PHYTOCHEMISTRY AND PHARMACOGNOSY II, UNIT III, ISOLATION IDENTIFICATION AND ANALYSIS OF GLYCOSIDES

1. SUB –
PHYTOCHEMISTRY AND
PHARMACOGNOSY II
UNIT III
ISOLATION
IDENTIFICATION AND
ANALYSIS OF
GLYCOSIDES
Prepared by : Snehal Uttam Kashid (Asso. professor)
Department of Pharmacognosy, SPMs college of pharmacy,
akluj

2. GLYCYRRHETINIC ACID

3. Glycyrrhetinic acid
 Synonyms: Liquorice, Sweet Wood, Regolizia (Italian),
Jeshtamadh, Yastimadhu
 Biological source - It is obtained from the roots and
stolons of Glycyrrhiza glabra
 Family – Leguminosae
 Chemical Constituents – A major C.C. is sweet
triterpenic saponin glycoside , glycyrrhizin (It is a
potassium and calcium salt of Glycyrrhizic acid
Glycyrrhetinic acid) is a Pentacyclic triterpenoid
aglycone.

4. Isolation of glycyrrhetinic acid
Method 1:
The Liquorice / Glycyrrhiza coarse powder is extracted
with chloroform.
Filter and discard the filtrate.
Extract the marc with 0.5 M Sulphuric acid for a few hours
Filter and extract the filtrate with three portions of
chloroform

5. Isolation of glycyrrhetinic acid
Separate and combine the chloroform layers
Distill off the chloroform extract to yield a dry residue of
glycyrrhetinic acid.
White crystalline powder, insoluble in water, soluble in
chloroform, benzene, ether etc

6. Isolation of glycyrrhetinic acid
Method :2 (Acid precipitating method)
weigh 50g liquorice powder add 300ml water in the beaker
and boil with stirring
decant the supernant liquid
filter the remaining residue and collect the filtrate
Adjust pH 2.8 by the addition of acid, Glycyrrhizin
precipitates out

7. Isolation of glycyrrhetinic acid
Filter and collect the ppt
Wash the ppt with cold water to make it free from acid
Transfer the ppt to china dish and heat gently to remove
the water content
shiny brown mass of glycerrhizin is observed

8. Identification of glycyrrhetinic
acid
Thin layer chromatography (TLC)
 Sample preparation – Dissolved about 1mg of
Glycyrrhetinic acid in 1ml of methanol- Chloroform
(1:1)
 Stationary phase - Silica gel –G
 Detecting agent – 1% vanillin- Sulphuric acid and
heat for 10 minutes at 1100 C
 Mobile phase – Toluene–Ethyl acetate-Glacial
acetic acid (12.5:7.5:0.5)
 Standard - Glycyrrhetinic acid
 RF Value – Purplish – 0.41

9. Thin layer chromatography
(TLC)

10. Identification of glycyrrhetinic
acid
Chemical tests –
 Liebermann- Burchard test
2 ml test extract with 1ml chloroform in a dry test tube.
Add several drops of acetic anhydride and then 2 drops
of concentrated H2SO4 and mix carefully. Blue green to
red orange color develops. Steroidal moiety present.
 Liebermann test –
2 ml of test extract is mixed with acetic anhydride boil the
solution and add 0.5 ml H2SO4 which develops blue
color, Phenol present.

11. Uses
 Glycyrrhizic acid is widely applied in foods as
a natural sweetener.
 anti-inflammatory
 anti-ulcer
 anti-allergic
 Antioxidant
 anti-tumor
 anti-diabetic
 hepatoprotective

12. Rutin

13. Rutin
 Rutin is flavonoidal compound obtained from buckwheat,
apricots, cherries, grapes, grapefruit, plums, and oranges.
 Biological sources and family -
1. it is also extracted from buck wheat of Fagopyrum
esculentum
Family - Polygonaceae
2. Ruta – Ruta graveolens
Family - Rutaceae

14. Rutin
 There are around 200 types of Quercetin, Flavanoid
glycosides, among this the rutin is the one of most
important type.
 It is chemically Quercetin-3- rutinoside . On hydrolysis ,
it yields the aglycone quercetin and the sugars glucose
and rhamnose.
 It is used as a Vitamin P OR Capillary fragility factor

15. Isolation of Rutin

16. Identification of Rutin
Thin layer chromatography (TLC)
Sample preparation – Dissolved about 1mg of Rutin in
1ml of methanol
Stationary phase - Silica gel –G
Mobile phase – Toluene:ethyl acetate:formic acid with
ratio (5:4:0.5, v/v/v)
Standard drug - Rutin
RF Value – Yellow spot – 0.43

17. Identification of Rutin
Chemical tests –
 Shinoda test –
A few fragments of magnesium ribbon and concentrated
hydrochloric acid were added to the ethanolic extract of
rutin. The appearance of red to pink color after few
minutes indicates the presence of flavonoids.

18. Uses
 antioxidant
 strengthen blood vessels
 use it for internal bleeding
 hemorrhoids
 prevent strokes due to broken veins or arteries
(hemorrhagic strokes).
 prevent a side effect of cancer treatment called
mucositis.

19. Bibliography
 Kokate, C.K., Purohit, A.P. and Gohkale, S.B.
(2019) Pharmacognosy. In Terpenoids, 57th Edition,
Nirali Prakashan, Pune.
 Pharmacognosy And Phytochemistry By Vinod Rangari.
2009 2nd edition, Career Publications.