Extraction Techniques

BY Melese Legesse

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 Introduction
• Extraction involves the separation of medicinally active
portions of plant or animal tissues from the inactive or
inert components by using selective solvents in
standard extraction procedures.
• A wide range of technologies is available for the
extraction of active components and essential oils from
medicinal and aromatic plants.
– The choice depends on the economic feasibility and suitability
of the process. 2
Extraction techniques
Extraction
 It is a process in which one or more components are separated
selectively from either a liquid or solid sample mixtures
 It is the transfer of a solute from one phase to another
 It is a way to separate soluble material from insoluble material
 The transfer of the components from the sample to the
solvent is controlled by the solubility behavior of each
component.

 It is now understood that extraction process is facilitated by

both
 Solubility
 Diffusion
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• The extract obtained may be
– ready for use as a medicinal agent in the form of tinctures
and fluid extracts,
– further processed to be incorporated in any dosage form
such as tablets or capsules, or it may be fractionated to
isolate individual chemical entities(modern drugs).

• Thus, standardization of extraction procedures contributes

significantly to the final quality of the herbal drug.

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 Extraction methods
 conventional extraction methods
Batch extraction

Continuous extraction

Countercurrent extraction

 Advanced methods of extraction

 Microwave

 Ultrasound-assisted extractions

 Pressurized liquid extraction (PLE).

 Supercritical fluids extraction

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 1. Batch extraction
Batch extraction: the solute is extracted from one phase by shaking/contact

it with a solvent.

Single process
 maceration,
 percolation,
 Infusion
 Decoction
 Digestion
 Distillation

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 Percolation (Exhaustive Extraction)
Process
• Organized vegetable drug in a suitably powdered form.
• Uniform moistening of the powdered vegetable drugs with
menstruum for a period
of 4 hrs in a separable
vessel.
• Packed evenly into the
percolator.

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 Percolation…..
After maceration, the outlet is opened and solvent is percolated
at a control rate with continuous addition of fresh volume.

75% of the volume of the finished product is collected.

Marc is pressed and expressed liquid is added to the percolate

giving 80% to 90% of the final volume.

Evaporation and concentration to get finished products by

applying suitable techniques and apparatus

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Percolation extraction
 Percolation
 concerns the movement and filtering of fluids through porous
materials
or
 Extraction and filtration occurs at the same time
e.g. Instant coffee
 Advantage of percolation extraction
Fast extraction (preliminary analysis)
Disadvantages
 Not suitable for unorganized drugs
 Excess solvents
 Drugs that have high mucus content
 Low yield (limited time contact) 9
Maceration – process that involves soaking
of the material in appropriate solvent with
intermittent shaking until soluble matter is
dissolved

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Modifications of the General Processes of

Maceration

Repeated maceration may be more efficient than a single

maceration,

– appreciable amount of active principle may be left

behind in the first pressing of the marc.

– in cases where active constituents are more valuable.

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 How to increase the yield for maceration

Multiple extraction

Agitation
 is method is best suitable for use in case of the thermolabile drugs
 Disadvantage:
1. Consumes too much solvent
2. Time taking
3. Don’t exhaustively extract the drug.

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 Infusion
 Infusions are dilute solutions containing the readily-
soluble constituents of crude drugs.
Formerly, fresh infusions, prepared by macerating the drug for a short
period in cold water or boiling water were used.

Now, infusions are usually prepared by diluting one volume of a

concentrated infusion to ten volumes with water.

Infusions are liable to fungus and bacterial growth, and it

is necessary to dispense them within twelve hours of their
preparation.
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• Preparation of Concentrated Infusions
The official monographs also recognize certain “concentrated
infusions” in which 25% alcohol is added during or
subsequent to the infusion process.

Concentrated infusions are especially prepared in which the

active and desirable principles of drug are equally soluble in
water or in the menstruum used for concentration.

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 Decoction

 used for the extraction of the water soluble and heat stable constituents
from crude drug by boiling it in water for 15 minutes, cooling, straining
and passing sufficient cold water through the drug to produce the
required volume.
 Advantage:
o Simplicity
o Quick method
 Disadvantage:
o Thermolabile compounds will be destroyed

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 Digestion

a kind of maceration in which gentle heat is applied during the

maceration extraction process.

used when moderately elevated temperature is not unpleasant and the

solvent efficiency of the menstruum is increased thereby.

 Advantage:
• Simplicity
• Preserve thermolabile compounds
 Disadvantage:
• Consumes too much solvent

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 Distillation
It is separation of the constituents of a mixture by partial vaporization of the mixture and include:
a. The separation of one liquid from non volatile impurities
b. The separation of one liquid from one or more other liquids

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 2.Continuous extraction

Continuous extraction: requires special apparatus like

Soxhlet extractor, where multiple extraction occurs

 Soxhlet extraction

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2. Continuous extraction
Multiple extraction

Soxhlet Extraction 20
(A) Soxhlet Apparatus (Hot Continuous Extraction)
designed for the extraction of a lipiq from solid material
The solvent is heated to reflux (condensed) and percolates the
solid material.
involves successive extraction with solvents of increasing
polarity from a non-polar (hexane) to a more polar solvent
(methanol) to ensure that a wide polarity range of compound
could be extracted.

only required for a particle with limited solubility in a

solvent, and the impurity is insoluble in that solvent.
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Its vapour passes up the larger right hand tube into the
upper part of the drug and then to the condenser where it
condenses and drops back on to the drug.

During its percolation, it extracts the soluble constituents.

When the level of the extracts reaches the top level of syphon
tube, the whole of the percolates syphon over into the flask.

The process is continued until the drug is completely

extracted and the extract in the flask is then processed.

This extraction is series of short maceration.

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 Facts about soxhlet extractor
 Using a soxhlet extractor is a way to separate soluble
material from insoluble material
 Soxhlet extraction method involves vaporization and
condensation occur throughout the apparatus
 A soxhlet extractor repeatedly extracts with fresh solvent
until it is stopped. Thus, it is possible to extract all of the
soluble material, if given enough time.
 Solubility is a function of how well a substance dissolves
in a solvent (temp. affects solubility)

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 Advantage of Soxhlet extraction over batch
extraction method

 Soxhlet method give a high yield

 Soxhlet reduces the extraction time (about 3 hrs)
 Soxhlet reduces solvent consumption significantly

 Limitation of Soxhlet extraction

 It is not a good method for extract which are labile to
heat and oxidation

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 3. Countercurrent extraction

Countercurrent extraction: extraction process

which separates two or more solutes with

different partition coefficients from each other by

a series of partitions between two immiscible

liquid phases.
• Advantages:

i) much smaller volume of solvent as compared to other

ii) CCE is commonly done at room temperature,

 spares the thermolabile constituents from exposure to heat.

iii) crushing of the drug is done under wet conditions,

 the heat generated is neutralized by water (spares the
thermolabile constituents )

iv) more efficient and effective than continuous hot

extraction.
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3. Countercurrent extraction
3.1. Liquid-liquid extraction

Liquid-liquid extraction is a useful method to separate

components (compounds) of a mixture

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Let's see an example.

 Suppose that you have a mixture of sugar in vegetable

oil (it tastes sweet!) and you want to separate the sugar
from the oil.
 You observe that the sugar particles are too tiny to filter
and you suspect that the sugar is partially dissolved in
the vegetable oil.

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 How about shaking the mixture with water
Will it separate the sugar from the oil?
 Sugar is much more soluble in water than
in vegetable oil, and, as you know, water is
immiscible (=not soluble) with oil.

 Did you see the result?

 The water phase is the bottom layer and
the oil phase is the top layer, because water
is denser than oil.

*You have not shaken the mixture yet, so

sugar is still in the oil phase.

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 By shaking the layers (phases) well,
you increase the contact area between
the two phases.
 The sugar will move to the phase in
which it is most soluble: the water layer

Now the water phase tastes sweet,

because the sugar is moved to the water
phase upon shaking.
**You extracted sugar from the oil with
water.**In this example
 water was the extraction solvent;
the original oil-sugar mixture was the
solution to be extracted;
and sugar was the compound extracted
from one phase to another.
 Separating the two layers
accomplishes the separation of the
sugar from the vegetable oil
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 Liquid-liquid extraction is based on the transfer of a solute
substance from one liquid phase into another liquid phase
according to the solubility

 Extraction becomes a very useful tool if you choose a

appropriate extraction solvent.You can use extraction to
separate a substance selectively from a mixture, or to
remove unwanted impurities from a solution.

 In the practical use, usually two phases

 one phase is a water or water-based (aqueous) solution
 Second phase is an organic solvent which is immiscible
with water.

The success of this method depends upon:

 the difference in solubility of a compound in various
solvents.
 For a given compound, solubility differences between
solvents is quantified as the "distribution coefficient"
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To facilitate the extraction process/increase the extracting
yield
 Multiple extraction process
 Salting-out: is the process where we saturate the
aqueous phase with NaCl salt so that any organic
dissolved in aqueous forced to move into organic phase

Organic molecules
Organic layer moving into organic layer

Aqueous layer Aqueous phase saturated

With NaCl

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3.2. Acid-base extraction
 Can you change the solubility property of a compound? How?

 Most organic compounds are more soluble in organic solvents

than in water, usually by the distribution coefficient K > 4
 However, specific classes of organic compounds can be
reversibly altered chemically to become more water-soluble.

 This is a powerful technique and allows you to separate

organic compounds from a mixture -- if they belong to
different solubility classes
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 What type of organic compounds can be made water-
soluble?
 Compounds belonging to the following solubility classes
can be converted to their water-soluble salt form
(1) Organic acids include
carboxylic acids (strong organic acids) and
phenols (weak organic acids).

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(2) Organic bases includes amines

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How can organic acids or bases be converted to a water-
soluble form?

 Organic Acids can be converted to their salt form when

treated with an

 aqueous solution of inorganic base e.g.,

 NaOH (sodium hydroxide) and
 NaHCO3 (sodium bicarbonate).

 Salts are ionic, and in general, ions are soluble in water

but not soluble in water-immiscible organic solvents.

 Remember: water is a very polar solvent thus salts (i.e.,

ionic species) are well dissolved in it.

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 Carboxylic Acids are converted to the salt form with 5% NaOH aqueous
solution. NaOH is a strong inorganic base.

 Carboxylic acids are strong organic acids (pKa = 3 to 4), so they can also be
ionized with weak inorganic bases (e.g., NaHCO3 (sodium bicarbonate))
aqueous solution.

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E.g. 1: Here is a mixture of naphthalene and benzoic acid, dissolved in
dichloromethane.

You want to separate these two

compounds. What will you do?

You may use an aqueous solution of

either 5% NaOH or sat. NaHCO3, to
extract benzoic acid as a salt form

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Phenols or substituted phenols are considered to be weak
organic acids.

 treatment with NaOH, a strong inorganic base, can change

phenol to its ionic (salt) form
Sodium bicarbonate (NaHCO3), a weak inorganic base, will
not deprotonate phenols to make it ionic, because it is not
strong enough.

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 Here is a mixture of:
e.g. 1: Benzoic acid and p-methoxyphenol, dissolved in
dichloromethane.

You want to separate these two compounds.

What will you do?

You cannot use 5% NaOH to separate these two

compounds.
NaOH will react with both benzoic acid and p-
methoxyphenol, thus both compounds will be extracted into
the aqueous layer.
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 NaOH was too strong a base, thus it does not
differentiate the strong and weak organic acids.
 Use of weak inorganic base such as NaHCO3 will
differentiate between the compounds

 Strong organic acids such as benzoic acid would be

deprotonated and ionized, while weak organic acids
such as phenols would NOT be deprotonated

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 Organic Bases (amines) can be converted to their salt
form when treated with an aqueous solution of an
inorganic acid such as HCl (hydrochloric acid).

 Recall that salts are ionic and generally soluble in water

but not soluble in water-immiscible organic solvents.

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Eg.1 : Here is a mixture of:
Benzoic acid and p-chloroaniline, dissolved in dichloromethane.

You want to separate these two compounds.

What will you do?

You may use an aqueous solution of 5% HCl,

 P-chloroaniline converts to a salt form and
 benzoic acid remains unchanged in the organic layer

You may treat the extract with 5% NaOH solution,

 benzoic acid convert sto a salt form and
 P-chloroaniline remains unchanged in the organic layer 43
E.g. 1: Separation of four different classes of compounds from a mixture based on
differing solubility properties. The four classes are:

1. Amines (organic base) 2. Carboxylic acids (strong acid)

3. Phenols (weak acid) 4. Neutral compounds.

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After the separation of the mixture of four components, we will have four
solutions: each solution contains one component.

The first three compounds are chemically altered, existing in their salt form
dissolved in aqueous solution. The fourth compound is not chemically altered,
but it is dissolved in an organic solvent.We now want to recover each compound
in its original state (i.e., in the non-ionic form) to complete the experiment. We
call this step isolation or recovery.

Let's see, one by one, how to recover each compound obtained from the
separation process
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• Assignment---Recovery
 Advanced methods of extraction

 Advanced methods of extraction

Pressurized liquid extraction (PLE).

Static headspace extraction

microwave

Supercritical fluids extraction

ultrasound-assisted extractions
Pressurized liquid extraction (PLE)

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Facts about PLE
 PLE is a recently adopted extraction method that uses
o high pressure and
o high temperature
o under a forced flow of solvent.
 PLE , known as accelerated solvent extractor
 Compressed nitrogen is usually used as a carrier to move
the solvent under high pressure.
 Increased temperature and elevated pressure accelerate
the extraction kinetics so as to enable rapid extraction.
 PLE is important extraction methods for those extract
liable to oxidation .
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Operating principle of PLE
 The PLE works similar to Soxhlet extraction, with the exception
that during the extraction process the solvents inside the PLE
extraction column are near their supercritical region which has
high extraction properties.

 the near supercritical region the

 high solubility due to high temperature and
 high diffusion rate while the high pressure keeps
the solvent below its boiling point.

 At high pressures and high temperatures the solvents penetrate

the solid samples at a much higher rate permitting a fast and
efficient extraction process with minimal solvent usage
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Phase diagram--- （ P---T ）

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Static headspace extraction/analysis

Extraction Factors:
- temperature and
- amount of sample
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 Microwave-assisted extraction (MAE)
 MAE involves heating solid sample-solvent mixtures in a
closed vessel with microwave energy under
 temperature-controlled and
 pressure-controlled conditions.
 This closed extraction system enables analyte extraction
with elevated temperatures and pressure accelerating
the extraction process
 MAE yields a performance comparable to the standard
Soxhlet method.

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MAE apparatus
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 supercritical fluids extraction

A supercritical fluid is any substance

above its critical temperature and
critical pressure. In the supercritical
area there is only one state-of-the-
fluid and
it possesses both gas- and liquid-like
properties.
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Phase diagram--- （ P---T ）

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 Properties of supercritical fluids
• A supercritical fluid exhibits
physicochemical properties intermediate
between those of liquids and gases.
• Characteristics of a supercritical fluid
are :
– Solubilities approaching liquid phase
– Diffusivities approaching gas phase.
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 Properties of supercritical
fluids
• Therefore, the properties of
- gas-like diffusivity,
- liquid-like density have provided
the impetus for applying supercritical
fluid technology to various problems.

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Supercritical fluid extractor
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 ultrasound-assisted extractions (Sonication)
involves the use of ultrasound with frequencies ranging from 20 kHz to
2000 kHz;
increases the permeability of cell walls and produces cavitation.
its large-scale application is limited due to the higher costs.
disadvantage
– 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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Variation in extraction methods
usually depends upon
Length of the extraction period,

Solvent used,

pH of the solvent,

Temperature,

Particle size of the plant tissues

The solvent-to-sample ratio

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 Factors Affecting Choice of Extraction Process

•The final choice of the process to be used for the

extraction of a drug will depend on a number of

factors, including:

Character of Drug

If hard and tough , use percolation.

If soft and parenchymatous ,use maceration.

If ‘unpowderable’ , use maceration. 62

If an ‘unorganized drug , use maceration.

If preferable to avoid powdering , use maceration.

Thus, knowledge of the pharmacognosy of the drug is

essential to selection of the extraction process that will give

the best results.

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 Parameters for Selecting an Appropriate
Extraction Method
1. Authentication of plant material
 Any foreign matter should be completely eliminated.

2. Use the right plant part &, for quality control purposes, record
the age of plant & the time, season and place of collection.
3. 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.
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 Parameters for………
4. Grinding methods should be specified & techniques that generate
heat should be avoided as much as possible.
5. Powdered plant material should be passed through suitable sieves
to get the required particles of uniform size.
6. Nature of constituents:
a) If the therapeutic value lies in non-polar constituents, a non-
polar solvent may be used.
b) If thermolabile constituent: cold maceration, percolation & CCE.
C) If thermostable constituents
• (Soxhlet extraction (if nonaqueous solvents are used)and
decoction (if water is the menstruum)).
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 Parameters for………
d) Suitable precautions should be taken when dealing with
constituents that degrade while being kept in organic
solvents.
– e.g. flavonoids and phenyl propanoids.
e) Higher than required temperature should be avoided (Some
glycosides).
f) Standardization of time of extraction
– Insufficient time (incomplete extraction).
– time is longer, unwanted constituents extracted.
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 Parameters for………
g) The number of extractions required for complete extraction is as
important as the duration of each extraction.

7. The quality of water or menstruum used should be specified and

controlled.

8. 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 is increasingly employed (expensive).

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 Parameters for………
9. Analytical parameters of the final extract, such as TLC and
HPLC fingerprints.
• Steps Involved in the Extraction of Medicinal Plants.

1. Size reduction

2. Extraction

3. Filtration

4. Concentration

5. Drying
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 Choice of solvents
 For Successful determination of biologically active
compounds from plant material is largely dependent on the
type of solvent used in the extraction procedure.

 A property of a good solvent in plant extractions includes:

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

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• Solvents, such as methanol, ethanol, acetone, ethyl acetate,
and their combinations have been used for the extraction of
phenolics from plant materials, often with different
proportions of water.
– Selecting the right solvent affects the amount and rate of polyphenols
extracted.

• A preliminary selection of solvents must be based on an

estimate of chemical interactions that could be important.
– mainly ones of solute–solvent and solvent–solvent.

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• The interaction is
– least when both are nonpolar;
– becomes stronger as the polarity of the molecules increases

– strongest when there are interactions caused by formation

of hydrogen bonds, charge transfer complexes, or
coordination complexes.

• In reactive solvent extraction, it is desirable to find a

solvent that can form strong, specific interactions with
the solute to be extracted.
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72
• The yields of antioxidative components extracted by
80% methanol varied significantly (p < 0.05) from
other solvents used.
• The amount of compounds extracted ranged from 9.6
for pure ethanol to 13.0% for 80% aqueous methanol,
indicating that added water, the most polar solvent
used, caused further extraction of polar components.

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Thank you!!
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