Abstract :

The crude methanolic extracts derived from the Tridax procumben leaves was screened in vitro for
possible phytochemical and biological. Crude plant powders were extracted sequentially with
methanol and the dried extracts obtained demonstrated the presence of significant pharmacological
activity on diabetes, microorganisms responsible for disease. The objective of this study is to
characterize the functional compounds that were extracted and separated from leaves of Tridax
procumben and were carried out using different methods using Thin Layer Chromatography (TLC),
Vacuum Liquid Chromatography (VLC), Column Chromatography. Under phytochemical analysis,
antioxidant test & Chemical screening was done. The antioxidant property found in crude methanolic
extracts derived from the leaves was very good. Tridax procumben leaves have also been found to
contain anthraquinones and anthracene derivatives of rhein, emodol, aloe-emodin, sennosides A and
B, 4,5- dihydroxy-1- hydroxymethylanthrone and 4,5-dihydroxy-2-
[Link] screening of the leaves and roots of Tridax procumben
evealed the presence of alkaloids, carbohydrates, tannins, saponins, phenols, flavonoids,
anthraquinones and cardiac glycosidess. Amongst the secondary metabolites are steroids, flavonoids,
anthraquinones, anthrones, and a few less common compounds such as ellagitannin, naphthalene,
phenolic acid, purine, and [Link] special interest are compounds such as kaempferol glycosides
and anthraquinones, already proven to have antimicrobial properties

RATIONALE AND OBJECTIVE OF THE WORK :

Medicinal plants have been identified and used throughout human history. Plants have the ability to
synthesize a wide variety of chemical compounds that are used to perform important biological
functions, and to defend against attack from predators such insects, fungi etc. Plants have been used
for health and medical purposes for several thousands of years. According to world health
organization, The number of higher plant species on earth is about 250 000 and It is estimated that
35 000 to 70 000 species have, at one time or another, been used in some cultures for medicinal
purposes. A majority of the world's population in developing countries still relies on herbal
medicines to meet its health needs. Herbal medicines are often used to provide firstline and basic
health service, both to people living in remote areas where it is the only available health service, and
to people living in poor areas where it offers the only affordable remedy. Even in areas where
modern medicine is available, the interest on herbal medicines and their utilization have been
increasing rapidly in recent years. Bangladesh is also a major country where people use a high
percentage of medicinal plants for various therapeutic activities. Use of volatile and penetrating plant
extracts in therapeutic applications for psychological and physical well being was in practice from
ancient times. Bangladesh is gifted by extraordinary natural resources which continuously help us in
many ways. One of the most beneficial natural resources is the plant resource which provides us with
food, shelter and medicine. According to the World Health Organization more than 80% of the world
population in developing countries depends on plant-based medicines for basic healthcare needs
Fabaceae, also called Leguminosae, pea family of flowering plants (angiosperms), within the order
Fabales. Fabaceae, which is the third largest family among the angiosperms after Orchidaceae
(orchid family) and Asteraceae (aster family), consists of more than 700 genera and about 20,000
species of trees, shrubs, vines, and herbs and is worldwide in distribution Tridax procumben plant
(family of Fabaceae) is grown in several tropical countries. It produces a large shrub with very large
once-compound leaves consisting of 8-14 pairs of leaflets and the very large leaflets (5-17 cm long
and 2-5 cm wide) have entire margins and rounded tips. Its golden yellow flowers are borne in dense
elongated clusters (30-60 cm long) near the tips of the branches, these flowers are interspersed with
yellow or orange floral bracts.

Its elongated pods (15-25 cm long) are somewhat four-angled and have papery wings Tridax
procumben plant has been used to skin problems, arthritis, HBP (high blood pressure), and laxative
or purgative, boils, wound, eye, urinary and gastrointestinal tract infections, diarrhoea and scarlet
fever. Recent reports have credited the use of Tridax procumben in the successful treatment of
haemorroids, constipation, inguinal hernia, intestinal parasitosis, blennorrhagia, syphilis and
diabetes. Tridax procumben leaves has several medicinally important phytochemical constituents
including, mineral elements: k, Zn, Cd, Na ,Mg ,Fe ,Ca. And the vitamin elements are βCarotene (IU),
Vitamin C (mg/L), Vitamin E (IU). Vitamin C anthraquinones and anthracene derivatives of rhein,
emodol, aloe-emodin, sennosides A and B, 4,5- dihydroxy-1- hydroxymethylanthrone and 4,5-
dihydroxy-2- hydroxymethylanthrone, alkaloids, carbohydrates, tannins, saponins, phenols,
flavonoids, anthraquinones and cardiac glycosidess, Amongst the secondary metabolites are steroids,
flavonoids, anthraquinones, anthrones, and a few less common compounds such as ellagitannin,
naphthalene, phenolic acid, purine, and xanthone etc. The aim of this research project was to carry
out the characterization of the functional molecules present in the methanolic extract of leaves of
and in Tridax procumben investigate their biological activities.

Introduction :

Plants are one of five big groups (kingdoms) of living things. They are autotrophic eukaryotes,
which means they have complex cells. Trees, herbs, bushes, grasses, vines, ferns, mosses, and
green algae are included in plant. The scientific study of plants, known as botany, has identified
about 350,000 extant (living) species of plants. Plants help maintain gaseous balance in the air also
prevent soil erosion. They help to reduce heat and prevent drying up of moisture. Thus they are
environmental savvy. Plants like blue green algae and bacteria are also extensively used to fix
nitrogen in the soil for agriculture (Ranga et al, 2015). A large group of plants used in medicine or
veterinary practice for therapeutic or prophylactic purposes. Medicinal plants have been identified
and used throughout human history. Plants have the ability to synthesize a wide variety of
chemical compounds that are used to perform important biological functions, and to defend
against attack from predators such as insects,fungi and herbivorous mammals. At least 12,000 such
compounds have been isolated so far; a number estimated to be less than 10% of the total(Tap and
sell et al, 2006). Although, there are no apparent morphological characteristics in the medicinal
plants growing with them, yet they possess some special qualities or virtues that make them
medicinally important. It has now been established that the plants which naturally synthesis and
accumulate some secondary metabolites, like alkaloids, glycosides, tannins, volatiles oils and
contain minerals and vitamins, possess medicinal properties termed as medicinal plants (Samy et
al, 2008).
Plants are the natural reservoir of many antimicrobial, anticancer agents, analgesics, antidiarrheal
as well as various therapeutic activities. Bangladeshi people have traditional medical practice as an
integral part of their culture. A lot of medicinal plants are available for the treatment of various
diseases. However, scientific studies have been conducted on only a relatively few medicinal
plants, and then only to a superficial extent (Faysal, 2008). The use of plants as medicines predates
written human history. Ethnobotany (the study of traditional human uses of plants) is recognized
as an effective way to discover future medicines. In 2001, researchers identified 122 compounds
used in modern medicine which were derived from "ethnomedical" plant sources; 80% of these
have had an ethnomedical use identical or related to the current use of the active elements of the
plant (Fabricant and Farnsworth, 2001) There are hundreds of drugs and biologically active
compounds developed from the traditional medicinal plants, a few of which are mentioned here;
the antispasmodic agent vasicin isolated from Justicia adhatoda, anticancer agents such as
vincristine, vinblastine and D-tubocurarine isolated from Catharanthus roseus (Gurib-Fakim, 2006),
antibacterial agents isolated from Diospyros melanoxylon (Mallavadhani et al, 1998), antimalarial
agent isolated from Sida acuta (Karou et al., 2006), steroid and lancamarone with cardiotonic
properties, lantamine with antipyretic and antispasmodic properties from Lantana camara
(Ghisalberti, 2000), antimicrobial agents isolated from Acorus calamus (Chowdhury et al, 1993),
antiviral, antibacterial and antiinflammatory agents isolated from Urtica dioica (Harborne and
Buxter, 1993), anticancer agents isolated from Aloe vera, Allium sativum, Andrographis paniculata,
Curcuma longa, Moringa oleifera, Phyllanthus amarus, Piper longum, Semecarpus anacardium,
Tinospora cordifolia and Withanica somnifera (Balachandran and Govindarajan 2005), promising
and potent antimalarial drug artemisinin isolated from Artemesia annua (Dhingra et al, 2000).

1.1 PHTYOCHEMICALS :

Phytochemistry is the study of phytochemicals produced in plants, describing the isolation,

• The nature of the plant material

• Its origin • Degree of processing

• Moisture content

• Particle size (Tiwari and Kumer, 2011).

All plants produce chemical compounds as part of their normal metabolic activities. These
phytochemicals are divided into (1) primary metabolites such as sugars and fats, which are found in
all plants; and (2) secondary metabolites—compounds which are found in a smaller range of plants,
serving a more specific function. For example, some secondary metabolites are toxins used to deter
predationand others are pheromones used to attract insects for pollination (Meskin and Mark,
2002).Carbon dioxide gas deals with the photosynthesis process in plants in the presence of light
energy. Photosynthesis and pentose pathway together pools the phosphate group present in the
sugar molecules of plants which leads to glycolysis process and which is accounted for producing
many of phytochemicals of plants, such as, shikimic acid, proteins, aliphatic and aromatic acids,
mevalonic acids, fatty acids, flavanoids, terpinoids, steroids etc. There are lots of medicinal plants
which contain a number of phytochemicals and those phytochemicals are used medicine purpose to
treat various kinds of diseases. In the following table a list is shown of phytochemicals having
medicinal values (Tiwari and Kumer, 2011).

1.2 NECESSITY OF STUDYING OF MEDICINAL PLANTS

The term of medicinal plants include a various types of plants used in herbalism and some of these
plants have a medicinal activities. These medicinal plants consider as a rich resources of ingredients
which can be used in drug development and synthesis. Besides that these plants play a critical role in
the development of human cultures around the whole world.

• Medicinal plants have played an essential role in the development of human culture, for example
religions and different ceremonies. (E.g. Dutura has long been associated with the worship of Shiva,
the Indian god).

• Many of the modern medicines are produced indirectly from medicinal plants, for example aspirin.

• Plants are directly used as medicines by a majority of cultures around the world, for example
Chinese medicine and Indian medicine.

• Many food crops have medicinal effects, for example garlic. Medicinal plants are resources of new
drugs. It is estimated there are more than 250, 000 flower plant species (Andrew, 2004)

• Studying medicinal plants helps to understand plant toxicity and protect human and animals from
natural poisons.

• Cultivation and preservation of medicinal plants protect biological diversity, for example metabolic
engineering of plants.

• Plant resources (E.g. Angiosperm, Gymnosperm, Seedless vascular plants, Bryophytes) for new
medicine.

• The medicinal plants find application in pharmaceutical, cosmetic, agricultural and food industry
( Andrew, 2004)

• With onset of scientific research in herbals, it is becoming clearer that the medicinal herbs have a
potential in todays synthetic era, as numbers of medicines are becoming resistant. According to one
estimate only 20% of the plant flora has been studied and 60%of synthetic medicines owe their
origin to plants. Ancient knowledge coupled with scientific principles can come to the forefront and
provide us with powerful remedies to eradicate the diseases.

• To identify alternative and complementary medicine.

• To reduce the toxicity of drug therapy especially toxicity reduction of synthetic and semi synthetic
drugs. • To find the lead compound diversification to treat various diseases (Andrew, 2004)
1.2.1 HISTORY OF TRADITIONAL HERBAL MEDICINE IN BANGLADESH

“Traditional medicine refers to health practices, approaches, knowledge and beliefs incorporating.
Plant, animal and mineral based medicines, spiritual, therapies, manual techniques and exercises,
applied singularly or in combination to treat, diagnose and prevent illnesses or maintain well‐
[Link] definition, ‘traditional’ use of herbal medicines implies substantial historical use, and this is
certainly true for many products that are available as ‘traditional herbal medicines’ .In many
developing countries, a large (Allison et al, 2001).

The earliest mention of traditional medicine is found in Rigveda, the oldest repository of knowledge
in this subcontinent. Later Ayurveda, developed from the Vedic concept of life,became the important
source of all systems of medical sciences. In course of time it became a part of culture and heritage
of the people of the Indian subcontinent . Traditional medicine involves the use of both material and
non-material components. The material components invariably comprise parts or organs of plants
and their products. They also consist of animal organs, minerals and other natural substances. The
non-material components, which constitute important items of religious and spiritual medicines,
include torture, charms, magic, incantations, religious verses, amulets and rituals like sacrifices,
appeasement of evil spirits, etc. Treatments in traditional medicine are carried out by internal and
external application of medicaments, physical manipulation of various parts of the body, performing
rituals, psychological treatment, and also by minor surgery. Ayurvedic medicinal preparations consist
mainly of plant materials in the form of powders, semi-solid preparations, decoctions, elixirs and
distillates. Many of them also contain inorganic chemical substances, minerals and animal products.
Alcoholic extracts and alcoholic solutions of the ingredients, tinctures and elixirs are also frequently
used in Ayurvedic medicine .Whole plants or their powders or pastes or products and their extracts,
infusions, decoctions and distillates constitute the major constituents of Unani medicine. Minerals,
inorganic chemicals and animal products are also frequently used in preparing these
medicines(Samy,Pushparaj & Gopalakrishnakone, 2008).

The desire to capture the wisdom of traditional healing systems has led to a resurgence of interest in
herbal medicines (Tyler, 2000), particularly in Europe and North America, where herbal products
have been incorporated into so-called ‘alternative’, ‘complementary’, ‘holistic’ or ‘integrative’ medical
system. The practice of Traditional medicine is deeply rooted in the cultural heritage of Bangladesh
and constitutes an integral part of the culture of the people of this country. Different forms of
Traditional medicines have been used in this country as an essential means of treatment of diseases
and management of various health problems from time immemorial. The practice of traditional
medicine in this country has flourished tremendously in the recent years along with that of modern
medicine. As a result, even at this age of highly advanced allopathic medicine, a large majority (75-
80%) of the population of this country, particularly in the rural and semiurban areas, still prefer to
use traditional medicine in the treatment of most of their diseases even though modern medical
facilities may be available in the neighbourhood. However, the concept, practice, type and method of
application of traditional medicine vary widely among the different ethnic groups living in different
parts of the country according to their culture, living standard, economic status, religious belief and
level of education. Thus traditional medicine practice in Bangladesh includes both the most primitive
forms of folk medicine (based on cultural habits, superstitions, religious customs and spiritualism) as
well as the highly modernised Unani and Ayurvedic systems (based on scientific knowledge and
modern pharmaceutical methods and technology). These various aspects of Traditional medicine
practice in Bangladesh, their current official status (acceptability, recognition, etc.) in the country as a
means of treatment, and their contribution to, and impact on, the overall health management
programmes of the country are described and discussed in this paper supported by documentary
evidences and scientific data (Ghani and Abdul, 1998). Today the pharmacologically active
ingredients of many Ayurvedic medicines are being identified and their usefulness in drug therapy
being determined. As only a certain percentage of plants are used in traditional medicines, it is
roughly estimated that of the discovered 17,000 species, nearly 3,000 species are used in medicinal
field.

1.3 Plant metabolite

Metabolites are compounds synthesized by plants for both essential functions, such as growth and
development (primary metabolites), and specific functions, such as pollinator attraction or defense
against herbivory (secondary metabolites). Metabolites are organic compounds synthesized by
organisms using enzyme-mediated chemical reactions called metabolic pathways. Primary
metabolites have functions that are essential to growth and development and are therefore present
in all [Link] contrast, secondary metabolites are variously distributed in the plant kingdom, and
their functions are specific to the plants in which they are found. Secondary metabolites are often
colored, fragrant, or flavorful compounds and they typically mediate the interaction of plants with
other organisms. Such interactions include those of plant-pollinator, plant-pathogen, and plant-
herbivore).

1.3.1 Primary metabolite

A plant produces primary metabolites that are involved in growth and metabolism. Primary
metabolites comprise many different types of organic compounds, including, but not limited
to, carbohydrates, lipids, proteins, and nucleic acids. They are found universally in the plant
kingdom because they are the components or products of fundamental metabolic pathways
or cycles such as glycolysis, the Krebs cycle, and the Calvin cycle. Because of the importance
of these and other primary pathways in enabling a plant to synthesize, assimilate, and
degrade organic compounds, primary metabolites are essential. Examples of primary
metabolites include energy rich fuel molecules, such as sucrose and starch, structural
components such as cellulose, informational molecules such as DNA (deoxyribonucleic acid)
and RNA (ribonucleic acid), and pigments, such as chlorophyll. In addition to having
fundamental roles in plant growth and development, some primary metabolites are
precursors (starting materials) for the synthesis of secondary metabolites.

1.3.2 Secondary metabolite

Secondary metabolites are those metabolites which are often produced in a phase of
subsequent to growth, have no function in growth (although they may have survival
function), are produced by certain restricted taxonomic groups of microorganisms, have
unusual chemicals structures, and are often formed as mixtures of closely related members
of a chemical family. The simplest definition of secondary products is that they are not
generally included in standard metabolic charts. A metabolic intermediate or product, found
as a differentiation product in restricted taxonomic groups, not essential to growth and the
life of the producing organism, and biosynthesis from one or more general metabolites by a
wider variety of pathways than is available in general metabolism. Secondary metabolites are
not essential for growth and tend to be strain specific. They have a wide range of chemical
structures and biological activities. They are derived by unique biosynthetic pathways from
primary metabolites and intermediates (David and Wang, 2014) Of the estimated 400,000 –
500,000 plant species around the globe, only a small percentage has been investigated
phytochemically and the fraction subjected to biological or pharmacological screening is
even lower. The ability to synthesize secondary metabolites has been selected through the
course of evolution in different plant lineage when such compounds address specific needs •
Floral scent volatiles and pigments have evolved to attract insect pollinators and thus
enhance fertilization.
• To synthesize toxic chemical has evolved to ward off pathogens and herbivores or to
suppress the growth of neighboring plants.
• Chemicals found in fruits prevent spoilage and act as signals (in the form of color, aroma,
and flavor) of the presence of potential rewards (sugars, vitamins and flavor) for animals that
eat the fruit and thereby help to disperse the seeds.
• Other chemicals serve cellular functions that are unique to the particular plant in which
they occur (e.g. resistance to salt or drought)( (David and Wang, 2014)