extraction ppt.ppt

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EXTRACTION PROTOCOLS

Extraction
• Is the separation of medicinally active
portions of plant (and animal) tissues using
selective solvents through standard
procedures.
• The products so obtained from plants are
relatively complex mixtures of metabolites,
in liquid or semisolid state or in dry powder
form (after removing the solvent), & are
intended for oral or external use.

Medicinal plants
• The Medicinal plants constitute an effective
source of both traditional and modern
medicines, herbal medicine has been shown
to have genuine utility and about 80% of
rural population depends on it as primary
health care. [WHO, (2005)]

Medicinal plants are the richest bio-resource
drugs of traditional systems of medicine,
modern medicines,
nutraceuticals,
food supplements,
folk medicines,
pharmaceutical intermediates,
chemical entities for synthetic drugs.

galenicals
These include classes of preparations viz.,
decoctions,
infusions,
fluid extracts,
tinctures,
pilular (semisolid) extracts,
powdered extracts.

Evidence for Ethnopharmacology
• De Materia Medica,
• Historia Plantarum,
• Species Plantarum.
have been variously published in
to provide scientific information on
medicinal uses of plants.

Standardized Extraction
The purpose of standardized extraction procedures
for crude drugs
(medicinal & aromatic plant parts)
 To attain the therapeutically desired portions
 To eliminate unwanted material by treatment
with a selective solvent known as “menstrum”

The extract thus obtained, after standardization,
may be used as medicinal agent
• as such in the form of tinctures
• fluid extracts
• further processed to be incorporated in any
dosage form such as tablets and capsules

The general techniques of medicinal plant extraction
 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).

Extraction techniques For aromatic plants
 Hydrodistillation techniques (water distillation,
steam distillation, water and steam distillation),
 Hydrolytic maceration followed by distillation,
expression and enfleurage (cold fat extraction)
 Headspace trapping,
 Solid phase micro-extraction,
 Protoplast extraction,
 Microdistillation,
 Thermomicrodistillation,
 Molecular distillation.

Choice of solvents
• Successful determination of biologically active
compounds depends on the type of solvent used in the
extraction procedure.
• The choice of solvent is influenced by what is
intended with the extract.

Properties of a good solvent in plant extractions
 low toxicity,
 ease of evaporation at low heat,
 promotion of rapid physiologic absorption of the
extract,
 preservative action,
 inability to cause the extract to complex or dissociate.

Solvents used for active component extraction

Water
• Water is universal solvent.
• used to extract plant products with antimicrobial
activity.
• Traditional healers use primarily water & consistent
antimicrobial activity is obtained.
Plant extracts: organic solvents >>> water extract.
• Water soluble flavonoids (mostly anthocyanins)
have no antimicrobial significance.
• only water soluble phenolics are important as
antioxidant compound.

Acetone
• Dissolves many hydrophilic and lipophilic
components.
• a very useful extractant, especially for
antimicrobial studies (phenolic group extract).
• extraction of tannins + phenolics:
aqueous acetone >>> aqueous methanol
• Both acetone and methanol were found to extract
saponins  antimicrobial activity.

Alcohol
• The identified components from plants (antimicrobial)
= aromatic or saturated organic compounds  most
often obtained through initial ethanol or methanol
extraction.
• Ethanol, found easier to penetrate the cellular
membrane to extract the intracellular
ingredients(polyphenols) from the plant material.
• Methanol is more polar than ethanol but due to its
cytotoxic nature.
• The higher concentrations of more bioactive flavonoid
compounds were detected with ethanol
70% due to its higher polarity than pure ethanol.

Chloroform
• Used to obtain tannins and terpenoids.
• Terpenoid lactones successive
extractions of dried barks with
chloroform.

Ether
• Commonly used selectively for the
extraction of coumarins and fatty acids.

Dichloromethanol
• Specially used for the selective extraction of
only terpenoids.

Steps Involved in the Extraction of Medicinal
Plants
1. Size reduction
2. Extraction
3. Filtration
4. Concentration
5. Drying

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

Extraction
Medicinal plants Aromatic plants
1. Cold aqueous percolation
2. Hot aqueous extraction
(decoction)
3. Solvent extraction
(cold / hot)
1. Essential oils
2. Concretes
3. Bsolutes
4. Pomades
5. Esinoids

parameters influencing the quality of an extract
• Plant part used as starting material
• Solvent used for extraction
• Extraction procedure

Effect of extracted plant phytochemicals depends on
• The nature of the plant material
• Its origin
• Degree of processing
• Moisture content
• Particle size

Selection of plant
• Plant based natural constituents can be derived
from any part of the plant like bark, leaves,
flowers, roots, fruits, seeds, etc.
• Plants are usually air dried to a constant weight
before extraction.
• oven drying: every part were cut into pieces
dried in an oven @ 60°C for 9 hrs. &
pulverized.
• Other method for drying the plants is the oven
drying at about 40°C for 72 h.

Filtration
• The extract so obtained is separated out from the
marc (exhausted plant material) by allowing it to
trickle into a holding tank through the built-in
false bottom of the extractor, which is covered
with a filter cloth.
• The marc is retained at the false bottom, and the
extract is received in the holding tank.
• From the holding tank, the extract is pumped into
a sparkler filter to remove fine or colloidal
particles from the extract.

Concentration
• The enriched extract from percolators or extractors, known as
miscella, is fed into a wiped film evaporator where it is
concentrated under vacuum to produce a thick concentrated
extract.
• The concentrated extract is further fed into a vacuum chamber
dryer to produce a solid mass free from solvent.
• The solvent recovered from the wiped film evaporator and
vacuum chamber dryer is recycled back to the percolator or
extractor for the next batch of plant material.
• The solid mass thus obtained is
pulverized and used directly for the
desired pharmaceutical formulations
or further processed for isolation
of its phytoconstituents.

Drying
• The filtered extract is subjected to spray drying
with a high pressure pump at a controlled feed rate
and temperature  to get dry powder.
• The desired particle size of the product is obtained
by controlling the inside temperature of the
chamber and by varying the pressure of the pump.
• The dry powder is mixed with suitable diluents or
excipients and blended in a double cone mixer to
obtain a homogeneous powder that can be straight
away used (for example, for filling in capsules or
making tablets).

Variation in extraction methods
• Length of the extraction period,
• Solvent used,
• pH of the solvent,
• Temperature,
• Particle size of the plant tissues,
• The solvent-to-sample ratio.

Parameters for Selecting an Appropriate Extraction Method
I. Authentication of plant material by botanist.
II. Use the right plant part + the age of plant + the time,
season & place of collection.
III. The nature of its chemical constituents.
IV. Grinding methods & powdering techniques.
V. Nature of constituents (polar/nonpolar).
VI. The quality of water / menstruum.
VII.The design & material of fabrication of the extractor.
VIII.Analytical parameters of the final extract,
(TLC/HPLC).

The general techniques of medicinal plant extraction
 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).

Maceration
• The whole / coarsely powdered crude drug is placed
in a stoppered container with the solvent.
• Allow to stand @ room temperature for a period of
at least 3 days with frequent agitation until the
soluble matter gets dissolved.
• The mixture then is strained, the marc (the damp
solid material) is pressed,
• The combined liquids are clarified by filtration or
decantation after standing.
• This method is best suitable
for use in case of the
thermolabile drugs.

Infusion
• Fresh infusions are prepared by macerating
the crude drug for a short period of time with
cold or boiling water.
• These are dilute solutions of the readily
soluble constituents of crude drugs.

Digestion
• This is a form of maceration in which gentle
heat is used during the process of extraction.
• It is used when moderately elevated
temperature is not objectionable.
• The solvent efficiency of the menstruum is
thereby increased.
Image=microwave
Digestion system

Decoction
• In this process, the crude drug is boiled in a
specified volume of water (1;4) for a defined time,
• Volume is reduced to 1/4th the original,
• It is then cooled and strained / filtered.
• This procedure is suitable for extracting  water-
soluble, heat-stable constituents.
• Typically used in preparation of Ayurvedic extracts
= “quath” / “kawath”

Percolation
• Used most frequently to extract active ingredients in the
preparation of tinctures and fluid extracts.
• The solid ingredients are moistened with an appropriate
amount of the specified menstruum,
• Allowed to stand for approximately 4 hours in a well
closed container, After stand time, the mass is packed
& the top of the percolator is closed.
• the mixture is allowed
• to macerate in the closed
percolator for 24 h.

,
• Additional menstruum is added as required, until the
percolate measures about three-quarters of the
required volume of the finished product.
• The marc is then pressed and the expressed liquid is
added to the percolate.
• Sufficient menstruum is added to produce the
required volume.
• The mixed liquid is
clarified by filtration or
by standing followed
by decanting.

Hot Continuous Extraction
(Soxhlet)
• The finely ground crude drug is placed in a
porous bag or “thimble” made of strong filter
paper, which is placed in chamber of the
Soxhlet apparatus.
• The extracting solvent in flask is heated, and its
vapors condense in condenser.
• The condensed extractant drips into the thimble
containing the crude drug & extracts it by
contact.

Soxhlet apparatus
• When the level of liquid in chamber rises to the
top of siphon tube, the liquid contents of chamber
siphon into flask
• This process is continuous and is carried out until
a drop of solvent from the siphon tube does not
leave residue when evaporated.

Aqueous Alcoholic Extraction by Fermentation
• Some medicinal preparations of Ayurveda (asava &
arista) adopt the technique of fermentation for
for extracting the active principles.
• The extraction procedure involves soaking the crude
drug, [powder / a decoction (kasaya)], for a specified
period of time
• Undergoes fermentation & generates alcohol in situ.
• This facilitates the extraction of the active
constituents contained in the plant material.
• The alcohol thus generated also serves as a
preservative.

,
• Some examples of Ayurvedic preparations:
karpurasava,
kanakasava,
dasmularista.
• If the fermentation is to be carried out in an earthen
vessel, it should not be new: water should first be
boiled in the vessel.
• In large-scale manufacture, wooden vats, porcelain
jars or metal vessels are used in place of earthen
vessels.

Counter-current Extraction
• Wet raw material is pulverized using toothed disc disintegrators to
produce a fine slurry.
• Material to be extracted is moved in one direction generally in the
form of a fine slurry within a cylindrical extractor where it comes in
contact with extraction solvent.
• The further the starting material moves, the more concentrated the
extract becomes.
• Complete extraction is thus possible when the quantities of solvent
& material. Their flow rates should be optimized.
• 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.

Ultrasound Extraction (Sonication)
• The procedure involves the use of ultrasound with
frequencies ranging from 20 kHz to 2000 kHz.
• This increases the permeability of
cell walls & produces cavitation.
Eg: extraction of rauwolfia root.
• Deleterious effect: Ultrasound energy (>20 kHz) on
the active constituents of medicinal plants through
formation of free radicals and consequently
undesirable changes in the drug molecules.

Supercritical Fluid Extraction
• Cylindrical extraction vessels are used.
• The collection of the extracted analyte following SFE is
another important step: significant analyte loss can occur
during this step.
• CO2 as the extracting fluid.
• Organic solvents are frequently added to the CO2
extracting fluid to alleviate the polarity limitations
• The component recovery rates generally increase with
increasing pressure/temperature.
• The highest recovery rates in case of argon:
@ 500 atm & 150° C.

Phytonics Process
• A new solvent based on hydrofluorocarbon-134a, a new
technology to optimize the extraction of plant materials.
• Advanced Phytonics Limited (Manchester, UK) has developed
 patented technology termed “phytonics process”.
• The products are fragrant components of essential oils &
Biological/phytopharmacological extracts.
• The core of the solvent is 1,1,2,2-tetrafluoroethane, (HFC-
134a).
• HFC-134a developed as a replacement for
chlorofluorocarbons. (Boiling Point -25° C).
• the solvents can be customized: by using modified solvents
with HFC-134a.
• The process can be made highly selective in extracting a
specific class of phytoconstituents.

Aromatic Plant Extracts
• essential oils,
• concretes,
• absolutes,
• pomades
• resinoids.

Essential oils
• Used in a wide variety of consumer goods viz.,
detergents, soaps, toilet products, cosmetics,
pharmaceuticals, perfumes, confectionery food
products, soft drinks, distilled alcoholic
beverages (hard drinks) and insecticides.
• Production technology is an essential element
to improve the overall yield & quality of
essential oil.

Presence of Essential Oils in plant parts.

Hetreogenous chemical group present in essential oils

The traditional methods for essential oil extraction
• Water distillation,
• Water and steam distillation,
• Steam distillation,
• Cohobation,
• Maceration,
• Enfleurage.

methods
1. Distillation: Originally introduced by Von Rechenberg.
• water distillation.
• water and steam distillation.
• direct steam distillation.
2. Hydrolytic maceration distillation.
3. Expression.
4. Cold fat extraction / Enfleurage.

• Distillation methods are good for powdered
almonds, rose petals and rose blossoms.
• Maceration is adaptable when oil yield from
distillation is poor.
• Solvent extraction is suitable for expensive,
delicate & thermally unstable materials like
jasmine, tuberose, and hyacinth.
• Water distillation is the most favored method of
production of citronella oil from plant material.

1. Headspace trapping techniques
- Static headspace technique
- Vacuum headspace technique
- Dynamic headspace technique
2. Solid phase micro-extraction (SPME)
3. Supercritical fluid extraction (SFE)
4. Phytosol (phytol) extraction
5. Protoplast technique
6. Simultaneous distillation extraction (SDE)
7. Microwave distillation
8. Controlled instantaneous decomposition (CID)
9. Thermomicrodistillation
10. Microdistillation
11. Molecular spinning band distillation
12. Membrane extraction
Modern (Non-traditional) Methods of Extraction

Concrete
• This is an extract of fresh flowers, herbs, leaves and the
flowering tops of plants obtained by the use of a hydrocarbon
solvent such as butane, pentane, hexane and petroleum ether.
• Concrete is rich in hydrocarbon soluble material & devoid of
water-soluble components.
• It is generally a waxy, semisolid, dark-colored material free
from the original solvent.
• concretes are produced in
static extractors.
• It is a normal practice to circulate fresh
solvent through a battery of extractors.
• The enriched solvent from the extractor is pumped into an
evaporator for solvent recovery & the solvent content is
reduced to about 1/10th the original volume.

Absolutes
• To make an absolute, the concrete is mixed with absolute alcohol
& agitated thoroughly in a vessel with an agitator.
• During agitation, the temperature is kept at 40°-60° C and the
concrete is immersed in the solution.
• The solution is cooled down to -5° to -10° C to precipitate out the
wax, since waxes are normally insoluble in alcohol below -1° C.
• The precipitated wax is removed by passing the solution through a
rotary filter.
• The filtrate from the rotary filter is pumped into a primary
evaporator, where it is concentrated to about 10% alcohol content.
• Finally, the concentrated extract is pumped into an
agitating-type evaporator, where the alcohol
is carefully removed under high vacuum.

Resinoids
• Resinoid is an extract of naturally resinous material,
made with a hydrocarbon solvent.
• Resinoids are usually obtained from dry materials.
• The extraction process is same as that of concrete
production, except that perforated discs are not used
for stacking the material;
• instead powder from dry plant
material is fed into the extractor.

Pomades
• Pomades are obtained by a process known as
enfleurage, which is a cold fat extraction method.
• The fat is spread out on glass plates contained in wooden
frames, leaving a clear margin near the edges.
• The absorptive surface of the fat is increased by surface
grooves made with a wooden spatula.
• Fresh flowers are spread out on the surface of the fat and the
frames are stacked in piles.
• After the perfume oils have been absorbed from the flowers,
the spent flowers are removed by hand.
• Fresh flowers are again spread on the fat surface.
• This is repeated until the fat surface is completely enriched
with perfume oils.
• The pomade so obtained is ready for cold alcoholic extraction

major constraints in sustainable industrial exploitation
• Poor agricultural practices for MAPs,
• Unscientific and indiscriminate gathering practices from the wild, poor
postharvest & post-gathering practices leading to poor quality raw
material,
• Lack of research for the development of high-yielding varieties of
medicinal and aromatic plants-MAP’s.
• Poor propagation methods.
• Inefficient processing techniques.
• Poor quality control procedures.
• Lack of research on process
& product development.
• Difficulty in marketing.
• Non-availability of trained personnel.
• Lack of facilities & tools to fabricate equipment locally.
• Finally lack of access to the latest technologies & market information.

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