CHAPTER ONE

1.0 INTRODUCTION

1.1 BACKGROUND OF STUDY

Medicines of plant origin have been used to combat human diseases since time

immemorial even before the history of their documentation. Medicines are regarded

actually the first awareness about plants after food and shelter. Presently plant based

drugs are being used worldwide. These are either used directly extracted from plants

or modified through further synthesis Das and Srivastav, (2014). These specific

chemicals belong to plant derived compounds called phytochemicals such as the

metabolites of primary and secondary metabolites. Primary metabolites are high

volume chemicals of low value. Secondary metabolites are biosynthetically derived

from primary metabolites in lower volume and have higher value compounds. The

secondary metabolites are much of interest because of their antimicrobial Das and

Srivastav, (2014).

Andrographis paniculata belongs to the Acanthaceae family and is popularly known

as king of bitters in English. Andrographis paniculata grows erect to a height of 30–

110 cm in moist, shady places. The slender stem is dark green, squared in cross-

section with longitudinal furrows and wings along the angles. The lance shaped leaves

have hairless blades mea ruing up to 8 centimeters long by 2.5 wide. The small

flowers are borne in spreading racemes. The fruit is a capsule around 2 centimeters

long and a few millimeters wide. It contains many yellow brown seeds. It is also

known as Bhui-neem, meaning “neem of the ground”, since the plant, though being a

small annual herb, has a similar strong bitter taste as that of the large Neem tree. It is

an annual herbaceous plant which is commonly cultivated in Southern Asia, in China,

and in some parts of South East Asia. A. paniculata has antimicrobial Nagajothi et al.,
(2018) and antisnake- bite activities. Diterpenoids and flavonoids are the main

antimicrobial constituents of A. paniculata. It is used to overcome sannipata type of

fever, difficulty in breathing, hemopathy burning sensation, cough, skin diseases,

fever, ulcer and worms. It is also useful in acidity and liver complaints Nagajothi et

al., (2018).

The present study was aimed to investigate the phytochemical constituents of A.

paniculata as its uses in the treatment of different ailments.

1.2 STATEMENT OF PROBLEM

Andrographis can cause side effects such as loss of appetite, diarrhea, vomiting, rash,

headache, runny nose, and fatigue. When used in high doses or long-term,

andrographis might cause swollen lymph glands, serious allergic reactions, elevations

of liver enzymes, and other side effects.

1.3 JUSTIFICATION

Andrographis is also used for a wide assortment of other conditions. It is used for

digestive complaints including diarrhea, constipation, intestinal gas, colic, and

stomach pain; for liver conditions including an enlarged liver, jaundice, and liver

damage due to medications; for infections including leprosy, pneumonia, tuberculosis,

gonorrhea, syphilis, malaria, cholera, leptospirosis, rabies, sinusitis, and HIV/AIDS;

and for skin conditions including wounds, ulcers and itchiness.

Some people use andrographis for sore throat, coughs, swollen tonsils, bronchitis, and

allergies. It is also used for “hardening of the arteries” (atherosclerosis), and

prevention of heart disease and diabetes.

Other uses include treatment of snake and insect bites, loss of appetite, kidney

problems (pyelonephritis), hemorrhoids, and an inherited condition called familial

Mediterranean fever.

Andrographis is also used as astringent, bacteria killing agent, painkiller, fever

reducer, and treatment for worms.

1.4 AIM AND OBJECTIVES

The Main Aim:

To investigate the Phytochemicals presents in Andrographis paniculata (King of

bitter) and therapeutic importance to health.

Specific Objectives

This study will be conducted to;

1. Identifying the main phytochemicals that can be found in Andrographis

paniculata (King of bitter)

2. To contribute with new information on health uses of Andrographis

paniculata (King of bitter)

3. To determine compound in methanol and water extract of Andrographis

paniculata (King of bitter)

 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 DESCRIPTION OF ANDROGRAPHIS PANICULATA (KING OF BITTER)

Andrographis Paniculata Nees (Kalmegh) belongs to the family of Acanthaceae and

is popular worldwide with the name of “King of Bitters” in English. It has no local

name because it isn't a local herb. But they give it a Yoruba name in Oke-Ogun area

of Oyo state: they call it "méje-méje" (seven-seven) - based on the erroneous belief

that women should eat seven leaves. It is an annual herbaceous plant which is widely

cultivated in Southern Asia, India, China and some parts of Europe Nyeem et al.,

(2017). It is an erect, annual herb and 30-90 cm tall with upper part of stem

quadrangular while the lower part nearly rounded stem. Leaves are opposite sessile or

subsessile, linear-lanceolateor lanceolate, 3-8 cm long, acute, glabrous or minutely

puberulous beneath and base cuneate, margin slightly undulate. Flowers are

pedicelled, biliped, white-purple or spotted purple and solitary. Pedicel is 2.5–10 mm

in size, slender and glandular pubescent. Bracts are acicular and 2.5 mm long. Calyx

lobes are subacute, 2.5-3.7 mm long and glandular Nyeem et al., (2017). Corolla is

7.5-12.5 mm in size, tube about half as long as the corolla. Filaments are hairy and

anthers are purple beared at base. Fruit is a capsule, oblong, 18-20X4.5–5.0 mm,

young ones sparsely glandular and hairy; when mature it is glabrous. Seeds are

subquadrate, yellow to brownish in colour and rugose. Flowering and fruiting occurs

in October – December (North India) Nyeem et al., (2017).

The species is a native of tropical South-East Asia and occurs throughout hotter parts

of India. The plant comes up well in tropical and subtropical regions all over India. It

is a hardy species, therefore, can be grown in medium fertile sandy loam to clay-loam
soils, possibly with irrigation. It can withstand partial shade of trees, say few hours,

but it is cultivated in open fields Rajalakshmi and Cathrine, (2016).

It can be easily raised through seed and vegetative methods. But in commercial

cultivation, propagation through seed is easy and economical. This crop is grown

during cooler climate and it remains for 120 days in field; usually, ratoon crop is also

taken all over north India. Cooler climate helps plants in synthesizing more bitter

ingredients Rajalakshmi and Cathrine, (2016).

Seeds are soaked in water for 24 hours and sown in the nursery beds in early

September. About 650-750 gm seeds are required for raising nursery for one hectare

of land. Nursery is prepared with soil, sand and organic matter in 1:1:1 ratio and sown

in early September at 5 cm spacing in rows and it takes 8-10 days for germination to

commence. Six weeks old seedlings are planted in field at 30X15 cm or 15X15 cm

spacing Dhiman et al., (2012). Direct sown crop is broadcasted thinly and has a seed

rate of 1.5 kg/ha. It matures early, but nursery raising is preferred. 10-25 cm long

seedlings raised in the nursery beds during September are transplanted in the main

field (after 6 weeks of sowing) at a distance of 30X15 cm between plant to plant and

row to row. 4-6 light irrigations are required till harvesting the crop. Since it is a

herbaceous plant, the field should be free from weeds. Two to three weeding are

essential during the crop season viz. at 20 days and 60 days after transplantation. It is

a hardy plant and not attacked by any pest and disease Dhiman et al., (2012).

The crop matures after 120 days of sowing. It is harvested when most plants are in

bloom. It is at this stage, the plants should be uprooted. However, a small lot of
healthy plants should be left in the field for seed production. When the fruits become

mature, these should be picked up and dried in the sun and seeds are collected. The

seeds should be kept in open sun for complete drying. After this, these are stored in

air-tight containers for next sowing Dhiman et al., (2012).

After uprooting the plant, first it should be dried in the sun for two days and

afterwards in the shade. This properly dried material should be packed in laminated

gunny bags, lest it absorbs moisture. The harvested dry material should be stored in

dark, airy and moisture-free places. One year of storage from the time of harvest. The

leaves contain three bitter principles; deoxyandrographolide, andrographolide and

neoandrographolide. These are also present in whole plant. The leaves should yield

2.5% chemical constituents on analysis. The yield (whole plant) is 2.5 t/ha. It has

sizeable demand and yields a reasonable profit to the growers. It is commercially

cultivated in several States of India. Rs. 25000/- is the cost of cultivation for one

hectare Dhiman et al., (2012).

The whole herb is bitter in taste and is source of several diterpenoids of which a bitter

water soluble lactone “andrographolide” is important. The plant is acrid, cooling,

laxative, antipyretic, antiperiodic, anti-inflammatory, expectorant, sudorific,

anthelmintic, digestive and stomachic. It is useful in burning sensation, chronic fever,

malaria and intermittent fever, inflammation, cough, bronchitis, skin diseases,

intestinal worm, dyspepsia, flatulence, colic, diarrhoea, dysentery, haemorrhoids and

vitiated condition of pitta Dhiman et al., (2012).

2.2 TAXONOMY OF ANDROGRAPHIS PANICULATA (KING OF

BITTER)

Kingdom : Plantae

Order : Lamiales

Family : Acanthaceae

Genus : Andrographis

Species : A.paniculata

FIGURE 1: ANDROGRAPHIS PANICULATA (KING OF BITTER)

2.3 TRADITIONAL USES OF ANDROGRAPHIS PANICULATA (KING OF

BITTER)

Andrographis paniculata is an important medicinal plant that is widely used around

the world. It is ethnobotanically used for the treatment of snake bite, bug bite,

diabetes, dysentery, fever and malaria Anil et al., (2012). The aerial parts of the plant

contain a large number of diterpenes, whilst the presence of flavones in the root has

also been reported. This plant exhibits various biological activities such as anti-

microbial, cytotoxicity, anti-protozoan, anti-inflammatory, anti-oxidant,

immunostimulant, anti-diabetic, anti-infective, anti-angiogenic, hepato-renal

protective, sex hormone modulatory, liver enzymes modulatory and insecticidal

activities. It also has anti-cancer, anti-diarrheal, anti-hepatitis, anti-HIV, anti-

hyperglycemic, anti-inflammatory, anti-microbial and anti-malarial activities Anil et

al., (2012).

The roots and leaves are primarily used to reduce fever, tone the stomach, increase

appetite and generally improve overall health condition. It also has a significant anti-

inflammatory effect and it inhibits oedema. Decoction of the leaves or roots can be

used as treatment for stomach pain, dysentery, typhus, cholera, influenza and

bronchitis. Infusion can be used for treating female disorders, dyspepsia,

hypertension, gonorrhoea, jaundice, rheumatism, amenorrhoea and torpid liver. When

used as a poultice, it cures swollen legs or feet, vitiligo and piles. It can also be a

remedy for diabetes when used together with Orthosiphon aristatus. It is used to treat
hepatitis, gastrointestinal tract and upper respiratory infections, fever, herpes and a

variety of other chronic and infectious diseases Anil et al., (2012).

Trials using the leaf extract have concluded that the powdered leaves have the

capacity to significantly shorten the duration of common colds and that the leaves can

be as effective as Paracetamol in relieving fever and sore throat of people with

pharyngotonsillitis. The leaves have also been shown to have significant anti-

inflammatory effects and also significantly inhibit oedema Anil et al., (2012). An

ethanol extract of the leaves stimulates both antigen-specific and nonspecific immune

responses more than the purified leaf extracts. An ethanol extract has also shown

significant antipyretic activity. A standardized leaf extract exhibits significant

antipyretic properties and is an effective analgesic. The crude water extract of the

leaves as well as the semi-purified n-butanol and aqueous fractions have shown

significant hypotensive activity. An extract of the leaves has also been shown to

lower cholesterol levels, whilst anti-thrombotic effects were also observed. It is

believed that these effects might be at least partially due to flavones present in the

extract Anil et al., (2012).

Various preparations and compound formulas of the herb have been used to treat

infectious and non-infectious diseases with significant efficacy reported in case of

epidemic encephalitis B, neonatal subcutaneous annular ulcer, vaginitis, cervical

erosion, pelvic inflammation, herpes zoster, chicken pox, mumps, neurodermatitis,

eczema, and burns. Administered intraperitoneally, the diterpenes andrographiside

and neoandrographolide (found in the leaves) have a significant protective effect on

the liver. An extract of the leaves has shown antidiarrhoeal activity Anil et al., (2012).

The diterpenes andrographolide and neoandrographolide isolated from the alcoholic

extract showed potent antisecretory activity against Escherichia coli enterotoxin

induced secretions. Various compounds have shown significant anti-ulcer properties.

It is suggested that this effect is due to the antisecretory activity and protective effect

on the gastric mucosa. Oral administration of 20mg of the dry leaf powder for 60 days

has an antifertility effect in males. The alcoholic extract of the rhizomes exhibits good

in vitro anthelmintic activity against Ascaris lumbricoides. Neoandrographolide,

isolated from the leaves, exhibits significant antimalarial activity. An infusion or sap

from the crushed leaves has been recommended for the treatment of fever, as a tonic

and for itching skin eruptions. A decoction of the leaves or roots is used against

stomach-ache, dysentery, typhus, cholera, influenza and bronchitis, as a vermifuge

and is considered a diuretic. Pills or infusions are also recommended to treat female

disorders, dyspepsia, hypertension, rheumatism, gonorrhoea, amenorrhoea, torpid

liver and jaundice. Another use is as a poultice on swollen legs or feet, vitiligo and

piles Anil et al., (2012).

2.4 HEALTH BENEFITS OF THE KING OF BITTERS

During the last two decades, the King of Bitters (Andrographis paniculata) has

become popular in the prevention and treatment of colds and respiratory infections,

and has been shown to reduce the symptoms.

The antioxidant compounds in the herb have a beneficial effect on the liver, improves

gall bladder function, aids digestion, is used to treat e.coli, and promotes a healthy

heart by preventing blood clots and blocked arteries Adefegha et al., (2022).

Overall, the herb is known to be a powerful immune booster. In recent history, the

global flu epidemic of 1919, killed millions in countries all over the world, but India
used the prophylatic benefits of the herb and were credited with stopping the

epidemic.

2.4.1 HOW HAS IT BEEN USED?

Ancient and Modern Chinese and Indian medicinal literatue has described

Andrographis paniculata as an important herbal remedy for a wide variety of

diseases. This includes disorders of the urinary, digestive and cardiovascular systems.

Andrographis paniculata has also been used to treat fever, upper respiratory

infections and sore throat. Herbalists use the herb to eliminate symptoms of malaria,

dysentery, and herpes Adefegha et al., (2022).

2.4.2 MODERN BENEFITS

Research into the benefits of andrographis paniculata has shown it to be useful in the

treatment of cancer, HIV/AIDs infections, heart attacks, blood clots, diabetes, fever,

flu and cold. Andrographis paniculata has a known reputation in Tribal folklore as

one of the best remedies for malaria; it is believed to be a better option than quinine.

2.4.3 HOW IS IT PREPARED?

The active phytochemicals are extracted from the stems and leaves. The main

medicinal component is andrographolide, which is colorless and crystalline in

appearance and has a very bitter taste. The herb was originally prepared as a tea,

tincture, and fresh juice beverage Adefegha et al., (2022).

The herbs can now be found on the market in capsule and tablet form, and as an

extract, gel and ointment.

2.4.5 WHAT IS ANDROGRAPHIS?

Andrographis is an herb or herbal recipe that has long been believed to be a natural

immune-booster. Also referred to as “Indian Echinacea,” the Andrographis herb is

most often taken alone or with Siberian ginseng – generally as an intended remedy for

flu, a cold or as a treatment for various cancers Adefegha et al., (2022).

With its white-purple flowers, the andrographis plant is native to Sri Lanka, India and

other nearby Asian countries. Andrographis paniculata is the widely considered the

most popular for this purpose

2.4.6 ANDROGRAPHIS IN AYURVEDIC MEDICINE

Noted for its unique bitter taste, this herb is sometimes given top honors for this

achievement and called the “King of Bitters.” Within Ayurveda medicine application,

the herb is sometimes referred to as “Kalamegha” or “Kalmegh” – meaning “dark

cloud.”

2.4.7 ANDROGRAPHIS BENEFITS

Herbalists use the underground stem and leaf of specific types of these plants to make

medicine or tonics. Some researchers believe that this plant may have the ability to

enhance blood cell counts, offer anti-oxidant benefits and lower blood -- among other

health-boosting enhancements. Various therapy options for this herb are often

available at health food stores. Tincture, tea and capsule formulations seem to be

among the most common options available.

In addition to the flu, cold and cancer applications, andrographis may also be applied

as a fever reducer, painkiller, astringent and as a sore throat or cough therapy.

Andrographolide is frequently considered to be the one of the main active element in

andrographis products. Credited with having possible future neuro-protective and

anti-inflammatory implications, research results following experiments with stroke

and cell-signaling impact is likely Adefegha et al., (2022).

2.4.8 ANDROGRAPHIS SIDE EFFECTS

Because of the intense bitter taste, many medical professionals recommending this

course of therapy first suggest the capsule form of this herb. Doing so also allows

patients to carefully monitor their dosage levels. Although Andrographis products are

widely considered to be generally safe for many people -- the herbal extracts may

cause negative conditions like nausea, stomach irritation, diarrhea, fatigue and

headaches, as well as allergic reactions. Patients taking blood pressure or blood-

thinning medications should consult with a doctor prior to using any andrographic

program. Chemotherapy drug users are also strongly recommended to seek health

consultations prior to using these products. Some herbalists also warn that

Andrographis could impair female and male fertility function for some patients

Adefegha et al., (2022).

 CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Sample Collection

The fresh Andrographis paniculata plant was purchased from local market of Oba

Adesola market, Lusada, Igbesa, in Ado – Local Government, Ogun State, Nigeria.

The plant were identified by a technologist in Biology Unit, Department of Science

and Technology Igbesa, Ogun State Institute of Technology, Igbesa, Ogun State.

3.2 Sample Preparation for Drying

Fresh Andrographis paniculata leaves were sorted of unwanted materials, destalked,

rinsed with distilled water to remove sand particles and drain with sieves to remove

adhering water. The leaves were first washed in tap water and then in double distilled

water to remove dirt. The leaves were then dried at room temperature for seven days,

finely powdered and used for phytochemical analysis.

This dried powdered material was also used for solvent extraction for the preparation

of sample required for the determination of phytochemical analysis.

3.3 Preparation of Samples for Phytochemical analysis Analysis

Two different solvents were used (ethanol and distilled water). 150 g of leaf powder

was soaked in 750 ml each of the solvents and mixed by stirring with a glass rod. The

mixtures were left at room temperature for 24 h. The extracts were filtered using a

sieve and finally with Whatman No one filter paper. The resulting solution from

ethanol were evaporated in a rotary evaporator while, the aqueous extract was

evaporated in a water bath at a temperature of 55°C until semi solid extract was

attained. The extracts were kept in a plastic until required for use.

3.4 Phytochemical Analysis: Qualitative Chemical Examination (Aqueous

and Ethanolic extract) :

The qualitative tests for phytochemicals were performed according to several

previously published standard protocols (Nagajothi et al., (2018); Rajalakshmi and

Cathrine, (2016); Singh et al., (2017); Balamurugan et al., (2019) and Murali et al.,

(2014).

3.5 QUALITATIVE ANALYSIS OF PRIMARY METABOLITES:

3.5.1 Test for Carbohydrates

1. Benedict’s test: About 0.5 ml of the filtrate was taken to which 0.5 ml of

Benedict’s reagent is added.

This mixture was heated for about 2 minutes in a boiling water bath. The

appearance of red precipitate indicates the presence of sugars

2. Molisch’s test: To about 2ml of the sample, 2 drops of alcoholic solution of α-

napthol was added and to the mixture after being shaken well. Few drops of
 conc.H2SO4 were added along the sides of the test tube. A violet ring indicates

the presence of sugars

3.5.2 Test for Proteins

1. Biuret test: 2ml of filtrate was taken to which 1 drop of 2% copper sulphate

solution was added; 1ml of 95% ethanol was added. Then it was followed by

excess addition of KOH. The appearance of pink colour indicates the presence

of protein.

2. 2ml of extract was mixed with 2ml of water and about 0.5% of conc. HNO3

was added. The appearance of yellow colour indicates the presence of

proteins.

3. To about 2 ml of the extract, 2ml of miller’s reagent was added white

precipitate which turns red on heating will confirm the presence of proteins.

3.5.3 Test for Amino Acids

1. To 1ml of the extract, few drops of ninhydrin reagent (10mg of ninhydrin in

200ml of acetone) was added. The appearance of purple colour indicates the

presence of amino acids.

2. To 2ml of extract few drops of nitic acid were added along the sides of the

tube the appearance of yellow colour indicates the presence of protein and free

amino acids.

3.5.4 Miscellaneous compounds

Test of Resins
1. Precipitation test: about 0.2 g of extract was extracted with 15ml of 95%

ethanol. The alcoholic extract was then poured into a beaker containing about

20ml of distilled water.

2. 1ml of extract was taken and to this few ml of acetic anhydride was added to

this 1ml of conc.H2SO4 was added. The appearance of orange to yellow colour

indicates the presence of resins

3.5.5 Gums and Mucilage

To 1ml of extract, distilled water, 2ml of absolute ethanol was added with

constant stirring white or cloudy precipitate indicates the presence of gums or

mucilage

3.6 QUALITATIVE ANALYSIS OF SECONDARY METABOLITES

3.6.1 Test for anthraquinones

To 5ml of extract, few ml of conc.H 2SO4 was added and 1ml of diluted

ammonia was added to it. The appearance of rose pink confirms the presence

of anthraquinones

3.6.2 Test for Quinones

To 1ml of extract, alcoholic KOH is added the presence of red to blue colour

indicates the presence of quinones

3.6.3 Test for Alkaloids

1. Mayer’s test: to a few ml of filtrate, 2 drops Mayer’s reagent was added a

creamy or white precipitate shows a positive result for alkaloids.

2. Wagner’s test (iodine – potassium iodine reagent): To about an ml of extract

few drops of Wagner’s reagent were added. Reddish – brown precipitate

indicates presence of alkaloids.

3. To 5ml of extract 2ml of HCl was added. Then 1 ml of Dragendroff‟s reagent

was added an orange or red precipitate shows a positive result for alkaloids.

3.6.4 Test for Glycosides

1. Borntrager’s test: to 2ml of filtrate, 3ml of chloroform is added and shaken.

The chloroform layer is separated and 10% ammonia solution was added. The

pink colour indicates the presence of glycosides

2. 5ml of extract was hydrolysed with 5ml of conc. HCl boiled for few hours in a

boiling water bath, small amount of alcoholic extract was dissolved in 2ml of

water and 10% of aqueous 10% NaOH was added the presence of yellow

colour was a positive result for the glycosides.

3. 2ml of extract is mixed with about 0.4 ml of glacial acetic acid containing

traces of ferric chloride and 0.5 of conc. H2SO4 was added the production of

blue colour is positive for glycosides.

3.6.5 Test for Cardiac Glycosides (Keller-Killani test)

1. 5ml of solvent extract was mixed with 2ml of glacial acetic acid and a drop of

ferric chloride solution was added followed by the addition of 1ml of conc.

H2SO4. A brown ring in the interface indicates the presence of deoxy sugars of
 cardenoloides. A violet ring may appear beneath the brown ring while acetic

acid layer a green ring may also form just gradually towards the layer.

3.6.6 Test for Phenol

1. Gelatine test: To 5ml of extract 2ml of 1% solution of gelatine containing

10% of NaCl is added.

Appearance of white precipitate indicates the presence of phenol

2. Lead acetate test: To 5 ml of extract 3ml of 10%lead acetate solution was

added and mixed gently. The production of bulky white precipitate is positive

for phenols.

3.6.7 Test for Tannins

1. To 5ml of extract few drops of neutral 5% ferric chloride solution was added,

the production of dark green colour indicates the presence of tannins

3.6.8 Test for Flavonoids

1. To the aqueous solution of the extracts 10% ammonia solution is added and is

heated. The production of fluorescence yellow is positive for flavonoids.

2. 1ml of extract was taken and 10% of lead acetate was added. The yellow

precipitate is positive inference for the flavonoids

3. The extract is treated with concentrated H 2SO4 resulting in the formation of

orange colour indicates the positive result for flavonoids.

4. To 5ml of dilute ammonia the plant extract is added and shaken well. The

aqueous portion is separated and concentrated H2SO4 is added. The yellow

colour indicates the presence of flavonoids.

3.6.9 Test for Phlobatannins

1. Aqueous extract was boiled with diluted HCl leading to the deposition of

reddish precipitate indicates the presence of phlobatannins

3.6.10 Test for Saponins

1. 0.5 mg of extract was vigorously shaken with few ml of distilled water. The

formation of frothing is positive for saponins

2. The froth from the above reaction is taken and few drops of olive oil is added

and shaken vigorously and observed for the formation of emulsion.

3.6.11 Test for Steroids

2ml of extract with 2ml of chloroform and 2ml of concentrated H2SO4 are

added, the appearance of red colour and yellowish green fluorescence

indicates the presence of steroids

3.6.12 Test for Terpenoids (Salkowski test)

3ml of the extract was taken and 1ml of chloroform and 1.5 ml of concentrated H 2SO4

are added along the sides of the tube. The reddish brown colour in the interface is

considered positive for the presence of terpenoids

3.6.13 Test for Coumarins

To 2 ml of the extract, 3 ml of 10% aqueous solution of NaOH is added. The

production of yellow colour indicates the presence of coumarins

3.7 QUALITATIVE ANALYSIS OF VITAMINS

3.7.1 Test for vitamin – C

In 5ml of distilled water, 1ml of the sample was diluted and a drop of 5% sodium

nitroprusside and 2ml of NaOH is added. Few drops of HCl are added dropwise, the

yellow colour turns blue. This indicates the presence of vitamin- C

 CHAPTER FOUR

4.0 RESULTS AND DISCUSSION

4.1 RESULTS

Phytochemical screening of the ethanolic extracts of A.Paniculata revealed the

presence of alkaloids, carbohydrates, saponin, protein, phytosterol, phenolic

compounds, flavonoid and glycoside (Table 4.1).

Table 1: THE ANALYSIS OF PHYTOCHEMICALS IN THE ETHANOLIC

AND AQUEOUS EXTRACTS OF ANDROGRAPHIS PANICULATA

S. No. Phytochemical Tests Ethanol Aqueous

Extract Extract

1. Alkaloids ( Mayer’s reagent) +++ +++

2. Carbohydrates (Sugar (Benedict’s reagent) - +++

3. Glycosides ( molisch’s test) ++ +

4. Saponin ( Foam test) ++ ++

5. Protein ( Millon’s test) - -

6. Phenolic compounds ++ +++

7. Tannin ( Ferric chloride test) ++ +++

8. Flavonoid (Alkaline reagent test) ++ +++

9. Tannins ++ +++

10. Phlobatannins +++ +++

11. Terpernoid +++ +++

12. Amino Acid - -

13. Steriods +++ +++

14. Resin + +

15 Gum + +
16. Vitamin C - -

17. Coumarin + ++

18. Quinones ++ ++

19. Terpernoid +++ +++

Keys: + = present; ++ = Moderate; +++ = Maximum - = absent

4.2 DISCUSSION

Plants are an important source of phytochemicals which are an important source of

drug and medicine. These phytochemicals have extraordinary properties like

antibacterial, antifungal, anti-cancerous, antioxidant, antiinflammatory, anti-diabetic

activities. Different phytochemicals have been found to possess a wide range of

activities, which may help in protection against chronic diseases. For example,

Alkaloids protect against chronic disease. Saponins protect against

hypercholesterolemia and antibiotic properties. Steroids and triterpenoids show the

analgesic properties Nyeem et al., (2017). The Steroids and saponins were responsible

for central nervous system activities, flavonoids have been referred to as nature’s

biological response modifiers, because of their inherent ability to modify the body’s

reaction to allergies and virus and they showed their anti-allergic, anti-inflammatory,

anti-microbial and anti-cancer activities.

The phenolic compounds are one of the largest and most ubiquitous groups of plant

metabolites. They possess biological properties such as antiapoptosis, antiaging,

anticarcinogen, antiinflammation, antiatherosclerosis, cardiovascular protection and

improvement of endothelial function, as well as inhibition of angiogenesis and cell

proliferation activities Singh et al., (2017). Several studies have described the
antioxidant properties of medicinal plants which are rich in phenolic compounds.

Natural antioxidant mainly come from plants in the form of phenolic compounds such

as flavonoid, phenolic acids, tocopherols etc. Tannins bind to proline rich protein and

interfere with protein synthesis Singh et al., (2017). Flavonoids are hydroxylated

phenolic substances known to be synthesized by plants in response to microbial

infection and they have been found to be antimicrobial substances against wide array

of microorganisms in vitro. Their activity is probably due to their ability to complex

with extracellular and soluble proteins and to complex with bacterial cell wall. They

also are effective antioxidant and show strong anticancer activities Rajalakshmi and

Cathrine, (2016).

Glycosides are known to lower the blood pressure according to many reports. The

results obtained in this study thus suggest the identified phytochemical compounds

may be the bioactive constituents and these plants are proving to be an increasingly

valuable reservoir of bioactive compounds of substantial medicinal merit Murali et

al., (2014).

Thus all these phytochemicals act synergistically and exhibit beneficial effects in

treatment of wide variety of diseases conditions ranging from pyrexia to cancer

Nagajothi et al., (2018). Hence, Andrographis paniculata is included as an ingredient

in several polyherbal preparations for its heaptoprotective, antiviral and

immunostimulant activity not only in human but also in animals.

 CHAPTER FIVE

5.0 SUMMARY, FINDINGS AND RECOMMENDATIONS

5.1 SUMMARY AND FINDINGS:

The leaf extract of Andrographis paniculata Nees have great potential as

phytochemical compounds against diseases. Andrographis paniculata Nees contains

large number of secondary metabolites like flavonoids, terpenoids, alkaloids, tannins,

saponins and phenolic compound with antibacterial activities and thus are sources of

natural bioactive molecules to control pathogens that cause disease in humans. Based

on the results we concluded that the aqueous extract was found to be more potent than

Ethanolic extract. Thus, they can be used in the treatment of infectious diseases

caused by the different microbes.

5.2 RECOMMENDATIONS

Additional investigation into the specific phytochemicals present in Andrographis

paniculata Nees extracts may lead to the development of effective therapeutics for the
prevention and/or treatment of these disease states. More research should be

conducted to evaluate its usefulness as a means of preventing oxidative stress, cancer,

ulcer, diabetes, inflammation, neuroinflammation, and microbial infections.

Andrographis paniculata Nees can be a potential source of useful drugs.

Incorporation of this plant in diet as nutraceuticals is worth recommendation.

Moreover, they are ubiquitous, can be grown or cultivated and is not even

endangered. We believed that the data provided by us will be helpful to explore more

medicinal plants.

Further researches are necessary to determine precisely the different constituents’

present especially for flavonoids, glycosides, saponins, terpeniods and phenols as

quantitative contents.
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