Experiment No.

1
Aim: To isolate and detect the active constituent, caffeine present in tea dust.
Caffeine is a purine base (1, 3, 7 – trimethylxanthine). This is mostly produced from the
tea leaves and leaf buds synthetically. As green tea it is available from China and Japan
and as black tea from Sri Lanka and India. Leaves are green in colour, apex is blunt. Tea
leaves are the rich source of caffeine which is a weak base. Caffeine is present 1-4% in
tea leaves while 1-2% in coffee seed.
REQUIREMENTS
Apparatus: Extraction unit,TLC plate,Evaporation unit,Funnel,Test
tube,Beaker,Measuring cylinder
Chemicals: Lead acetate,H2SO4,Charcoal,Chloroform,Alcohol,Ethanol,MgO,Acetic
acid,Silica,HNO3,NaOH,Methanol.
PROCEDURE
Isolation of caffeine
1. Tea leaves (coarse powder form) is extracted with boiling water. In that hot
condition the aqueous extract is filtered. To precipitate tannin compound, the
warm extract sample is treated with lead acetate. By using dilute H2SO4
precipitation of excess lead acetate present in sample is done. To remove the
colouring matter, the filtered sample is boiled with charcoal and again filtered to
remove the charcoal. By using chloroform the filtered decolourized sample is
again extracted. After evaporation the combined chloroform extract gives caffeine
as a white colour material and by using alcohol again it is re crystallize.
2. In soxhlet extractor, by using ethanol the coarsely powdered tea leaves are
extracted. The extracted sample contains caffeine and it is adsorbed on MgO.
After treatment of 10% H2SO4 caffeine is disorbed. Finally by using chloroform
the sample is extracted and re-crystallized2.
Detection of caffeine
Murexide test: Caffeine are treated with a few crystals of HNO3 (3-4 drops) in a porcelain
dish and subjected to evaporation for drying. 2 drops of NaOH solution is added to the
residue, which gives a purple colour1.
TLC of caffeine: 1 mg of caffeine is dissolved in 1 ml of methanol or chloroform. Using
the solvent (acetate:methanol:acetic acid, 80:10:10) the sample is eluted and spot on the
TLC plate. By exposing to vapour of iodine, the dried plates are visualized at an Rf value
of 0.40-0.42.2
CONCLUSION
The active constituent caffeine was isolated and detected from the leaves of tea. It can be
used as CNS stimulant, specific analgesic in migraine.
REFERENCES
1. Kokate CK, Purohit AP, Gokhale SB. Druds containing alkaloid (caffeine). Text
book of Pharmacognosy, 51 edition. Nirali Prakashan, Pune. 2015: 15-74.
2. Shah B, Seth AK. isolation of phytopharmaceuticals. Text book of
Pharmacognosy and Phytochemistry, 2nd edition. CBS Publishers & Distributors
pvt ltd. 2014: 448.
------------------------------------------------------------------------------------------------------------
Experiment No.2
Isolation and detection of active principles, Sennoside from Senna
Aim: To isolate and detect the active constituent, sennoside present in senna.
Synonym of Indian Senna is Tinnevelly senna, Cassia senna. It consists of dried
compound leaflets of Cassia angustifolia belongs to family Leguminosae. Mostly its
cultivation is done in Tinnevelly, Madurai, Rajasthan, Gujurat and Andhra Pradesh. The
plant is a small shrub having 3-7 pairs of leaflets. The drug should be protected from light
during its storage. The major chemical constituent is sennoside-A and sennoside-B which
are anthraquinone glycoside. Both show stereoisomerism. Borntrager’s test confirms the
presence of anthraquinone.
REQUIREMENTS
Apparatus: Electronic shaker,Distillation unit,Extraction unit,Dessicator,Test
tube,Conical flask,Funnel,Filter paper,Water bath
Chemicals: Benzene,70% methanol,HCl, calcium chloride,Ammonia,P2O5,Oxalic
acid,Methanol, Triethylamine,Calcium chloride
PROCEDURE
Isolation of sennoside
Method-1:
Make dried and powdered sample. For two hours the sample is shaken with benzene on
an electronic shaker. The solvent is filtered and distilled followed by drying at room
temperature. With 70% methanol, the sample is extracted for 5-6 hours and the extract is
filtered under vacuum followed by re-extraction again for 2 hour and filtrate is collected.
The methanolic extract sample is combined and concentrated in to 1/8th portion of its
initial volume. Using HCl the concentrated sample is acidified to a pH of 3.0-3.2. At 5ºC
temperature, the acidified sample is kept aside. The sample is filtered and anhydrous
calcium chloride is added by dissolving in 25 ml of denatured spirit with vigorous
stirring. By using ammonia the pH is adjusted to 8 and it is kept aside for 1-2 hour.
Finally the sample is filtered and the obtained precipitate is dried by using P2O5 in a
dessicator.
Method-2:
By shaking with ethanolic chloroform for 30 minutes, the powdered drug sample is
extracted (chloroform: ethanol = 93:7). Again the extraction is carried out by using acidic
methanol. Acidic methanol is prepared by using 1.2 g of oxalic acid per litre of methanol.
Both the extracted sample is combined and concentrated. This sample is allowed to stand
for overnight at room temperature. Then sennoside-B remains in solution and sennoside-
A is precipitated. Re-crystallization of sennoside-A is carried out using triethylamine and
precipitation of sennoside-B is carried out by 10% methanolic solution of calcium
chloride. Again the sample is separated by methanolic ammonia solution. This solution is
prepared by adding 60 ml of methanol and 40 ml of ammonia. The sample is washed and
dried with water and allowed to stand for one day followed by re-crystallization using
glycolmonoethylether.
Detection of sennoside
Different organic solvent like benzene, ether, chloroform is added to the sample and
shaken. The organic layer is separated and ammonia solution is added to it. Pink or red
colour is produced by confirming the ammonical layer which shows the presence of
anthraquinone.
TLC of sennoside
The solvent system is ethyl acetate:methanol:water having ratio 100:16.5:13.5. The
obtained purified sample is spotted in the plate of silica gel and developed in ethyl
acetate:methanol:water .When the dried plates of silica gel is sprayed with HNO3 (25%),
red coloured spots will be seen. After drying with alcoholic KOH red colour is converted
in to yellow colour.
CONCLUSION
The active constituent sennoside was isolated and detected from senna. It is used as
purgative, stimulant laxatives.
REFERENCES
1.Kokate CK, Purohit AP, Gokhale SB. Druds containing alkaloid (belladonna), Text
Book of Pharmacognosy, 51 edition, Nirali Prakashan, pune. 2015: 9.24.
2.Shah B, Seth AK. Isolation of phytopharmaceuticals, Text book of Pharmacognosy
and Phytochemistry, 2nd edition. CBS Publishers & Distributors pvt ltd. 2014: 460.
---------------------------------------------------------------------------------------------------------
Experiment No.3
Isolation and detection of active principles, atropine from belladonna
Aim: To isolate and detect the active constituent, atropine present in belladonna.
Atropine is an alkaloid containing tropane belongs to family Solanaceae. The herb of
belladonna consists of dried leaves of Atropa belladonna Linn. Mainly, it is cultivated in
England, European countries, India (Himachal Pradesh, Kashmir). For the cultivation it
needs an altitude of 1400 m above the sea level. The chemical constituents of the drug
includes 1% alkaloid, l-hyoscyamine. Atropine is the racemate form of l-form and d-form
of hyoscyamine.
REQUIREMENTS
Apparatus: Test tube,Extraction unit,Measuring cylinder,Filter paper,Funnel,Beaker
Chemicals: Ether,Benzene,Na2CO3,Acetic acid,Sodium
sulphate,Alcohol,NaOH,HNO3,KOH
PROCEDURE
Isolation of atropine
From the juice of the powdered form of drug, atropine is isolated. By using the aqueous
solution of Na2CO3, the powdered drug material is thoroughly moistened and extracted by
using ether and benzene. From the solvent, the free base of alkaloid is extracted with
acetic acid. To remove the colouring matter the acid solution is shaken with ether and
precipitation of alkaloid is carried out by using Na2CO3. Then the solution is filtered,
washed and dried. Again the dried sample is dissolved in acetone or ether and dehydrated
by using anhydrous sodium sulphate before filtration. A crude crystal of atropine is
obtained after the concentration of filtrate following cooling of the solution. For the
complete racemization of hyoscyamine to atropine, we have to dissolve the crude
crystalline sample in alcohol and NaOH solution.2
Detection of atropine
The sample solution is diluted and treated with concentrated HNO3. Evaporation of the
mixture takes place for the drying of sample on the water bath and it produce a pale
yellow colour residue. A drop of freshly prepared KOH is added to the yellow colour
residue and the colour changes the violet (Vitali-Morin reaction).
CONCLUSION
The active constituent atropinewas isolated and detected from belladonna plant. It is used
as antidote for anticholinesterase inhibitor toxicity, treatment of bradysystolic cardiac
arrest, pupil dilator, for uveitis treatment.

REFERENCES
3. Kokate CK, Purohit AP, Gokhale SB. Druds containing alkaloid (belladonna),
Text Book of Pharmacognosy, 51 edition, Nirali Prakashan, pune. 2015:15-44.
4. Shah B, Seth AK. Isolation of phytopharmaceuticals, Text book of
Pharmacognosy and Phytochemistry, 2nd edition. CBS Publishers & Distributors
pvt ltd. 2014: 447.
 Experiment No.4
Isolation and detection of active principles, diosgenin from dioscorea
Aim:To isolate and detect the active constituent, diosgenin present in dioscoria.
Diosgenin is obtained from dried tubers of Dioscorea deltoidea (Family- Diascoreaceae).
it is mainly found growing in North Western Himalayas from Kashmir and Punjab to
Nepal and China. Also found in U.A.S. and Mexico. Synonym is Yam, Rheumatism root.
Generally it is a climber containing alternate root. Chemically it contains 75% of starch
including diosgenin, sapogenase an enzyme. It is used for the treatment of rheumatic
arthritis and also used as oral contraceptives.
REQUIREMENTS
Apparatus: Test tube,Extraction unit,Refluxtion unit,Measuring cylinder,Filter
paper,Funnel,TLC plate,Mesh
Chemicals: H2SO4,HCl,Alcohol,Ethanol,Hydrocarbon
solvent,Acetone,Heptanes,Toluene,Ethyl acetate,Silica,Anisaldehyde.
PROCEDURE
Isolation of diosgenin
 Alcoholic extraction method:
The tubers are cut in to small pieces and dried under sun light. Then the dried tuber is
subjected to make a powder form and extracted with ethyl alcohol or methyl alcohol two
times for six-eight hours. This solution is filtered. The filtrate is concentrated to make a
syrupy liquid form and hydrolysed using HCl and H2SO4 for approximately 2-12 hours.
About 85% of the crude diosgenin is precipitated. Then precipitates are subjected to
filtration following water washing and purified by alcohol.
 Acid hydrolysis method:
The powder form of sample was meshed to get 100-200 meshes following refluxtion and
heated with 2-4 N mineral acid in autoclave for 2-6 hours. The solution was filtered and
the obtained hydrolate is washed with water till it become neutral. By using hydrocarbon
solvent, again it is dried and extracted for six hours. The solution is concentrated in to
about 25 ml. In refrigerator, it is kept for some time (1 hour). The crystals of diosgenin
are filtered out and then it is washed with acetone.2
 Fermentation cum acid hydrolysis method:
Fresh green leaves is collected and smashed by using a hammer mill. In the fermentation
bin, the mesh is placed and allowed for fermentation for 2 days. To reduce the moisture
content up to 7-8%, the fermented mesh should be dried. At reduced temperature, by
using mineral acid it is subjected to hydrolysis. Finally by using heptanes, the resulting
solution is extracted to obtain diosgenin.2
 Incubation cum acid hydrolysis method:
At 37 degree centigrade, the fresh sample plant material is incubated in water for few
days. Acid hydrolysis of the incubated sample was carried out and the obtained
hrdrolysed liquid is concentrated. Finally to obtain diosgenin, the solution is extracted by
using hydrocarbon solvent. (M.P.=204-207 ºc).2
Detection of diosgenin
TLC method
The obtained purified sample is dissolved in methyl alcohol is spotted in the plate of
silica gel and developed in toluene: ethyl acetate (7:3). When the dried plates of silica gel
is sprayed with anisaldehyde and sulphuric acid reagent, a dark green spot of diosgenin
will appear (Rf=0.34-0.37).
CONCLUSION
The active constituent diosgenin was isolated and detected from the root of dioscorea. It
is used as cancer chemopreventive, natural alternative to estrogens, rheumatoid arthritis,
menstrual disorders.
REFERENCES
1.Kokate CK, Purohit AP, Gokhale SB. Druds containing alkaloid (belladonna), Text
Book of Pharmacognosy, 51 edition, Nirali Prakashan, pune. 2015: 9-48.
2.Shah B, Seth AK. Isolation of phytopharmaceuticals, Text book of Pharmacognosy
and Phytochemistry, 2nd edition. CBS Publishers & Distributors pvt ltd. 2014: 451.
===============================================================
Experiment No.5
Morphology, Histology and Powder Characteristics of Clove
Aim-The present study is aimed to evaluate the morphology, histology and powder
characteristics of clove.

These are the dried flower buds of Eugenia caryophyllus belonging to the family
Myrtaceae.

REQUIREMENTS
Apparatus: Microscope,Cover slip,Blotting paper,Spirit lamp,Brush,Watch glass,Needle
Glass slide,Blade
Chemicals: Potassium,Phoroglucinol,Sodium hydroxide,Hydrochloric acid,Alcohol
Glycerin,Ferric chloride
PROCEDURE
Preparation of mount
The thinnest possible section of plant part is taken on a slide and mounted in a solution of
chloral hydrate on slightly warming the solution of chloral hydrate cover the section
carefully with cover slip. Care must be taken so that no air bubble should produce within
the cover slip. Glycerin also added to the mounted medium.
Morphology
1. Plump is heavy, about 16-20 mm long.
2. Reddish brown in colour.
3. Lower stalk-like portion called ‘hypanthium’ upper ‘crown’ or ‘cap’
4. Hypanthium subcylindrical, slightly flattened and tapering below, 10-13 mm
long, 4 mm wide, 2 mm thick.
5. Upper portion shows inferior bilocular ovary and oil glands.
6. Crown consists of calyx, corolla, stamen and style.
7. Calyx – 4 thick walled spreading sepals
8. Corolla – dome shaped with yellow, imbricate membranous petals.
9. Stamens-numerous, free, introse, tetradelphous.
10. Gynaecium-inferior bilocular ovary with numerous ovules and axile
placentation, style in the centre, nectar disc at the base.
11. Strong, spicy, aromatic odour
12. Pungent aromatic taste
13. Should sink when put in freshly boiled and cooled water
14. Exudes volatile oil when pressed between the fingers.

Identification test
hick section of hypanthium with 5% potassium hydroxide; needle shaped crystals of
potassium eugenolate are observed under microscope. A drop of chloroform extract with
3% sodium hydroxide; crystals of sodium eugenolate are formed. Alcoholic extract of
drug treated with solution of ferric chloride shows blue colour.
Histology
Transverse section through hypanthium of clove shows the following characteristics.

 Epidermis with thick cuticle with stomata.

 Collenchymatous parenchyma containing numerous schizolysigenous oil glands.
 Calcium oxalate clusters in parenchyma.
 Zone of slightly thick walled cells embedding a ring of about 15 bilateral vascular
bundles.
  Meristele enclosed by lignified pericyclic fibres.
 Xylem of 5-6 lignified vessels.
 Parenchyma containing air spaces or lacuna or aerenchyma.
 Central columella- Thick walled parenchymatous cells containing cluster
crystals of calcium oxalate and 20-25 small vascular bundles.
Transverse section through region of ovary shows the following characteristics.
 Ovary and with ovarian wall.
 Parenchymatous dessepiment
 Ovules with axile placentation.
 Starch, prisms of calcium oxalate and stone cells are absent.
Powder characteristics
 Epidermis which stomata.
 Pollen grains.
 Lignified fibres with parenchyma.
 Oil glands.
 Aerenchyma.
 Fibrous layer of anther wall.
Important constituents
Volatile oil: Eugenol
Use: antiseptic, Carminative.
CONCLUSION
Morphology, histology and powder characteristics of clove is evaluated.
REFERENCES
1.Kokate CK. Practical Pharmacognosy, 4th edition, Nirali Prakashan, Pune; 1994: 48-49.
2.http://www.yourarticlelibrary.com/biology/plants/clove-sources-cultivation-and-uses-
with-diagram.

Experiment No.6
Morphology, Histology and Powder Characteristics of Senna
Senna is obtained from dried leaf-lets of Cassia angustifolia Vahl. (Indian senna)
belonging to family Leguminosae.

Aim-The present study was aimed to evaluate the morphology, histology and powder
characteristics of senna.

REQUIREMENTS
Chemicals: Glycerine water,Phloroglucinol,Chloral hydrate,HCl,Benzene,Sulphuric acid

Apparatus: Microscope,Microscope slide

PROCEDURE
Preparation of mount

The thinnest possible section of plant part is taken on a slide and mounted in a solution of
chloral hydrate on slightly warming the solution of chloral hydrate cover the section
carefully with cover slip. Care must be taken so that no air bubble should produce within
the cover slip. Glycerin also added to the mounted medium. 1

Morphology

Paripinnately compound leaf about 2.5 to 6.0 cm long.

Leaflet lanceolate with entire margin, reticulately pinnate venation, acute and
mucronate apex, asymmetrical base.
Outer surface pubescent with pressure markings
 Pale green in colour.
Mucilaginous and slightly bitter taste.

Histology

A transverse section shows

Upper and lower epidermis- polygonal tubular cells with straight anticlinal walls.

Mucilage in the inner periclinal walls, stains red with ruthenium red.
Epidermal trichomes- unicellular, conical, thick walled with warty cuticle, curved at
base.
Palisades – a single layer below upper and lower epidermis (isobilateral) continuous
over the meristele but absent underneath.
Rubiaceous or paracytic stomata.
Large veins accompanied by calcium oxalate crystals.
Cluster crystals of calcium oxalate in palisade and spongy tissue, crystal sheath in
mid-rib region.
Spongy tissue of parenchymatous cells
 Meristele of radiate xylem and phloem with an arc of pericyclic fibres below and
sclerenchyma above.
Powder characteristics
1. Epidermis with paracytic stomata
2. Calcium oxalate prisms
3. Trichomes
4. Palisade and spongy cells
5. Xylem vessels with annular thickening
6. Crystal sheath
Important constituents
Anthracene glycosides: sennosides A & B.

Identification test

Borntrager test: Boil a few leaves with dilute sulphuric acid, filter hot and to the cooled
filtrate add organic solvent like benzene or there (5 ml), shake well. Separate the organic
solvent layer and add equal volume of dilute ammonia solution, shake well. Ammoniacal
layer acquire rose pink colour.
Use: Laxative
CONCLUSION
Morphology, histology and powder characteristics of senna is evaluated and identified.
REFERENCES
1. Kokate CK. Practical Pharmacognosy, 4th edition, Nirali Prakashan, Pune; 1994:
40-41.
2. http://www.yourarticlelibrary.com/biology/glycoside/tinnevelly-senna-leaves-
sources-collection-and-uses.

Experiment No.7
Morphology, Histology and Powder Characteristics of CORIANDER
Synonyms Fructus coriandri, Coriander fruits, Cilantro, Chinese parsley.
Biological Source Coriander consists of dried ripe fruits of Coriandrum sativum Linn.,
belonging to family Umbelliferae.
Geographical Sources
Cultivated in Central and Eastern Europe, particularly in Russia, Hungary, in Africa and
India. In India it is cultivated in Maharashtra, U.P., Rajasthan, Jammu, and Kashmir. It is
also found in a antiwild state in the east of England.
Cultivation and Collection
 The coriander seeds are sown in dry weather either in March or in early autumn. Shallow
drills, about 1/2 inch deep and 8 inches apart are made and the seeds are sown in it, the
rate of germination is slow. The plants are annual herb, which grow to a height of 1 to 3
feet high, slender, and branched. The flowers are in shortly stalked umbels with five to
ten rays. The seeds fall as soon as ripe and when the seeds are ripe (about August), the
disagreeable odour is produced. Plant is then cut down with sickles; the fruits are
collected and dried. During drying fruits develop aromatic smell and the unpleasant
odour disappears.
Characteristics
The fruit is a cremocarp, subspherical in shape, Yellowish-brown in colour. The size of
the fruit is 3 to 4 mm in diameter, with aromatic odour, and spicy, aromatic taste.

Coriandrum sativum
Powder characteristics
Sclerenchymatous layer: Groups of fusiform fibres of sclerenchyma running way and at
times crossing with each other or with thin walled lignified cells of the mesocarp.
Endocarp: Fragments of parquetry arrangement of thin walled lignified cells with the
polygonal cells of mesocarp.
Microscopy
The transverse section of coriander shows the presence of a dorsal surface and a
commissural surface. The dorsal surface consists of two vittae and a carpophore. The
dorsal surface has five primary ridges and four secondary ridges. The epicarp consists of
a single row of small thick-walled cells with calcium oxalate crystals. The mesocarp has
an outer loosely arranged tangentially elongated parenchyma cells and the middle layer
consisting of sclerenchyma. The middle layer is again divided into; the outer region of
sclerenchyma is represented by longitudinally running fibres, whereas the inner region
has tangentionally running fibres. The vascular bundles are present below the primary
ridges. The inner layer has polygonal, irregularly arranged parenchyma cells. The
endocarp has the parquetry arrangement. In the testa it has single-layered, yellowish cells,
and the endosperm is thick, polygonal, colourless parenchyma with fixed oil and aleurone
grains.
 Transverse section of coriander fruit (mericarp)

Chemical Constituents
Coriander consist of about 1% of volatile oil the chief volatile components are D-(+)-
linalool (coriandrol), along with other constituents like, borneol, p-cymene, camphor,
geraniol, limonene, and alpha-pinenes. The fruits also contain fatty oil and
hydroxycoumarins. The fatty oils include acids of petroselic acid, oleic acid, linolenic
acid, whereas the hydroxycoumarins include the umbelliferone and scopoletine.

Uses
Aromatic, carminative, stimulant, alterative, antispasmodic, diaphoretic and flavouring
agent. It is also used as tonic, appetizer, diuretic, aphrodisiac, and stomachic. Coriander
can be applied externally for rheumatism and painful joints. The infusion of decoction of
dried fruit of cardamom is useful for the treatment of sore-throat, indigestion, vomiting,
flatulence, and other intestinal dis-orders.
Experiment No.8
Morphology, Histology and Powder Characteristics of Fennel
Synonyms Fructus foeniculli, Fennel fruit, Fenkel, Florence fennel, Sweet fennel, Wild
fennel, Large fennel.
Biological Source-Fennel consists of the dried ripe fruits of Foeniculum vulgare Miller.,
belonging to family Umbelliferae.
Geographical Source
Fennel is indigenous to Mediterranean countries and Asia; it is largely cultivated in
France, Saxony, Japan, Galicia, Russia, India, and Persia.
Characteristics
The fruit is an entire cremocarps with pedicels, oval-oblong and 5 to 10 mm long, 2 to 4
mm broad. It has greenish-brown to yellowish brown colour with five prominent primary
ridges and a bifid stylopod at the apex.

Foeniculum vulgare
Microscopy
The transverse section of mericarp region of fennel shows two prominent surfaces, the
dorsal and the commissural surface. The commisural surface has a carpophore and two
vittae, and the dorsal surface has a total of five ridges. The mericarp is divided into
pericarp, consisting of the epicarp and mesocarp; the testa and the endocarp. Epicarp
consists of polygonal cells of epidermis which are tangentially elon-gated and covered by
the cuticle. Mesocarp has parenchyma cells with five bicollateral vascular bundles; below
each primary ridge a lignified reticulate parenchyma surrounds the vascular bundles.
There are four vittae on dorsal surface and two vittae on commisural or the ventral
surface. Inner Epidermis or Endocarp shows parquetry arrangement (a group of four to
five cells arranged parallelly at acute angles with groups of similar cells in different
direction). Testa is a single-layered tangentially elongated cell with yellowish colour.
Endosperm consists of thick-walled, wide polyhedral, colourless cells. Cells contain fixed
oil, aleurone grains, and rosette crystals of calcium oxalate.
 Transverse section of Fennel fruit (Mericarp)

Powder characteristics
 Mesocarp: Lignified and reticulate nature of the parenchyma.
 Endocarp: Cells showing parquetry arrangement.
 Endosperm: Polyhedral, thick walled cells containing aleurone grains, minute calcium
oxalate crystals and oil globules.
 Vittae: Many in the form of yellowish brown fragments.
Chemical Constituents
The best varieties of Fennel contain 4 to 5% of volatile oil. The primary constituents of
volatile oil are 50 to 60% of anethole, a phenolic ester; and 18 to 22% of fenchone, a
ketone. Fenchone is chemically a bicyclic monoterpene which is a colourless liquid and
the odour and taste is pungent and camphoraceous. The oil of Fennel has β-pinene, anisic
acid, phellandrine, and anisic aldehyde. Fennel also contains about 20% fixed oil and
20% proteins.
Uses
Fennel is used as stomachic, aromatic, diuretic, carminative, diaphoretic, as a digestive,
pectoral, and flavouring agent. Anethole may have estrogen-like activity and inhibit
spasms in smooth muscles. Fennel can increase production of bile, used in the treatment
of infant colic, to promote menstrua-tion in women, can increase lactation, act as
antipyretic, antimicrobial and anti-inflammatory.
Adulterants
Fennel is generally adulterated with exhausted fennel and due to improper caring during
harvesting they are also adulterated with sand, dirt, stem, weed seeds, etc in which part of
volatile oil is removed either by extraction with alcohol or steam distillation. Fruits
exhausted by water or steam are darker in colour, contain less essential oil and sink in
water, but those exhausted by alcohol still hold 1 to 2% of oil in them.

===============================================================

Experiment No.9
Morphology, Histology and Powder Characteristics of CINCHONA
Synonyms Cortex Cinchonae, Countess, Peruvian or Jesuit’s bark, Cinchona
Biological Source Cinchona is the dried bark of the stem or of the root of Cinchona
calisaya Wedd., Cinchona ledgeriana Moens., Cinchona officinalis Linn., and Cinchona
succirubra Pavon., or hybrids of any of the first two species with any of the last two
species, belonging to family Rubiaceae.

Powder characteristics of cinchona shows

1. Crystals of calcium oxalate are present in the cortex.

2. The phloem fibers are spindle shaped. However, the distribution of phloem fibres
varies among the different species of Cinchona.
3. Starch grains are about 6-10 cm diameter.

Characteristics
 Twig and bark of Cinchona ledgeriana

Microscopic Characters
Transverse section of bark shows cork composed of uniformly arranged several layers of
thin-walled cells, containing amorphous reddish-brown matter. Below cork is a redion of
cortex, composed of tangentially elongated parenchymatous cells with red-brown and
thin walls, containing small starch grains. Idioblasts, containing micro-crystals of calcium
oxalate (2–6 μ long), and secretion cells are scattered in the cortex. Phloem consists of
compressed and collapsed sieve tubes, phloem parenchyma similar to cortex, and
irregularly arranged, large spindle-shaped lignified fibres. Medullary rays are narrow,
two to three cells wide and almost straight. Longitudinal section of bark shows brick-
shaped cells of medullary rays, longitudinally elongated cells of phloem parenchyma, and
fibres with conspicuous pits.

Transverse section of Cinchona bark

Chemical Constituents
More than 30 alkaloids have been reported in cinchona. The chiefly identified alkaloids
are quinidine, quinine, cinchonine and cinchonidine. These constituents are the
stereoisomers of each other like quinine is stereoisomer of quinidine and cinchonine is
stereoisomer of cinchonidine. The other constituents available are quiniarnine,
cinchotine, hydroquinine, hydrocinchonidine, cinchotannic acid, etc. Quinine and
quinidine has a methoxy group in it but cinchonine and cinchonidine do not have a
methoxy group. Other than these it also consist of bitter glycoside, starch grains, calcium
oxalate crystals and crystalline acid like quinic acid.
Chemical Test
1)Thalleioquin test: To the extract of cinchona powder add one drop of dilute sulphuric
acid and 1 ml of water. Add bromine water drop wise till the solution acquires per-
manent yellow colour and add 1 ml of dilute ammonia solution, emerald green colour is
produced.
2) The powdered drug when heated with glacial acetic acid in dry test tube, evolves red
fumes, which con-dense in the top portion of the tube.
3)Cinchona bark, when moistened with sulphuric acid and observed under ultraviolet
light shows a blue fluorescence due to the methoxy group of Quinine and quinidine.
Uses
It is mainly employed as antimalarial drug, but it is also used as analgesic, antipyretic,
protoplasmic, bitter stomachic and tonic. Quinidine is cardiac depressant and
Cinchonidine is used in rheumatism and neuralgia.
Substitutes
Cuprea Bark (Remijia pedupiculato); Family: Rubiaceae, it differs in its morphological
character with cinchona but consist of constituents like Quinine, quinidine, cinchonine,
cinchonamine, etc., the other species of Remijia, that is, R. purdieana (false Cuprea bark)
does not contain quinine.
===============================================================

Experiment No.10
Morphology, Histology and Powder Characteristics of CINNAMON
Synonyms-Cortex cinnamoni, Ceylon cinnamon, Saigon cinnamon, Chinese
cassia, Cinnamomum aromaticum, Cinnamomum laurus.
Biological Source -Cinnamon is the dried inner bark of the coppiced shoots
of Cinnamomum zeylanicum Nees., belonging to family Lauraceae.

Geographical Sources Cinnamomum zeylanicum is widely cultivated in Ceylon, Java,

Sumatra, West Indies, Brazil, Mauritius, Jamaica, and India.

Morphological Characteristics
Cinnamon are either in single- or double-compound quills, with a size of 1 m length, 0.5
mm thickness, and 6 to 10 mm diameter. The outer surface has yellowish brown colour
having longitudinal lines of pericyclic fibre and scars and holes representing the position
of leaves or the lateral shoots. The inner surface is darker than the outer. Cinnamon has a
fragrant perfume; taste aromatic and sweet.

Leaf and bark of Cinnamomum zeylanicum

Microscopy
The transverse section shows the presence of three to four layers of sclereids which are
horse shoe shaped consisting of starch grains. The pericyclic fibres (6 to 15) are present
on the outer margin. It consists of sieve tubes which are completely collapsed and are
arranged tangentially; lignified phloem fibres, arranged as tangential rows of four to five
cells; biseriate medullary rays with needle-shaped calcium oxalate crystals; longitudinally
elongated idioblast consisting of volatile oil; sub-rectangular parenchyma cells with
starch grains and calcium oxalate crystals.
 T.S. (schematic) Cinnamon bark

Transverse section of Cinnamon bark

Chemical Constituents
Cinnamon contains about 10% of volatile oil, tannin, mucilage, calcium oxalate and
sugar. Volatile oil contains 50 to 65% cinnamic aldehyde, along with 5 to 10% eugenol,
terpene hydrocarbons and small quantities of ketones and alcohols.
Powdered cinnamon contains numerous thick-walled and pitted phloem in fibres,
isolated or in groups, abundant starch grains and acicular microcrystals of calcium
oxalate in parenchymatous cells, and a few sclerenchymatous cells.

Chemical Tests
1. A drop of volatile oil is dissolved in 5 ml of alcohol and to it a drop of ferric chloride is
added, A pale green colour is produced. Cinnamic aldehyde gives brown colour with
ferric chloride, whereas eugenol gives blue colour.
2. The alcoholic extract is treated with phenylhydrazine hydrochloride, it produces red
colour due to the formation of phenylhydrazone of cinnamic aldehyde.
Uses
It is used as an alterative, aromatic, carminative, flavouring agent, analgesic, antiseptic,
antirheumatic, antispasmodic, demulcent, digestive, expectorant, stomachic, diaphoretic,
antibacterial, antifungal, etc. It stops vomiting, relieves flatulence and is given with chalk
and as astringents for diarrhoea and haemorrhage of the womb. It is also used in the
treatment of bronchitis, colds, palpitations, nausea, congestion, and liver problems.
Other Species
Cinnamon cassia is often used as a substituent. C. culiawan is native of Amboyna and
the bark has the flavour of clove, C. iners, Cassia burmarin, Saigon cinnamon, and C.
nitidum are also used.