Extraction methods

Pharmacognosy and phytochemistry

 Content

• Extraction
• Type of extraction
• Reference

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 Extraction
• 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 of crude drug to be extracted.
• It relies on the principle of “like dissolves like” when a mixture is exposed to
a solvent the target compound preferentially dissolves in that solvent while
the unwanted impurities are left behind . If two phase are used, they must be
immiscible so they can be easily separated after the transfer.
• The solvent used for extraction is known as Menstruum and the inert
insoluble material that remain after extraction is called Marc.
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Step of extraction

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 Factor affecting extraction

• Extraction solvent
• Particle size of raw material
• Solvent – solid ration
• Extraction temperature
• Extraction duration

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 Type of extraction techniques

Conventional method

Soxhlet
Maceration Decoction Infusion Percolation Digestion
extraction

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 Modern method

Microwave- Ultrasound- Enzyme- Pressurized hot

Supercritical
assisted assisted assisted water
fluid extraction
extraction extraction extraction extraction

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 Maceration

• Maceration is a simple extraction method that

involves soaking the plant prepared raw material in a
coarse or powder form in a solvent of interest at
room conditions for at least three days with
intermittent agitation.
• After the extraction is completed, the mixture is
strained either through sieves or a net with tiny holes.
Subsequently, the marc is pressed, and the liquid
extract is cleaned using either filtration or decantation
after standing.
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 Decoction
• In this process, the crud drug is boiled in a specified
volume of water for a defined time.
• It is then cooled and strained or filtered.
• This process is suitable for extracting water soluble,
heat stable constituents.
• The extract produced using the decoction technique
is likely to have many undesirable products. It may
also be noted that it is not the ideal method for
thermolabile compounds.

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 Infusion
• Infusion is described as a dilute solution of easily
soluble constituents of the plant material.
• It is an extraction technique in which the plant
material is immersed in boiling solvent,
particularly water, and left to stand in a stoppered
container for about 15 min, after which time the
extract is poured off and separated from the marc
using a filter.
• Tea may be considered as the best example of an
infusion.
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 Percolation
• Percolation is the most popular procedure for preparing
fluid extracts such as tinctures.
• Percolation means "to pass a liquid through a solid
material drop by drop."
• During percolation, the solvent, commonly ethyl
alcohol, is slowly passed through the plant material,
gradually packing itself with phytochemicals, and is
gradually propelled down by another fresh solvent
added from the top.
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 Digestion

• In this method, gentle heat is applied during the extraction. The preferred
solvent and powdered drug compound is added into a sterile holder. This
combination is then put into a water bath or into an oven that is heated to a
temperature of nearly 50°C.
• The viscosity of extraction solvent is reduced by continuously heating it
throughout the procedure of extraction, and this also helps increase the
elimination of the secondary plant constituents.

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 Soxhlet extraction

• Soxhlet extraction is a continuous extraction of phytochemicals using a

hot solvent.
• The Soxhlet extraction method uses a small amount of solvent and is
very cost-effective. The Soxhlet extraction uses the solvent reflux and
siphon principle to continuously extract the solid matter by pure
solvent, which saves the solvent extraction efficiency.
• The solid sample is placed on a thimble-shaped filter paper, positioned
into Soxhlet extractor, and the device is assembled. The solvent is added to
the solvent reservoir flask and mounted onto a heating mantle.
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• After heating, the condensed vapors of the solvent come in contact with
the sample powder, and the soluble part of the powder gets mixed with the
solvent for extraction.
• When the solvent surface exceeds the maximum height of the siphon, the
solvent containing the extract is siphoned back. The flask is repeated,
extracting a portion of the material each time so that the solid material is
constantly used as a pure solvent and the extracted material is
concentrated in the flask.
• [Link]

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 Limitation of Conventional method

• High solvent consumption

• Prolong extraction time
• Thermal degradation
• Low selectivity
• Low extraction efficiency

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 Microwave-assisted extraction

• Microwave-assisted extraction (MAE), also known as microwave

extraction, is a recent way of extracting natural products that incorporate
microwaves and solvents during the extraction process.

Principle:
• Microwaves are electromagnetic radiations with a frequency from 0.3 to
300 GHz. Microwave-assisted extraction (MAE) is an extractive method
based on the utilization of microwave energy.
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• 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.

Procedure:
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• 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 vapour
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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Advantages:
Disadvantages:
• Less time consuming
• Need specialized set up
• Low solvent consumption
• More expensive
• Improve extraction yield
• Not suitable for thermolabile
• Provide better accuracy and compound
precision

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 Application:
• Extracting phenolic molecules, quacertines, isoflavines compound.
• Extract nutraceutical products from plant
• Extraction of taxanes from taxus brevifolia
• A high microwave temperature and short extraction time are more effective in
extracting anti oxidant phenolic compound from tomato.
• [Link]

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 Ultrasound-assisted extraction

• Ultrasounds are mechanical waves with

higher frequencies than sound waves audible
to the human ear.
• The range of ultrasound utilized is from 20
kHz to 2000 kHz. It travels through a
medium involving expansions and
contractions following the wave nature. The
mechanical effect of acoustic cavitation from
the ultrasound increases the surface area of
contact between solvents and plant samples
and the permeability of cell walls. 24
• The bubble formation, its growth and collapse is termed as cavitation.
Some studies observed that frequency used can modify and favorably
influence the extraction of compounds from the sample.
• Ultrasonic-assisted extraction technique extracts bioactive compounds by
relying on ultrasound's mechanical action on plant cell walls. The
mechanical action of ultrasound enhances the surface contact between
solvent molecules and the plant sample matrix.
• Thus, ultrasound modifies and disrupts plant materials' physical and
chemical characteristics, expedites the release of phytochemicals, and
further reinforces the solvent system's mass movement into plant cells.

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Advantages: Disadvantages:
• Less time consuming • Lower efficiency
• Low solvent consumption • Large scale application is limited
• Applicable for small sample due to the higher cost
• Increase yield • Not useful for commercial use
• Applicable for thermolabile and
unstable compound

 Application:
• Extraction of pectin from cell wall
• Extracting of polyphenols, dietary fibers, polysaccharides and oils from plant
material. 26
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 Supercritical fluid extraction

• Each substance has a critical temperature (Tc) and pressure (Pc) beyond
which no applied pressure can force the substance into its liquid phase. If both
the temperature and pressure of a substance are greater than the Tc and Pc for
that particular substance, it is referred to as a supercritical fluid.
• Supercritical fluid extraction (SFE) is another sample preparation method
with over-all goals of decreased use of organic solvents and improved output
of sample.

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• Supercritical fluid extraction (SFE) uses
supercritical fluid (SF) as the extraction solvent.
SF has similar solubility to liquid and similar
diffusivity to gas, and can dissolve a wide
variety of natural products.
• Their solvating properties dramatically changed
near their critical points due to small pressure
and temperature changes.

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• Supercritical carbon dioxide (S-CO2) was widely used in SFE because
of its attractive merits such as low critical temperature (31 °C),
selectivity, inertness, low cost, non-toxicity, and capability to extract
thermally labile compounds.
• Fluid extraction technology that uses carbon dioxide above its critical
temperature (31°C) and pressure (73.8 bar), where it exhibits both gas-
like diffusivity and liquid-like solvating power.
• In this state, CO₂ selectively dissolves target compounds with high
efficiency while leaving no solvent residues.
• The mixture to be fractionated is passed in the extraction column along the
length of which the heater is located.
• CO2 is purged through the column.
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• Once the extraction column is pressurized, drug material gets saturated in
the supercritical fluid which moves along the length of the column.
• The operating conditions, i.e. pressure and temperature, are selected.
• In the pressure controlled type of extraction, the solution is just expanded
in the separation stage to precipitate the extract and then again the gas is
recompressed for recycle.
• In temperature control type operation, the extract is precipitated by heating
the solution which lowers the solvent density. The density is then
increased by isobaric cooling for recycling.
• Operation of supercritical fluid extraction system is controlled from a PC.
PC is used to set the operating conditions like pressure, temperature and
flow rate.
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Advantages: Disadvantages:
• Leaves no toxic chemical residues • High initial capital cost for
• Retains the delicate taste, aroma, equipment.
and therapeutic properties of natural • Requires high-pressure engineering
products. and strict safety measures.
• Highly automated and efficient, • Higher energy consumption
with easy solvent recovery. required to maintain high pressures.

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 Application:
• Extracting essential oils, antioxidants, and Omega-3 fatty acids from algae or
plants.
• Extracting active pharmaceutical ingredients (APIs) and purifying delicate
organic compounds.
• Preparing complex samples for chemical testing and chromatography
• [Link]

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 Enzyme-assisted extraction
• Enzyme-Assisted Extraction (EAE) is a green extraction technology that
employs hydrolytic enzymes to degrade plant or animal cell wall
components, facilitating the release of intracellular bioactive compounds.
• Plant cell walls consist mainly of cellulose, hemicellulose, pectin, lignin,
structural proteins. Which act as barriers that restrict the release of
intracellular compounds.
• Hydrolytic enzymes degrade these structural polymers through enzymatic
hydrolysis, resulting in cell wall disruption, increased porosity, reduced
diffusion resistance, enhanced mass transfer and improved extraction
efficiency.
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• The released compounds dissolve into the extraction solvent and are
subsequently separated by filtration or centrifugation.

Factor affecting EAE

• Enzyme type and specificity
• Enzyme concentration
• pH of the reaction medium

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• Temperature
• Incubation time
• Particle size
• Solid-to-liquid ratio
• Agitation speed
• Solvent composition

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Advantages: Disadvantages:
• High extraction efficiency • High cost of commercial enzyme
• Increased yield of bioactive • Activity depend on strong pH and
compound temperature
• Reduced solvent consumption • Long incubation time
• Eco friendly • Enzyme denatured under unstable
• Lower energy requirement condition
• Additionl purification required
 Application:
• Food industry
• Pharmaceutical industry 40

• Essential oil extraction

 Pressurized hot water extraction
• Pressurized
Hot Water Extraction (PHWE), also known as Subcritical
Water Extraction (SWE) or Pressurized Liquid Extraction (PLE).
• This technique uses hot water extractant at enhanced pressure. It has
drawn considerable attention as a likely alternative to conventional
extractive techniques mainly because it is an environmentally favourable
technique affording considerable yields.
• The technique is based on the principle that the liquid state of water is
maintained under pressure. A pressure of 5 MPa ascertains a liquid
water state between 100 and 250 °C.

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 Procedure :
• The extraction process involves several steps:
• Sample preparation by drying and grinding.
• Loading the sample into a stainless-steel extraction vessel.
• Heating water to the desired temperature (100–250°C or higher).
• Applying pressure to maintain water in the liquid state.
• Passing hot pressurized water through the sample.
• Dissolution of bioactive compounds into the water.
• Cooling the extract to room temperature.
• Collection and purification of the extract.
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Advantages:  Disadvantages :
• Short extraction time • High cost
• Better quality extract • High temperature may degrade
• Cheap solvent heat sensitive compound

 Application:
• Food industry
• Pharmaceutical industry
• Biotechnology
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 References
• Bitwell C, Indra SS, Luke C, Kakoma MK. A review of modern and conventional
extraction techniques and their applications for extracting phytochemicals from
plants. Scientific African. 2023;19.
• Godhi B, Rudraswamy S, Shankar HJ, Kenganora M, Sumana MN. Detailed
understanding of different extraction methods for the research on medicinal plants.
Indian Journal of Oral Health and Research. 2021;7(1).
• Shen L, Pang S, Zhong M, Sun Y, Qayum A, Liu Y, et al. A comprehensive review of
ultrasonic assisted extraction (UAE) for bioactive components: Principles,
advantages, equipment, and combined technologies. Ultrason Sonochem.
2023;101:106646.
• Zhang QW, Lin LG, Ye WC. Techniques for extraction and isolation of natural
products: a comprehensive review. Chin Med. 2018;13:20.
• Shah BN, Seth AK. Textbook of Pharmacognosy and Phytochemistry: elsiver; 2010.45
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