PHYTOCHEMICAL ANALYSIS:

Phytochemicals are chemically active compounds that occur naturally in plants. Different types
of chemical tests were performed for the screening and identification of different medicinally
active substances found in ethanolic extract, such as terpenoids, phenols, steroids, flavonoids,
saponins, diterpenes, phlobotannins, carboxylic acid.

1- Detection of Terpenoids
Salkowski test: This test is also referred as Salkowski test. Mix 1 mL of chloroform (CHCL3) to
2.5 mL of ethanolic extract solution carefully added 1.5 ml of concentrated sulphuric acid
(H2SO4). The formation of red coloration specifies the presence of terpenoids.

2- Detection of Phenols
The ferric chloride (FeCl3) test used to the determination of phenols in a given sample or
compound. Phenol was confirmed by the presence of bluish black color formation.

3- Detection of Steroids
Take 1 ml of plant extract in a test tube then adds 2 mL of acetic acid mixed in it and stirred
with a glass rod then add 2 mL of concentrated sulphuric acid (H2SO4) to the test tube and
shake gradually. Presences of steroids were confirmed by the color change from violet to green
or blue.

4- Detection of Flavonoids
Alkaline reagent test: Alkaline reagent test for the detection of flavonoids. Take 1 mL of plant
extract then added a few drops of 10% NaOH. Intense yellow color was observed in the extract,
by the addition of diluted HCL color disappeared and this indicates the presence of flavonoids.

5- Detection of Saponins:
About 1g of the powdered sample was boiled with 10 mL of water for a few minutes and then
filtered. The filtrate was mixed with 5 ml of distilled water and shaken vigorously until the
formation of froth. Saponins were confirmed by the formation of froth formation.

Detection of Diterpenes
Copper acetate test: Take 0.1g of copper acetate dissolved in 3 ml of aqueous filtrate of sample
and then mixed virgorously. Diterpenes was confirmed by the appearance of green coloration.
 6- Detection of Phlobotannins
This test was used to determination of phlobotannins in the sample. Take 5 ml of plant extract
in a test tube then adds 5 ml of 1% HCL was mixed in it. After adding of HCL then the mixture
boiled for 5 minutes. The formation of red precipitates confirmed the presence of
phlobotannins.

7- Detection of Carboxylic Acid

Take 1 ml of extract then adds a few ml of sodium bicarbonate (NaHCO3) solution added in the
extract. Effervescence was confirmed by the presence of carboxylic acid.

RESULTS

Phytochemical constituents of ethanolic extract of Moringa oleifera seeds.

S.No Constituents Reagent/Test Results: Seeds Fruit Pulp

1 Terpenoids Salkowski test - +

2 Phenols Ferric acid - -

Acetic anhydride and
3 Steroids
H2SO4
-

4 Flavonoids Alkaline reagent test +

5 Saponins Froth test +

6 Diterpenes Copper acetate + +

7 Phlobatannins 1% HCL - -

8 Carboxylic acid Sodium bicarbonate + +