
1.6 Justification of the Study ……………………………………………………………………………………7
1.7 Aim and Objectives of the Research ………………………………………………………………….8
CHAPTER TWO ……………………………………………………………………………………………………..9
2.0 LITERATURE REVIEW …………………………………………………………………………………9
2.1 Man and Parasite ……………………………………………………………………………………………..9
x
2.2 Pathogenesis and Pathology of Ascariosis ………………………………………………………….9
2.3 Economical Importance of Ascariosis ………………………………………………………………11
2.4 Control of Ascariosis ………………………………………………………………………………………12
2.5 Nematocidal Activity of Plants ………………………………………………………………………..12
2.6 Chemical Constituents …………………………………………………………………………………….13
2.6.1 Alkaloids …………………………………………………………………………………………………….14
2.6.2 Flavonoids and polyphenols ………………………………………………………………………….16
2.6.3 Glycosides and saponins ……………………………………………………………………………….19
2.6.4 Condensed tannins and sesquiterpene lactones…………………………………………………20
2.6.5 Triterpenes ………………………………………………………………………………………………….25
2.6.5.1 Naturally-ocurring pentacyclic triterpenes ……………………………………………………..27
2.7 Fabaceae ……………………………………………………………………………………………………28
2.7.1 Mucuna ……………………………………………………………………………………………………….29
2.7.2 Taxonomy and botanical descriptions ……………………………………………………………..29
2.7.3 Mucuna pruriens …………………………………………………………………………………………29
2.7.4 Species distribution ………………………………………………………………………………………31
2.7.5 Ethno botanical description of Mucuna pruriens………………………………………………31
2.7.6 Traditional uses of Mucuna pruriens ………………………………………………………………31
2.8 In vitro and in vivo Pharmacological Activities of Mucuna pruriens ………………31
2.8.1 Anti-parkinson’s activity ………………………………………………………………………………31
2.8.2 Antiglycaemic effect …………………………………………………………………………………….32
2.8.3 Hypoglycemic activity ………………………………………………………………………………….32
2.8.4 Antioxidant activity ……………………………………………………………………………………..32
2.8.5 Antivenom activity ………………………………………………………………………………………33
2.8.6 Aphrodisiac activity ……………………………………………………………………………………..33
2.8.7 Antimicrobial activity …………………………………………………………………………………..33
xi
2.8.8 Anthelmintic ……………………………………………………………………………………………….34
2.9 Isolated Compounds from Mucuna spp ……………………………………………………….34
2.10 Burseraceae Kunth Family …………………………………………………………………………40
2.11 The Genus Canarium L. ……………………………………………………………………………..40
2.11.1 Species distribution ……………………………………………………………………………………..41
2.12 Botanical Features of Canarium L. ……………………………………………………………..41
2.13 Traditional and Medicinal Uses of Canarium L. ………………………………………….42
2.14 Canarium schweinfurthii Engl. ……………………………………………………………………42
2.15. In vitro and In vivo Pharmacological Activities of Canarium L …………………….43
2.15.1 Antioxidant ………………………………………………………………………………………………..43
2.15.2 Antibacterial and antifungal activities ……………………………………………………………44
2.15.3 Hepatoprotective activities……………………………………………………………………………45
2.15.4 Anti-diabetic activity …………………………………………………………………………………..45
2.15.5 Antitumor potential ……………………………………………………………………………………..45
2.15.6 Other biological activities ……………………………………………………………………………..46
2.16 Isolated Compounds from Canarium spp ……………………………………………………46
2.17. Microbial Agents ……………………………………………………………………………………….48
2.17.1 Infectious disease ………………………………………………………………………………………..48
2.17.2 Chemotherapeutic agents: Factors affecting their effectiveness …………………………48
2.17.3 Antimicrobial drug resistance ……………………………………………………………………….49
CHAPTER THREE ………………………………………………………………………………………………..50
3.0 MATERIALS AND METHODS ………………………………………………………………..50
3.1 Chemicals/Reagents …………………………………………………………………………………..50
3.1.1 Equipment/Materials …………………………………………………………………………………..50
3.1.2 Parasite ……………………………………………………………………………………………………..50
3.1.3 Microorganisms …………………………………………………………………………………………51
xii
3.2 Plant Materials ………………………………………………………………………………………..51
3.3 Reference Drug ……………………………………………………………………………………….51
3.4 Extraction of Crude Plant Active Ingredients …………………………………………..55
3.5 Preliminary Phytochemical Screening ……………………………………………………….55
3.5.1 Detection of alkaloids …………………………………………………………………………………55
3.5.2 Detection of glycosides ……………………………………………………………………………….55
3.5.3 Detection of saponins ………………………………………………………………………………….55
3.5.4 Detection of phytosterols …………………………………………………………………………….55
3.5.5 Detection of flavonoids ……………………………………………………………………………….55
3.5.6 Detection of phenols …………………………………………………………………………………..55
3.5.7 Detection of tannins ……………………………………………………………………………………55
3.6 Antimicrobial Evaluation ………………………………………………………………………….55
3.6.1 Preparation of inoculum ………………………………………………………………………………55
3.6.2 Antibacterial/fungal susceptibility test ………………………………………………………….57
3.6.3 Determination of minimum inhibitory concentration ………………………………………57
3.6.4 Determination of minimum bactericidal/fungicidal concentration …………………….58
3.7 Anthelmintic Activity of Crude Plant Extracts ……………………………………………58
3.7.1 Collection and identification of worms …………………………………………………………58
3.7.2 Culture of Ascaris suum eggs ……………………………………………………………………..58
3.7.3 Egg hatch assay ………………………………………………………………………………………….59
3.7.4 Larval development assay ……………………………………………………………………………59
3.8 Column Chromatography …………………………………………………………………………63
3.9 Isolation of Bioactive Compounds ……………………………………………………………..63
3.10 Preparative Thin-Layer Chromatography …………………………………………………63
3.11 Statistical Analysis ……………………………………………………………………………………68
CHAPTER FOUR …………………………………………………………………………………………………..69
xiii
4.0 RESULTS ………………………………………………………………………………………………..69
4.1 Extraction ………………………………………………………………………………………………..69
4.2 Column Chromatographic Separation ………………………………………………………69
4.3 Phytochemical Screening of Extracts …………………………………………………………69
4.4 Antimicrobial Screening …………………………………………………………………………….69
4.5 Minimum Inhibitory Concentration ……………………………………………………………69
4.6 Minimum Bactericial /Fungicidal Concentration …………………………………………79
4.7 In-vitro Anthelmintic Activity ……………………………………………………………………..79
4.7.1 Egg hatch assay ……………………………………………………………………………………………79
4.7.2 Larval hatch assay ………………………………………………………………………………………..79
4.8 UV-Visible Spectrophotometer ……………………………………………………………………79
4.9 Isolation of Bioactive Compounds and NMR Spectroscopy ………………………….79
4.10 Spectral Results ………………………………………………………………………………………….92
CHAPTER FIVE ………………………………………………………………………………………………….119
5.0 DISCUSSION …………………………………………………………………………………………..119
5.1 Extraction ………………………………………………………………………………………………..119
5.2 Phytochemical Screening of Crude Extracts ………………………………………………119
5.3 Antimicrobial Susceptibility Assasy of the Crude Extracts …………………………120
5.4 Zone of Inhibition of Crude Extracts ………………………………………………………..122
5.5 Minimum Inhibitory Concentration of the Crude Extracts ………………………..122
5.6 Minimum Bactericial/Fungicidal Concentration of the Crude Extracts ………123
5.7 Zone of Inhibition of the Isolated Compounds ………………………………………….123
5.8 Minimum Inhibitory Concentration of the Isolated Compounds ……………….124
5.9 Minimum Bactericidal/Fungicidal Concentration of the Isolated
Compounds ……………………………………………………………………………………………..125
5.10 Effect of Extracts on the Eggs and Larval of Ascaris suum …………………………126
5.11 Ovicidal Activity of the Isolated Compound ………………………………………………128
CHAPTER ONE
1.0 INTRODUCTION
1.1 Natural Products and the Earliest Known Medicines to Man
Natural products have been the most successful sources of potential drug leads (Butler, 2004;
Cragg and Newman, 2005; Haefner, 2003; Mishra and Tiwari, 2011; Rey-Ladino et al., 2011).
However, their recent implementation in drug discovery and development efforts have
somewhat demonstrated a decline in interest (Mishra and Tiwari, 2011). Nevertheless, natural
products continue to provide unique structural diversity in comparison to standard
combinatorial chemistry, which presents opportunities for discovering mainly novel low
molecular weight lead compounds. Since less than 10% of the world’s biodiversity has been
evaluated for potential biological activity, many more useful natural lead compounds await
discovery with the challenge being how to access this natural chemical diversity (Cragg and
Newman, 2005). The earliest records of natural products were depicted on clay tablets in
cuneiform from Mesopotamia (2600 B.C.) which documented oils from Cupressus
sempervirens (cypress) and Commiphora species (myrrh) which are still used today to treat
coughs, colds and inflammation (Cragg and Newman, 2005). The Ebers Papyrus (2900 B.C.) is
an Egyptian pharmaceutical record, which documents over 700 plant-based drugs ranging from
gargles, pills, infusions, to ointments. The Chinese Materia Medica (1100 B.C.) (Wu Shi Er
Bing Fang, contains 52 prescriptions), Shennong Herbal (~100 B.C., 365 drugs) and the Tang
Herbal (659 A.D., 850 drugs) are documented records of the uses of natural products (Cragg
and Newman, 2005). The Greek physician, Dioscorides, (100 A.D.), recorded the collection,
storage and the uses of medicinal herbs, whilst the Greek philosopher and natural scientist,
Theophrastus (~300 B.C.) dealt with medicinal herbs. During the Dark and Middle Ages the
monasteries in England, Ireland, France and Germany preserved this Western knowledge whilst
the Arabs preserved the Greco-Roman knowledge and expanded the uses of their own
resources, together with Chinese and Indian herbs unfamiliar to the Greco-Roman world
2
(Cragg and Newman, 2005). It was the Arabs who were the first to privately own pharmacies
(8th century) with Avicenna, a Persian pharmacist, physician, philosopher and poet,
contributing much to the sciences of pharmacy and medicine through works such as the Canon
Medicinae (Cragg and Newman, 2005).
1.2. Medicinal Plants in Folklore
The use of natural products as medicines has been described throughout history in the form of
traditional medicines, remedies, potions and oils with many of these bioactive natural products
still being unidentified. The dominant source of knowledge of natural product uses from
medicinal plants is as a result of man experimenting by trial and error for hundreds of centuries
through palatability trials or untimely deaths, searching for available foods for the treatment of
diseases (Kinghorn, et al., 2011). One example involves the plant genus Salvia which grows
throughout the south-western region of the United States as well as north-western Mexico and
which was used by Indian tribes of southern California as an aid in childbirth (Sam, 1989) Male
new-born babies were “cooked” in the hot Salvia ashes as it was believed that these babies
consistently grew to be the strongest and healthiest members of their respective tribes and are
claimed to have been immune from all respiratory ailments for life (Sam, 1989). The plant,
Alhagi maurorum Medik (Camels thorn) secretes a sweet, gummy material from the stems and
leaves during hot days (Duke et al., 2008). This gummy sap is called “manna” and consists
mostly of melezitose, sucrose and invert sugar and it has been documented and claimed by the
Ayurvedic people that the plant aids in the treatment of anorexia, constipation, dermatosis,
epistaxis, fever, leprosy, and obesity (Duke et al., 2008). It was also used by the Israelis who
boiled the roots and drank the extract as it stopped bloody diarrhea. The Konkani people
smoked the plant for the treatment of asthma, whilst the Romans used the plant for nasal polyps
(Duke et al., 2008). The plant Ligusticum scoticum Linnaeus found in Northern Europe and
Eastern North America was eaten raw first thing in the morning and was believed to protect a
person from daily infection (Dillenius, 1724); the root was a cure for flatulence (Beith, 1999;
3
Lightfoot, and Scotica, 1977; Martin, 1934), an aphrodisiac (Beith, 1999) and was used as a
sedative in the Faeroe Islands (Martin, 1934; Svabo, 1959). Atropa belladonna Linnaeus
(deadly nightshade) is found in Central and Southern Europe, Western Asia, North Africa,
North America and New Zealand. Its notoriously poisonous nature (three berries are sufficient
to kill a child) firmly excluded it from the folk medicine compilation and seemed to have been
accepted as dangerous to handle or to experiment with (Allen and Hatfield, 2004).
1.3. Historically Important Natural Products
Traditional medicinal practices have formed the basis of most of the early medicines followed
by subsequent, clinical pharmacological and chemical studies (Butler, 2004). Probably the most
famous and well known example to date would be the synthesis of the anti-inflammatory agent,
acetylsalicyclic acid (aspirin) derived from the natural product, salicin isolated from the bark
of the willow tree Salix alba L. (Der Marderosian and Beutler, 2002). Investigation of
Papaver somniferum L. (opium poppy) resulted in the isolation of several alkaloids including
morphine, a commercially important drug, first reported in 1803. It was in the 1870s that crude
morphine derived from the plant P. somniferum, was boiled in acetic anhydride to yield
diacetylmorphine (heroin) and found to be readily converted to codeine (painkiller).
Historically, it is documented that the Sumerians and Ancient Greeks used poppy extracts
medicinally, whilst the Arabs described opium to be addictive (Der Marderosian and Beutler,
2002). Digitalis purpurea L. (foxglove) had been traced back to Europe in the 10th century but
it was not until the 1700s that the active constituent digitoxin, a cardiotonic glycoside was
found to enhance cardiac conduction, thereby improving the strength of cardiac contractibility.
Digitoxin and its analogues have long been used in the management of congestive heart failure
and have possible long term detrimental effects and are being replaced by other medicines in
the treatment of “heart deficiency” (Der Marderosian and Beutler, 2002). The anti-malarial
drug quinine approved by the United States FDA in 2004, isolated from the bark of Cinchona
succirubra Pav. ex Klotsch, had been used for centuries for the treatment of malaria, fever,
4
indigestion, mouth and throat diseases and cancer. Formal use of the bark to treat malaria was
established in the mid-1800s when the British began the worldwide cultivation of the plant (Der
Marderosian and Beutler, 2002). Pilocarpine found in Pilocarpus jaborandi (Rutaceae) is an Lhistidine-
derived alkaloid, which has been used as a clinical drug in the treatment of chronic
open-angle glaucoma and acute angle-closure glaucoma for over 100 years. In 1994, an oral
formulation of pilocarpine was approved by the FDA to treat dry mouth (xerostomia) which is
a side effect of radiation therapy for head and neck cancers and also used to stimulate sweat
glands to measure the concentrations of sodium and chloride (Aniszewski, 2007). In 1998, the
oral preparation was approved for the management of Sjogren’s syndrome, an autoimmune
disease that damages the salivary and lacrimal glands.
1.4. Natural Products from Plants
Plants have been well documented for their medicinal uses for thousands of years. They have
evolved and adapted over millions of years to withstand bacteria, insects, fungi and weather to
produce unique, structurally diverse secondary metabolites. Their ethnopharmacological
properties have been used as a primary source of medicines for early drug discovery (Fellows
and Scofield, 1995; McRae, et al., 2007). According to the World Health Organization (WHO),
80% of people still rely on plant-based traditional medicines for primary health care
(Farnsworth et al.,1985) and 80% of 122 plant derived drugs were related to their original
ethnopharmacological purpose (Fabricant and Farnsworth, 2001). The knowledge associated
with traditional medicine (complementary or alternative herbal products) has promoted further
investigations of medicinal plants as potential medicines and has led to the isolation of many
natural products that have become well known pharmaceuticals. The most widely used breast
cancer drug is paclitaxel (Taxol®), isolated from the bark of Taxus brevifolia (Pacific Yew). In
1962 the United States Department of Agriculture (USDA) first collected the bark as part of
their exploratory plant screening program at the National Cancer Institute (NCI) (Cragg, 1998).
The bark from about three mature 100 year old trees is required to provide 1 g of Paclitaxel
5
given that a course of treatment may need 2 g of the drug. Current demand for Paclitaxel is in
the region of 100–200 kg per annum (i.e 50,000 treatments/year) and is now produced
synthetically (Dewick, 2002). The first of several FDA approvals for various uses for Taxol®
was announced in 1992 (Cseke, et al., 2006). Taxol® is present in limited quantities from
natural sources; its synthesis (though challenging and expensive) has been achieved (Nicolaou
et al., 1994). Baccatin III present in much higher quantities and readily available from the
needles of T. brevifolia and associated derivatives is an example of a structural analogue that
can be efficiently transformed into Taxol®(Dewick, 2002).
Other examples of antitumor compounds currently in clinical trials include ingenol 3-Oangelate
a derivative of the polyhydroxy diterpenoid ingenol isolated from the sap of
Euphorbia peplus (known as “petty spurge” in England or “radium weed” in Australia) which
is a potential chemotherapeutic agent for skin cancer is currently under clinical development by
Peplin Biotech for the topical treatment of certain skin cancers (Kedei et al., 2004; Ogbourne
et al., 2004). PG490-88 (14-succinyl triptolide sodium salt), a semisynthetic analogue of
triptolide is a diterpene-diepoxide isolated from Tripterygium wilfordii which is used for
autoimmune and inflammatory diseases in the People’s Republic of China (Kiviharju et al.,
2002 and Fidler et al., 2003). Combretastatin A-4 phosphate a stilbene derivative from the
South African Bush Willow, Combretum caffrum acts as an anti-angiogenic agent causing
vascular shutdowns in tumors (necrosis) and is currently in Phase II clinical trials (Cragg and
Newman, 2005; Holwell et al., 2002).
The Acquired Immune Deficiency Syndrome (AIDS) pandemic in the 1980s forced the
National Cancer Institute (NCI) and other organizations to explore natural products as sources
of potential drug candidates for the treatment of AIDS. Over 60,000 extracts of plants and
marine organisms were tested against lymphoblastic cells infected with HIV-1. The most
important result of these tests is the class of compounds known as the calanolides. In particular
the isolation of calanolide A and calanolide B from the Calonphyllum species, along with
6
prostratin from Homalanthus nutans, have now progressed into clinical and preclinical
development (Cox, 2001; Gustafson et al., 1992; Kashman et al., 1992). Calanolide A was
licensed and evaluated to Phase II clinical trials by Sarawak Medichem Pharmaceuticals,
however there has been no subsequent announcement for further drug development. In 2010,
Phase I human clinical trials of prostratin were carried out by the AIDS Research Alliance in
Los Angeles, California
Arteether, introduced in 2000, as Artemotil derived from artemisinin (introduced in 1987 as
Artemisin) which was first isolated from the plant Artemisia annua as approved antimalarial
drugs (Cragg and Newman, 2007). The plant was originally used in traditional Chinese
medicine as a remedy for chills and fevers. Other derivatives of artemisinin are in various
stages of clinical development as antimalarial drugs in Europe (Cragg, 1998 and Dewick,
2002). To date, a synthetic trioxolane modeled on the artemisinin pharmacophore, is being
assessed in combination with piperaquine (a synthetic bisquinoline antimalarial drug) in an
effort to treat malaria (Davidson et al., 2009).
Grandisines A and B are two indole alkaloids which were isolated from the leaves of the
Australian rainforest tree, Elaeocarpus grandis. Grandisine A contains a unique tetracyclic
skeleton, while Grandisine B possesses an unusual combination of isoquinuclidinone and
indolizidine ring systems. Both Grandisine A and Grandisine B, exhibit binding affinity for the
human δ-opioid receptor and are potential leads for analgesic agents (Carroll et al., 2005).
Galantamine hydrobromide is an Amaryllidaceae alkaloid obtained from the plant Galanthus
nivalis and has been used traditionally in Turkey and Bulgaria for neurological conditions and
is used for the treatment of Alzheimer’s disease (Heinrich and Teoh, 2004; Howes et al., 2003).
Apomorphine is a derivative of morphine isolated from the poppy (Papaver somniferum) and
is a short-acting dopamine D1 and D2 receptor agonist, as well as a potent dopamine agonist,
used to treat Parkinson’s disease (Deleu et al., 2004). “Curare” is the arrow poison of the
South American Indians and is prepared in the rain forests of the Amazon and Orinoco.
7
Tubocaurarine isolated from the climbing plant, Chondrodendron tomentosum
(Menispermaceae) is one of the active constituents used as a muscle relaxant in surgical
operations, reducing the need for deep anesthesia. The limited availability of tubocurarine has
led to the development of a series of synthetic analogues which are now preferred over the
natural product (Dewick, 2002).
1.5 Statement of the Problem
Helminth infections are among the most widespread infections in humans, distressing a huge
population of the world. The majorities of infections due to helminthes are generally restricted
to tropical regions and cause enormous hazards to health and contribute to the prevalence of
malnourishment, anemia, eosinophilia and pneumonia (Bundy, 1994).
Gastrointestinal nematode infections of livestock, which are bred for the production of meat,
milk, or wool all around the world, lead to enormous economic losses. The control of these
parasites has relied on the use of chemical anthelmintics, resulting in development of drugresistant
strains.
Alternative control methods are biological control, vaccination, and traditional medicinal
plants, which are the focus of examination over the world. The evidence of anthelmintic
properties of plants is gained primarily from ethnoveterinary and ethnomedical knowledge.
Novel approaches to use the plants for control of gastrointestinal nematodes in small ruminants
are outlined in the excellent reviews of Githiori et al., (2006) and Hoste and Torres-Acosta
(2011).
1.6 Justification of the Study
The gastrointestinal helminthes are becoming resistant to both the benzimidazoles and
macrocyclic lactones which are the available anthelminthic (Gill et al., 1991). Therefore, there
is a foremost problem in treatment of helminthes diseases (Sondhi et al., 1994) and there is an
increasing demand towards new anthelminthics, to treat drug-resistance helminth infections.
Anthelmintics of the macrocyclic lactone family (e.g. ivermectin, doramectin) have a
8
significantly longer residual effect in comparison with the other anthelmintics. Over time,
questions have been raised concerning the long-term impact of the massive application of these
highly efficacious, broad-spectrum anthelmintic compounds on the environment. Due to the
bioavilability of some of these drugs, high percentages of these substances are being excreted
unchanged after oral or systemic administration (Farkas et al., 2003; Plumb, 2008).
Mucuna pruriens (DC) and Canarium schweinfurthii (Engl) have a wide reputation among
ancient India of being curative for helminthiases like elephantiasis (Oudhia, 2002), intestinal
worms, genito-urinary diseases, black tongue, round worm, gonorrhea and stomach disorders
(Warrier et al.,1996).
1.7 Aim and Objectives of the Research
The aim of this study is to isolate and characterise triterpenoids from Mucuna pruriens (DC)
and Canarium schweinfurthii (Engl) and, evaluate the medicinal potentials.
The objectives of this work are to:
(i). extract the plants M. pruriens and C. schweinfurthii,
(ii) test the activities of the crude extracts against selected micro-organisms and Ascaris
suum,
(iii) purify and isolate and the phytochemicals from M. pruriens and C. schweinfurthii,
(iv) characterise and structurally elucidate the isolated compounds using spectral techniques
such as U.V, 1HNMR, 13CNMR, DEPT, COSY, NOESY, HSQC, HMBC) and
(v) test the activities of the pure compounds against on the selected micro-organisms and A.
suum
9
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