Introduction

Phytochemistry, the branch of chemistry deals with the chemical process

associated with plant life and the chemical compounds produced by plants.
Phytochemistry deals with the analysis of plant chemicals called natural
products, and with changes occurring in such chemicals due to alterations in
environmental conditions. Ecology nowadays includes an increased amount of
chemistry because communication between a plant and its environment
depends to a large extent on secondary metabolites (Zobel and Clarke, 1999).

Today we can witness explosive growth of herbal drug industry.

Standardized herbal extracts and phytochemicals are in high demand for applied
research as well as commercial use. The phytochemistry course taught in
pharmacy schools deals with process of isolation, purification and pre-clinical
studies of herbal extracts and phytochemical (Singh, 2007).

In the present era, a lot of approaches are adopted for the determination
of new biologically active compounds in the medicinal plants for the synthesis
of safe and effective drugs for the welfare of mankind. About 80% of
individuals from developed countries use traditional medicines from medicinal
plants. Therefore, these plants should be investigated for the better
understanding of the properties, safety and efficiency of these medicinal plants
derived drugs. According to the World Health Organization (WHO), medicinal
plants are the best source for extraction of a variety of drugs. About 61% of new
drugs developed (1981 to 2002) were based on natural products and they proved
very effective against certain diseases, especially in infectious diseases and
cancer. (Ajaib et al., 2015). Plants have been a source of medicine for thousands
of years, and phytochemicals continue to play an essential role in medicine
(Aggarwal, 2003).

1
 Medicinal plants are in greater demand due to their increased popularity;
thus, it is being suggested by a large number of conservation groups, that wild
medicinal plants should be brought into cultivation. Numerous medicinal
plants, as well as, their purified components have shown beneficial therapeutic
potentials. Various herbs and other plant species are reported to show
antioxidant activity. Majority of the antioxidant potential is due to the presence
of flavones, flavonoids, isoflavones, anthocyanin, lignans, coumarin, catechins
and is catechins. Plants are rich in a wide variety of secondary metabolites for
example tannins, terpenoids, alkaloids, flavonoids, glycosides, etc. (Aqil et al.,
2004).

Phytochemicals:

Phytochemicals literally mean ―plant chemicals‖ Scientists have

identified thousands of different phytochemicals found in vegetables, fruits,
whole grains, nuts and seeds. Eating lots of plant foods rich in phytochemicals
may help to prevent at least one in every five cases of cancer, as well as other
serious aliment such as heart diseases.

All over the world, medicinal plants serves as a potent weapon in the
diagnosis of diseases, making of traditional and modern medicines and in the
treatment of various diseases which are caused by microbial agents such as
bacteria, viruses and fungi. These diseases if not treated appropriately using
suitable herbs or drugs could degenerate into severe medical health
complications. The therapeutic value of plants used in traditional medicine is
derived from the presence of phytochemical principles, such as alkaloids,
tannins and phenols which are found in parts of plants.
Phytochemicals are plant derived chemicals which are beneficial to human
health and disease prevention. The term is generally used to refer to those
chemicals that may have biological significance, for example antioxidants, but

2
are not established as essential nutrients which when in excess could be
detrimental. It is pertinent to state that in as much as they help to strengthen the
body defence mechanism serving mostly as antioxidants; moderate
consumption is highly recommended so that they will not serve as anti-nutrient
to the body. Scientists estimate that there are thousands of known
phytochemicals having the potentials to affect disease such as cancer, stroke
and metabolic syndrome and those caused by microorganisms. Potential
phytochemicals in freshly harvested plant foods are often destroyed or removed
by local and modern processing techniques. Industrially processed foods likely
contain fewer phytochemicals and may thus be less beneficial than unprocessed
foods. Absence of deficiency of phytochemicals in processed foods may
contribute to increased risk of preventable diseases. The most important of these
bioactive compounds of plants are alkaloids, flavonoids, tannins, and phenolic
compounds. This is not say that others are not important but the mentioned few
occur commonly in plants, hence the review will focus on nine phytochemicals;
flavonoids, cardiac glycosides, alkaloids, phytate, haemagglutinin, saponins,
tannins, oxalate, and phenols.

Phytochemicals are the chemicals that present naturally in plants. Now-

adays these phytochemicals become more popular due to their countless
medicinal uses. Phytochemicals play a vital role against number of diseases
such as asthma, arthritis, cancer etc. unlike pharmaceutical chemicals these
phytochemicals do not have any side effects. Since the phytochemicals cure
diseases without causing any harm to human beings these can also be
considered as ―man- friendly medicines‖.

Phytochemicals are biologically active, naturally occurring chemical

compounds found in plants, which provide health benefits for humans further
than those attributed to macronutrients and micronutrients. They protect plants
from disease and damage and contribute to the plant ‘s colour, aroma and

3
flavour. 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. 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.
Phytochemicals can be classified into 10 categories (Li, 2010) according
to their different structures and properties:
1. Water-soluble organic acids, straight-chain alcohols, aliphatic aldehydes,
and ketones;
2. Simple lactones;
3. Long-chain fatty acids and polyacetylenes;
4. Quinines (benzoquinone, anthraquinone and complex quinines);
5. Phenolics;
6. Cinnamic acid and its derivatives;
7. Coumains;
8. Flavonoids;
9. Tannins;
10.Steroids and terpenoids (sesquiterpene lactones, diterpenes,
and triterpenoids).
Terpenoids:

The terpenes, or isoprenoids, are one of the most diverse classes of

metabolites. The Dictionary of Natural Products lists over 30000, mainly of
plant origin, encompassing flavours and fragrances, antibiotics, plant and
animal hormones, membrane lipids, insect attractants and antifeedants, and
mediators of the essential electron-transport processes which are the
senergygenerating stages of respiration and photosynthesis. Terpenoids are the

4
largest and most diverse family of natural products, ranging in structure from
linear to polycyclic molecules and in size from the five-carbon hemiterpenes to
natural rubber, comprising thousands of isoprene units. All terpenoids are
synthesized through the condensation of isoprene units (C5) and are classified
by the number of five-carbon units present in the core structure. Many flavour
and aromatic molecules, such as menthol, linalool, geraniol and caryophyllene
are formed by monoterpenes (C10), with two isoprene units, and sesquiterpenes
(C15), with three isoprene units. Other bioactive compounds, such as diterpenes
(C20), triterpenes (C30) and tetraterpenes (C40) show very special properties.
(Harborne, 1984)

Phenolic Compounds:

Phenolics are characterized by having at least one aromatic ring with one or
more hydroxyl groups attached. In excess of 8000 phenolic structures have been
reported and they are widely dispersed throughout the plant kingdom.
Phenolics range from simple, low molecular-weight, single aromatic-ringed
compounds to large and complex tannins and derived polyphenols. They can be
classified based on the number and arrangement of their carbon atoms and are
commonly found conjugated to sugars and organic acids. Phenolics can be
classified into two groups: the flavonoids and the non-flavonoids.( Harborne,
1984)

Flavonoids:

Flavonoids are poly phenolic compounds comprising fifteen carbons,

with two aromatic rings connected by a three-carbon bridge. They are the most
numerous of the phenolics and are found throughout the plant kingdom. They
are present in high concentrations in the epidermis of leaves and the skin of
fruits and have important and varied roles as secondary metabolites. They are
the most numerous of the phenolics and are found throughout the plant

5
kingdom. They are present in high concentrations in the epidermis of leaves and
the skin of fruits and have important and varied roles as secondary metabolites.
In plants, flavonoids are involved in such diverse processes as UV protection,
pigmentation, stimulation of nitrogen-fixing nodules and disease resistance.
(Harborne, 1993)

Alkaloids:

True alkaloids derive from amino acid and they share a heterocyclic ring
with nitrogen. These alkaloids are highly reactive substances with biological
activity even in low doses. All true alkaloids have a bitter taste and appear as a
white solid, with the exception of nicotine which has a brown liquid. True
alkaloids form water-soluble salts. Moreover, most of them are well-defined
crystalline substances which unite with acids to form salts. True alkaloids may
occur in plants (1) in the Free State, (2) as salts and (3) as N-oxides. These
alkaloids occur in a limited number of species and families, and are those
compounds in which decarboxylated amino acids are condensed with a
nonnitrogenous structural moiety. The primary precursors of true alkaloids are
such amino acids as L-ornithine, L-lysine, L-phenylalanine/ L-tyrosine, L-
tryptophan and L-histidine. Examples of true alkaloids include such
biologically active alkaloids as cocaine, quinine, dopamine, morphine and
usambarensine.
(Pelletier, 1983).
The alkaloids are a large and structurally diverse group of compounds;
some of them are not entirely distinguishable from amines (e.g., ephedrine).
The names of these molecules tend to end in the suffixes –ine or –in. Many are
derived from amino acids, but others result from modification of various classes
of molecules including polyphenols, terpenes, or steroids. With a few notable
exceptions, alkaloids are most soluble in hydroethanolic media and they
generally occur as salts (e.g., chlorides or sulfates) and or as N-oxides in the

6
plants. Most of them have a heterocyclic nitrogenous ring or ring system and a
basic (alkaline) character. They are often dramatic compounds, having
noticeable physiological effects – whether therapeutic or detrimental. Among
the alkaloids we find potent medicinal molecules as well as toxic or even
potentially fatal ones.

Tannins

Tannin is the name derived from French ―Tanin‖ (tanning substance)

and used for a range of natural polyphenols. The dark brown or black substance
has been tested and found to be tannin. Tannins are naturally occurring plant
polyphenols and are found in bark, wood, leaves, fruit, roots and seeds. Tannins
bond readily with proteins and have been used medicinally, in the food industry
and in the making of leather. Most plants, including edible fruits and nuts,
naturally contain tannins. For example, tannins are found in grapes, blueberries,
chocolate, red wine, coffee and tea. They produce a bitter or astringent taste or
reaction, particularly in unripe fruit. As the fruit ripens, its tannin content
diminishes. With regard to the constituents of wood, tannins are considered to
be an extractive since they can be removed from the wood with water or other
solvents. Tannins are typically found in higher concentrations in bark and
leaves compared to the wood. Tannins vary by plant part, species, growth
conditions and other factors. (Haslam, 1989)
The tannins are the phenolic compounds that precipitate proteins. They
are composed by a very diverse group of oligomers and polymers. They can
form the complex with proteins, starch, cellulose and minerals. They are
synthesized via shikimic acid pathway, also known as the phenylpropanoid
pathway. The same pathway leads to the formation of other phenolics such as
isoflavones, coumarins, lignins and aromatic amino acids. Tannins are water
soluble compounds with exception of some high molecular weight structures.
They are usually subdivided in two groups: HT (hydrolysable tannins) that

7
includes gallotannins, elligatannins, complex tannins, and PA, also known as
condensed tannins. The tannins also constitute the active principles of
plantbased medicines. According the literature, the tannins containing plants
are used as astringents against diarrhea, adiuretic against stomach and duodenal
tumors and anti-inflammatory. (Miller, 1999).

Antioxidants:

Antioxidants are an inhibitor of the process of oxidation, even at

relatively small concentration and thus have diverse physiological role in the
body. Antioxidant constituents of the plant material act as the radical
scavengers, and helps in converting the radicals to less reactive species. A
variety of free radical scavenging antioxidants is found in dietary sources like
fruits, vegetables and tea, etc. (Hall, 2001). Oxygen is absolutely essential for
the life of aerobic organism but it may become toxic if supplied at higher
concentrations. Di oxygen in its ground state is relatively unreactive; its partial
reduction gives rise to active oxygen species (AOS) such as single oxygen,
super oxide radical anion, hydrogen peroxide etc. This is partly due to the
oxidative stress that is basically the adverse effect of oxidant on physiological
function. Free oxygen radicals plays cardinal role in the etiology of several
diseases like arthritis, cancer, atherosclerosis etc. The oxidative damage to
DNA may play vital role in aging (Satyavati et al., 1976) and the presence of
intracellular oxygen also can be responsible to initiate a chain of inadvertent
reaction at the cellular level and these reaction cause damage to critical cell
biomolecules. These radicals are highly toxic and thus generate oxidative stress
in plants. Plants and other organism have in built wide range of mechanism to
combat with these Free Radical problems. Free radicals are an atom or molecule
that bears an unpaired electron and is extremely reactive, capable of engaging
in rapid change reaction that destabilize other molecules and generate many
more free radicals. In plants and animals these free radicals are deactivated by

8
antioxidants. These antioxidants act as an inhibitor of the process of oxidation,
even at relatively small concentration and thus have diverse physiological role
in the body. Antioxidant constituents of plat materials act as radical scavengers,
and convert the radicals to less reactive species. (Devi and Ganasoundari,
1999). Plants have developed an array of defence strategies (antioxidant
system) to cope up with oxidative stress. The antioxidative system includes both
enzymatic and non-enzymatic systems. The non-enzymatic system includes
ascorbic acid (vitamin C); ά-tocopherol, cartenes etc. and enzymic system
include superoxide dismutase (SOD), catalase (CAT), peroxidase (POX),
ascorbate peroxidase (APX), glutathione reductase (GR) and polyphenol
oxidase (PPO) etc. The function of this antioxidant system is to scavenge the
toxic radicals produced during oxidative stress and thus help the plants to
survive through such conditions. Spices and herbs in food as medicine is a
current hot trend that is capturing everyone ‘s imagination with images of a new
magic bullet or fountain of youth. The intake of antioxidant compounds present
in food is an important health-protecting factor. Natural antioxidants present in
foods and other biological materials have attracted considerable interest
because of their presumed safety and potential nutritional and therapeutic
effects. Because extensive and expensive testing of food additives is required
to meet safety standards, synthetic antioxidants have generally been eliminated
from many food applications. The increasing interest in the search for natural
replacements for synthetic antioxidants has led to the antioxidant evaluation of
a number of plant sources. (Satyavati et al., 1976)
Most Commonly Known Antioxidants & their food sources:
• Carotenoids: (A form of vitamin A) the substance that gives fruits and
vegetables their deep rich colours. May be effective allies against prostate
cancer. Apricots, peaches, broccoli, pumpkin, cantaloupes, carrots, spinach
and sweet potatoes

9
• Vitamin C: Enhances the immune response and protects against infection.
Citrus fruits like oranges and lime etc, green peppers, broccoli, green leafy
vegetables, strawberries and tomatoes. Vitamin E: May help prevent the
oxidation of LDL or ―bad‖ cholesterol which contributes to plaque buildup
in the arteries. Nuts & seeds, whole grains, green leafy vegetables, vegetable
oil and liver oil
• Selenium: Fish & shellfish, red meat, grains, eggs, chicken and garlic some
common phytochemicals & their food sources:
• Flavonoids / polyphenols: soy, red wine, purple grapes or Concord grapes,
pomegranate, cranberries, tea,
• Lycopene: Tomato and tomato products, pink grape fruit, watermelon
• Lutein: dark green vegetables such as kale, broccoli, kiwi, brussels sprout and
spinach
• Lignan: flax seed, oatmeal, barley, rye
Antioxidant enzymes made by the body: superoxide dismutase (SOD),
catalase, glutathione peroxidase (American Dietetic Association, 2010).
Alternanthera sessilis
Alternanthera sessilis commonly known as ‘sessile joy weed’ or
‘dwarf copperleaf’ is found throughout the hotter parts of India up to an
altitude of 1200 m2. In Assam, the plant has been traditionally used in the
treatment of jaundice along with other ailments. The study focuses
primarily on the evaluation of hepatoprotective activity of the plant with
special references to its putative protective role in carbon tetrachloride
induced liver injury on Wister albino rat. The in vivo hepatoprotective
activity of the methanolic extract at the dose of 250 mg/kg body weight
was highly effective in controlling SGPT, SGOT, ALP, serum cholesterol and
serum bilirubin level as compared to silymarin. The said extract also
significantly lowered the lipid profile caused by CCl 4. The activity shown

10
by methanolic extract of the whole plant of A. sessilis is of considered
importance and thus justified its use in controlling hepatic diseases in
traditional treatment system.
In the treatment of human diseases and ailments use of natural
products from plant, animal and minerals have always been of first
preference due to toxicity and side effects of allopathic medicines. Drugs
from various sources are still in the traditional medicine for treating a
number of diseases and it has been estimated that about 80% of people in
developing countries still relays on traditional medicine for their primary
health care [3]. Herbal medicines are far safer than allopathic medicines in
concern of side effects, acute and severe allergic reactions and liver or
kidney diseases. Now a days the use of herbal medicine is becoming
popular compared to allopathic medicines leading to sudden increase in
the number of herbal formulations in the market [4]. As per the World
Health Organization (WHO) list of 21,000 medicinal plants of the world,
2500 species are found in India. Among these plants around 150 species
are used commercially in large scale making India one of the largest
producers of medicinal herbs in the world Liver disease is a worldwide
problem irrespective of age, sex, region or race. The allopathic system of
medicines used in the treatment of liver diseases is sometimes not enough
and can have serious adverse effects. As compared to allopathic drugs,
herbal drugs are more widely used for the treatment of hepatic disorder
because of their better compatibility and acceptability with minimal/no
side effects. In India, more than 40 polyherbal formulations are available
in the market claiming to have hepatoprotective activity. The scrutiny of
literature indicates that 160 phytoconstituents from 101 plants have
hepatoprotective activity The plant is commonly known as ‘sessile
joyweed’ or ‘dwarf copperleaf’. This is a perennial herb often found in and

11
near ponds, canals and reservoirs. In Assam, this plant is commonly known
as ‘Matikanduri’. Alternanthera sessilis possibly originates from tropical
America but is now widespread in the tropics and subtropics of the world
including the whole of tropical Africa. It is a weed in tropical lands and can
grow in all soil types. Shoots and leaves are often eaten as vegetable.
Depending on location, many other common names exist: ‘Daun tolod’ in
Indonesia, ‘Keremak’ in Malaysia, ‘Brede chevrette’ in French, ‘Lian zi cao’
in China, ‘bunga-bunga’ in Philippines, ‘Angelica’ in Spain, ‘Lupo’ in Ilonggo
etc. It is used as a local medicine often in mixtures with other medicinal
plants, to treat hepatitis, tight chest, bronchitis, asthma and other lung
troubles. The leaves and shoots boiled and drunk as antihypertensive
remedy

Alternanthera sessilis

12
Aim and Objective
Medicinal plants are a rich source of bioactive phytochemicals or bio
nutrients. Studies carried out during the past 2–3 decades have shown that these
phytochemicals have an important role in preventing chronic diseases like
cancer, diabetes and coronary heart disease. The major classes of
phytochemicals with disease-preventing functions are dietary fibre,
antioxidants, anticancer, detoxifying agents, immunity-potentiating agents and
neuro pharmacological agents. Each class of these functional agents consists of
a wide range of chemicals with differing potency. Some of these
phytochemicals have more than one function. There is, however, much scope
for further systematic research in screening Indian medicinal plants for these
phytochemicals and assessing their potential in protecting against different
types of diseases. Natural antioxidants, particularly in fruits and vegetables
have gained increasing interest among consumers as antidotes to aging and
associated chronic diseases. Flavonoids are phenolic substances isolated from
a wide range of vascular plants, and more than 8150 different types have been
reported. They act in plants as antioxidants, antimicrobials, photoreceptors,
visual attractors, feeding repellents, and for light screening. The main goal of
this study is to evaluate the phytochemicals constituents nutritional profiling
and biological efficiency of the methanolic extracts of Alternanthera sessilis.
The main objectives are

1. Preparation of alternanthera sessilis is extract using soaking extraction

method.
2. Quantitative estimation of protein, carbohydrate, amino acid, tannin,
polyphenol, alkaloid, flavonoid, glycosides, and terpene
3. Analysis of nutrient profiling like moisture, ash, vitamin E, vitamin C,
sodium and potassium.
4. Estimation of biological activities like antioxidant, antibacterial and
antidiabetic activity.

13
Review of Literature

Alternanthera sessilis commonly known as ‘sessile joy weed’ or ‘dwarf

copperleaf’ is found throughout the hotter parts of India up to an altitude of
1200 m2. In Assam, the plant has been traditionally used in the treatment of
jaundice along with other ailments. The study focuses primarily on the
evaluation of hepatoprotective activity of the plant with special references to
its putative protective role in carbon tetrachloride induced liver injury on
Wister albino rat. The in vivo hepatoprotective activity of the methanolic
extract at the dose of 250 mg/kg body weight was highly effective in controlling
SGPT, SGOT, ALP, serum cholesterol and serum bilirubin level as compared
to silymarin. The said extract also significantly lowered the lipid profile caused
by CCl4. The activity shown by methanolic extract of the whole plant of A.
sessilis is of considered importance and thus justified its use in controlling
hepatic diseases in traditional treatment system.
In the treatment of human diseases and ailments use of natural products from
plant, animal and minerals have always been of first preference due to toxicity
and side effects of allopathic medicines. Drugs from various sources are still in
the traditional medicine for treating a number of diseases [1, 2] and it has been
estimated that about 80% of people in developing countries still relays on
traditional medicine for their primary health care [3]. Herbal medicines are far
safer than allopathic medicines in concern of side effects, acute and severe
allergic reactions and liver or kidney diseases. Now a days the use of herbal
medicine is becoming popular compared to allopathic medicines leading to
sudden increase in the number of herbal formulations in the market [4]. As per
the World Health Organization (WHO) list of 21,000 medicinal plants of the
world, 2500 species are found in India. Among these plants around 150 species
are used commercially in large scale making India one of the largest producers
of medicinal herbs in the world.

14
 Liver disease is a worldwide problem irrespective of age, sex, region or race.
The allopathic system of medici nes used in the treatment of liver diseases is
sometimes not enough and can have serious adverse effects. As compared to
allopathic drugs, herbal drugs are more widely used for the treatment of hepatic
disorder because of their better compatibility and acceptability with minimal/no
side effects. In India, more than 40 polyherbal formulations are available in the
market claiming to have hepatoprotective activity. The scrutiny of literature
indicates that 160 phytoconstituents from 101 plants have hepatoprotective
activity.
The plant is commonly known as ‘sessile joyweed’ or ‘dwarf copperleaf’.
This is a perennial herb often found in and near ponds, canals and reservoirs.
In Assam, this plant is commonly known as ‘Matikanduri’. Alternanthera ses
silis possibly originates from tropical America but is now widespread in the
tropics and subtropics of the world including the whole of tropical Africa. It is
a weed in tropical lands and can grow in all soil types. Shoots and leaves are
often eaten as vegetable. Depending on location, many other common names
exist: ‘Dauntolod’ in Indonesia, ‘Keremak’ in Malaysia, ‘Brede chevrette’ in
French, ‘Lian zi cao’ in China, ‘bunga-bunga’ in Philip pines, ‘Angelica’ in
Spain, ‘Lupo’ in Ilonggo etc.
A decoction of A. sessilis is recommended as an herbal remedy to treat
wounds, flatulence, nausea, vomiting, cough, bronchitis, diarrhea, dysentery
and diabetes. Its root can relieve inflamed wounds [9]. It is used as a local
medicine often in mixtures with other medicinal plants, to treat hepatitis, tight
chest, bronchitis, asthma and other lung troubles. The leaves and shoots boiled
and drunk as antihypertensive remedy
The main objective of this study is to focus on the evaluation of
hepatoprotective activity of the whole plant of A. sessilis with special
references to its putative protective role in carbon tetrachloride induced liver
injury on Wister albino rat.
15
 The whole plant of A. sessilis were collected from Siva sagar district, Assam
in the month of July, 2013. The plant is identified and authenticated
taxonomically by Dr. A A Mao at Botanical Survey of India, Shillong
The freshly collected whole plant of A. sessilis were air dried for 25–30 days.
Then it was grinded to make fine powder and stored in a desiccator to protect
the content from moisture
Male Wistar rats weighing about 100–200 gm was used for the study. All
studies were performed in accordance with the guidance for the care and use of
laboratory animals, as adopted and promulgated by the Institutional Animal
Care Committee, CPCSEA, India (Approval No. IACE/DU/64 Dated
24/9/2013).
The animal house was well ventilated and animals were kept under conditions
of 24 ± 20 C and RH 44–56% and 12 height: 12 h dark cycles each day for 1
week before and during the experiments. Animals described as fasted were
deprived of food for 16 h but allowed free access to water. Preparation of Plant
Extract of Alternanthera sessilis the crude drug was extracted with methanol
using hot continuous process and concentrated to dryness in rotary vacuum
evaporator.
Acute toxicity study was performed for methanolic extract according to
OECD guidelines. Female albino rats were kept fasting for overnight providing
only water before administering the extract orally at dose level of 250, 1000
and 2500 mg/kg body weight [11]. The animals were observed continuously
for 2 h and then occasionally for further 4 h and finally any mortality behaviour
of the animals and any other toxic symptoms also observed for 72 h. The
animals were kept under observation up to 14 days.
The animals are divided randomly into six groups of six rats as mentioned in
Table 1. The hepatoprotective activity of the plant extracts was tested using
three days CCl4 model. The rats of group (I) received double doses of 5% gum
acacia mucilage at 24 h intervals. The rats of group (II) received two doses of
16
vehicle at 12 h intervals and a single dose of Carbon tetrachloride diluted in
liquid paraffin. The rats of group (III) received double doses of Silymarin
followed by CCl4 after 30 min of last dose received. While the rats of group
(IV), (V) and (VI) received double doses of methanolic extract of A. sessilis at
0 and 24 h followed by CCl4 after 30 min of last dose received.
Liver tissues were collected from each experimental rat, washed and
homogenized in 1.15% w/v solution of KCl and centrifuged at 1200 rpm at 4 C
for 10 min. The same procedure was repeated twice with increasing centrifugal
force at 10,000 rpm and at 20,000 at 4 C for 10 and 60 min respectively. The
microsomal fraction was isolated and stored at-20 C.
Liver tissues were collected, washed, weighed and stored at-20 C. Tissue
fragments (200 mg) were thawed and homogenized on ice in 1 ml of
homogenizing buffer (250 Mm sucrose, 20 mg Tris–HCl, 1 mg dithiothreitol,
pH 7.4), using glass-Teflon homogenizer. The homogenates were centrifuged
at 75,000 rpm at 40 C for 2 h and stored at-20 C.
Each 0.1 ml of tissue sample was kept on ice bath with 2.4 ml of EDTA
solution. After 10 min 2 ml of distilled water and 0.5 ml of 50% TCA were
added and centrifuged at 3000 rpm for 15-min at 4 C. To 1.0 ml of supernatant
2.0 ml of Tris buffer and 0.05 ml of DTNB solution (Ell man’s reagent) was
added and vortexed thoroughly. OD was read (within 2–3 min after the addition
of DTNB) at 412 nm in Shimadzu UV 160. Appropriate standards were run
simultaneously.
Catalase activity was measured based on the ability of the enzyme to break
down H2O2.10ll samples were taken in tube containing 3.0 ml of H2O2 in
phosphate buffer pH 4.2. Time required for 0.05 optical density changes was
observed at 240 nm against a blank containing the enzyme source in H2O2 free
phosphate buffer (0.16 ml H2O2 is 30% w/v was diluted to 100 ml of phosphate
buffer pH 4.2). Reading was taken at every 3 s interval. Catalase activity is
expressed as follows. Isolated liver samples were preserved at 2–3 C with
17
Bouin’s solution. For the histopathological examination samples were prepared
using Rapid process. The samples were cut into thin ribbon using rotary
microtome and strips placed in glass slides. These slides were then incubated
overnight at a temperature of 37 C. After treatment with xylene and alcohol the
slides were finally washed with water for 20 min. The slides were stained with
haematoxylin and eosin, after which it was fixed and observed under
microscope (Leica Photomicroscope DM1000).
Liver enzymes are liberated into blood whenever liver cells are damaged and
enzyme activity in the plasma is increased. The whole plant extracts of A.
sessilis at doses of 50, 200 and 250 mg/kg body weight showed varied effect
on biochemical parameters. Increased serum concentration of enzymes such as
SGPT, SGOT and ALP were observed in CCl4 induced rats indicating an
altered liver function and/or liver mitochondrial injury in comparison to normal
control rats receiving only the vehicle. Liver injury con- tributes to increased
serum level of transaminase enzymes due to easy availability of amino acids.
On treatment with test extract of varied doses it was found that the dose 250
mg/kg body weight significantly reversed the elevated marker enzymes i.e.
SGOT, SGPT, ALP indicating its hepatoprotective effect.
The lipid profile of CCl4 induced rats showed important indicator for
metabolic disturbance including hepatic dis-ease. The higher concentration of
serum cholesterol, serum bilirubin may be attributed to liver disease such as
hep-atitis, cirrhosis, obstructive jaundice etc. The treatment with methanolic
extract of A. sessilis, particularly with that of the higher dose showed marked
improvement in diminishing the higher levels of these markers with respect to
that of hepatic control group.
The oxidative stress induced by CCl4 may lead to imbalance of in vivo
antioxidant system which was also evaluated by this study. The oxidative stress
in the hepatic animals measured by free radical (CH3+) measurement is
difficult due to their very short half-life and their low concentration. Moreover,
18
oxidative stress markers are lipid peroxidation and total antioxidant status.
Elevated lipid-peroxidation was also studied in CCl4 induced hepatic animal.
The 250 mg/kg body weight dose of methanolic extracts of A. sessilis and
silymarin treated groups significantly reduced the oxidation of lipids in liver.
Total antioxidant status was measured by amount of enzymatic GSH. GSH
plays the important role in balance the oxidative stress. In hepatic control
groups, the decreased GSH may be due to reduction in GSH synthesis or
degradation of GSH by oxidation stress in hepatic animal. CCl4 induced group
produce less glutathione reductase activity (20.141) whereas the standard drug
showed the activity of 36.093. The A. sessilis treated groups; particularly the
higher dose groups have showed prominent activity in the increment of
reductase power.
The pharmacological parameters of A. sessilis have been evaluated with the
objective to validate the traditional hepatoprotective uses. In vivo
hepatoprotective activities were carried out and measurement of different
biochemical parameters were performed after the 36 h of induction of CCl4
followed by sacrifice of animals by cervical dislocation. From the results
obtained, we can conclude that the dose of 250 mg/kg body weight was more
effective than the dose of 50 mg/kg body weight. The methanolic extract of the
whole plant of A. sessilis also significantly normalized the lipid profile caused
by CCl4 induction of rats. The activity shown by methanolic extract of the
whole plant of A. sessilis is of considerable importance and thus justified its
use in controlling hepatic diseases in traditional treatment system.
Further studies are required to identify the bio-active chemical constituents
present in the crude extracts of this plant and development of herbal drug
formulations for the treatment of hepatic diseases.

19
Result and Discussion
Medicinal plants are a rich source of bioactive phytochemicals or bio
nutrients. Studies carried out during the past 2– 3 decades have shown that these
phytochemicals have an important role in preventing chronic diseases like
cancer, diabetes and coronary heart disease. The major classes of
phytochemicals with disease-preventing functions are dietary fibre,
antioxidants, anticancer, detoxifying agents, immunity-potentiating agents and
neuropharmacological agents. Each class of these functional agents consists of
a wide range of chemicals with differing potency. Some of these
phytochemicals have more than one function. There is, however, much scope
for further systematic research in screening Indian medicinal plants for these
phytochemicals and assessing their potential in protecting against different
types of diseases.

1. Estimation of Primary and Secondary Metabolites

Table -1 Estimation of Primary and Secondary Metabolites

metabolites Concentration %
Protein 6.48
Carbohydrate 4.01
Amino Acid 1.10
Tannin 4.8
Polyphenol 5.57
Alkaloid 2.13
Flavonoid 2.36
Terpene 4.05
Glycosides 6.78

20
 Table -1 showed the result of primary and secondary metabolites of
alternanthera sessiles, from the result protein content is 6.48%, carbohydrates
is 4.01 % , amino acid is 1.104%, tannin is 4.8%, polyphenol is 5.57%, alkaloids
is 2.13%, flavonoids is 2.36%, glycosides is 6.78%, terpene is 4.05%.

6 6.78
6.48
5 5.57
CONCENTRATION %
4.8
4
4.01 4.05
3

2 2.36
2.13
1
1.1
0

Series 1

Figure-1 Estimation of Primary and Secondary Metabolites

21
 2 Estimation of Nutrient Profiling

Table-2 Estimation of Nutrient Profiling

Nutrient Concentration %
Moisture 88.25
Ash 1.29
Vitamin C 0.27
Vitamin E 0.97
Sodium
Potassium

Table-2 showed the result of nutrient profiling of alternanthera sessiles from

the result Moisture content is 88.25%, Ash content is 1.29%, Vitamin C content
is 0.27%, Vitamin E content is 0.97%, Sodium content is%, Potassium content
is %,

100

90
88.25
80

70

60

50
Concentration%
40

30

20

10
1.29 0.27 0.97
0
Moisture Ash Vitami C vitami E
Series 1

Figure-2 Estimation of Nutrient profiling

22
3 Estimation of Biological Activity

Table-3 Estimation of Biological Activity

Biological Activity Concentration %

Antioxidant 0.165

Antidiabetic 34.2

Table-3 showed the result of Biological activity of alternanthera sessiles from

the result Antioxidant content is 0.165%, Antidiabetic content is 34.2%.

40

34.2
35

30

25

Concentration% 20
15

10

5
0.165
0
Antioxidant Antidiabetic

Series 1

Figure-3 Biological activity

23
 4. Estimation of Antibacterial activity

Table - 4. Estimation of Antibacterial activity

Zone of inhibition (cm)

Organism used
Antibiotic Sample

E coli 3.3 cm 1.8

Staphylococcus 3.1 cm 1.6

Plate-1: Activity against E.coli Plate-2: Activity against

Staphylococcus

24
Discussion.

The weed Alternanthera sessilis, having Green (ASG) and Red (ASR)
cultivars, is a member of the Amaranthaceous. Both cultivars are known to
possess high medicinal and nutraceutical values as they are widely utilized in
traditional medicine. Populations from various Asian regions commonly
consume this medicinal weed as part of diet and as therapeutic agent. In general,
both cultivars have been employed for treating a broad range of human ailments
including cardiovascular diseases, diabetes, hypertension, and inflammatory
disorders such as asthma, bronchitis and hepatitis apart from its administration
as a painkiller. Following its ethnomedicinal uses, an attempt was made to
review and consolidate the phytochemical constituents of AS Gand ASR
discovered from previous studies that utilized advanced technologies,
respective traditional application among different ethnicities of Asia and
their established
pharmacological properties. All the information in this study was acquired
through a literature search of research databases, namely ScienceDirect,
Scopus, PubMed and Google Scholar using keywords such as Alternanthera
sessilis, Alternanthera sessilis red, dwarf copperleaf and sessile joy weed. The
results of each study were reviewed to identify its relevance and the cultivar
that was being investigated. All relevant studies were collected and its
subsequent results were reported and discussed to provide insights into the
health enhancing qualities of ASG and ASR. Inspection of phytoconstituents
in ASG and ASR showed the presence of polyphenol, flavonoid, carotenoid,
terpenes and alkaloid with polyphenol being the major chemical group found
in both cultivars.
It was also revealed that whilst ASR is traditionally consumed in China,
Malaysia, Singapore, and Taiwan, indigenous usage of ASG is more common
in nations such as Sri Lanka, India, and Indonesia.
25
 Both ASG and ASR showed potent anti-oxidant, anti-hyperglycaemic,
antimicrobial, antifungal and hepatoprotective effects while the anti-
inflammatory, anti-cancer and analgesic properties were disclosed in ASG
only. Further processing and commercialization of this remedial weed as a
product benefiting the wellbeing of humans is recommended as Alternanthera
sessilis has high potential to be utilized as a curative agent while being a vital
ingredient that can be included in daily consumption.

Summary and Conclusion.

A sessilis exists in two deferent colours, one with green aerial parts known
as green cultivar and the other having red aerial parts known as red cultivar.
Both plants have various active phytoconstituents including polyphenol,
terpenes, alkaloid, avoids, and carotenoids that exhibit a variety of
pharmacological activities. the plants also have gallic acid, which may
contribute to anti-cancer properties. the other bioactive compounds in the plants
exhibit strong pro-inflammatory, antioxidant, wound healing, analgesic, and
anti-angiogenesis properties. Extracts from deferent portions of A. sessilis such
as leaves, aerial and stem as well as the whole plant can be exploited to develop
novel antioxidants, anti-diabetic, anti-bacterial, anti - inflammation, anti-
cancer, hepatoprotective and analgesic agents. However, it is necessary to
understand that A. sessilis properties require more in-depth studies to detect the
mechanism action and safety of its phytoconstituents since in vitro studies are
still in its early phases. Further in vivo and clinical studies are needed,
particularly to investigate its anti-cancer properties over a wider range of
concentrations. Despite the fact that a lot of scientific publications tend to focus
on the green cultivar, presumably because it is so widely available, further
research on the red cultivar is necessary since it displays characteristics that are
comparable to those of the green cultivar.

26
 Further processing and commercialization of this remedial weed as a
product benefiting the wellbeing of humans is recommended as A sessilis has
high potential to be utilized as a curative agent while being a vital ingredient
that can be included in daily consumption. Companies that manufacture
products containing. A sessilis should include relevant information on its
bioactive components and any additional nutritional elements added to the
products and their concentration to raise awareness among consumers about the
safety of nutraceutical products.

27
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