EXTRACTION METHODS

Extraction

• Is the separation of medicinally active

portions of plant (and animal) tissues using
selective solvents through standard
procedures.

• The products so obtained from plants are

relatively complex mixtures of metabolites,
in liquid or semisolid state or in dry
powder form (after removing the solvent),
& are intended for oral or external use.
 Medicinal
plants
• The Medicinal plants constitute an
effective source of both traditional and
modern medicines, herbal medicine has
been shown to have genuine utility and
about 80% of rural population depends
on it as primary health care. [WHO, (2005)]
 Medicinal plants are the richest bio-resource

 drugs of traditional systems of medicine,

 modern medicines,
 nutraceuticals,
 food supplements,
 folk medicines,
 pharmaceutical intermediates,
 chemical entities for synthetic drugs.
 Galenicals

These include classes of preparations

viz.,
 decoctions,
 infusions,
 fluid extracts,
 tinctures,
 pilular (semisolid) extracts,
 powdered extracts.
 Standardized Extraction

The purpose of standardized extraction

procedures for crude drugs
(medicinal & aromatic plant parts)

 To attain the therapeutically desired

portions
 To eliminate unwanted material by treatment
with a selective solvent known as
“menstrum”
The extract thus obtained, after standardization,
may be used as medicinal agent
• as such in the form of tinctures
• fluid extracts
• further processed to be incorporated in any
dosage form such as tablets and capsules
The general techniques of medicinal plant extraction
 Infusion,
 Decoction,
 Digestion,
 Maceration,
 Percolation,
 Hot continuous extraction (Soxhlet),
 Aqueous-alcoholic extraction by fermentation,
 Counter-current extraction,
 Microwave-assisted extraction,
 Ultrasound extraction (sonication),
 Supercritical fluid extraction,
 Advanced Phytonic extraction (with hydrofluorocarbon solvents).
Extraction techniques For aromatic plants

 Hydrodistillation techniques (water distillation,

steam distillation, water and steam
distillation),
 Hydrolytic maceration followed by distillation,
expression and enfleurage (cold fat extraction)
 Ecuelle method
 Enfleurage method
 Sponge method
 Absolutes and Concret
 Choice of solvents
• Successful determination of biologically
active compounds depends on the type of
solvent used in the extraction procedure.

• The choice of solvent is influenced by what is

intended with the extract.
 Properties of a good solvent in plant extractions

 low toxicity,
 ease of evaporation at low heat,
 promotion of rapid physiologic absorption of
the extract,
 preservative action,
 inability to cause the extract to complex or
dissociate.
Solvents used for active component extraction
 Water
• Water is universal solvent.
• used to extract plant products with antimicrobial
activity.
• Traditional healers use primarily water &
consistent antimicrobial activity is obtained.
• Water soluble flavonoids (mostly anthocyanins)
have no antimicrobial significance.
• only water soluble phenolics are important as
antioxidant compound.
 Aceton
e
• Dissolves many hydrophilic and lipophilic
components.
• a very useful extractant, especially for
antimicrobial studies (phenolic group extract).
•extraction of tannins + phenolics:
• Both acetone and methanol were found to
extract saponins  antimicrobial activity.
 Alcohol
• The identified components from plants
(antimicrobial) = aromatic or saturated organic
compounds  most often obtained through initial
ethanol or methanol extraction.
• Ethanol, found easier to penetrate the cellular
membrane to extract the intracellular
ingredients(polyphenols) from the plant material.
• Methanol is more polar than ethanol and its
cytotoxic in nature.
• The higher concentrations of more bioactive
flavonoid compounds were detected with ethanol
70% due to its higher polarity than pure ethanol.
 Chloroform

• Used to obtain tannins and

terpenoids.

• Terpenoid lactones are

obtained by successive
extractions of dried barks
with chloroform.
 Ether

• Commonly used selectively for the

extraction of coumarins and fatty
acids.
 Dichloromethanol

• Specially used for the selective extraction of

only terpenoids.
Steps Involved in the Extraction of Medicinal
Plants
1. Size reduction
2. Extraction
3. Filtration
4. Concentration
5. Drying
 1. S Iize
Reduction
Objective:
• To rupture plant organ, tissue & cell structures so that its
medicinal ingredients are exposed to the extraction
solvent.
• Size reduction maximizes the surface area, which in turn
enhances the mass transfer of active principle from
plant material to the solvent.
The 30-40 mesh size is optimal.
Hammer mill or a disc pulverizer which has built in
sieves controlled by varying the speed of the rotor
clearance b/w the hammers & the lining of the grinder.
 Extraction

Medicinal Aromatic plants

plants 1. Essential oils
1.Cold aqueous 2. Concretes
percolation
3. Absolutes
2.Hot aqueous extraction
4. Pomades
(decoction)
5. Esinoids
3.Solvent extraction
(cold / hot)
 parameters influencing the quality of an extract

• Plant part used as starting material

• Solvent used for extraction
• Extraction procedure
Effect of extracted plant phytochemicals depends on

• The nature of the plant material

• Its origin
• Degree of processing
• Moisture content
• Particle size
 Selection of plant
• Plant based natural constituents can be derived from
any part of the plant like bark, leaves, flowers, roots,
fruits, seeds, etc.
• Plants are usually air dried to a constant weight before
extraction.
• oven drying: every part were cut into pieces
dried in an oven @ 60°C for 9 hrs. &
pulverized.
• Other method for drying the plants is the oven drying
at about 40°C for 72 h.
 Filtration
• The extract so obtained is separated out from the marc
(exhausted plant material) by allowing it to trickle into
a holding tank through the built- in false bottom of the
extractor, which is covered with a filter cloth.
• The marc is retained at the false bottom, and the
extract is received in the holding tank.
• From the holding tank, the extract is pumped into a
sparkler filter to remove fine or colloidal particles
from the extract.
 Concentration
• The enriched extract from percolators or extractors, known as
miscella, is fed into a wiped film evaporator where it is
concentrated under vacuum to produce a thick concentrated
extract.
• The concentrated extract is further fed into a vacuum
chamber dryer to produce a solid mass free from solvent.
• The solvent recovered from the wiped film evaporator and
vacuum chamber dryer is recycled back to the percolator or
extractor for the next batch of plant material.
The solid mass thus obtained is pulverized and used directly for
•
the desired pharmaceutical formulations or further processed for
isolation of its phytoconstituents.
 Drying
• The filtered extract is subjected to spray drying
with a high pressure pump at a controlled
feed rate and temperature  to get dry
powder.
• The desired particle size of the product is obtained
by controlling the inside temperature of the
chamber and by varying the pressure of the
pump.
• The dry powder is mixed with suitable diluents or
excipients and blended in a double cone mixer to
obtain a homogeneous powder that can be
straight away used (for example, for filling in
capsules or making tablets).
 Variation in extraction methods

• Length of the extraction period,

• Solvent used,
• pH of the solvent,
• Temperature,
• Particle size of the plant tissues,
• The solvent-to-sample ratio.
 Parameters for Selecting an Appropriate Extraction Method

I. Authentication of plant material by botanist.

II. Use the right plant part + the age of plant + the
time, season & place of collection.
III. The nature of its chemical constituents.
IV. Grinding methods & powdering techniques.
V. Nature of constituents (polar/nonpolar).
VI. The quality of water / menstruum.
VII.The design & material of fabrication of the extractor.
VIII. Analytical parameters of the final extract
(TLC/HPLC).
The general techniques of medicinal plant extraction
 digestion,
 decoction,
 infusion,
 maceration,
 percolation,
 hot continuous extraction (Soxhlet),
 aqueous-alcoholic extraction by fermentation,
 counter-current extraction,
 microwave-assisted extraction,
 ultrasound extraction (sonication),
 supercritical fluid extraction,
 Advanced phytonic extraction (with hydrofluorocarbon solvents).
 Infusion
• Fresh infusions are prepared by macerating the
crude drug for a short period of time with cold or
boiling water.
• These are dilute solutions of the readily soluble
constituents of crude drugs.
 Decoction
• In this process, the crude drug is boiled in a
specified volume of water (1:4) for a defined time,
• Volume is reduced to 1/4 th the original,
• It is then cooled and strained / filtered.
• This procedure is suitable for extracting  water-
soluble, heat-stable constituents.
• Typically used in preparation of Ayurvedic
extracts
= “quath” / “kawath”
 Digestion
• This is a form of maceration in which gentle heat is
used during the process of extraction.
• It is used when moderately elevated
temperature is not objectionable.
• The solvent efficiency of the menstruum is thereby
increased.
 Maceration
• The whole / coarsely powdered crude drug is
placed in a stoppered container with the solvent.
• Allow to stand @ room temperature for a period of
at least 3 days with frequent agitation until the
soluble matter gets dissolved.
• The mixture then is strained, the marc (the damp
solid material) is pressed,
• The combined liquids are clarified by filtration or
decantation after standing.
• This method is best suitable
for use in case of
thermolabile drugs.
 Percolation
• Used most frequently to extract active
ingredients in the preparation of
tinctures and fluid extracts.
• The solid ingredients are moistened
with an appropriate amount of the
specified menstruum,
• Allowed to stand for approximately 4
hours in a well closed container, After
stand time, the mass is packed & the top
of the percolator and is closed, the
mixture is allowed to macerate in the
closed percolator for 24 h.
• Additional menstruum is added as required, until
the percolate measures about three-quarters of the
required volume of the finished product.
• The marc is then pressed and the expressed liquid is
added to the percolate.
• Sufficient menstruum is added to produce the
required volume.
• The mixed liquid is
clarified by filtration or
by standing followed by
decanting.
Hot Continuous Extraction
(Soxhlet)
• The finely ground crude drug is placed in a
porous bag or “thimble” made of strong filter
paper, which is placed in chamber of the Soxhlet
apparatus.
• The extracting solvent in flask is heated, and its
vapors condense in condenser.
• The condensed extractant drips into the thimble
containing the crude drug & extracts it by contact.
 Soxhlet apparatus
• When the level of liquid in chamber rises to the
top of siphon tube, the liquid contents of
chamber siphon into flask
• This process is continuous and is carried out
until a drop of solvent from the siphon tube does
not leave residue when evaporated.
 Aqueous Alcoholic Extraction by Fermentation
• Some medicinal preparations of Ayurveda (asava &
arista) adopt the technique of fermentation for
extracting the active principles.
• The extraction procedure involves soaking the
crude drug, [powder / a decoction (kasaya)], for a
specified period of time
• Undergoes fermentation & generates alcohol in situ.
• This facilitates the extraction of the active
constituents contained in the plant material.
• The alcohol thus generated also serves as a
preservative.
 • Some examples of Ayurvedic preparations:
karpurasava,
kanakasava,
,
dasmularista
.

• If the fermentation is to be carried out in an earthen

vessel, it should not be new: water should first be boiled
in the vessel.
• In large-scale manufacture, wooden vats, porcelain
jars or metal vessels are used in place of earthen
vessels.
 Counter - ccurrent Extraction
• Wet raw material is pulverized using toothed disc
disintegrators to produce a fine slurry.
• Material to be extracted is moved in one direction
generally in the form of a fine slurry within a
cylindrical extractor where it comes in contact with
extraction solvent.
• The further the starting material moves, the more
concentrated the extract becomes.
• sufficiently concentrated extract comes out at one
end of the extractor while the marc, practically
free of visible solvent falls out from the other end.
CRAIG’S APPARATUS
 Ultrasound Extraction (Sonication)
• The procedure involves the use of ultrasound waves
with frequencies ranging from 20 kHz to 2000 kHz.
•This increases the permeability of cell
walls & produces cavitation.
Eg: extraction of rauwolfia root.
Sonication achieves a complete extraction
and thereby superior extract yields are
obtained in a very short extraction time.
• Deleterious effect: Ultrasound energy (>20 kHz) on the
active constituents of medicinal plants causes formation
of free radicals and consequently undesirable changes
in the drug molecules.
MECHANISM:
• Highly energetic ultrasound waves travel through the
liquid creating alternating high-pressure / low-pressure
cycles, which results in the phenomenon of acoustic
cavitation. Acoustic or ultrasonic cavitation leads locally
to extreme temperatures, pressures, heating/cooling rates,
pressure differentials and high shear forces in the medium.
• When cavitation bubbles implode on the surface of solids
(such as particles, plant cells, tissues etc.), micro-jets and
interparticlular collision generate effects such as surface
peeling, erosion, particle breakdown, sonoporation (the
perforation of cell walls and cell membranes) and cell
disruption. Additionally, the implosion of cavitation
bubbles in liquid media create macro-turbulences and
micro-mixing.
 Applications
 Sonication can be used for the production of
nanoparticles, such as nano emulsions, nano crystals,
liposomes and wax emulsions, as well as for waste
water purification
 extraction of plant oil
 production of biofuels
 crude oil desulphurization, cell disruption,
 polymer and epoxy processing, adhesive thinning, and
many other processes.
 It is applied in pharmaceutical, cosmetic, water, food, ink,
paint, coating, wood treatment, metalworking, nano
composite, pesticide, fuel, wood product and many other
industries.
 Sonication can also be used to initiate crystallization
processes and even control polymorphic crystallizations.
 It is used to intervene in anti-solvent precipitations
(crystallization) to aid mixing and isolate small crystals.
 Supercritical Fluid Extraction (SFE) (principle)
 Supercritical Fluid Extraction (SFE) is an alternative sample preparation
method with general goals of reduced use of organic solvents and increase
sample throughput.
 The factors to consider include temperature, pressure, sample volume, analyte
collection, modifier ( co- solvent) addition, flow and pressure control, and
restrictors.
PHASE DIAGRAM of CO2
 Supercritical Fluid Extraction
• Cylindrical extraction vessels are used.
• The collection of the extracted analyte following SFE is
another important step: significant analyte loss can
occur during this step.
• CO2 as the extracting fluid.
• Organic solvents are frequently added to the CO2
extracting fluid to alleviate the polarity limitations
• The component recovery rates generally increase with
increasing pressure/temperature.
• The highest recovery rates in case of argon:
@ 500 atm & 150° C.
 Advanced Phytonics
• Process
A new solvent which is non-chlorinated fluorocarbons based on
hydrofluorocarbon-134a, and are known as Phytosols are
used.
• Advanced Phytonics Limited (Manchester, UK) has developed
 patented technology termed “phytonics process”.
• The products are fragrant components of essential oils &
Biological/phytopharmacological extracts.
• The core of the solvent is 1,1,2,2-tetrafluoroethane, (HFC-
134a).
• HFC-134a developed as a replacement for
chlorofluorocarbons. (Dichlorodifluromethane)
(Boiling Point -25° C).
• the solvents can be customized: by using modified solvents with
HFC-134a.
• The process can be made highly selective in extracting a
 Unlike chlorofluorocarbons, it does not deplete the
ozone layer. It has a vapor pressure of 5.6 bar at
ambient temperature. By most standards this is a
poor solvent. For example, it does not mix with
mineral oils or triglycerides and it does not dissolve
plant wastes.
 The biological products made by this process have
extremely low residual solvent, LESS THAN 20 PPB
 These solvents are neither acidic nor alkaline and,
therefore, have only minimal potential reaction
effects on the botanical materials.
 The technique is being used in the extraction of high-quality
essential oils, oleoresins, natural food colours, flavors and
aromatic oils from all manner of plant materials.
 The technique is also used in refining crude products
obtained from other extraction processes.
 Advantages of Phytosols/TECHNIQUE
 Nonflammable, non-corrosive, non-toxic, odorless, inert,
highly selective
• Easily modifiable by addition of co-solvents
• Requires low pressure for Liquefaction
• Low boiling point means no thermal degradation
• Readily recyclable so no need for solvent recovery
Does not require high pressure equipment's for
application of CO2
Phytosols require much low pressure for Liquefaction
Low operating cost
 INTRODUCTION
• Microwave-assisted extraction is an efficient method
which involves deriving natural compounds from raw
plants.

• Microwave extraction allows organic compounds to be

extracted more rapidly, with similar or better yield as
compared to conventional extraction methods.
 INTRODUCTION

• MAE of anthraquinone in pure alcohol at

60°C for 30mins gave a recovery of 65%
which resulted from 3 days maceration in pure
ethanol at room temperature.
• The advantages that MAE has over Soxhlet
are:
– Reduction in extraction time
– Improved yield
– Better accuracy
– Suitable for thermolabile substances
INTRODUCTION
 MICROWAVE THEORY

• Microwaves are non-ionizing electromagnetic waves

of frequency between 300MHz to 300 GHz and
positioned between the X-ray and infrared rays in the
electromagnetic spectrum.

• Microwaves serves two major purposes:

– Communication
– Energy vectors
 MICROWAVE THEORY

• The energy vector application is the direct action of

waves on materials that has the ability to convert a part
of the absorbed electromagnetic energy to heat energy.

• Microwaves are made up of two oscillating

perpendicular field’s i.e.
– Electric field
– Magnetic field
 MICROWAVE THEORY

• In MAE, heating occurs in a targeted and selective

manner with practically no heat being lost to the
environment as the heating occurs in a closed
system.

• This unique heating mechanism significantly reduces

the extraction time (usually less than 30min) as
compared to Soxhlet.
 EXTRACTION PRINCIPLE
• Dried plants contain a minute microscopic trace of
moisture which serves as target for microwave
heating.

• High temperature and pressure is generated inside

the oven.

• High temperature causes dehydration of cellulose

which accounts for its reduction of mechanical
strength.
 EXTRACTION PRINCIPLE
• The MAE process is as follows:
– Microwave radiation
– Moisture get heated up
– Moisture evaporates
– Generation of tremendous pressure
on cell wall
– Swelling of plant cell
– Rupture of the cell
– Leaching out of phyto-constituents

• This phenomenon can be intensified if the plant

matrix is impregnated with solvents with higher
heating efficiency under microwave.
 INSTRUMENTS

• There are two types of commercially available

MAE systems:
– Closed extraction vessels
– Focused microwave ovens

• The former performs extraction under

controlled pressure and temperature.

• In the latter, only a part of the extraction

vessel containing the sample is irradiated with
microwave.
Open and Closed-vessel system
 Aromatic Plant Extracts

• essential oils,
• concretes,
• absolutes,
• pomades
• resinoids.
 Essential oils
• Used in a wide variety of consumer goods viz.,
detergents, soaps, toilet products, cosmetics,
pharmaceuticals, perfumes, confectionery food
products, soft drinks, distilled alcoholic
beverages (hard drinks) and insecticides.
• Production technology is an essential element
to improve the overall yield & quality of
essential oil.
Presence of Essential Oils in plant parts.
Hetreogenous chemical group present in essential oils
 The traditional methods for essential oil extraction

• Water distillation,
• Water and steam distillation,
• Steam distillation,
• Cohobation,
• Maceration,
• Enfleurage
• Ecuelle method
 methods

1. Distillation: Originally introduced by Von Rechenberg.

• water distillation.
• water and steam distillation.
• direct steam distillation.
2. Hydrolytic maceration distillation.
3. Expression.
4. Cold fat extraction / Enfleurage.
• Distillation methods are good for powdered
almonds, rose petals and rose blossoms.
• Maceration is adaptable when oil yield from
distillation is poor.
• Solvent extraction is suitable for expensive,
delicate & thermally unstable materials like
jasmine, tuberose, and hyacinth.
• Water distillation is the most favored method of
production of citronella oil from plant material.
 Concrete
• This is an extract of fresh flowers, herbs, leaves and the flowering
tops of plants obtained by the use of a hydrocarbon solvent such
as butane, pentane, hexane and petroleum ether.
• Concrete is rich in hydrocarbon soluble material & devoid of
water-soluble components.
• It is generally a waxy, semisolid, dark-colored material free
from the original solvent.
• concretes are produced in
static extractors.
• It is a normal practice to circulate fresh
solvent through a battery of extractors.
• The enriched solvent from the extractor is pumped into an evaporator
for solvent recovery & the solvent content is reduced to about 1/10th
the original volume.
 A b so lu te s
• To make an absolute, the concrete is mixed with absolute alcohol
& agitated thoroughly in a vessel with an agitator.
• During agitation, the temperature is kept at 40°-60° C and the
concrete is immersed in the solution.
• The solution is cooled down to -5° to -10° C to precipitate out the
wax, since waxes are normally insoluble in alcohol below -1° C.
• The precipitated wax is removed by passing the solution through
a rotary filter.
• The filtrate from the rotary filter is pumped into a primary
evaporator, where it is concentrated to about 10% alcohol
content.
• Finally, the concentrated extract is pumped into an
agitating-type evaporator, where the alcohol is
carefully removed under high vacuum.
 R esin o id s
• Resinoid is an extract of naturally resinous material,
made with a hydrocarbon solvent.
• Resinoids are usually obtained from dry materials.
• The extraction process is same as that of concrete
production, except that perforated discs are not used
for stacking the material;
•instead powder from dry plant
material is fed into the extractor.
 P omades
•Pomades are obtained by a process known as
enfleurage, which is a cold fat extraction method.
• The fat is spread out on glass plates contained in wooden
frames, leaving a clear margin near the edges.
• The absorptive surface of the fat is increased by surface
grooves made with a wooden spatula.
• Fresh flowers are spread out on the surface of the fat and the
frames are stacked in piles.
• After the perfume oils have been absorbed from the flowers,
the spent flowers are removed by hand.
•Fresh flowers are again spread on the fat surface.
•This is repeated until the fat surface is completely enriched with
perfume oils.
•The pomade so obtained is ready for cold alcoholic extraction