1. Isolation of Important Phyto Drug
Term Paper leading to Thesis
By
Milantirtha Mete
Exam. Roll no.-M2PHA18023
Class roll no.-001711402026
Under the guidance of
Prof. (Dr.)Subhash C. Mandal
Pharmacognosy & phytotherapy Research Laboratory
Division of pharmacognosy
Department of Pharmaceutical Technology
Jadavpur University
Kolkata-32
2018
2. Isolation
Isolation is the crucial first step in the analysis of
medicinal plants, because it is necessary to extract the
desired chemical components from the plant materials
for further separation and characterization. The basic
operation included steps, such as pre-washing, drying of
plant materials or freeze drying, grinding to obtain a
homogenous sample and often improving the kinetics of
analytic extraction and also increasing the contact of
sample surface with the solvent system.
3. A brief summary of the experimental conditions for
various methods of extraction for plants material
Soxhlet
extraction
Sonification Maceration
Common
Solvents
used
Methanol,
ethanol, or
mixture of
alcohol and
water
Methanol,
ethanol, or
mixture of
alcohol and
water
Methanol,
ethanol, or
mixture of
alcohol and
water
Temperature
(°C)
Depending on
solvent used
Can be
heated
Room
temperature
Time
required
3–18 hr 1 hr 3–4 days
Volume of
solvent
150–200 50–100 Depending on
the sample
size
4. Identification and
characterization
It is a common practice in isolation of these
bioactive compounds that a number of different
separation techniques such as TLC, column
chromatography, flash chromatography and HPLC,
should be used to obtain pure compounds. The pure
compounds are then used for the determination of
structure and biological activity.
5. . EXTRACTION OF STARCH FROM POTATOES
BIOLOGICAL SOURCE-
Starch is a polysaccharide obtained commercially from wheat, maize, potato and
rice. It is present in the form of grains in different parts of the plant and chemically
consists of amylopectin and amylose
PROCEDURE OF EXTRACTION
. Wash starch 2-3 times with
distilled water with constant
stirring , centrifuge the milky liquid
& dry it in oven
Allow the
milky liquid to
settle down.
Decant the
supernatant
liquid
the slurry
through
shaking
sieves in
order to
remove the
cell debris
. Wash
potatoes
thoroughly
with water
to remove
adhering soil
STANDARDIZATION OF
STARCH
1.An Iodine test
2.Microscopic evaluation
of powdered starch
CONCLUSION-After performing this experiment ,2.75%yield
was obtained
6. EXTRACTION OF CALCIUM CITRATE FROM LEMONS
Citric acid is one of the commonly distributed plant acids.
The lemon fruit (family Rutaceae)
about 50 ml of lemon juice ,add 15% sodium
hydroxide with constant stirring until the mixture is
slightly alkaline ,as indicated by colour change from
yellow to brown
Strain the mixture through muslin cloth to remove
pulp particles and then through buechner funnel .If
the filter paper gets clogged ,change it as required to
complete filtration
Add 5 ml of 10% calcium chloride solution for each
10 ml of filtrate ,heat to boiling and filter off
while hot copious precipitate of calcium citrate
through Buechner funnel
Resuspend the precipitate in minimum quantity of
cold water, heat to boiling ,filter and allow the
salt to air dry .
METHOD FOR
EXTRACTION
CONCLUSION-After performing this experiment ,11.02%yield was obtained
7. IDENTIFICATION OF CALCIUM
CITRATE
To a neutral solution of the substance under examination
add a solution of calcium chloride, no precipitate is
produced.
Boil the solution, a white precipitate is formed which is
soluble in 6M Acetic acid
USES IN PHARMACEUTICAL INDUSTRY –
Antioxidant
8. EXTRACTION OF PIPERINE FROM BLACK PEPPER
Extract the powdered drug (20 g)
with 250 ml of ethanol(95%) ina
soxhlet apparatus for 3 hours
Filter the solution and concentrate
under vacuum on a water bath at 60
degree C.
Add 20 ml of 10 % alcoholic potassium
hydroxide with constant stirring to
the concentrated extract and filter
PROCEDURE
USES IN PHARMACEUTICAL
INDUSTRY –
Aromatic. Stomachic. Stimulant.
Carminative.
Anti tussive agent
CONCLUSION-After performing this experiment ,2.3%yield was obtained
BIOLOGICAL SOURCE-
It consists of dried unripe fruits of
Pipper nigrum Linn.
FAMILY-
Piperaceae
9. STANDARDIZATION OF
PIPERINE
TLC was done to standardize obtained sample against
standard Rf of 0.45.
Mobile phase used = toluene:methanol in the ratio 85:15
Solvent was run and then the TLC plate was introduced
in the UV chamber.
Fluorescent Spots were observed and solute front and
solvent front calculated.
Distance travelled by solute = 2.5cm
Distance travelled by solvent= 5.6 cm
Therefore,
Rf = 0.446
10. EXTRACTION OF CAFFEINE FROM TEA Scientific name-
Camellia sinensis1.6 tea bags weighed and taken in a
beaker containing 90ml of water.
2.6 gm Sodium Carbonate added and
mixed thoroughly
3.Kept on water bath to boil for
10mins
4. 10mins the concentrated tea
extract was transferred to a flask.
5.The tea bags were further
squeezed to produce some more
extract and again boiled in 60 ml
water which was again transferred to
the flask. The tea bags are discarded
6.Filter the tea extract
The filtrate is taken in a separating
funnel. 15 ml Dichloromethane(DCM)
added and the funnel is shaken
gently.
7.The filtrate was dried and crystals
of caffeine were obtained
PROCEDURE
USES IN PHARMACEUTICAL INDUSTRY –
Central nervous system stimulant, due
to its vasoconstrictor effect
CONCLUSION-After performing this
experiment ,0.76%yield was obtained
11. STANDARDIZATION OF
CAFFEINE
TLC was done to standardize obtained sample against
standard Rf of 0.03
Mobile phase used= ethyl acetate: acetic acid in the
ratio 20:1
Tlc plate was then introduced in the prepared iodine
chamber and spots were observed under uv lamp.
Distance travelled by solute= 0.29 inch
Distance travelled by solvent= 7.8 inch
Therefore,
Rf of extracted caffeine= 0.037
12. EXTRACTION OF CURCUMIN FROM TURMERIC
botanical name - Curcuma longa
family- Zingiberaceae.
PROCEDURE –
1.Extract Powdered drug (50g) and add 95%of ethanol
2.Distill off alcoholic extract to a semi solid brown
coloured mass.
3.Dissolve crude extract in 50 ml of benzene, Extract
twice with equal volume of 0.1% NaOH solution.
4.Combined the alkaline extracts with diluted HCL
5.Concentrate the extract by boiling on water bath &At
the same time dissolve ,the precipitation in boiling
water.
6.Filter the solution & cool it.Get the yield of curcumin .
USES IN PHARMACEUTICAL INDUSTRY –
Curcumin is used in the treatment of tumors, arthritis, gastric ailments and viral infections
CONCLUSION-After performing this
experiment,2%yield was obtained
13. STANDARDISATION OF CRUDE DRUG (CURCUMIN)
mobile phase used= ethyl acetate:hexane in the ratio 7:3
Distance travelled by solvent = 7.3 inch
Distance travelled by solute = 5.4 inch
Therefore,
Rf= 0.74
14. EXTRACTION OF CAPSAICIN FROM RED CHILLI POWDER
Capsicum frutescens plants have smooth, medium-sized, elliptical
leaves, and slender branches which are 30–120 cm long
Family Solanaceae
Procedure:
10gm red chilli powder taken in a glass container. 200 ml
ethanol added and tightly fitted lid.
The container is kept like this for 7days.
Sedimentation occurs.
After 7 days the suspension is filtered. And the filtered
solid residue is discarded
The filtrate is heated on a water bath until an oily
composition of capsaicin is left
USES IN PHARMACEUTICAL INDUSTRY
Capsicum oleoresin is the primary
form of peppers used for
pharmaceutical purposes
CONCLUSION-After
performing this experiment,
7.2%yield was obtained,
15. Standardization of Capsaicin
TLC was done to standardize obtained sample against a
standard Rf value of 0.60
Mobile phase used= Chloroform: Methanol: Acetic Acid in
the ratio 9.5: 0.5:0.1
After running the solvent, the TLC pate was introduced
in the Iodine chamber.
Spots were observed and solute front and solvent front
was calculated.
Distance travelled by solute= 4.9cm
Distance travelled by solvent= 7.8 cm.
Therefore, Rf= 4.9/7.8= 0.62
16. CONCLUSION
All samples were collected from local markets.
Turmeric, Red Chilli, Black pepper and Tea were
collected as powder form which are used
commercially. Phytochemical analysis of the individual
extracts were performed to confirm the presence of
claimed secondary metabolite in the plant extracts
REFERENCE
• INTERNATIONALE PHARMACEUTICA SCIENCIA | Jan-March 2011 | Vol. 1
| Issue 1 |
• Agarwal OP. 2010. Chemistry of organic natural products. Meerut,
India: Goel Publishing House
• Practical pharmacognosy fifth edition | C.K. Kokate| 2018| Extractin
of Phytochemicals
• amrita.olabs.edu.in,. (2015). Detection of Carbohydrates, Proteins
and Fats. Retrieved 10 June 2018, from amrita.olabs.edu.in
