ADVANCED SEPARATION PROCESSES (3160507)

Ch 6 : Chromatographic separation
CONTENT
🞭 Chromatographic separation
🞭 Principle and operation
🞭 Column chromatography
🞭 Ion exchange chromatography
🞭 Gel filtration and affinity chromatography
🞭 Thin layer and paper chromatography
🞭 Liquid chromatography
🞭 Advantages and disadvantages of
chromatographic separations.
HISTORY AND BACKGROUND
🞭 The term chromatography and its principle
were first developed by Mikhail Tswett- a
Botanist (1872-1919).
🞭 In 1906 Tswett used to chromatography to
separate plant pigments
🞭 In his experiment, a leaf extract sample in
petroleum ether was allowed to pass
through a column of 𝐶𝑎𝐶𝑂3.
🞭 Pure ether was continued to flow through
the column, as a result of which various
chlorophyll pigments, etc., were separated
into a series of colored and easily
distinguished zones.
🞭 He called the new technique
chromatography because the result of the
analysis was 'written in color' along the
length of the adsorbent column
🞭 Chroma means “color” and graphein
means to “write”
Thin layer chromatography is used to separate the
colorful components of a plant extract
HISTORICAL DEVELOPMENT IN CHROMATOGRAPHY

Investigator(s) Year Contribution

Way and Thompson 1848 Recognized the phenomenon of ion exchange in solids.
Runge, Schoenbein, and Goeppelsroeder 1850-1900 Studied capillary analysis on paper
Lemberg 1878 Illustrated the reversibility and stoichiometry of ion exchange in
aluminum silicate minerals.
Reed 1892 First recorded column separation: tubes of kaolin used for
separation of FeCI3 from CuSO4.
Tswett 1903-1906 Invented chromatography with use of pure solvent to develop
the chromatogram; devised nomenclature; used mild
adsorbents to resolve chloroplast pigments.

Karrer, Kuhn and Starin 1930-1932 Used activated lime, alumina and magnesia absorbents
Holmes and Adams 1935 Synthesized synthetic organic ion exchange resins.
Reichstein 1938 Introduced the liquid or flowing chromatogram, thus extending
application of chromatography to colorless substances.

Izmailov and Schraiber 1938 Discussed the use of a thin layer of unbound alumina spread
on a glass plate.
Brown 1939 First use of circular paper chromatography.
Tiselius 1940-1943 Devised frontal analysis and method of
displacement development.
Martin and Synge 1941 Introduced column partition chromatography.
HISTORICAL DEVELOPMENT IN CHROMATOGRAPHY

Investigator(s) Year Contribution

Consden, Gordon, and Martin 1944 First described paper partition chromatography.

Boyd, Tompkins, et al 1947-1950 Ion-exchange chromatography applied to various analytical

problems.

M. Lederer and Linstead 1948 Applied paper chromatography to inorganic compounds.

Kirchner 1951 Introduced thin-layer chromatography as it is practiced

today.
James and Martin 1952 Developed gas chromatography.

Sober and Peterson 1956 Prepared first ion-exchange celluloses

Lathe and Ruthvan 1956 Used natural and modified starch molecular sieves for
molecular weight estimation.

Porath and Flodin 1959 Introduced cross-linked dextran for molecular sieving.

J. C. Moore 1964 Gel permeation chromatography developed as a practical

method.
PRINCIPLE AND OPERATION
🞭 Chromatography may be regarded as an analytical technique employed for purification and
separation of organic and inorganic substances.
🞭 Chromatography is a technique for separating the components, or solutes of a mixture on
the basis of the relative amounts of each solute distributed between a moving fluid stream,
called the mobile phase and a contiguous stationary phase.
🞭 The mobile phase may be either a liquid or a gas, while the stationary phase is either a solid
or a liquid.
🞭 Chromatography is the ability to separate molecules using partitioning characteristics of
molecule to remain in a stationary phase versus a mobile phase. Once a molecule is
separated from the mixture, it can be isolated and quantified.
🞭 The stationary phase is a substance that binds and shortly releases the molecules moving
through the system. The particles can move through the system due to the mobile phase.
🞭 The compounds that are more like the stationary phase have higher affinity towards it,
move slower than the compounds that are more like the mobile phase.
🞭 Chromatography may be viewed as a series of equilibrations between the mobile and
stationary phase.
🞭 The relative interaction of a solute with these two phases is described by the partition (K) or
distribution (D) coefficient (ratio of concentration of solute in stationary phase to
concentration of solute in mobile phase).
 PRINCIPLE AND OPERATION
Chromatography

Based on types of stationary phase & Principle

Adsorption chromatography Partition chromatography Ion exchange Permeation

(competition b/w solid (competition b/w liq chromatography chromatography
adsorbent and the mobile stationary phase and (competition b/w ion (competition b/w a
phase mobile phase exchange resin stationary polymer matrix and
phase and liq mobile phase liq mobile phase

Type of Mobile Type of Mobile Type of Mobile Type of Mobile

phase phase phase phase

Gas Liquid Gas Liquid Liquid Liquid

LC,
GSC HPLC
TLC GLC LLC, Ion exchange Gel permeation
PC SFC HPLC Chromatography Chromatography

*GSC-Gas Solid chromatography , LC-liq column chromatography, HPLC- High performance liquid chromatography, TLC-Thin layer
chromatography, PC- Paper chromatography , GLC-Gas liq chromatography , SFC- Super critical fluid chromatography, LLC-liq-liq
chromatography,
PRINCIPLE AND OPERATION
🞭 Chromatography is an extremely powerful analytical tool for separating and analyzing complex
mixture.
🞭 The principle of chromatography is elaborated now. It constitutes a bed of particles through which a
gas or liquid stream flows.
🞭 The flowing phase is also known as carrier gas (in case of flowing phase is gas) or solvent (in case of
flowing phase is liquid).
🞭 A feed pulse (a small amount of feed) containing various solutes is introduced into the system.
🞭 The solutes in the feed pulse are then separated by difference in their velocities.
🞭 Therefore, the solutes emerging from the chromatographic column (the housing of the packed bed)
are detected by refractive index or ultraviolet absorbance and that measurement is directly related to
concentration of solutes. Based on this basic principle, chromatographic separation occurs.

It contains the following features:

1. For liquid system, a pump is used to
push the fluid through the column.
2. A pulse of feed is injected into the
system.
3. The column is often enclosed in an
oven to control temperature.
4. Detector analyzes the stream for some
properties like RI, UV absorbance,
those can be related to concentration.
Schematic of an analytical high pressure liquid chromatograph
 COLUMN CHROMATOGRAPHY
🞭 Column chromatography is the most useful method of separating compounds in a mixture.
🞭 Fractionation of solutes occurs as a result of differential migration through a closed tube of stationary phase, and
analytes can be monitored while the separation is in progress.
🞭 In column chromatography, the mobile phase is liquid and the stationary phase can be either solid or liquid
supported by an inert solid.
🞭 The prepared stationary phase (resin, gel or packing material) is packed into a column (usually glass), the length
and diameter of which are determined by the amount of sample to be loaded, the separation mode to be used,
and the degree of resolution required.
🞭 The sample to be fractionated, dissolved in a minimum volume of mobile phase, is applied in a layer at the top
(or head) of the column.
🞭 low pressure chromatography utilizes only gravity flow or a peristaltic pump to maintain a flow of mobile phase
(eluent or eluting solvent) through the column.
🞭 The process of passing the mobile phase through the column is called elution, and the portion that emerges from
the outlet end of the column is sometimes called the eluate
🞭 As elution proceeds, components of the sample are selectively retarded by the stationary phase based on the
strength of interaction with the stationary phase, and thus they are eluted at different times.
🞭 The column eluate may be directed through a detector and then into tubes, changed at intervals by a fraction
collector.
🞭 The detector response, in the form of an electrical signal, may be recorded (the chromatogram), using either a
chart recorder or a computerized software, and used for qualitative or quantitative analysis,
🞭 The fraction collector may be set to collect eluate at specified time intervals or after a certain volume or number
of drops has been collected.
🞭 Components of the sample that have been chromatographically separated and collected then can be further
analyzed as needed.
 COLUMN CHROMATOGRAPHY
Methodology:
🞭 The chromatographic column consists of glass or
metal column filled with porous sorbent. Surface
of the sorbent acts as a stationary phase.
🞭 Stationary phase is held in a narrow tube through
which the mobile phase is forced under pressure
or under the effect of gravity
🞭 A detecting device must follow the column in
order to detect the separated substances.
🞭 Simple photometer can serve as a detector
🞭 At time 𝑡0 the analytes are introduced at the top
of the column. As the mobile phase flows, it
carries the substances with it.
🞭 Every substance has its characteristic velocity,
which depends on the time that particles spend
in mobile phase vs. the time they spend on
stationary phase.
🞭 The graph on the lower part of Figure shows the
Column chromatography setup
absorbance, measured by the photometer, during
the elution process. Such graphs are called
chromatograms
 COLUMN CHROMATOGRAPHY
🞭 At time 𝑡1 components A and B have moved some
distance in the column.
🞭 At the same time, the solvent used to introduce those
substances has passed the column and reached the
detector.
🞭 The change in detector output signal when a substance
passes it, is called chromatographic peak.
🞭 A peak is characterized by the time at its maximum.
🞭 The moment at which the maximum of solvent peak exits
is called column dead time (𝑡𝑚).
🞭 Dead time is time it takes for the unretained species
(move at the same velocity as eluent) to reach the
detector.
🞭 At 𝑡2 the substances have moved along the column. The
further they move the more they separate from each
other.
🞭 Substance A reaches the end of column at 𝑡3 .
Principle of column chromatography and the chromatogram.
🞭 The detector registers a peak which is maximum time is
called the retention time of substance A – 𝑡𝐴. Substance
B reaches the detector at 𝑡4 – retention time of B is 𝑡𝐵.
🞭 So knowing the retention time of a substance under
specific experimental conditions enables to identify the
peaks.
ION EXCHANGE CHROMATOGRAPHY
🞭 Ion-exchange chromatography is a versatile, high resolution chromatography
techniques to purify the protein from a complex mixture.
🞭 Principle: This chromatography distributes the analyte (sample to be separated)
molecule as per charge and their affinity towards the oppositively charged matrix.
🞭 The analytes bound to the matrix are exchanged with a competitive counter ion to
elute. The interaction between matrix and analyte is determined by net charge, ionic
strength and pH of the buffer.
🞭 For example, when a mixture of positively charged analyte (M, M+,M-1, M-2) loaded
onto a positively charged matrix, the neutral or positively charged analyte will not
bind to the matrix where as negatively charged analyte will bind as per their relative
charge and needed higher concentration of counter ion to elute from matrix

Affinity of analytes (M, M+,M-1, M-2) towards positively charged matrix.

 ION EXCHANGE CHROMATOGRAPHY
🞭 Here are two types of ion exchange chromatography.
1. Cation exchange chromatography: In cation
exchange chromatography, matrix has a negatively
charged functional group with a affinity towards
positively charged molecules.
 The positively charged analyte replaces the
reversible bound cation and binds to the matrix.
 In the presence of a strong cation (such as Na+) in
the mobile phase, the matrix bound positively
charged analyte is replaced with the elution of
analyte.
2. Anion Exchange chromatography- In anion
exchange chromatography, matrix has a positively
charged functional group with a affinity towards
negatively charged molecules.
 The negatively charged analyte replaces the
reversible bound anion and binds to the matrix.
 In the presence of a strong anion (such as Cl-) in Ion exchange chromatography
the mobile phase, the matrix bound negatively
charged analyte is replaced with the elution of
analyte.
ION EXCHANGE CHROMATOGRAPHY
🞭 Isoelectric point and charge on a protein:
🞭 Protein is a polymer made up of amino acids with
ionizable side chain.
🞭 At a particular pH, these amino acid side chain
ionizes differentially to give a net charge
(positive/negative) to the protein.
🞭 The pH at which the net charge on a protein is zero
is called as Isoelectric point (pI).
🞭 The protein will have a net positive charge below
the pI where as it has net negative charge above
the pI value .
ION EXCHANGE CHROMATOGRAPHY
🞭 Choice of a Ion-exchange column matrix:
1. pI value and Net charge- a cation exchange chromatography can
be use below the pI where as an anion exchange chromatography
can be use above the pI value.
2. Structural stability-3-D structure of a protein is maintained by
electrostatic and vander waal interaction between charged amino
acid, Π-Π interaction between hydrophobic side chain of amino
acids.
 As a result, protein structure is stable in a narrow range around its
pI and a large deviation from it may affect its 3-D structure.
3. Enzymatic activity-Similar to structural stability, enzymes are
active in a narrow range of pH and this range should be consider
for choosing an ion-exchange chromatography.
ION EXCHANGE CHROMATOGRAPHY
🞭 Parameters to be considered while performing ion exchange chromatography.
1. Column material and stationary phase-Column material should be chemically inert
to avoid destruction of biological sample. It should allow free low of liquid with
minimum clogging. It should be capable to withstand the back pressure and it
should not compress or expand during the operation.
2. Mobile Phase-The ionic strength and pH are the crucial parameters to influence
the property of the mobile phase.
3. Sample Preparation- The sample is prepared in the mobile phase and it should be
free of suspended particle to avoid clogging of the column. The most
recommended method to apply the sample is to inject the sample with a syringe.
4. Elution- There are many ways to elute a analyte from the ion-exchange column. (1)
Isocratic elution (2) Step-wise gradient (3) Continuous gradient either by salt or pH
(4) affinity elution (5) displacement chromatography
5. Column Regeneration- After the elution of analyte, ion-exchange chromatography
column require a regeneration step to use next time. column is washed with a salt
solution with a ionic strength of 2M to remove all non-specifically bound analytes
and also to make all functional group in a iononized form to bind fresh analyte.
ION EXCHANGE CHROMATOGRAPHY
🞭 The basic process of chromatography using ion
exchange can be represented in 5 steps
(assuming a sample contains two analytes A & B):
1. eluent loading
2. sample injection,
3. Separation of sample
4. Elution of analyte A
5. Elution of analyte B
🞭 Elution is the process where the compound of
interest is moved through the column.
ION EXCHANGE CHROMATOGRAPHY
🞭 Step 1: The eluent loaded onto the column displaces any anions bonded to the resin and saturates
the resin surface with the eluent anion.
🞭 Step 2: A sample containing anion A and anion B are injected onto the column.
🞭 Step 3: After the sample has been injected, the continued addition of eluent causes a flow through
the column. As the sample elutes (or moves through the column), anion A and anion B adhere to the
column surface differently. The sample zones move through the column as eluent gradually
displaces the analytes.
🞭 Step 4: As the eluent continues to be added, the anion A moves through the column in a band and
ultimately is eluted first.
🞭 Step 5: The eluent displaces anion B, and anion B is eluted off the column.
GEL FILTRATION CHROMATOGRAPHY
🞭 It is also called molecular sieve chromatography or size
exclusion chromatography, separates substance on the
basis of molecular size.
🞭 A solution to be separated is passed over a column
made up of small beads composed of cross linked
polymers.
🞭 The degree of cross-linking will defined a pore size.
🞭 Solutes are larger than this pore size are excluded from
the matrix and pass through the column unrestricted.
🞭 Smaller solutes will enter the gel matrix and are retained
on the column longer. The retention time is inversely
proportional to the size of the solute. Unlike other
chromatographic methods the solute does not bind to
the stationary phase during chromatography.
🞭 Hence, gel filtration is generally carried out in buffers
containing 0.15-1.0 M salt to prevent interactions of
proteins with the support matrix.
🞭 since protein do not bind to the stationary phase, the
resolution is dependent upon loading the smallest
possible volume of sample.
 GEL FILTRATION CHROMATOGRAPHY
🞭 Gel filtration can also be used to determine
the molecular weight of a protein if the
columns are calibrated by using MW
standards.
🞭 Proteins of known size are passed over the
column and the 𝐾𝑎𝑣 is determined as follow:
🞭 𝐾𝑎 𝑣 = (𝑉𝑒 − 𝑉𝑜 )/(𝑉𝑡 − 𝑉𝑜 )
🞭 Where, 𝑉𝑜= void volume/excluded volume (is
the elution volume of a substance which is two
large to enter the matrix of the support
medium). This determined experimentally and
represents the solvent between the beads.
🞭 𝑉𝑡= the total volume is calculated from the
volume of the column bead.
🞭 𝑉𝑒 depends on total volume as shown in figure.
🞭 The 𝐾𝑎𝑣 values are plotted against the log of
the molecular weight for each protein
standard.
🞭 The molecular weight of the unknown protein
can then be determined from its 𝐾𝑎𝑣 and the
standard curve.
 AFFINITY CHROMATOGRAPHY
🞭 Affinity chromatography is unique in that separation
is based on the specific, reversible interaction
between a solute molecule and a ligand immobilized
on the chromatographic stationary phase.
🞭 Affinity chromatography could be viewed as the
ultimate extension of adsorption chromatography.
🞭 The principles of affinity chromatography are
illustrated in fig.
🞭 A ligand, chosen based on its specificity and
strength of interaction with the molecule to be
isolated (analyte), is immobilized on a suitable
support material.
🞭 As the sample is passed through this column,
molecules that are complementary to the bound
ligand are adsorbed while other sample components
are eluted.
🞭 Bound analyte is subsequently eluted via a change
in the mobile-phase composition.
🞭 After reequilibration with the initial mobile phase,
the stationary phase is ready to be used again.

(a) The support presents the immobilized ligand to the analyte to be isolated (b) the analyte
makes contact with the ligand and attaches itself (c) the analyte is recovered by the
introduction of an eluent, which dissociates the complex holding the analyte to the ligand (d)
the support is regenerated
AFFINITY CHROMATOGRAPHY
🞭 The ideal support for affinity chromatography should be a porous, stable,
high-surface-area material that does not adsorb anything itself.
🞭 Thus, polymers such as agarose, cellulose, dextran, and polyacrylamide are
used, as well as controlled-pore glass.
🞭 Ligands for affinity chromatography may be either specific or general (i.e.,
group specific). Specific ligands, such as antibodies, bind only one
particular solute.
🞭 Elution methods for affinity chromatography may be divided into nonspecific
and (bio)specific methods.
🞭 Nonspecific elution involves disrupting ligand analyte binding by changing
the mobile-phase pH, ionic strength, dielectric constant, or temperature.
🞭 If additional selectivity in elution is desired, for example, in the case of
immobilized general ligands, a biospecific elution technique is used.
🞭 Affinity chromatography has been useful especially in the separation and
purification of enzymes and glycoproteins
LIQUID CHROMATOGRAPHY
🞭 There are several liquid chromatography techniques
applied in food analysis, namely paper chromatography,
thin layer chromatography (TLC) (both of these
techniques may be referred to as planar
chromatography).
🞭 Column liquid chromatography, all of which involve a
liquid mobile phase and either a solid or a liquid
stationary phase.
🞭 However, the physical form of the stationary phase is
quite different in each case.
🞭 Separation of the solutes is based on their
physicochemical interactions with the two phases.
THIN LAYER CHROMATOGRAPHY
🞭 TLC is a method for identifying substances and testing the purity of compounds.
🞭 TLC is a useful technique because it is relatively quick and requires small quantities
of material.
🞭 Separations in TLC involve distributing a mixture of two or more substances between
a stationary phase and a mobile phase.
🞭 The stationary phase is a thin layer of adsorbent (usually silica gel or alumina)
coated on a plate.
🞭 The mobile phase is a developing liquid which travels up the stationary phase,
carrying the samples with it.
🞭 Components of the samples will separate on the stationary phase according to how
much they adsorb on the stationary phase versus how much they dissolve in the
mobile phase.
🞭 Principle: The separation principle of the TLC procedure is based on the given
compound’s relative affinity towards the mobile and the stationary phase. The
process begins here by moving the mobile phase over the stationary phase’s
surface. During this movement, the higher affinity compounds gain less speed as
compared to the lower affinity compounds. This results in their separation.
THIN LAYER CHROMATOGRAPHY
🞭 TLC utilizes a thin (250 μm thick) layer of
sorbent or stationary phase bound to an inert
support in a planar configuration.
🞭 The support is often a glass plate (traditionally,
20 cm × 20 cm), but plastic sheets and
aluminum foil also are used.
🞭 Four frequently used TLC sorbents are silica
gel, alumina, diatomaceous earth, and
cellulose
THIN LAYER CHROMATOGRAPHY
🞭 General procedure (TLC separation of mixture x)
🞭 Step1: The starting line is drawn close to the one end of the
chromatographic plate. Spots of the substances to be
determined (a, b, c) and the mixture x are transferred onto the
starting line
🞭 Step 2: The plate is placed in the elution chamber with small
amount of eluent (mobile phase) in it.
🞭 Step 3: The lower edge of the plate is immersed in the eluent
so that eluent does not reach the spots on the starting line. To
avoid evaporation of eluent away from the plate the elution Spots of substances on chromatographic plate; elution chamber.
chamber is covered with a lid. Eluent starts moving upward
through the solvent layer due to capillary forces
🞭 Step 4: As the eluent flows, it carries the analytes with it. The
velocity of substance depends on their affinity towards
stationary phase. substances which “like” to be in liquid phase
move faster and the substances which prefer stationary phase
move slower.
🞭 Step 5:The substances usually move slower than the eluent
front. Before the eluent front reaches the upper edge of the
chromatographic plate, the plate is removed from the elution
chamber and is placed in horizontal position to let the eluent
evaporateAs a result of movement at different speeds, at the
end of the experiment, the substances are placed at different
distances between the starting and the stop lines. Comparison
of distances of spots of analyte and known substances from Eluent movement
the starting line reveals the substances present in the mixture.
THIN LAYER CHROMATOGRAPHY
🞭 Analytes are very often colorless substances and thus
invisible on the chromatographic plate.
🞭 In such cases the plate has to be developed after elution
process. For developing different chemicals can be used
which form colored substances upon reaction with analytes.
Some substances which have no color appear as colored
spots when observed under ultraviolet lamp.
🞭 In this course two practical works have set up which use TLC.
“Determination of amino acids by TLC” employs paper as
stationary phase and visualization is achieved by chemical
reaction. “Determination of water soluble vitamins by TLC”
uses aluminum foil coated with silica as stationary phase
and the spotsare analyzed under ultraviolet lamp.
PAPER CHROMATOGRAPHY
🞭 A method of partition
chromatography using filter
paper strips as carrier or inert
support.
🞭 The factor governing separation
of mixtures of solutes on filter
paper is the partition between
two immiscible phases.
🞭 One is usually water adsorbed on
cellulose fibers in the paper
(stationary phase).
🞭 The second is the organic
solvent flows past the sample on
the paper (stationary phase).
PAPER CHROMATOGRAPHY
🞭 The dissolved sample is applied as a small spot or streak one half inch or
more from the edge of a strip or square of filter paper (usually cellulose),
which is then allowed to dry.
🞭 The dry strip is suspended in a closed container in which the atmosphere is
saturated with the developing solvent (mobile phase), and the paper
chromatogram is developed.
🞭 The end closer to the sample is placed in contact with the solvent, which then
travels up or down the paper by capillary action, separating the sample
components in the process.
🞭 When the solvent front has traveled the length of the paper, the strip is
removed from the developing chamber and the separated zones are detected
by an appropriate method.
🞭 The stationary phase in paper chromatography is usually water
🞭 In paper and thin-layer chromatography, components of a mixture are
characterized by their relative mobility (𝑅𝑓) value, where:
Distance moved by component
𝑅𝑓 =
Distance moved by solvent
ADVANTAGES AND DISADVANTAGES OF
CHROMATOGRAPHIC SEPARATIONS.
Advantages
🞭 Chromatographic method gives a precise and accurate result than the other technique.
🞭 More efficient technique than the other
🞭 Chromatographic technique can separate the mixture contain more than one component.
🞭 It requires the small amount of sample for the analysis.
🞭 Most widely used method in pharmaceutical industry.
🞭 Continuous analysis is possible in chromatography.
🞭 There are many types of chromatography technique available from them we can choose the
appropriate chromatographic method.
🞭 The chromatography system is versatile. Wide ranges of a sample can analysis by using
chromatography technique.
🞭 Capable of separating complex mixtures at low operating temperature
🞭 Large scale batch or continuous operation possible
🞭 Capable of separating materials according to size and/or chemical properties
🞭 Can be used to separate delicate or heat labile compounds
🞭 Separation can be achieved by a variety of methods
🞭 Very pure products can be recovered
ADVANTAGES AND DISADVANTAGES OF
CHROMATOGRAPHIC SEPARATIONS.
 Disadvantages:
 Process scale-up is a problem (hence low
🞭

throughput)
 🞭 Irreversible adsorption of materials creates
🞭 Over-loading of columns with feed material may cause incomplete
problems.
separation.
🞭  🞭 To
Feed achieve
material efficient
is diluted separations,
by flowing mobile phase.high operating
🞭 Non-uniform
pressurescolumnmay bepacking can lead to a significant decrease in
required.
separation performance.
🞭 Pre-filtration of feed material is usually required
Periodic column re-packing / regeneration required.
🞭 Only trained and experienced person can operate the system.
🞭 Expensive method. Cost is high of the system.
🞭 The equipment need to handle with the care.
Applications Continue…..