AMLA

BOTANICAL DESCRIPTION
Plant profile Amla is the medium size deciduous plant. It grows to the height of 8 -18
meters. It has a crooked trunk and spreading branches. Its flower is yellow greenish in
colour. The fruit is spherical pale yellow with six vertical furrows. The mature fruits are
hard and do not fall for the gentle touch. The average weight of the fruit is 60 -70 g. It
has a gray bark and reddish wood. Its leaves are feathery, linear oblong in shape and
smell like lemon. Its wood is hard in texture. It wraps and splits when exposed in the
Sun or in the excessive heat
TAXONOMICAL CLASSIFICATION TAXON
Kingdom Plantae
Division Angiospermae
Class Eudicots
Order Malpighiales
Family Phyllanthaceae
Genus Phyllanthus
Species P. Emblica

GEOGRAPHICAL CLASSIFICATION
Amla is said to have been the first tree to be grown in this universe, according to
Indian mythology . E. officinalis is found in many tropical and subtropical
nations, including southeast Asia, China, India, and Indonesia . The plant has
been used for thousands of years to treat a wide range of conditions, including
diabetes, hyper-lipidemia and others, have been shown to have antioxidant, anti-
inflammatory, anticancer, adaptogenic, and antibacterial properties in order to
combat oxidative damage. Increased intake of foods high in antioxidants has been
linked to lower illness risk as well as lower morbidity and death rates .

FIGURE NO.7: IMAGE OF AMLA

Phytoconstituents of amla –

Phytoconstituents found in E. officinalis are known for their nutritious properties and consist
of tannins, mucic acid, amino acids, alkaloids, flavone glycosides, phenolic glycosides, flavonol
glycosides, phenolic acids, sesquiterpenoids, norsesquiterpenoids, and carbohydrates. Gallic
acid, chlorogenic acid, ellagic acid, and quercetincan be extracted from the fruit of
[Link] .

Fig – ascorbic acid

Materials and method

1. . Collection of plant material

Fruits of Phyllanthus emblica were purchased from Gollamari local market.

2. Preparation of extract
Before extraction, the amla fruits were cut into pieces and washed thoroughly with
distilled water and shade dried for four weeks at ambient temperature. Then fruits of
Amla were ground into fine powder and stored in a dry, cool, and dark area in an
airtight container. Nearly 20 g of the powdered fruits were extracted using 200 mL of
methanol. These mixtures were kept at 25 C for 72 h with intermittent stirring. Samples
were filtered and dried at 55 C in a rotary evaporator with lower pressure to obtain
methanolic crude extract (extract yield 2.53% of the dry plant material).

PHYSICAL PROPERTY
1. Determination Of Total Ash :-

Ashing involves an oxidation of the components of the product. A high ash

value is indicative of contamination, substitution, adulteration or carelessness in
preparing the crude drug for marketing. Total ash is designed to measure the
total amount of material produced after complete incineration of the ground drug
at as low temperature as possible (about 450°C) to remove all the carbons.

Place about 2-4 g of ground air dried material, accurately weighed in a

previously ignited and tared crucible of platinum or silica. Spread the material in
an even layer and ignite it by gradually increasing the heat to 500-600°C until it
is white, indicating the absence of carbon. Cool in a dessicator and weigh.
 2. Determination Of Acid Insoluble Ash :-

Ash Insoluble in hydrochloric acid is the residue obtained after extracting the
sulfated or total ash with HCL, Calculate with the reference to 100g of drug.

Boil the ash with 25 ml of 2M hydrochloric acid for 5 minutes, collect the
insoluble matter in a Gooch crucible or on an ashless filter paper, wash with hot
water, ignite, cool in a dessicator and weigh. Calculate the percentage of acid-
insoluble ash with reference to the air dried drug.

IMAGE: Determination of Total Ash & Acid Insoluble Ash

CHEMICAL TEST

1. TESTS FOR ALKALOIDS :-

Evaporate the aqueous, alcoholic and chloroform extracts separately. To residue,

add dilute HCI. Shake well and filter. With filtrate, perform following tests:
Mayer's test: 2-3 ml filtrate with few drops Mayer's reagent gives ppt.

2. TEST FOR FLAVONOIDS :-

Flavonoids contain conjugated aromatic systems therefore show

prominent/intense absorption bands in visible and UV regions of the spectrum.
e.g. 470 nm-560 nm (principal maxima) and 275 nm (subsidiary maxima)
indicate presence of anthocyanin.
Colour properties of Flavonoids in visible and UV light also helps in their
identification.
 e.g. most flavonol glycoside have very pale yellow colour in visible light while
in UV light they show dark brown colour (alone), and with ammonia light
yellow/yellow brown colouration.

3. TESTS FOR GLYCOSIDES :-

Determine free sugar content of the extract. Hydrolyse the extract with mineral
acid (dil. HCl/dil. H:SO₂). Again determine the total sugar content of the
hydrolised extract. Increase in sugar content indicates presence of glycoside in
the extract.
Glycoside - Aglycon (genin) + Glycon (sugar)

SAPONIN GLYCOSIDES:-
Foam test: Shake the drug extract or dry powder vigorously with water.
Persistent foam observed.

4. TESTS FOR TANNINS AND PHENOLIC COMPOUNDS :-

To 2-3 ml of aqueous or alcoholic extract, add few drops of following reagents.

Lead acetate solution: white ppt
5% FeCl3 Solution : deep blue-black Colour
Sr. Test Observation Results
no.
1 TESTS FOR ALKALOIDS :- Orange colour ppt
Mayer's test: 2-3 ml filtrate with is present
few drops Mayer's reagent gives Positive
ppt.

2 TEST FOR FLAVONOIDS :- Orange reddish

Sulphuric Acid Test: On addition colour is present
of sulphuric acid (66% or 80%) Positive
Flavanes give orange to red
colours.

3 TESTS FOR SAPONIN Stable Foam are

GLYCOSIDES :- present
Foam test: Shake the drug extract Positive
or dry powder vigorously with
water. Persistent foam observed.
4 TESTS FOR TANNINS AND
PHENOLIC COMPOUNDS :-
1. Lead acetate solution: white 1. White ppt is
ppt present Positive
2. 5% FeCl3 Solution : deep 2. deep blue-black
blue-black Colour Colour is
present
ASSAY OF ASCORBIC ACID

Principle of assay of ascorbic acid –

Ascorbic acid reduces iodine to iodide and itself gets oxidized to dehydroascorbic acid.
The liberated iodine is titrated with the help of starch as an indicator in an acidic
medium, where dilute sulfuric acid is used instead of acetic acid.

 Requirements:
[Link]: burette stand ,pipette, conical falsk, beakers, measuring cylinder, funnel,
weighing balance.
[Link]: extraction of Amla, H2SO4, starch indicator, Iodine.

 Preparation of chemicals :

1)Preparation of 1ml H2SO4:

 molecular weight of H2SO4 is 98.07 g/mole.
 Weighed 98.07g of H2SO4 and dissolve into the 1000ml of distilled water and
dissolve it properly until clear solution occurs .
2)Preparation of 0.05 M iodine solution –
 First take about 14 gram of iodine
 Then dissolve it in a solution containing 36 gram of potassium iodide in 100ml
of distilled water
  Add 03 drops of hydrochloride acid
 Dilute the solution with more distilled water until you have a total of 1000ml
3)Preparation of starch indicator:
 Weigh 1 gram of soluble starch powder.
 Add it to 100 mL of distilled water in a beaker. Stir to make a smooth paste using
a small amount of cold water (about 10–20 mL) to avoid clumping.
 Add the paste into about 80–90 mL of boiling distilled water slowly while stirring.
 Continue boiling and stirring the solution until it becomes clear (usually 5–10
minutes).
 Allow the solution to cool to room temperature.

ASSAY procedure:

 Dissolve 0.1 g in a mixture of 100 ml of freshly boiled and cooled distilled

water .
 And add 25ml of 1M sulphuric acid
 Immediate titrate with 0.05M iodine using starch solution as indicator until
the a persistent blue-violet colour is obtained.


Mechanism of action amla antioxidant property -
Quenching ROS and Induction of Antioxidant Enzymes:
Amla exhibits antioxidant properties by neutralizing reactive oxygen species
(ROS) and stimulating the production of antioxidant enzymes like superoxide
dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT).

Reducing Oxidative Damage:

Studies have shown that amla extracts can suppress alcohol-induced
oxidative damage by reducing lipid peroxidation and restoring the activity of
antioxidant enzymes.

Uses of Amla

Boosts Immunity

 Strengthens your defence system: The extremely high vitamin C content

fortifies your white blood cells, which play a vital role in fighting infections and
viruses.
 Antioxidant protection: Flavonoids and polyphenols neutralise free radicals,
reducing oxidative stress and inflammation, which are root causes of many
chronic illnesses.

Improves Digestion

 Stimulates digestive enzymes: It encourages the secretion of gastric juices

that improve food breakdown and nutrient absorption.
 Alleviates acidity and bloating: With its cooling properties, amla soothes
inflammation and discomfort in the digestive tract.

Enhances Heart Health

 Reduces bad cholesterol: Studies show amla helps lower LDL (bad
cholesterol) while improving HDL (good cholesterol).
 Improves blood circulation: Iron and antioxidants work together to maintain
flexible, healthy blood vessels.

Manages Blood Sugar Levels

 Lowers fasting glucose: It has been found to reduce blood sugar levels
naturally over time.
 Improves insulin sensitivity: Polyphenols in amla improve the way your body
uses insulin.
  Reduces sugar cravings: Amla’s bitter-sour taste helps balance sweet
cravings and supports dietary control.

Promotes Hair Growth & Reduces Hair Fall

 Stimulates hair follicles: Vitamin C boosts collagen production which

strengthens roots.
 Reduces dandruff and itchiness: Its antifungal properties help soothe the
scalp and improve overall hair health.