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Extraction

Involving the separation of the medicinally active

constituent of plants or animal tissues from the inactive
or inert component by using solvent(s) and by using one
of the standard extraction procedures.
• Mechanism of extraction
1. Penetration of the solvent into the plant cells and
swelling of the cells.
2. Solubilisation of active constituents from drug.
Diffusion of the dissolved extractive substances out of
the cell.
 Steps Involved in the Extraction of Medicinal Plants
In order to extract medicinal ingredients from plant material, the following sequential steps are involved:
1. Size reduction
2. Extraction
3. Filtration
4. Concentration
5. Drying and further purification

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 Properties of ideal solvent
1.Be highly selective for the compound to be extracted.
2.Have a high capacity for extraction in terms of coefficient of saturation of the
compound in the medium.
3.Not react with the extracted compound or with other compounds in the plant
material. (inert and non toxic)
4. Must be easily available and economic.
5.Be harmless to human being and to the environment. (Important)
6.Be completely volatile.

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 Infusion
• Fresh infusions are prepared by treating the crude drug for a short period
of time with cold or boiling water. The drug is not boiled with menstruum.
• These are dilute solutions of the readily soluble constituents of crude
drug
Types of Infusion
Fresh Infusion: e.g. Infusion of orange
Concentrated Infusion: These use alcohol as menstruum or it is
used as preservative. e.g. Concentrated infusion of Chirata

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 Decoction
• In this process, the crude drug is
boiled in a specified volume of
water for a defined time.
• It is then cooled and strained or
filtered.
• This procedure is suitable for
extracting water-soluble, heat
stable constituents. e.g. Tea ,
Coffee.

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 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.
e.g. extraction of morphine

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 Maceration
• In this process solid ingredients are
placed in a stoppered container with
the whole of the solvent and allowed
to stand for a period of at least 3
days with frequent agitation, until
soluble matter is dissolved.
• The mixture is then strained (through
sieves / nets), the marc pressed and
the combined liquids clarified
(cleaned by filtration) or by
decantation, after standing.

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 Types of maceration
• Simple maceration: Process used for making tinctures from organized drugs
E.g. roots, leaves, rhizomes.
• Modified maceration: Process used for making tinctures from unorganized
drugs e.g. tincture of tolu, tincture of benzoin.
• Double maceration: drug is macerated twice.
• Triple maceration: drug is extracted three times by the menstrum divided by
three parts.

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 Process of maceration
• Plant material crushed or cut small or moderately coarse powder, placed in a
closed vessels.
• Whole of the selected solvent (menstruum) is added.
• Allowed to stand for 3 to 7 days, shaking occasionally.
• Liquid strained off solid residue (marc) pressed, strained and expressed
liquids mixed.
• Clarified by subsidence or filtration evaporation and concentration

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 Percolation
• It is continuous downward
displacement of the solvent through
the bed of crude drug material to get
extract.
• Most frequently used to extract
active ingredients in the preparation
of tinctures and fluid extracts.
• It is the method of short successive
maceration or process of
displacement.
• A percolator a narrow, cone-shaped
vessel open at both ends is
generally used.

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 Steps in percolation
• Size reduction: The drug to be extracted is subjected to suitable degree of size reduction,
usually from coarse powder to fine powder.
• Imbibition: During imbibition the powdered drug is moistened with a suitable amount of
menstruum and allowed to stand for four hours in a well closed container.
• Packing: After imbibition the moistened drug is evenly packed into the percolator.
• Maceration: After packing sufficient menstruum is added to saturate the material. The
percolator is allowed to stand for 24 hours to macerate the drug.
• Percolation: The lower tap is opened and liquid collected therein is allowed to drip slowly
at a controlled rate until 3/4th volume of the finished product is obtained.

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 Soxhlet Extraction

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 Soxhlation
• Material coarsely powered and placed inside the thimble.
• Thin layer of cotton to be placed at the bottom of thimble in order to avoid the entry of
drug particles into siphon tube.
• Top layer of drug also covered with cotton in order to avoid disturbance to drug by
solvent addition
• A portion of the apparatus together with side tube, thimble and siphon tube is known
as soxhlet extractor.
• This assembly is attached to round bottom flask.
• From top solvent is added slowly.
• Condensor is attached to the soxhlet assembly.
• The whole assembly is set on heating mantle and mantle is switched on.

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 • Once the solvent begins to boil, the solvent vapour travels to condenser and
falls into the thimble dissolving out the desired component from drug.
• When the thimble is almost full, it gets emptied into the RBF through the
siphon tube. The cycle is repeated several times.

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 Merits and demerits
Merits
• Large amount of drug can be extracted with much smaller quantity of solvent.
• Tremendous economy in terms of time, energy & ultimately financial inputs.
• Small scale used a batch-process.
• Becomes more economical when converted into continuous extraction.

Demerits
• Not fit for all drugs.
• Not fit for environment vs supercritical fluid extraction

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The industrial processing of medicinal and aromatic plants starts with the extraction of the active
components using various technologies. The general techniques of medicinal plant extraction include, but
not limited to:
maceration,
• infusion
• percolation
• digestion
• decoction
• hot continuous extraction (Soxhlet)
• aqueous-alcoholic extraction by fermentation
• counter-current extraction
• microwave-assisted extraction
• ultrasound extraction (sonication)
• supercritical fluid extraction
• phytonic extraction (with hydrofluorocarbon solvents)

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 For aromatic plants
hydro-distillation techniques (water distillation, steam distillation, water
steam distillation),
hydrolytic maceration followed by distillation, expression and enfleurage
(cold fat extraction) may be employed.

Some of the latest extraction methods for aromatic plants include

headspace trapping
solid phase micro-extraction
protoplast extraction
microdistillation
thermomicrodistillation
molecular distillation

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 Microwave-assisted extraction (MAE)
• Microwave is a form of electromagnetic radiation with wavelengths
ranging from about 1 meter to 1 millimeter corresponding to
frequencies between 300 MHz and 300 GHz respectively.
• The prefix micro indicates that microwaves are shorter wavelengths
compared to radiowaves.
• In order to avoid interferences with radio communications,
domestic and industrial microwaves generally operate at 2.45 GHz .
• Owing to their electromagnetic nature, microwaves possess electric
and magnetic fields which are perpendicular to each other.
• The electric field causes heating via two simultaneous mechanisms,
namely, dipolar rotation and ionic conduction.

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Vision: To Pursue Excellence in Pharmaceutical Education & Research to Develop Competent Professionals
Vision: To Pursue Excellence in Pharmaceutical Education & Research to Develop Competent Professionals
 • Microwave-assisted extraction offers a rapid delivery of energy to a
total volume of solvent and solid plant matrix with subsequent heating
of the solvent and solid matrix, efficiently and homogeneously.
• Components of the sample absorb microwave energy in accordance to
their dielectric constants.
• When plant material is immersed inside a microwave transparent
solvent, the heat of microwave radiation directly reaches to the solid
without being absorbed by the solvent, resulting in instantaneous
heating of the residual moisture in the solid.

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 • Heating causes the moisture to evaporate and creates a high vapor
pressure that breaks the cell wall of substrate and releases the
content into solvent.
• The extracting selectivity and the ability of the solvent to interact
with microwaves can be modulated by using mixtures of solvents.
• One of the most commonly used mixtures is hexane-acetone.

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 A a programmable laboratory microwave oven extraction system
(RTP-Plus MARS-S model, CEM Corporate Matthews, NC, USA)

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• MAE techniques can be classified according to the pressure through which
they operate: higher than the atmospheric pressure (closed MAE system) and
lower than the atmospheric pressure (open MAE system).

• As regards closed systems, the temperature is set over the boiling point of the
solvent and the pressure is under control to avoid an excessive development.

• Among the closed systems we can enumerate high pressure microwave

assisted extraction (HPMAE) which uses high pressure and temperature in
order to enhance the capacity of the solvent to incorporate the energy from
radiation and to avoid large amount of solvent for the extraction.

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 In case of thermolabile molecules, soft conditions are needed and so the choice will fall
on an open system or the vacuum microwave assisted extraction (VMAE) that allows
the reduction of the boiling point of the solvent.

For compounds that are susceptible of oxidation it has been developed the nitrogen-
protected microwave-assisted extraction operating under pressurized inert gas.

When the bioactive compounds are susceptible of hydrolysis such as the essential
oils, solvent-free microwave-assisted extraction (SFME) is used to avoid the
loss/degradation of these products.

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 Another parameter to consider in MAE is the power of extraction: an
increased power boosts the temperature reducing the solvent viscosity
and leading to a better efficiency, except in case of thermolabile
molecules.

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 Applications of microwave-assisted extraction

• MAE can extract nutraceuticals products from plant sources in a faster

manner than conventional solid–liquid extractions.
• MAE (80% methanol) could dramatically reduce the extraction time of
ginseng saponin from 12 h using conventional extraction methods to a few
seconds.

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 • Extraction of taxanes from Taxus brevifolia
• Azadiractin related limonoids from Azadirachta indica seed kernels
• Extraction of glycyrrhizic acid from Glycyrrhizia glabra roots
• Extraction of artemisinin from Artemisia annua
• A higher microwave temperature and a short extraction time are more effective in
extracting anti-oxidative phenolic compounds from tomato
• It has been proven as a potential alternative to traditional methods for extraction
of phenols, such as chlorogenic acids, from green coffee beans.

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 Advantages of microwave-assisted extraction
• It reduces solvent consumption.
• It has a shorter operational time.
• Has a good reproducibility and minimal sample manipulation for extraction process.
• Microwave-assisted extraction gives several advantages with respect to classical
extractive processes such as Soxhlet. MAE allows a gain of time, higher quality and
yields.
• It is also cheaper than supercritical fluid extraction (SFE) and faster than ultrasonic-
assisted extraction (UAE).

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 Disadvantages of microwave-assisted extraction
• It requires specialized setup.
• An additional filtration or centrifugation is necessary to remove the solid residue during
MAE.
• Furthermore, the efficiency of microwaves can be very poor when either the target
compounds or the solvents are non-polar, or when they are volatile.
• It is more expensive than UAE.
• Less ecofriendly than SFE due to the use of organic solvents.
• Not suitable for thermolabile compounds because the irradiation could promote chemical
reactions with the loss of the desirable products, and not efficient when the target
molecules and/or the solvent of extraction are non-polar because they do not absorb
energy from the source.

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 Ultrasonication-assisted extraction (UAE)
• The procedure involves the use of ultrasound waves,
which have frequencies higher than 20 KHz, have great (means the spontaneous
formation of bubbles in a
effects on extraction yield and kinetics.
liquid below its boiling point
• UAE involves ultrasonic effects of acoustic cavitations. resulting from strong
Under ultrasonic action solid and liquid particles are dynamic stressing),
vibrated and accelerated and, because of that solute
quickly diffuses out from solid phase to solvent.
• Ultrasound assisted extractors are ultrasonic baths or
closed extractors fitted with an ultrasonic horn
transducer. The mechanical effects of ultrasound induce
a greater penetration of solvent into cellular materials
and improve mass transfer.

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 Sonication is the act of applying sound energy to agitate particles in a
sample, for various purposes such as the extraction of multiple compounds
from plants, microalgae and seaweeds.

Ultrasonic frequencies (>20 kHz) are usually used, leading to the process
also being known as ultrasonication or ultra-sonication.

In the laboratory, it is usually applied using an ultrasonic bath or

an ultrasonic probe, colloquially known as a sonicator.

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 UAE is based on the production of ultrasound waves and their transmission throughout the solvent with a
resulting cavitation. When the cavitation bubbles collapse, there is a generation of liquid circulation currents and
turbulence that improve the mass transfer rate.

The fractures formed in the cell wall enhance its permeability and so a bigger amount of solvent can enter into the
plant tissues to extract the bioactive metabolites.

In order to perform an extraction based on sonochemistry, the choice of solvent becomes an important parameter
because its physical properties like polarity, viscosity, vapor pressure and surface tension influence the cavitation
phenomena.

Ethanol, methanol and hexane are widely used in UAE, and sometimes water could be added to ethanol, even if its
amount must not be too much in order to avoid a decrease in extraction efficiency, probably due to the generation
of radicals from the ultrasonic dissociation of water.

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 An ultrasound sonicator UP200S manufactured by Dr. Hielscher,
Teltow, Germany

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 Other parameters to be considered are the frequency and the power: often the
former ranges from 20 to 100 kHz and the latter from 100 to 800 W.

Also the power dissipation is a critical factor, because the generation of

physical effects like turbulence is directly proportioned to the power dissipated
as heat.

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 Applications of ultra sonicated extraction
• Used to extract nutraceuticals from plants such as essential oils and lipids
dietary supplements. e.g. oils from almond, apricot and rice bran
• Extraction of saponin from ginseng, the observed total yield and saponin yield
increased by 15 and 30%, respectively.
• It was found that rice bran oil extraction can be efficiently performed in 30 min
under high-intensity ultrasound either using hexane or a basic aqueous
solution.
• Extraction rates of carvone and limonene by ultrasound-assisted extraction
with hexane were 1.3–2 times more rapid than those by the conventional
extraction depending on temperature.

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 Advantages of ultra sonicated extraction
• It increases extraction yield and has faster kinetics.
• It reduce the operating temperature allowing the extraction of thermolabile compounds.
• Compared with other novel extraction techniques such as microwave-assisted extraction,
the ultrasound apparatus is cheaper and its operation is easier.
• UAE is less expensive than the traditional extractive techniques; it can give high quantities
of products without spending time and without using large amounts of solvent.
• For a better performance, UAE can also be used in combination with other techniques like
supercritical fluid process.

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 Disadvantages of ultra sonicated extraction
• Lower efficiency as compared to other techniques.
• Although the process is useful in some cases, like extraction of rauwolfia root, its large-
scale application is limited due to the higher costs.
• This method is not useful for commercial purpose.
• Another problem that currently limits the use of UAE at large scales is the erosion of
transducers and their continuous replacement to avoid a decrease in the transmitted
energy.
• For the future, the design of reactors based on multiple transducers is needed in order to
operate at multiple frequencies and improve the efficacy of UAE.
• One disadvantage of the procedure is the occasional but known deleterious effect of
ultrasound energy (more than 20 kHz) on the active constituents of medicinal plants
through formation of free radicals and consequently undesirable changes in the drug
molecules.

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 Concentrations of main phenolic compounds in nettle extracts obtained by
different methods with1:30 solid to solvent ratio and detected by HPLC (mg/g
dry material)

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 42

Counter-Current Extraction
• In counter-current extraction, wet raw material is pulverized using toothed
disc disintegrators to produce fine slurry.
• The material to be extracted is moved in one direction (generally in the form
of 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.
• Finally, 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.

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 Phytonics Process
A new solvent based on hydrofluorocarbon-134a and a new technology to
optimize its remarkable properties in the extraction of plant materials offer
significant environmental advantages and health and safety benefits over
traditional processes for the production of high quality natural fragrant oils,
flavors and biological extracts.

Advanced Phytonics Limited (Manchester, UK) has developed this patented

technology termed “phytonics process”. The products mostly extracted by this
process are fragrant components of essential oils and biological or
phytopharmacological extracts which can be used directly without further
physical or chemical treatment.

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 Advantages
• Unlike other processes that employ high temperatures, the phytonics process is cool and gentle
and its products are never damaged by exposure to temperatures in excess of ambient.
• No vacuum stripping is needed which, in other processes, leads to the loss of precious volatiles.
• The process is carried out entirely at neutral pH and, in the absence of oxygen, the products
never suffer acid hydrolysis damage or oxidation.
• The technique is highly selective, offering a choice of operating conditions and hence a choice of
end products.
• It is less threatening to the environment.
• It requires a minimum amount of electrical energy.
• It releases no harmful emissions into the atmosphere and the resultant waste products (spent
biomass) are innocuous and pose no effluent disposal problems.
• The solvents used in the technique are not flammable, toxic or ozone depleting.
• The solvents are completely recycled within the system.

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 Applications
The phytonics process can be used for extraction in biotechnology (e.g for the production of
antibiotics), in the herbal drug industry, in the food, essential oil and flavor industries, and in the
production of other pharmacologically active products.

In particular, it is used in the production of top quality pharmaceutical-grade extracts,

pharmacologically active intermediates, antibiotic extracts and phytopharmaceuticals.

However, the fact that it is used in all these areas in no way prevents its use in other areas. The
technique is being used in the extraction of high-quality essential oils, oleoresins, natural food colors,
flavors and aromatic oils from all manner of plant materials.

The technique is also used in refining crude products obtained from other extraction processes. It
provides extraction without waxes or other contaminants. It helps remove many biocides from
contaminated biomass.

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 Enzyme-assisted extraction
An alternative approach to classical solvent extraction techniques is the enzyme-assisted
extraction. This method is innovative and convenient thanks to the fact that the enzymes
catalyze reactions in a specific way without operating under strong conditions that could
lead to the degradation of the desired products.

In addition, proteins like cellulases, hemicellulases and pectinases disrupt cell wall with the
hydrolysis of its components leading to a major permeability and allowing an easier release
of the metabolites from plants.

The application of enzymes such as lipases, proteases, phospholipases, permits to reduce

the use of the solvent for the extraction. For oil extraction from plants, cellulase, α-amylase
and pectinase are the most used enzymes.

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 These proteins can be obtained from fungi, bacteria, animals, and vegetables or
from genetic engineering methods and, thanks to their selective catalysis, they
can be used to recover a specific bioactive compound in high yields and in a
“green” approach, without wasting too much energy.

Nevertheless, there are some limitations due to the cost of the enzymatic
approach, the incomplete disruption of the cell wall and the complicated
application in a commercial scale because of the different behavior of the
enzymes according to the environmental circumstances such as the amount of
oxygen, the variety of nutrients and the operating temperature.

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 Counter current extraction
• Counter current extraction is a method of multiple
liquid- liquid extraction.
• Separation of components having variable solubility
in two immiscible liquid phases is achieved.
• In the counter current extraction two immiscible
solvents flow in an opposite direction in multiple
stages, (after several stages pure A and B solvents can
be obtained)
• In liquid- liquid extraction the solvent is used to
extract another liquid phase.

• To distribution of active principles between water and

organic solvent depends on the hydrophilic groups
present in the constituent molecules.

• If hydrophilic groups are ionisable ,PH will be an

important factor.
• If ionisation constants of isomers are different then
separation can be achieved.
 Instrumentation

A method of multiple liquid-liquid extractions

is countercurrent extraction, which permits the
separation of substances with different
distribution coefficients (ratios). A clever
design known as Craig apparatus is used for
this purpose (Lyman C. Craig, 1943).
A simple types of apparatus scale is a Craig apparatus.
Craig apparatus consists of a series of glass tubes (r: 0, 1,
2..) that are designed and arranged such that the lighter
liquid phase is transferred from one tube to the next.
The lower (heavier) phase of the two-phase solvent system (e.g.
water) is the "stationary phase", whereas the upper (lighter) phase
(e.g. hexane,) is the "mobile phase".

This tube has provision for separation upper layer and transferring to
next tube , where heavier solvents is placed. Fresh solvent is added
to tube 1.
Models are available to contain about 20-25 tubes, which can be
connected in sequence .

• Solvent selection.
Factors • Operating conditions.
affecting • Mode of operation.
• Extractor type.
extraction:
• Design criteria.
Other factors affecting solvent selection are :
:

Boiling point , stability.

Density,, Interfacial tension.

toxicity

cost

viscosity,

compatibility with product

 Disadvantages
1. Organic solvents are costly and inflammable.

2. Ideal solvent must have high partition coefficient. i.e. the

solvent must have high affinity for the required products.
Aqueous phases has negligible solubility.

3. Normally aqueous phase is large and the quantity of valuable

products is low .
 Application
• Isolation of antibiotics (penicillin G) from the aqueous
fermentation broth using immiscible solvent (amyl acetate or
butyl acetate)
• Isolation of chemical compounds from the aqueous systems
using small quantities of organic solvents in the production of
synthetic drugs and intermediates.
• In petroleum industry products having different chemical
structure but about the same boiling range are separated.
• Aromatic compounds are recovered from paraffin fraction of
the petroleum oil.
• Separation of components from synthetic
mixtures.
• Separation of components from plant extract.

• Purification of compounds (removal of

impurities)
 Supercritical fluid extraction

In thermodynamics, the triple point of a substance is the temperature and pressure at

which the three phases (gas, liquid, and solid) of that substance coexist in thermodynamic
equilibrium.

critical point ---A critical point or critical state is the endpoint of the phase
equilibrium curve. It is the highest temperature and pressure at which a pure substance can
exist in vapour/liquid equilibrium.
Supercritical fluid A supercritical fluid is any substance at a temperature and pressure above its
critical point.
Supercritical fluid extraction is the most effective and efficient way to extract
valuable constituent botanicals.
Supercritical Fluid Extraction (SFE) is the process of separating one component
(the extractant) from another (the matrix) using supercritical fluids that is CO2
as the extracting solvent.
Extraction conditions for supercritical CO2 are above the critical temperature of
31°C and
critical pressure of 74 bar.
S upercritical fluids are highly compressed gases, which have combined
properties of gases and liquids in an intriguing manner.
Supercritical fluids can lead to reactions, which are difficult or even impossible
to achieve in conventional solvents. I
t is a fast process completed in 10 to 60 minutes.
A supercritical fluid can be separated from analyte by simply releasing pressure,
leaving almost no trace and yields a pure residue.
Steps involved