PHYTOCHEMISTRY

(INTRODUCTION, PLANT
METABOLITES AND
PHYTOCHEMICAL SCREENING

Uswatun Khasanah, S.Farm.,M.Farm.,Apt

Farmasi Bahan Alam
Prodi Farmasi FKUB
REFERENCES
CONTENT
definition

SAROYA, 2011
PLANT METABOLITES

HOW and WHY

these
compounds
synthesized?

Kaufman et al.,1998
METABOLITES
PATHWAY
The jacket that encloses this cell is
the cell wall. This is the primary site for
polymerization of cell wall
polysaccharides (cellulose,
hemicellulose, and pectic
polysaccharides), lignin, and
amorphous silica gel in plants that
accumulate this polymer.

Kaufman et al.,1998
METABOLITES
PATHWAY
Primary Metabolites
LIPID
PROTEIN AND
NUCLEOTIDS
CELLULOSE
Primary metabolites
function
Secondary Metabolites
Secondary Metabolites
classification
ALKALOID
Fenol, FLAVONOID
terpenoid
secondary metabolites
function
PHYTOCHEMICAL SCREENING
purpose
method
SCREENING
SCREENING
ALKALOID SCREENING
ALKALOID SCREENING
ALKALOID SCREENING
ALKALOID SCREENING
PROCEDURE
1. Mayers test: Filtrates were treated with Mayers reagent (Potassium
Mercuric iodide). Formation of yellow coloured precipitate indicates the
presence of alkaloids.

2. Wagners test: Extracts when treated with Wagners reagent (iodine in

potassium iodide). Formation of brown /reddish
precipitate indicates the presence of alkaloids.

3. Hagers test: Filtrates were treated with Hagers reagent (saturated picric
acid solution). Presence of Alkaloids confirmed by the formation of yellow
coloured precipitate.
 ALKALOID SCREENING

1. Mayers Test 2. Wagners test 3.Hagers test

ALKALOID SCREENING USING TLC

Alkaloid spot in TLC

using dragendorf
reagents
FLAVONOID SCREENING
FLAVONOID SCREENING
FLAVONOID SCREENING

1. Alkaline reagent test: The Extracts were treated with few

drops of sodium hydroxide solution. Formation of intense
yellow colour, which becomes colour less on addition of
dilute , indicates the presence of flavanoids.

2. Lead acetate Test: The Extracts were treated with few drops
of Lead acetate solution. Formation of yellow colour
precipitate indicates the presence of flavanoids.

3. Ferric chloride test: 1 ml of the extract was treated with 1

ml of ferric chloride.Formation of brown colour precipitates
indicates the presence of flavanoids
1. Alkaline reagent test 3. Ferric chloride test

2. Lead acetate Test

FLAVONOID SCREENING
FLAVONOID SCREENING

apigenin fluorescent spots

when observed at 254 nm
after derivatisation with NP-
PEG reagent.
FLAVONOID SCREENING
SAPONIN DETECTION
Foam test: o.5gm of extract was shaken
with 2 ml of water. It foam produced
persists for ten minutes it indicates the
presence of saponins.
TANNIN DETECTION
1. Gelatin Test: To the extract, 1% gelatine solution containing
sodium chloride was added. Formation of white precipitate
indicates the presence of Tannins.

2. Modified Prussian blue test: To 1 ml of extract, add 1 ml .008M

potassium ferric cyanide and 1 ml of .oo2 M ferric chloride in 0.01
M HCl. Presence of blue colour indicates the presence of tannins.

3. Lead acetate Test: To the extract few drops of Aqueous basic lead
acetate solution were added, reddish brown bulky precipitate
indicates the presence of tannin.

4. Ferric chloride Test: To the extract, few drops of 1%natural ferric

chloride solution was added formation of blackish blue colour
indicates the presence of tannins
 Gelatin test ferrichloride test
PHENOL DETECTION
Ferric chloride Test: Extracts were
treated with 3-4 drops of ferric chloride
solution. Formation of bluish black
colour indicates the presence of phenols.
TANIN SCREENING
ANTHRAQUINONE DETECTION
Borutragers Test: To 1 ml of the extract , add 1 ml of 10 % ferric chloride
and .5 ml of concentrate hydrochloric acid. Boil in a water bath for few
minutes. Filter it and the filtrate is treated with 1 ml of Diethyl ether and
concentrate ammonia. Appearance of pink or deep red colour.
Detection of Cardiac glycosides

1. Legals test: Extracts

were treated with
sodium nitropruside
and pyridine and
sodium hydroxide.
Formation of pink to
red colour indicates the
presence of cardiac
glycosides.
TERPENOID AND STEROID
DETECTION
1. Salkowskis test :5ml of each extract was made mixed in 2 ml of chloroform and
concentrate sulphuric acid was carefully added to form a layer. A reddish brown
coloration of the interface was formed to show positive result for presence of terpenoids.

2. Copper acetate Test: Extract were dissolved in water and treated with 3-4 drops of copper
acetate solution. Formation of emerald green indicates the presence of terpenoids.

1.Salkowskis Test 2. Libermann Burchards Test

SAPONIN
(STEROID/TERPENOID) SCREENING

SPRAY USING
anisaldehide REAGENTS
SAPONIN
(STEROID/TERPENOID) SCREENING