PHYTOCHEMICAL INVESTIGATION OF LEAF OF

NYCTANTHES ARBOR-TRISTIS

A REPORT SUBMITTED TO THE

PURBANCHAL UNIVERSITY SCHOOL OF HEALTH SCIENCES
SUNDARHARAINCHA-8, GOATHGAUN, MORANG

Presented by Submitted to
Nirajan Dhakal Sushil Regmi
B. Pharmacy, 4rth Semester Assistant Professor
PU Reg. No.: 152-6-2-05392-2021 Department of Pharmacy
Monograph
Nyctanthes arbor-tristis popularly known as Parijat or Night jasmine belongs to
family Oleaceae. Latin name Nyctanthes has been coined from two Greek words
Nykhta (Night), and Anthos (flower).
1.1.COLLECTION AND PROCESSING OF SAMPLES:
Samples of Nyctanthes arbor-tristis (Leaf) were obtained from
Sundarharaincha-8, Salakpur, Morang. Freshly collected leaves of
Nyctanthes arbor-tristis were cleaned and dried under the shade at
normal room temperature for 5 days. Upon drying leaf was blended to
powder using an electric blender.

1.2.DETERMINATION OF ASH VALUE:

1.2.1. Determination of total ash
i. About 2 gm of the powdered drug was accurately weighed, taken
into a tared porcelain dish & incinerated on a Muffle Furnace.
ii. Powder was heated strongly until the carbon is burnt off.
iii. Cooled in a desiccator.
iv. The percentage of ash was calculated by using formula.
Weight of ash
Percentage of total ash= × 100
Weight of Sample

1.2.2. Determination of acid- insoluble ash value

i. The total ash obtained above was used for determining acid –
insoluble ash & boiled for 5 min with 25 ml dil. HCL.
ii. The content was filtered by using filter paper.
iii. Residue was washed twice with hot water. Then cooled &
weighed.
iv. Then the residue transferred into porcelain dish & ignite until the
vapours cease to be evolved & then more strongly until all the
carbon has been removed.
 v. Cool & weigh the residue & calculate the percentage of acid –
insoluble ash.
Weight of residue
Percentage of Acid−insoluble ash= × 100 %
Weight of Sample taken

1.2.3. Determination Water –Soluble Ash Value:

i. The total ash obtained after for determining acid-insoluble ash
above was used & boiled for 5 min with 25 ml distilled water.
ii. Then filter by using Whatman filter paper.
iii. Residue was washed twice with hot water.
iv. Then the residue transferred into porcelain dish & ignite until the
vapours cease to be evolved & then more strongly until all the
carbon has been removed.
v. Cool & weigh the residue & calculate the percentage of water -
insoluble ash value.
Weight of residue after incineration
Percentage of water−soluble ash= ×100
Weight of Sample

1.3.LOSS ON DRYING:
i. The % of LOD was determined by using gravimetric
method.
ii. About 1.5 gm of crude drug was weighed into a weighed
porcelain dish.
iii. Dried in the oven at 100 0C until two consecutive weighing
do not differ by more than 0.5 mg.
iv. Cooled in desiccator & weighed. The loss in weight is
usually recorded as a moisture.
( Weight of sample−Weight of dried sample )
Loss on Drying (LoD %)= × 100 %
Weight of sample

1.4.EXTRACTION:
 i. Around 5g of Nyctanthes arbor-tristis leaves powder were taken in a
beaker.
ii. Powder was soaked overnight in 100ml of 2 solvents (Aqueous and
methanol solvents) each for 24 hours. The solvents dissolve the
active biomolecules. The leaves powder remain as precipitate and
the active biomolecules were present in the solvent.
iii. The mixture was filtered and the following phytochemical assays
were conducted to
iv. The presence of secondary metabolites on each filtrate was tested.
1.5.PHYTOCHEMICAL ASSAYS
Phytochemical analysis was carried out on different extract of the whole
plant using standard procedure to identify the bioactive compounds.
Test for Tannins
2ml of extract was taken in a test tube and equal amount of bromine
water was added. Discoloration of bromine water confirm presence of
tannins.
Test for Protein
2ml of extract was taken in a test tube and 1ml of Millon's reagent was
added. Red colour precipitate is indication of presence of soluble protein.
Test for Saponin
1ml of the extract was boiled in 10ml of distilled water and filtered. 5ml
of filtrate was mixed with 2 ml of normal distilled water and shaken
vigorously. Occurrence of stable persistent froth indicates the presence
of saponins.
Test for Flavonoids
To 1 ml of the extract, few drops of dilute sodium hydroxide were added.
Presence of flavonoids is indicated upon production of an intense yellow
colour in the plant extract which became colourless on addition of 2-3
drops of 50% dilute acid.
 Test for Terpenoid
0.5 gm of plant extract was mixed with 2 ml of chloroform and equal
volume of concentrated sulfuric acid was added. Terpenoids presence is
confirmed by a reddish brown colouration of interface.
Test for Steroid
2 ml of plant extract was dissolved in 5 ml chloroform and then 5 ml of
concentrated sulfuric acid was added. Formation of 2 phases (upper red
and lower yellow with green fluorescence) indicates the presence of
steroid.
Test for Alkaloids
5ml of plant extract was mixed with 3 ml of aqueous HCl on water bath
and then filtered. 1 ml of Dragendorff’s reagent was added in the filtrate.
The occurrence of orange-red precipitate indicates the presence of
alkaloids in the sample extract.
Test for Reducing sugar
0.5 g of plant extract was dissolved with distilled water and filtered. The
filtrate was boiled with 2 drops of Fehling’s solution A and B for 5
minutes. An orange-red precipitate obtained indicates the presence of
reducing sugar.

S.N Pharmacopeial parameter Observation Inference

.
1. Description
Flower Snow white petal and
Snow white petal and orange red orange red center
center
Leaf Rough, light green, Complies with
Green, rough, heart shaped heart shaped with Pharmacopoeia
entire margined with reticulate reticulate venation
venation
 Bark Grey, rough bark
A small tree with a grey or
greenish, rough and peeling bark
2. Identification
a. Macroscopy
Flower Snow white petal and
Snow white petal and orange orange red center
red center
Leaf Green, rough, heart
Green, rough, heart shaped shaped entire
entire margined with margined with
reticulate venation reticulate venation

Bitter and astringent taste Bitter and astringent

taste

Bark Grey bark with Complies with

Outer surface of bark is grey, creamy colored inner Pharmacopoeia
fresh inner bark creamy bark.
colored that turns deep
brown on drying.
b. Microscopy Lack of stomata in
Leaf upper epidermis
Higher epidermis devoid of Two rows of
stomata epidermis seen.
Two rows of epidermis are Barrel shaped cell
seen in the lamina near the seen.
midrib
Uniseriate epidermis with
barrel shaped cells
3. Ash Values
a. Total Ash value 20% Complies with
b. Acid Insoluble ash value 7% Pharmacopoeia
 c. Water soluble ash value 17.5%

4. Loss on Drying 55% Complies with

Pharmacopoeia

5. Phytochemical Assay Aqueous Methanolic

a. Tannins + + Complies with
b. Protein - - Pharmacopoeia
c. Saponin + +
d. Flavonoids + -
e. Terpenoids - +
f. Steroids - -
g. Alkaloids - -
h. Reducing sugar + -