Modern methods of extraction by Dr. Amit Gangwal

Vision: To Pursue Excellence in Pharmaceutical Education & Research to Develop Competent Professionals
Basics of
Phytochemistry
Dr. Amit Gangwal
(Associate Professor)
SVKM’s Institute of Pharmacy, Dhule
B. Pharm. Vth Semester
Pharmacognosy &
Phytochemistry II
Modern methods of
extraction
(Other than supercritical fluid
extraction)

Disclaimer
Entire content in this video has been taken from textbooks and online
resources using Google as search engine. Author has no claim whatsoever on
the images or infographics taken from internet. This video is not made for
commercial interest, but just to share the educational stuff, purely from
academies point of view.

Continue….
The flow chart of
medicinal plant
study and
position of
extraction
techniques

Extraction is essential for isolation of different chemical constituent from
crude drug material.
Extraction depends on properties of material to be extracted.
Extraction is defined as the process of isolation of soluble material from an
insoluble residue, which may be liquid or solid by treatment with a solvent
on the basis of the physical nature (solid or liquid) of crude drug to be
extracted.

Menstruum: The solvent capable of penetrating the cell wall
of crude drugs and dissolves all types of active constituents
from it is called as Menstruum. There are two types of
menstruum.
Marc: The inert insoluble substance left after extraction is
called as Marc.

Factors affecting extraction process
• Nature of drug
• Nature of extracting solvent
• Solvent temperature
• Ratio of solvent to plant material
• Particle size
• Duration of extraction

Mechanism of Extraction
• Therefore, solvent used for extraction must diffuse into the cell to dissolve the desired
compounds whereupon the solution must pass the cell wall in the opposite direction and
mix with the surrounding liquid
• An equilibrium is established between the solute inside the cells and the solvent
surrounding the fragmented plant tissues
• Dissolution of extractive substances out of disintegrated cells.
• Dissolution of extractive substances out of intact plant cell by diffusion (requires steeping
and swelling)
• Penetration of the solvent into the plant cells and swelling of the cells.
• Diffusion of the dissolved extractive substances out of the cell.
• Plant constituents are usually contained inside the cells.

Parameters for Selecting an Appropriate Extraction Method
• Authentication of plant material should be done before performing extraction. Any foreign
matter should be completely eliminated.
• Use the right plant part and, for quality control purposes, record the age of plant and the
time, season and place of collection.
• Conditions used for drying the plant material largely depend on the nature of its chemical
constituents. Hot or cold blowing air flow for drying is generally preferred. If a crude drug
with high moisture content is to be used for extraction, suitable weight corrections should
be incorporated.
• Grinding methods should be specified and techniques that generate heat should be avoided
as much as possible.
• Powdered plant material should be passed through suitable sieves to get the required
particles of uniform size.

Nature of constituents: If the therapeutic value lies in non-polar constituents, a
non-polar solvent may be used. For example, lupeol is the active constituent of
Crataeva nurvala and, for its extraction, hexane is generally used.
Likewise, for plants like Bacopa monnieri and Centella asiatica, the active
constituents are glycosides and hence a polar solvent like aqueous methanol may
be used.
If the constituents are thermolabile, extraction methods like cold maceration,
percolation and CCE are preferred.

For thermostable constituents, Soxhlet extraction (if nonaqueous solvents
are used) and decoction (if water is the menstruum) are useful.
Suitable precautions should be taken when dealing with constituents that
degrade while being kept in organic solvents, e.g. flavonoids and
phenylpropanoids.
In case of hot extraction, higher than required temperature should be
avoided. Some glycosides are likely to break upon continuous exposure to
higher temperature.

Standardization of time of extraction is important for example
insufficient time means incomplete extraction.
If the extraction time is longer, unwanted constituents may also be
extracted. For example, if tea is boiled for too long, tannins are
extracted which impart astringency to the final preparation.

• The number of extractions required for complete extraction is as important as the
duration of each extraction.
• The quality of water or menstruum used should be specified and controlled.
• Concentration and drying procedures should ensure the safety and stability of the
active constituents. Drying under reduced pressure (e.g. using a Rotavapor) is widely
used.
• Lyophilization, although expensive, is increasingly employed.
• The design and material of fabrication of the extractor are also to be taken into
consideration.
• Analytical parameters of the final extract, such as TLC and HPLC fingerprints, should
be documented to monitor the quality of different batches of the extracts.

Types of extraction
• Solid extraction : extraction of solid from solid by using appropriate solvent
• Solvent extraction : process in which constituents that are extracted by solid
extraction process are partitioned between two immiscible solvents.

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

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.

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

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.

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

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.

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.

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

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.

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.

Soxhlet Extraction

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.

• 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.

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

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)

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

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.

• 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.

• 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.

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

• 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.

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.

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.

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.

• 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.

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).

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.

Ultrasonication-assisted extraction (UAE)
• The procedure involves the use of ultrasound waves,
which have frequencies higher than 20 KHz, have great
effects on extraction yield and kinetics.
• UAE involves ultrasonic effects of acoustic cavitations.
Under ultrasonic action solid and liquid particles are
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.
(means the spontaneous
formation of bubbles in a
liquid below its boiling point
resulting from strong
dynamic stressing),

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.

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.

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

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.

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.

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.

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.

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)

• 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.
42
Counter-Current Extraction

Advantages
• A unit quantity of the plant material can be extracted with much smaller
volume of solvent as compared to other methods like maceration, decoction,
and percolation.
• It is commonly done at room temperature, which spares the thermolabile
constituents from exposure to heat which is employed in most other
techniques.
• As the pulverization of the drug is done under wet conditions, the heat
generated during comminution is neutralized by water. This again spares the
thermolabile constituents from exposure to heat.
• The extraction procedure has been rated to be more efficient and effective
than Continuous hot extraction.

It is a continuous process of extraction which facilitates extraction of
active constituents from two immiscible solvents. All heat stable
Phytoconstituents can easily extracted by this method.
It requires more solvent than the other mode of extraction techniques.
Disadvantages

The applications of counter-current extraction covered many fields of plant
chemistry, including alkaloids, amino acids, antibiotics, antitumour
compounds, phenols including anthraquinone derivatives, cardiac
glycosides, essential oils, fatty acids, plant auxins, prostaglandins,
steroids and vitamins.
Other more recent developments involving the counter-current Principle
are high-speed counter-current chromatography (planet coil centrifugal
CCC), droplet counter-current chromatography (DCCC) and centrifugal
droplet CCC.

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.
Phytonics Process

• 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.
Advantages

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.
Applications

Ionic liquid Mediated Extraction (Ils)
The use of ionic liquid for analytical purposes has been developed in modern
times with advantages in terms of quality and efficacy of extraction.
In particular, an ionic liquid consists of a liquid organic salt that selectively
interacts with specific polar and non-polar compounds thanks to ion-exchanges,
π-stacking interactions, hydrophobic interactions or hydrogen bonds, improving
the selectivity of the extractive method.

For example, the interaction of 1-butyl-3-methylimidazolium hexafluorophosphate
([BMIM][PF6]) with hydrophilic amino acids enhances the extraction efficiency with
respect to a traditional organic solvent extraction.
Being ILs applied in several extractive processes such as MAE and UAE, they
should be stable enough to resist to the high temperatures through which the
technique works.
About this matter, tetraalkylammonium cations and imidazolium ions like
tetrafluoroborate [BF4]−, bis(trifluoromethylsulfonyl) imide [NTf2]−,
trifluoromethanesulfonate [CF3SO3]−, and hexafluorophosphate [PF6]−, possess a
great thermal stability; moreover, the organic anions have shown to be more stable
than the inorganic ones when operating at elevated temperatures.

In conclusion, extraction based on ILs represents a good choice to
recover in high yields organic and inorganic and metal ions as bioactive
components of plants and herbs.
ILs are liquid molten salts at temperatures below 100 °C and are typically composed of large
and unsymmetrical organic cations and organic or inorganic anions. Beyond the excellent
chemical, thermal, and electrochemical stability, no flammability, and negligible volatility
displayed by most aprotic ILs, ILs are also recurrently recognized by their excellent solvation
ability for a wide range of compounds and materials, from synthetically produced to natural
extracted ones, and as good stabilizing media for proteins, enzymes, nucleic acids, among
others.

Pressurized liquid/fluid extraction (PLE)
Pressurized liquid extraction (PLE or PFE in case of a general fluid) is a novel and
eco-friendly approach for the recovery of bioactives from plants.
This method often requires water as the solvent and so it can keep away from the
environmental and health risks due to the use of organic solvents.
Operating at high temperature (till 200°C) and pressure (from 35 to 200 bar), PLE
improves the quality of the extraction that can be carried out in a dynamic mode
in which the solvent is incessantly pumped through the vessel, or in a static mode
in which there are more cycles with a continuous replacement of the solvent.

At elevated temperatures there is a reduction of the viscosity of the solvent that can better
penetrate the matrix extracting the analytes of interest, even if this approach cannot be
used for the thermally unstable compounds and it could lead to a co-extraction of other
compounds because of the decreased selectivity of extraction at higher temperatures.
The elevated pressure permits maintaining the solvent in the liquid phase and the
disruption of plant cells wall exerting pressure on the matrix. In place of organic solvents, it
is possible to use additives like non-ionic surfactants, antioxidants like ascorbic acid, CO2
to drop aqueous pH or drying agents.
Furthermore, PLE apparatus protects light sensitive and oxygen sensitive products from
degradation, and it could be hyphenated with other modern extractive techniques like UAE
to improve its efficacy.

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.

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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Modern methods of extraction by Dr. Amit Gangwal

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