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

[email protected]

[email protected]

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


Introduction

nature of the material

yield and properties of the extracts

marketability of the products

Choice of

extraction

technique


Conventional Extraction Techniques

Mechanical extraction

Solvent extraction

Cold fat extraction

Distillation


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


Essential Oil Extraction Techniques


Distillation

Expression

Extraction

Extraction Techniques


Mechanic

al

Extr

acti

on

Expression

Scratch



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.



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



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


• 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


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.


• 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).


Liquified Gas Extraction Technique: Phytosol


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


Liquified Gas Extraction Technique: Phytosol

Supercritical Fluid Extraction Technique


• 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 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



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


• 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


• 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


• 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


• 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


Static headspace sampling

Vacuum headspace sampling

Dynamic headspace sampling

Static Headspace Sampling Technique


• 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


• 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


• 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


• Direct Sampling Method

Solid Phase Micro-Extraction (SPME)


• 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)


• 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)


• 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)


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)


• 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)


• 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)


• 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


• 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 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



Two Heating Model

M. Letellier and H. Budzinski*, Microwave assisted extraction of organic compounds, Analusis, 27, 259-271 (1999).



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.



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-assisted Hydrodistillation (MWHD) System

Milestone ETHOS E



DryDist Microwave-assisted

Hydrodistillation

(MWHD) System

Milestone



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).



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


• 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


• 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


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


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


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


What are we Expecting from New Techniques ?


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

[email protected]

[email protected]

Extraction, Separation and Analysis Techniques for MAPs ... · application in environmental as well...

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