Extraction, Separation and Analysis Techniques for MAPs ... · application in environmental as well...
Transcript of Extraction, Separation and Analysis Techniques for MAPs ... · application in environmental as well...
Extraction, Separation and
Analysis Techniques for MAPs:
from Field to Market
Prof. Dr. Temel ÖZEK Anadolu University, Faculty of Pharmacy, 26470-Eskişehir / TURKEY
Introduction
Conventional techniques Modern
techniques
MAPs Extraction, Separation and Analysis Techniques
Regional Expert Consultation on Medicinal and Aromatic Plants, 13-15 November 2013, Antalya 2
Introduction
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Introduction
nature of the material
yield and properties of the extracts
marketability of the products
Choice of
extraction
technique
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Conventional Extraction Techniques
Mechanical extraction
Solvent extraction
Cold fat extraction
Distillation
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Modern Extraction & Separation Techniques
• Liquified gas extraction
• Supercritical fluid extraction
• Protoplast technique
• Membrane extraction
• Solid phase micro-extraction (SPME)
• Headspace trapping techniques
— Static headspace
— Dynamic headspace
— Vacuum headspace
• Controlled instantaneous decomposition (CID)
• Simultaneous distillation extraction (SDE)
• Microwave extraction & distillation
• Microdistillation
• Molecular spinning band distillation
• Thermomicrodistillation
• Preparative fractionation
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Essential Oil Extraction Techniques
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Distillation
Expression
Extraction
Extraction Techniques
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Mechanic
al
Extr
acti
on
Expression
Scratch
Extraction Techniques
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Mechanic
al
Extr
acti
on
Expression
Scratch
• Expression or Cold Pressing is confined
to citrus oils.
• It involves inducing physical damage to
the essential oil glands on the surface
of citrus fruits to release the oil.
• It is simultaneously washed by the
passing water and recovered using an
oil-water separator.
Extraction Techniques
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Mechanic
al
Extr
acti
on
Expression
Scratch
• Today, there are four major processes
that are used to extract citrus oils at
commercial scale. • Pellatrice
• Sfumatrice
• Brown Peel Shaver
• FMC in-line extractor
Extraction Techniques
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Mechanic
al
Extr
acti
on
Expression
Scratch
• For natural rubber, workers called
tappers cut a shallow groove in the
bark of a tree and collect the drops of
latex in a cup.
• For opium production, the skin of the
ripening pods of these poppies is
scored by a sharp blade at a time
carefully and then dry latex collected.
Solvent Extraction Technique
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• The extraction is done by organic solvent.
• After the extraction, solvent is evaporated.
• This “concentrated solution” is then directed, after filtering,
into a vacuum evaporator.
• Vacuum distillation to remove the last traces of solvent is
necessary to produce an acceptable product.
• The product is called “Concrete”
if the material used is fresh floral.
• If it has been extracted from dry or
viscous products it is more commonly
called “Resinoid”.
Extraction with Cold Fats
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Enfleurage
• This technique used to be very popular.
• It is very rarely used nowadays.
• It used to be the method of choice for extracting flowers like
Jasmin and Tuberose.
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• The method called “advanced Phytonics” use 1,1,1,2-
tetrafluoroethane with or without modifiers.
• The solvents are named “Phytosols®”.
• Phytosol A consists only of the gas
• Phytosol B is a mixture of the gas with butane/isobutane
• Phytosol C consists of the gas with dimethylether as modifier
• Boiling point of 1,1,1,2-tetrafluoroethane or HFC 134 as known in
trade is –26.2°C.
• It is a non-toxic, odourless gas whose pressure in liquid state at room
temperature is 5 bar which can be compared to the pressure of a
bottle of Champagne (6 bar).
Modern Extraction & Separation Techniques
Liquified Gas Extraction Technique: Phytosol
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Applications: The process is aptly applicable to
those compounds that are
• sensitive to thermal or UV energy
• oxygen and acidic conditions
• pharmaceutical
• cosmetic
• medical
• food
• flavors and fragrances industries
Modern Extraction & Separation Techniques
Liquified Gas Extraction Technique: Phytosol
Supercritical Fluid Extraction Technique
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• The critical point of a pure substance is defined as
the highest temperature and pressure at which it
can exist in vapour-liquid equilibrium.
• At pressures and temperatures above this point, a
single homogeneous fluid which forms is said to be
supercritical.
• A substance in the supercritical phase is
neither a true liquid nor a true gas and
has some properties of each.
Supercritical Fluid Extraction Technique
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• Supercritical fluids possess superior mass transfer properties by
virtue of their low viscosities and high solute diffusivities along
with the ability to penetrate microporous materials.
• Among the variety of gasses that can be
rendered supercritical, carbondioxide
has become the most popular and most
widely used since it is • harmless
• non-flammable
• cheap
• abundantly available
• non-corrosive
• has a low boiling point
Supercritical Fluid Extraction Technique
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Applications:
• Food
• Pharmaceutical
• fine chemical industries
• Decaffeinating of coffee and tea
• Extraction of essential oils (vegetable and fish oils)
• Extraction of flavors from natural resources (nutraceuticals)
• Extraction of ingredients from spices and red peppers
• Extraction of fat from food products
• Fractionation of polymeric materials
• Extraction from natural products
• Photo–resist cleaning
• Precision part cleaning
Poroplast Extraction Technique
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• Liquid-liquid extraction can be defined as a partition technique
where the constituents of a solution are separated due to their
different partition coefficients between the two immiscible
liquids.
• Porous support which holds the stationary phase is teflon powder
made into a sponge through a special heat treatment.
• Aqueous dispersion is pumped into the column.
• Aromatic constituents are transferred from the
aqueous phase to the organic phase.
Membrane Extraction Technique
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• In all living systems the transfer of solutes is achieved through
membrane processes.
• In membrane technology, the membrane used is not always a
solid.
• Liquid Membrane Extraction consists of three liquid systems
• Feed
• Membrane
• Sripping
Membrane Extraction Technique
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• There are several membrane extraction and distillation techniques.
• Pervaporation
• Liquid membrane extraction
• The basic principle of pervaporation for the
removal of volatile organic compounds
(VOC) from aqueous media is as follows:
• Hot aqueous feed flows alongside a non-
porous hydrophobic membrane which
consists of a thin film of silicone rubber
on teflon.
Headspace Trapping Techniques
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• Headspace trapping techniques are used to capture
the odour emitted by aromatic materials.
• Odour can be sampled either directly or trapped on
an adsorbent material.
• The trapped odorous components can be freed by
solvent extraction or thermal desorption prior to
analysis by modern instrumental techniques.
Headspace Trapping Techniques
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Static headspace sampling
Vacuum headspace sampling
Dynamic headspace sampling
Static Headspace Sampling Technique
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• In the static headspace
sampling technique,
analyte is kept in a closed
vial and the air (headspace
air) above the solid or a
liquid sample is sampled by
a gas syringe or directed on
to the gas chromatography
column or more usually first
concentrated on an
adsorbent trap.
Vacuum Headspace Sampling Technique
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• Vacuum headspace sampling
technique involves suction of
the headspace air via a vacuum
pump through condensers
cooled with liquid nitrogen to
condense odorous principles.
• This technique is also used by
some perfumery companies for
commercial scale production of
fragrances.
Dynamic Headspace Sampling Technique
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• Dynamic headspace sampling
involves sweeping the analyte
with a stream of air or gas and
adsorption of the volatiles from
the gas stream on an adsorbent
trap.
• Dynamic headspace sampling
techniques can be applied in one
of the following ways:
• Closed-loop Stripping Method
• Direct Sampling Method
Dynamic Headspace Sampling Technique
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• Direct Sampling Method
Solid Phase Micro-Extraction (SPME)
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• Solid Phase Micro Extraction (SPME) is a micro
sampling technique which has found wide
application in environmental as well as flavour
and fragrance research.
• It is a solvent-free method which is used to trap
flavours and fragrances either from aqueous
samples (immersion SPME) or from the vapour
space above a liquid or a solid sample
(headspace SPME).
Solid Phase Micro-Extraction (SPME)
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• SPME can be efficiently used for headspace
sampling since contaminations or impurities
mentioned above do not occur and highly
comparable results are obtained.
• Several polar coatings are commercially
available such as
• polydimethylsiloxane (PDMS)
• polyacrylate (PA)
• polydimethylsiloxane/divinylbenzene (PDMS/DVB)
• carbowax (CW)
• carbowax/divinylbenzene (CW/DVB)
• carboxene/polydimethylsiloxane (C/PDMS)
Stir-Bar Sorptive Extraction (SBSE)
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• Stir-bar sorptive extraction (SBSE) has recently
been developed for aqueous samples.
• SBSE is an equilibrium technique like SPME.
• It is faster than with conventional techniques,
omitting time-costly preparation steps and
solvents and more sensitive than SPME.
Stir-Bar Sorptive Extraction (SBSE)
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Advantages of SBSE Lower detection limits than SPME
Quantitative applications with large linear dynamic range
Several samples may be extracted Simultaneonsly
Less time and labor consuming
Large application area
Applications Food and beverages
Flavors and perfumes
Environmental analysis of water or waste water
Biomedicine
Quality control
Residue analysis
Likens-Nickerson Simultaneous Distillation-Extraction (SDE)
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• Allows the simultaneous countercurrent
extraction of the distillate with an
immiscible solvent.
• In this ingenious design, aromatic plant
material is distilled with water in the
distillation flask and the immiscible solvent
is boiled in the receiver flask.
• Both vapours condense on the same
condenser in the middle of the assembly
and mix in a central port.
Controlled Instantaneous Decompression (CID)
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• The process involves subjecting the partially
humidified plant material for a short period of time
to a steam pressure varying from 0.5 to 3 bar
followed by a rapid decompression to a vacuum
(about 15 mbar) for 200 msec each time.
• The vapour in the plant material created by
autovaporization produces a mechanical strength
which ruptures the oil cells.
Controlled Instantaneous Decompression (CID)
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• Due to pressure differences in the extractor and
vacuum chamber, in each opening of the pressure
valve oil-rich vapour is instantaneously sucked into
the vacuum chamber where it instantly condenses
Microwave Extraction & Distillation
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• Microwaves are a form of electromagnetic radiation.
• They are very short waves that travel at the speed
of light.
• Microwave region is between
infrared and radio waves on the
electromagnetic spectrum.
Microwave Extraction & Distillation
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• Microwave region is also within the “non-ionizing”
region on the electromagnetic spectrum.
• There are two type of radiation.
• They are completely different from each other • Ionizing Radiation: HARMFUL
• Non-Ionizing Radiation: NON-HARMFUL
Microwave Extraction & Distillation
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Two Heating Model
M. Letellier and H. Budzinski*, Microwave assisted extraction of organic compounds, Analusis, 27, 259-271 (1999).
Microwave Extraction & Distillation
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Solvent Extraction Heating Mechanisms
Type-I : The sample could be immersed in a single solvent or mixture of solvents that
absorb microwave energy strongly.
Type-II : The sample could be extracted in a combined solvent containing solvents
with both high and low dielectric losses mixed in various proportions.
Type-III : Samples that have a high dielectric loss can be extracted with a microwave
transparent solvent.
Microwave Extraction & Distillation
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Heating Mechanism of Apolar Solvents
• Non-polar solvents such as hexane, pentane, xylene etc. will not heat up when
exposed to microwaves.
• These are called as transparent to microwave.
• These kind of solvents can be heated up by means of special inert materials.
Commercialy evailable materials for these purposes are:
MILESTONE : Weflon
CEM : Carboflon
These are patented materials
Microwave Extraction & Distillation
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Microwave-assisted Hydrodistillation (MWHD) System
Milestone ETHOS E
Microwave Extraction & Distillation
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DryDist Microwave-assisted
Hydrodistillation
(MWHD) System
Milestone
Microwave Extraction & Distillation
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Matching of HD and MW-HD Essential Oil Chromatograms of
Heracleum crenatifolium
T. Özek, G. Özek, K.H.C. Baser, A. Duran, Comparison of the essential oils of three endemic Turkish Heracleum species obtained by different isolation techniques,
J. Essent. Oil Res., 17, 605-610 (2005).
Microwave Extraction & Distillation
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MWHD-Classical Distillation
M. Koşar, T. Özek, F. Göger, M. Kürkçüoğlu, K.H.C. Başer, Comparison of Microwave-Assisted Hydrodistillation and Hydrodistillation Methods for the Analysis of
Volatile Secondary Metabolites, Pharm. Biol. 43(6) 491-495 (2005).
Material : Anethum graveolens L. and Coriandrum sativum L.
Material quantity : 100 g
Clevenger MWHD#
Time (min.) : 180 60
Yield (%)
Anethum graveolens L. : 2.1 2.5
Carvone+ (%) : 45.7 69.3
Coriandrum sativum L. : 0.4 0.5
Linalool* (%) : 79.6 81.2
# : Milestone ETHOS E + : Full fruit * : Crushed fruit
Microdistillation
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• Microdistillation is a micro scale distillation technique which allows • rapid • programmable distillation
• The system has 6 heating and 6 cooling units for
parallel operations.
• Small amount of material to be distilled (100-500
mg) is placed in a 15 mL capacity vial with water
(10 mL) and sealed with a cap having a small slit
on top
• requires less than 1 gr or 1 ml of material, which makes it excellent for working with
herbarium samples.
• fast preparation and set-up.
• fast isolation and readiness for analysis.
• cheaper and more efficient in long term use
• easy operation
Molecular Spinning Band Distillation
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• Spinning band distillation uses a rotating helical band to
create a high number of theoretical plates
• The spinning bands can be made of Teflon or metal
• Teflon spinning bands are used for distillations below 225 °C
• Metal bands are used for higher temperature distillations
where Teflon would become soft
• spinning band distillation gives a very efficient
separation in a short distillation column
• High Efficiency
• Low Column Hold Up
• Low pressure drop
Thermomicrodistillation
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1 Seal
2 Indicator silica
3 Glass tube (Pasteur pippette)
4 Oven (220oC)
5 Sample
6 Glass-wool
7 TLC Plate
TAS OVEN METHOD
Preparative Fractionation
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PREPARATIVE FRACTIONATION BY GC-FRACTION COLLECTOR - PFC -
• Essentially, GC is based on differential partitioning of
solutes between the mobile and stationary phases.
• Gas is used as a mobile phase which passes through the
column.
• This particular technique is quite simple, fast, reliable
and applicable to the separation of volatile materials
which are stable at a temperature up to 350-400°C.
• This fractionation is performed by switching the flow
passing through the column and the fractionation
collector traps.
• In both techniques, the traps are available in small
and/or medium size
Preparative Fractionation
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PREPARATIVE FRACTIONATION BY GC-FRACTION COLLECTOR - PFC -
• PFC permits reliable collection of
individual compounds that are closely
resolved.
• The reliability and reproducibility of the
system makes it possible to trap
compounds over the course of hundreds of
injections.
• PFC fractionation makes it possible to
obtain the fractions for further analysis
such as NMR, IR etc.
IR LC
GC-FID
NMR
GC-FT/IR
UV
HP-TLC
LC/MS GC/MS
GC-AED
UV
Analysis Techniques for MAPs
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What are we Expecting from New Techniques ?
Regional Expert Consultation on Medicinal and Aromatic Plants, 13-15 November 2013, Antalya 51
short processing time
less solvent consumption
min. environmental damage
lower energy consumption
less input / more output – max. yield
simplicity of application
no-risk or danger
no need to toxic and / or radioactive chemical input
cheapest and less input – low process cost
flexibility and functional usage
adaptation to different field easily
automation
validation
52
THANK YOU FOR YOUR
KIND ATTENTION
Prof. Dr.Temel ÖZEK Anadolu University, Faculty of Pharmacy,
Department of Pharmacognosy,
26470-Eskişehir