Useful for B.Pharm students

2. QUININE
Synonym – Quinine
It is a quinoline alkaloid of cinchona bark. The other
important alkaloids of this drug are quinidine,
cinchonine, cinchonidine, cinchonamine etc.,
Biological sources : It consists of dried inner bark of
C.Calisaya, C.succirubra, C.officinalis, C.ledgeriana
and hybrids of this. Family – Rubiaceae

3. Quinine and quinidine are stereo-isomers .
Quinine is levorotatory and quinidine is dextrorotatory
Uses :
Quinine is antimalarial
Quinidine is a cardiac depressant therefore used in
cardiac arrhythmias.

4. ISOLATION
1. The dry powder bark material is first well mixed
with about 30% of its weight of calcium hydroxide
or calcium oxide and sufficient quantity of sodium
hydroxide solution to make a paste. It is allowed to
stand for few hours.
2. The mass is then transferred to a Soxhlet
apparatus and extraction is carried out with
benzene.

5. 3.Subsequently the benzene extract is shaken with
successive portions of 5% sulphuric acid.
4.The aqueous acid extract is adjusted the pH 6.5 with
dilute sodium hydroxide, cool. Crystals of neutral
quinine sulphate are formed.
5.These crystals are freed from cinchonine and
cinchonidine by repeated recrystallization from hot
water.

6. 6.Colouring matter is removed by activated charcoal.
7.Quinine sulphate crystals are dissolved in dilute
sulphuric acid and made alkaline with ammonia.
Initially amorphous quinine is formed , which
becomes crystalline.
8.Finally washed to remove sodium and ammonium
salts and dried to 45- 55 o C.

7. IDENTIFICATION AND ANALYSIS
1. Chemical tests
2. Thin layer chromatography (TLC)

8. T.L.C Method
Sample preparation – Dissolved about 1mg of Quinine
or
Cinchona alkaloid in 1ml of methanol
Stationary phase - Silica gel –G
Detecting agent – Dragendroffs reagent
Mobile phase – Chloroform – Diethylamine (9:1)
RF Value – Quinine – 0.17, Quinidine -0.26

10. GLYCYRRHETINIC ACID
Biological source - It is obtained from the roots and
subterranean stems of Glycyrrhiza glabra
Family – Leguminosae
Chemical Constituents – A major component is sweet
triterpenic saponin glycoside , glycyrrhizin
Glycyrrhizin – It is a potassium and calcium salt of
Glycyrrhizic acid
Glycyrrhetinic acid is a Pentacyclic triterpenoid
aglycone. It is used as an antiulcer.

11. ISOLATION
• 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

12. • Filter and extract the filtrate with three portions of
chloroform
• 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

13. IDENTIFICATION AND ANALYSIS
1. Chemical tests – Liebermann test and
Liebermann – Burchard test
2. Thin layer chromatography (TLC)

14. T.L.C Method
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)
Reference drug - Glycyrrhetinic acid
RF Value – Purplish – 0.41

16. 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

17. SOURCES OF RUTIN
• 1. Fagophyrum esculentum- ( Polygonaceae )
• 2. Rhubarb – ( Rheum emodi- ( Polygonaceae )
• 3. Tobacco – (Nicotiana tobaccum –( Solanaceae )
• 4. Ruta – (Ruta graveolens – ( Rutaceae )
• 5. Tea – Thea sinensis – ( Theaceae )
• 6. Eucalyptus macroryncha ( Myrataceae )

18. ISOLATION
• Source -
• Eucalyptus macroryncha ( Myrataceae )
• Boil the powder drug with boiling water.Filter while
hot and collect the filtrate . Cool for the precipitation
of the rutin.
• Recrystallize it from boiling water , dry the product.
• Greenish yellow crystalline powder

19. IDENTIFICATION AND ANALYSIS
1. Chemical tests – Shinoda test
2. Thin layer chromatography (TLC)

20. T.L.C Method
Sample preparation – Dissolved about 1mg of
Rutin in 1ml of methanol
Stationary phase - Silica gel –G
Mobile phase – 10 % aqueous sodium chloride
solution
Standard drug - Rutin
RF Value – Yellow spot – 0.43

22. PODOPHYLLOTOXIN
• Podophyllotoxin is the Lactone resin present in the
root and rhizome of Podophyllum hexandrum
• Family – Berberidaceae
• It is used as anti-proliferative agent (Anti cancer
agent)

23. EXTRACTION
• Take the weighed quantity of rhizomes or roots of
Podophyllum emodi with methanol. Filter and
evaporate to semisolid mass.
• Dissolve semisolid mass into acidic water. Precipitate
is formed which should be allowed for at least for 2hrs

24. • Filter and wash filtrate with cold water. Collect the
residue, wash with acidified water and dry to
obtain dark brown amorphous powder.
• Extract the residue with hot alcohol. Filter and
evaporate to dryness.
• Re- crystallise the residue in benzene to yield
podophyllotoxin

25. IDENTIFICATION AND ANALYSIS
• 1. Chemical test –
Treat podophyllotoxin with 50% Sulphuric acid it will
show violet – blue colour.
• 2.Thin layer chromatography ( TLC )

26. T.L.C Method
Sample preparation – Dissolved about 1mg of
podophyllotoxin in 1ml of methanol
Stationary phase - Silica gel –G
Mobile phase –
Chloroform : Methanol (90:10) for about 6 cm (Only
glycosides are separated but aglycone like
podophyllotoxin
remains in the region of the front.

27. The same plate is again eluted with more weakly polar
Solvent
Chloroform : Acetone (65:35) upto 12cm
Standard drug – Podophyllotoxin
Detection – Spray with methanol Sulphuric acid and
heat
10 minutes at 1100 C
RF Value – Yellow spot – 0.65