TYPES of extraction in

phytochemicals of
medicinal plants
Submitted by
Abhijit Padhi
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.
How
antioxidant
works in
human body
 polyphenol Phenolic acids, flavonoids, stilbenes, lignans,
cucuminoids

carotenoids Beta-carotene, lycopene, lutein and zeaxanthin

Monoterpenes, sesquiterpenes, diterpenes,

Terpenoids triterpenes and steroids

Organosulfu Allicin, diallyl sulfide, glycosinolates, and

indole-3-carbinol

r compound Ginsenosides, soyasaponins, and diosgenin

Saponins
phytoche Chelates minerals

micals Phytic acids Beta-sitosterol, campesterol and stigmasterol

phytosterols

Classification of phytochemical
 Process of extraction of
phytochemicals
Size Concentrati
Extraction Filtration Drying
reduction on

Filtered extract is
To rupture plant To extract the subjected to
Used to
organ, tissue & phytochemical & The extract so spray drying
produced thick
cell structure so medicinal obtained is with a high-
concentrated
that its medicinal ingredient from separated out pressure pump at
extract by
ingredients are rupture plant from the marc a controlled feed
vacuum
exposed organ rate and
temperature
Steps involves in phytochemicals extraction
 Extraction
• This process is mainly used for separation of medicinal active
compounds from plant potions like tissue using a standard procedure
through solvents.
Techniques Maceration
used for extraction:
Extraction 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)
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.
 infusion
Select a suitable plant material that contains the chemical
compounds of interest and chop it into small pieces

Place the plant material in a vessel and add enough boiling water to
cover it. Also use other solvents such as oil or alcohol

Allow the mixture to steep for a specified

period, usually 15 minutes to several hours

Filter the mixture through a filter or filter paper to separate

the solid plant material from the liquid extract

Store the extract in a cool and dry place in a dark glass bottle. You
can also use the extract immediately for your purpose.
percolation
The plant material is chopped or powdered and moistened with a
suitable solvent

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 percolator is filled with the solvent and left to stand for 24 hours
in closed condition

The bottom outlet is opened and the liquid extract is collected in a

container.

The remaining plant material is pressed to remove and residual

solvent.

The percolate is concentrated by evaporating the solvent or using

other methods, such as freeze-drying or spray-drying.

The percolate can be further purified or isolated by using techniques

such as chromatography or crystallization.
 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
decoction
The decoction
The decoction
can be further
Just like is concentrated
purified or
maceration the After cool by evaporating
isolated by
plant material down the liquid the solvent or
using
placed in a extract using other
techniques
container and separated by methods, such
such as
boil it for 15- filtration as freeze-
chromatograph
60 minutes drying or
y or
spray-drying
crystallization
Hot Continuous extraction (soxhlet)
Fill a round bottom
Place the solid sample
flask with a suitable
that contains the Connect a condenser to Heat the round bottom
solvent that can
compound of interest in the upper end of the flask with a heating
dissolve the compound
a porous paper thimble extractor and provide a mantle or a hot plate to
of interest at high
and insert it into the cooling water supply to boil the solvent and
temperature and attach
main chamber of the it generate solvent vapors
it to the lower end of
soxhlet extractor
the extractor
The solvent vapors rise
When the solution
up through a distillation The condensed solvent The siphon tube also
reaches a certain level
tube and enter the main dissolves the compound creates an air gap that
in the main chamber, it
chamber of the of interest from the breaks the vacuum and
triggers a siphon tube
extractor, where they solid sample and forms allows fresh solvent
that drains the solution
condense and come in a solution in the main vapors to enter the main
back into the round
contact with the solid chamber chamber again
bottom flask
sample
Repeat steps 5 to 8 until
the extraction is Distill the solvent from
Stop the heating and
complete, which can be the round bottom flask
remove the round
determined by the color to recover the
bottom flask from the
change of the solvent in compound of interest
extractor
the main chamber or by and the solvent
a test of the thimble
 Select a suitable plant Distill the liquid extract to
material that contains the remove the excess water Store the extract in a cool
bioactive compounds of and ethanol and obtain the and dry place in a dark
interest and chop it into concentrated extract glass bottle
small pieces compound

After the fermentation is

complete, filter the mixture
Add water and sugar to the
to separate the solid plant
plant material and mix well
material from the liquid
extract

Add yeast to the mixture

Keep the vessel in a dark
and stir well and Transfer
and warm place for a
the mixture to a closed
specified period, usually a
vessel and seal it with an
few days to a few weeks
airlock

Aqueous-alcoholic extraction by fermentation

 Select a suitable plant Prepare a cylindrical
Counter-current material that contains the
bioactive compounds of
interest and chop it into
extractor that has two
immiscible liquids, such
as water and oil, flowing

extraction small pieces. in opposite directions

Transfer the solution to

Introduce the plant
the other end of the
material in the form of a
extractor, where it meets
fine slurry into the
the nonsolvent. The
extractor, where it comes
nonsolvent extracts the
in contact with the solvent
bioactive compounds
the bioactive compounds
from the solution and
forms a solution
forms a concentrate

Collect the concentrate

from the extractor and
Discard the remaining
separate the nonsolvent
plant material and the
from the bioactive
solvent from the extractor
compounds by distillation
or evaporation
 Select a suitable solvent that
can absorb microwave Place the sample and the

Microwave- radiation and heat up quickly

& compatible with the
sample and the analyte of
solvent in a microwave-safe
vessel. The ratio of sample to
solvent depends on the type

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

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 1:20extraction
Choose an appropriate ultrasound system, either open or closed. An open system operates at atmospheric
(sonication)
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 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

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.

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
 Select a suitable supercritical

Supercritica
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
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

l fluid
cost, and can be directly vented it
into the atmosphere

extraction Heat the round bottom flask with

a heating mantle or a hot plate to
raise the temperature and
pressure of the supercritical fluid
The supercritical fluid vapors rise
up through a distillation tube and
enter the main chamber of the
extractor, where they come in
The solvent-sample solution then
flows to the other end of the
extractor, where it meets the
above its critical point. The
contact with the sample and condenser and cools down.
critical point of CO2 is 31.1°C
dissolve the target compounds.
and 73.8 bar.

The cooling of the solution

Stop the heating and remove the
reduces the density and solubility
Repeat steps 5 and 6 until the round bottom flask and the
of the supercritical fluid, causing
extraction is complete, which can collecting vessel from the
the target compounds to
be determined by the color extractor. The supercritical fluid
precipitate out of the solution and
change of the solvent in the main can be vented into the
collect in the collecting
chamber or by a test of the atmosphere or recovered for
vessel. The supercritical fluid is
thimble. The extraction time can reuse. The target compounds can
then recycled back to the round
vary from 10 to 60 minutes be further concentrated, purified,
bottom flask for further
or analyzed
extraction.
 Select a suitable plant material that contains the essential
oils or other compounds of interest and chop it into small
Phytonic pieces and solvents also that can dissolve. Select a
suitable solvent that can dissolve the essential oils or other
extraction (with The phytonicand
compounds process uses hydrofluorocarbon
is compatible (HFC)
with the phytonic process
hydrofluorocarbo solvents, which are non-chlorofluorocarbons (non-CFCs)
that have low toxicity, low flammability, and low ozone
n solvents 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
Cool the vessel, solvent will become a liquid that can be
compounds
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
 TRAPPING / SOLID
ESSENTI PHASE MICRO-
AL OILS DISTILLATION
• PROTOPLAST
CONCRE

PLANTS
TES EXTR •
EXTRACTION
SOLVENT-FREE
PRODUC
TS
ABSOLU
TES ACTIO MICROWAVE
EXTRACTION
POMADE
S
N •
(SFME)
THEROMICRODISTI
LLATION
RESINOI • MOLECULAR
DS DISTILLATION

Other extraction PROCESS for various plants products

Hydro-distillation techniques
 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.

Hydro-
distillation

Steam distillation Water & steam

Water distillation
(Clevenger) distillation
clevenger Prepare the plant material by
cutting or grinding it into small
pieces
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.
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.
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.
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.
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.
Headspace tapping / Place the sample in a sealed vial and heat it to a desired
temperature. This will cause the volatile compounds to
solid phase evaporate and form a gaseous phase above the sample,
microdistillation 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.
 Transfer the
filtrate to a
Select the plant After a certain
centrifuge tube
tissue that you period of time,
Cut or peel off and spin it at a
want to extract depending on Count the
the outer layer Incubate the Filter the low speed to
protoplasts from, the tissue type number and the
of the tissue to tissue in a solution through sediment the
such as leaves, and the enzyme viability of the
expose the inner solution a sieve or a protoplasts at
roots, or stems. concentration, protoplasts using
cells. This will containing mesh to remove the bottom.
Wash and the cell walls a
make it easier enzymes that any undigested Discard the
sterilize the will be dissolved hemocytometer
for the enzymes can break down tissue fragments supernatant and
tissue to remove and the and a staining
to digest the cell the cell walls and debris. resuspend the
any protoplasts will dye
walls protoplasts in a
contaminants be released into
fresh solution of
and pathogens the solution
the same
osmotic agent

Protoplast extraction
Solvent-Free 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.
THERMOMICRO-
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.
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
Thank you