This document discusses various techniques for extracting phytochemicals from medicinal plants, including maceration, infusion, percolation, digestion, decoction, hot continuous extraction, aqueous-alcoholic extraction, counter-current extraction, microwave-assisted extraction, and ultra-sound extraction. It provides detailed step-by-step explanations of each extraction technique. The goal of extraction is to separate medicinal active compounds from plant materials using solvents and standard procedures.

1. TYPES of extraction in
phytochemicals of
medicinal plants
Submitted by
Abhijit Padhi

2. phytochemical
• Phytochemical are bioactive chemical compounds that occur naturally in plant-
based food. They have antioxidant properties, which means they can neutralize
harmlful free radicals.
• EXAMPLES: mainly founds in fruits, vegetables and some foods.
• In phytochemicals mainly found vitamin- ‘C’ and ‘E’, anthocyanidins,
carotenoids, catechins, beta-carotene, flavonoids, isoflavones, polyphenols.
Antioxidant:
Antioxidants are substances that can protect your cells from radicals.
Free radicals:
These are unstable molecules that can cause harm to human DNA, cell membrane,
and other parts of the cells.

3. How
antioxidan
t works in
human
body

4. Classification of phytochemical
phytoch
emicals
polyphenol
carotenoids
Terpenoids
Organosulfur
compound
Saponins
Phytic acids
phytosterols
Phenolic acids, flavonoids, stilbenes,
lignans, cucuminoids
Beta-carotene, lycopene, lutein and
zeaxanthin
Monoterpenes, sesquiterpenes,
diterpenes, triterpenes and steroids
Allicin, diallyl sulfide, glycosinolates,
and indole-3-carbinol
Ginsenosides, soyasaponins, and
diosgenin
Chelates minerals
Beta-sitosterol, campesterol and
stigmasterol

5. Steps involves in
Process of extraction of
phytochemicals
Size
reduction
To rupture plant
organ, tissue & cell
structure so that its
medicinal
ingredients are
exposed
Extraction
To extract the
phytochemical &
medicinal
ingredient from
rupture plant organ
Filtration
The extract so
obtained is
separated out from
the marc
Concentra
tion
Used to produced
thick concentrated
extract by vacuum
Drying
Filtered extract is
subjected to spray
drying with a high-
pressure pump at a
controlled feed rate
and temperature

6. Extraction
• This process is mainly used for separation of medicinal active
compounds from plant potions like tissue using a standard
procedure through solvents.
Techniques used for extraction:
Extractio
n
Maceration
infusion
percolation
digestion
Decoction
Hot continuous extraction (soxhlet)
Aqueous-alcoholic extraction by fermentation
Counter-current extraction
Microwave-assisted extraction
Ultra-sound extraction (sonication)
Supercritical fluid extraction
Phytonic extraction (with hydrofluorocarbon solvents)

7. Maceration
Chop or powder the plant material.
Soak the plant material in a suitable solvent for at least
three days.
Strain or filter the liquid extract from the solid residue.
Concentrate the extract by evaporating the solvent or
using other methods.
Purify or isolate the target compound by using
techniques such as chromatography or crystallization.

8. infusion
1.Select a suitable plant material that contains the chemical
compounds of interest and chop it into small pieces
1.Place the plant material in a vessel and add enough boiling
water to cover it. Also use other solvents such as oil or alcohol
1.Allow the mixture to steep for a specified
period, usually 15 minutes to several hours
1.Filter the mixture through a filter or filter paper to
separate the solid plant material from the liquid extract
1.Store the extract in a cool and dry place in a dark glass bottle.
You can also use the extract immediately for your purpose.

9. percolation
• The percolate can be further purified or isolated by using
techniques such as chromatography or crystallization.
•The percolate is concentrated by evaporating the solvent or using
other methods, such as freeze-drying or spray-drying.
The remaining plant material is pressed to remove and residual
solvent.
The bottom outlet is opened and the liquid extract is collected in a
container.
The percolator is filled with the solvent and left to stand for 24
hours in closed condition
The moistened plant material is placed in a percolator. The
opening is covered with perforated lid. The bottom outlet is closed
with an adjustable valve that control the flow rate of the liquid.
The plant material is chopped or powdered and moistened with a
suitable solvent

10. digestion
This is just like maceration in which gentle heat
at 35° to 50°C is used during extraction
Container is left for 24 hours. The duration is
depend on plants types
Liquid extract filtered to separate it from the
solid residue
The digestate is concentrated by evaporating
the solvent or using other methods, such as
freeze-drying or spray-drying.
The digestate can be further purified or isolated
by using techniques such as chromatography
or crystallization

11. decoction
Just like
maceration the
plant material
placed in a
container and boil
it for 15-60
minutes
After cool down
the liquid extract
separated by
filtration
•The decoction is
concentrated by
evaporating the
solvent or using
other methods,
such as freeze-
drying or spray-
drying
•The decoction
can be further
purified or
isolated by using
techniques such
as
chromatography
or crystallization

12. Hot Continuous extraction
(soxhlet)
1.Place the solid
sample that contains
the compound of
interest in a porous
paper thimble and
insert it into the main
chamber of the
soxhlet extractor
1.Fill a round bottom
flask with a suitable
solvent that can
dissolve the
compound of interest
at high temperature
and attach it to the
lower end of the
extractor
1.Connect a
condenser to the
upper end of the
extractor and provide
a cooling water
supply to it
1.Heat the round
bottom flask with a
heating mantle or a
hot plate to boil the
solvent and generate
solvent vapors
1.The solvent vapors
rise up through a
distillation tube and
enter the main
chamber of the
extractor, where they
condense and come
in contact with the
solid sample
1.The condensed
solvent dissolves the
compound of interest
from the solid sample
and forms a solution
in the main chamber
1.When the solution
reaches a certain
level in the main
chamber, it triggers a
siphon tube that
drains the solution
back into the round
bottom flask
1.The siphon tube
also creates an air
gap that breaks the
vacuum and allows
fresh solvent vapors
to enter the main
chamber again
1.Repeat steps 5 to 8
until the extraction is
complete, which can
be determined by the
color change of the
solvent in the main
chamber or by a test
of the thimble
1.Stop the heating
and remove the
round bottom flask
from the extractor
1.Distill the solvent
from the round
bottom flask to
recover the
compound of interest
and the solvent

13. Aqueous-alcoholic extraction
1.Select a suitable plant
material that contains
the bioactive compounds
of interest and chop it
into small pieces
1.Add water and sugar
to the plant material and
mix well
1.Add yeast to the
mixture and stir well and
Transfer the mixture to a
closed vessel and seal it
with an airlock
1.Keep the vessel in a
dark and warm place for
a specified period,
usually a few days to a
few weeks
1.After the fermentation
is complete, filter the
mixture to separate the
solid plant material from
the liquid extract
1.Distill the liquid extract
to remove the excess
water and ethanol and
obtain the concentrated
extract compound
1.Store the extract in a
cool and dry place in a
dark glass bottle

14. Counter-
current
extraction
1.Select a suitable
plant material that
contains the bioactive
compounds of interest
and chop it into small
pieces.
1.Prepare a cylindrical
extractor that has two
immiscible liquids,
such as water and oil,
flowing in opposite
directions
1.Introduce the plant
material in the form of
a fine slurry into the
extractor, where it
comes in contact with
the solvent the
bioactive compounds
forms a solution
1.Transfer the solution
to the other end of the
extractor, where it
meets the
nonsolvent. The
nonsolvent extracts
the bioactive
compounds from the
solution and forms a
concentrate
1.Collect the
concentrate from the
extractor and separate
the nonsolvent from
the bioactive
compounds by
distillation or
evaporation
1.Discard the
remaining plant
material and the
solvent from the
extractor

15. Microwave-
assisted
extraction
that can absorb
microwave radiation and
heat up quickly &
compatible with the
sample and the analyte of
interest. Ex, solvents are
acetone, acetonitrile,
dichloromethane, hexane,
and methanol
•Place the sample and the
solvent in a microwave-
safe vessel. The ratio of
sample to solvent
depends on the type and
size of the sample, but
usually ranges from 1:5 to
1:20
•Choose an appropriate
microwave system, either
open or closed. An open
system operates at
atmospheric pressure and
has a vent to release the
vapors. A closed system
operates at higher
pressure and has a valve
to control the pressure
•Set the microwave
parameters, such as
power, time, and
temperature. Generally,
higher power and
temperature can increase
the extraction efficiency,
but also increase the risk
of degradation and
decomposition of the
analyte. The extraction
time can vary from a few
seconds to several
minutes
•Start the microwave
extraction and monitor the
process. The solvent will
heat up and penetrate the
sample matrix, dissolving
the analyte. The analyte
will then diffuse into the
solvent and be
extracted. The extraction
process can be observed
•Stop the microwave
extraction and cool down
the vessel. Carefully open
the vessel and filter the
extract to remove any
solid residues. The extract
can then be concentrated,
purified, or analyzed by
various techniques, such
as chromatography,

16. Ultra-sound
extraction
(sonication)
•The extraction process can be observed by the color change of the solvent or the temperature
change of the vessel. Stop the ultrasound extraction and cool down the vessel. Carefully open the
vessel and filter the extract to remove any solid residues. The extract can then be concentrated,
purified, or analyzed by various techniques, such as chromatography, spectroscopy, or mass
spectrometry
•Start the ultrasound extraction and monitor the process. The ultrasound waves will create bubbles in
the solvent that collapse and generate high pressure and temperature, breaking the cell walls and
membranes of the plant material and releasing the bioactive compounds. The bioactive compounds
will then diffuse into the solvent and be extracted.
•Set the ultrasound parameters, such as power, time, and temperature. Generally, higher power and
temperature can increase the extraction efficiency, but also increase the risk of degradation and
decomposition of the analyte. The extraction time can vary from a few seconds to several minutes
•Choose an appropriate ultrasound system, either open or closed. An open system operates at
atmospheric pressure and has a vent to release the vapors. A closed system operates at higher
pressure and has a valve to control the pressure
•Place the plant material and the solvent in a vessel that is compatible with ultrasound. The ratio of
plant material to solvent depends on the size and type of the plant material, but usually ranges from
1:5 to 1:20
•Select a suitable plant material that contains the bioactive compounds of interest and chop it into
small pieces. Select a suitable solvent that can absorb ultrasound waves and dissolve the bioactive
compounds

17. Supercritic
al fluid
extraction
•Select a suitable supercritical
fluid(CO2) that can dissolve the
target compounds from the
sample matrix. CO2, which has
moderate critical pressure and
temperature, low toxicity and
reactivity, high purity and low
cost, and can be directly vented
into the atmosphere
•Place the sample that contains
the target compounds in a
porous paper thimble or a fine
slurry and insert it into the main
chamber of the supercritical fluid
extractor
•Fill a round bottom flask with
the supercritical fluid and attach
it to the lower end of the
extractor. Connect a condenser
to the upper end of the extractor
and provide a cooling water
supply to it
•Heat the round bottom flask
with a heating mantle or a hot
plate to raise the temperature
and pressure of the supercritical
fluid above its critical point. The
critical point of CO2 is 31.1°C
and 73.8 bar.
•The supercritical fluid vapors
rise up through a distillation tube
and enter the main chamber of
the extractor, where they come
in contact with the sample and
dissolve the target compounds.
•The solvent-sample solution
then flows to the other end of
the extractor, where it meets the
condenser and cools down.
•The cooling of the solution
reduces the density and
solubility of the supercritical
fluid, causing the target
compounds to precipitate out of
the solution and collect in the
collecting vessel. The
supercritical fluid is then
recycled back to the round
bottom flask for further
extraction.
•Repeat steps 5 and 6 until the
extraction is complete, which
can be determined by the color
change of the solvent in the
main chamber or by a test of the
thimble. The extraction time can
vary from 10 to 60 minutes
•Stop the heating and remove
the round bottom flask and the
collecting vessel from the
extractor. The supercritical fluid
can be vented into the
atmosphere or recovered for
reuse. The target compounds
can be further concentrated,
purified, or analyzed

18. Phytonic
extraction (with
hydrofluorocarb
on solvents
essential oils or other compounds of interest and
chop it into small pieces and solvents also that can
dissolve. Select a suitable solvent that can dissolve
the essential oils or other compounds and is
compatible with the phytonic process
•The phytonic process uses hydrofluorocarbon
(HFC) solvents, which are non-chlorofluorocarbons
(non-CFCs) that have low toxicity, low flammability,
and low ozone depletion potential
•Place the plant material and the solvent in a closed
vessel that is equipped with a heating and cooling
system, a pressure control system, and a collecting
system.
•Heat the vessel to raise the temperature and
pressure of the solvent above its boiling point. The
solvent will become a superheated vapor that can
penetrate the plant material and dissolve the
essential oils or other compounds
•Cool the vessel, solvent will become a liquid that
can be separated from the essential oils or other
compounds by gravity or centrifugation. The solvent
can be recycled back to the vessel for further
extraction
•Collect the essential oils or other compounds from
the vessel and store them in a cool and dry place in
a dark glass bottle

19. Other extraction PROCESS for
EXTRACTION
• HYDRODISTILLATION
TECHNIQUES
• HEADSPACE TRAPPING /
SOLID PHASE MICRO-
DISTILLATION
• PROTOPLAST EXTRACTION
• SOLVENT-FREE MICROWAVE
EXTRACTION (SFME)
• THEROMICRODISTILLATION
• MOLECULAR DISTILLATION
PLANTS
PRODUCTS
ESSENTIAL
OILS
CONCRETES
ABSOLUTES
POMADES
RESINOIDS

20. Hydro-distillation techniques
Hydro-
distillation
Water
distillation
Steam
distillation
(Clevenger)
Water & steam
distillation
 Hydro-distillation is a method of extracting essential oils from plant materials using water or steam.
 It is a type of steam distillation, but the plant materials are immersed in water and boiled together
with the water. The steam that carries the essential oils is then condensed and separated from the
water.
 Hydro-distillation is a traditional and simple technique that can be performed using a Clevenger
apparatus or a simple alembic.
 Some advantages of hydro-distillation are that it can extract essential oils from dried or fresh plants,
and it does not require any solvents or chemicals.
 However, some disadvantages are that it can cause thermal degradation or hydrolysis of some
components, and it can be time-consuming and energy-intensive.

21. clevenger •Prepare the plant material by
cutting or grinding it into small
pieces
•Fill a round-bottom flask with
water and add the plant
material. Attach the flask to a
heating source, such as a hot
plate or a Bunsen burner.
•Connect the flask to a
Clevenger apparatus, which is
a piece of specific glassware
that consists of a condenser, a
graduated burette, and a
diagonal conduit.
•Heat the flask until the water
boils and produces steam. The
steam will carry the volatile
compounds of the plant
material to the condenser,
where they will be cooled and
condensed into a liquid.
•The liquid will then fall into the
burette, where the essential oil
will float on top of the water.
The water will be gradually
returned to the flask through
the diagonal conduit, while the
essential oil will remain in the
burette.
•After 2 hours of extraction,
measure the volume of the
essential oil collected in the
burette. You can calculate the
yield of the extraction by
dividing the volume of the
essential oil by the mass of the
plant material used.

22. tapping /
solid phase
microdistillat
ion
Place the sample in a sealed vial and heat it to a
desired temperature. This will cause the volatile
compounds to evaporate and form a gaseous phase
above the sample, called the headspace.
Tap the headspace with a syringe or a needle and
withdraw a fixed volume of the gas. This is the
headspace sample that contains the volatile
compounds of interest.
Inject the headspace sample into the GC column for
separation and analysis. The GC column will
separate the volatile compounds based on their
boiling points and polarity, and the detector will
measure their concentrations.

23. Protoplast extraction
•Select the
plant tissue
that you want
to extract
protoplasts
from, such as
leaves, roots,
or stems.
Wash and
sterilize the
tissue to
remove any
contaminants
and pathogens
•Cut or peel off
the outer layer
of the tissue to
expose the
inner cells.
This will make
it easier for the
enzymes to
digest the cell
walls
•Incubate the
tissue in a
solution
containing
enzymes that
can break
down the cell
walls
•After a certain
period of time,
depending on
the tissue type
and the
enzyme
concentration,
the cell walls
will be
dissolved and
the protoplasts
will be
released into
the solution
•Filter the
solution
through a sieve
or a mesh to
remove any
undigested
tissue
fragments and
debris.
•Transfer the
filtrate to a
centrifuge tube
and spin it at a
low speed to
sediment the
protoplasts at
the bottom.
Discard the
supernatant
and resuspend
the protoplasts
in a fresh
solution of the
same osmotic
agent
•Count the
number and
the viability of
the protoplasts
using a
hemocytomete
r and a staining
dye

24. Microwave
Extraction
(SFME)
Place the plant material in a microwave oven
with a glass flask and a condenser attached to
it.
Turn on the microwave and adjust the power
and time according to the type and amount of
plant material.
Collect the condensed vapors in a receiver flask.
Separate the essential oil from the hydrosol by
decantation or using a separatory funnel.

25. O-
DISTILLATION
extraction
Place the plant material in a round-bottom flask
with a heating mantle and a condenser attached
to it.
Turn on the heating mantle and adjust the
temperature and time according to the type and
amount of plant material.
Collect the condensed vapors in a receiver
flask.
Separate the essential oil from the hydrosol by
decantation or using a separatory funnel.

26. Application of extraction
• Phytochemical analysis: Extraction is a principal method for isolating compounds from plant
materials. Extraction moves compounds from one liquid to another, so that they can be more easily
manipulated or concentrated.
• Medicinal and nutraceutical products: Extraction is used to obtain the bioactive compounds from
medicinal plants and herbs, such as alkaloids, flavonoids, terpenoids, phenolics, etc. These
compounds have various pharmacological effects, such as antioxidant, anti-inflammatory,
antimicrobial, anticancer, etc. Extraction methods can affect the yield, quality, and stability of the
extracted compounds.
• Natural dyes and pigments: Extraction is used to obtain the natural dyes and pigments from plant
sources, such as anthocyanins, carotenoids, chlorophylls, etc. These compounds have various
applications in food, textile, cosmetic, and pharmaceutical industries.
• Essential oils and fragrances: Extraction is used to obtain the essential oils and fragrances from
aromatic plants, such as lavender, rose, mint, etc. These compounds have various applications in
perfumery, aromatherapy, and flavoring industries.
• DNA and RNA: Extraction is used to obtain the DNA and RNA from plant cells, which are the genetic
materials that store the information for the synthesis of proteins and other molecules. DNA and RNA
extraction can be used for various purposes, such as genetic engineering, molecular breeding, gene
expression analysis, etc.

27. Thank you