Adsorption chromatography is a technique for separating components in a mixture based on differential adsorption of the components onto a stationary solid phase. It works by passing a mobile liquid or gas phase over an adsorbent stationary phase in a column, which causes components to separate as they are differentially retained on the surface of the adsorbent. Common types include thin layer chromatography, paper chromatography, and column chromatography. Adsorption chromatography has various applications such as separating amino acids, isolating antibiotics, and identifying carbohydrates.
ION EXCHANGE CHROMATOGRAPHY
By M.Vharshini
B.Sc. Bio Medical Science
Sri Ramachandra University
ION EXCHANGE CHROMATOGRAPHY
Ion-exchange chromatography is a process that allows the separation of ions and polar molecules based on their affinity to the ion exchanger.
It can be used for almost any kind of charged molecule including large proteins, small nucleotides and amino acids.
Cations or Anions can be separated using this method.
PRINCIPLE
It is based on the reversible electrostatic interaction of ions with the separation matrix (i.e.)
The separation occurs by reversible exchange of ions between the ions present in the solution and those present in the ion exchange resin.
CLASSIFICATION OF RESINS
According to the chemical nature they classified as-
1. Strong cation exchange resin
2. Weak cation exchange resin
3. Strong anion exchange resin
4. Weak anion exchange resin
According to the Source they can -
Natural resins : Cation - Zeolytes, Clay
Anion - Dolomite
Synthetic resins: Inorganic & Organic resins
◘Organic resins are polymeric resin matrix.
The resin composed of –
Polystyrene (sites for exchangeable functional groups)
Divinyl benzene(Cross linking agent)-offers stability.
Ion exchange resin should have following requirements
»It must be chemically stable.
»It should be insoluble in common solvents.
» It should have a sufficient degree of cross linking.
»The swollen resin must be denser than water.
»It must contain sufficient no. of ion exchange groups.
Physical properties of ion exchange resins
Cross linking:
It affects swelling & strength & solubility
Swelling:
When resin swells, polymer chain spreads apart
Polar solvents → swelling
Non-polar solvents → contraction
Swelling also affected electrolyte concentration.
Particle size and porosity
Increase in surface area & decrease in particle size will increase the rate of ion exchange.
Regeneration
Cation exchange resin are regenerated by treatment with acid, then washing with water.
Anion exchange resin are regenerated by treatment with NaOH, then washing with water until neutral.
EXPERIMENTAL SETUP OF ION EXCHANGE CHROMATOGRAPHY
Metrohm 850 Ion chromatography system
Instrumentation of ion exchange chromatography
PRACTICAL REQUIREMENTS
1.Column
» glass, stainless steel or polymers
2.Packing the column
» Wet packing method:
A slurry is prepared of the eluent with the stationary phase powder and then carefully poured into the column. Care must be taken to avoid air bubbles.
3.Application of the sample
After packing, sample is added to the top of the stationary phase, use syringe or pipette.
This layer is usually topped with a small layer of sand or with cotton or glass wool to protect the shape of the organic layer from the velocity of newly added eluent.
4.Mobile phase
Acids, alkalis, buffers…
6.Stationary phase
The ionic
ION EXCHANGE CHROMATOGRAPHY
By M.Vharshini
B.Sc. Bio Medical Science
Sri Ramachandra University
ION EXCHANGE CHROMATOGRAPHY
Ion-exchange chromatography is a process that allows the separation of ions and polar molecules based on their affinity to the ion exchanger.
It can be used for almost any kind of charged molecule including large proteins, small nucleotides and amino acids.
Cations or Anions can be separated using this method.
PRINCIPLE
It is based on the reversible electrostatic interaction of ions with the separation matrix (i.e.)
The separation occurs by reversible exchange of ions between the ions present in the solution and those present in the ion exchange resin.
CLASSIFICATION OF RESINS
According to the chemical nature they classified as-
1. Strong cation exchange resin
2. Weak cation exchange resin
3. Strong anion exchange resin
4. Weak anion exchange resin
According to the Source they can -
Natural resins : Cation - Zeolytes, Clay
Anion - Dolomite
Synthetic resins: Inorganic & Organic resins
◘Organic resins are polymeric resin matrix.
The resin composed of –
Polystyrene (sites for exchangeable functional groups)
Divinyl benzene(Cross linking agent)-offers stability.
Ion exchange resin should have following requirements
»It must be chemically stable.
»It should be insoluble in common solvents.
» It should have a sufficient degree of cross linking.
»The swollen resin must be denser than water.
»It must contain sufficient no. of ion exchange groups.
Physical properties of ion exchange resins
Cross linking:
It affects swelling & strength & solubility
Swelling:
When resin swells, polymer chain spreads apart
Polar solvents → swelling
Non-polar solvents → contraction
Swelling also affected electrolyte concentration.
Particle size and porosity
Increase in surface area & decrease in particle size will increase the rate of ion exchange.
Regeneration
Cation exchange resin are regenerated by treatment with acid, then washing with water.
Anion exchange resin are regenerated by treatment with NaOH, then washing with water until neutral.
EXPERIMENTAL SETUP OF ION EXCHANGE CHROMATOGRAPHY
Metrohm 850 Ion chromatography system
Instrumentation of ion exchange chromatography
PRACTICAL REQUIREMENTS
1.Column
» glass, stainless steel or polymers
2.Packing the column
» Wet packing method:
A slurry is prepared of the eluent with the stationary phase powder and then carefully poured into the column. Care must be taken to avoid air bubbles.
3.Application of the sample
After packing, sample is added to the top of the stationary phase, use syringe or pipette.
This layer is usually topped with a small layer of sand or with cotton or glass wool to protect the shape of the organic layer from the velocity of newly added eluent.
4.Mobile phase
Acids, alkalis, buffers…
6.Stationary phase
The ionic
This presentation contains all the topics related to column chromatography. That includes introduction, principle,apparatus, experimental aspects of column chromatography, application of column chromatography, advantage and disadvantage of column chromatography with reference.
This presentation contains all the topics related to column chromatography. That includes introduction, principle,apparatus, experimental aspects of column chromatography, application of column chromatography, advantage and disadvantage of column chromatography with reference.
Chromatography is an important biophysical technique that enables the separation, identification, and purification of the components of a mixture for qualitative and quantitative analysis.
The Russian botanist Mikhail Tswett coined the term chromatography in 1906.
The first analytical use of chromatography was described by James and Martin in 1952, for the use of gas chromatography for the analysis of fatty acid mixtures.
A wide range of chromatographic procedures makes use of differences in size, binding affinities, charge, and other properties to separate materials.
It is a powerful separation tool that is used in all branches of science and is often the only means of separating components from complex mixtures.
hromatography is based on the principle where molecules in mixture applied onto the surface or into the solid, and fluid stationary phase (stable phase) is separating from each other while moving with the aid of a mobile phase.
The factors effective on this separation process include molecular characteristics related to adsorption (liquid-solid), partition (liquid-solid), and affinity or differences among their molecular weights.
Because of these differences, some components of the mixture stay longer in the stationary phase, and they move slowly in the chromatography system, while others pass rapidly into the mobile phase, and leave the system faster.
Three components thus form the basis of the chromatography technique.
1. Stationary phase: This phase is always composed of a “solid” phase or “a layer of a liquid adsorbed on the surface solid support”.
2. Mobile phase: This phase is always composed of “liquid” or a “gaseous component.”
3. Separated molecules
Types of Chromatography
Substances can be separated on the basis of a variety of methods and the presence of characteristics such as size and shape, total charge, hydrophobic groups present on the surface, and binding capacity with the stationary phase.
This leads to different types of chromatography techniques, each with their own instrumentation and working principle.
For instance, four separation techniques based on molecular characteristics and interaction type use mechanisms of ion exchange, surface adsorption, partition, and size exclusion.
Other chromatography techniques are based on the stationary bed, including column, thin layer, and paper chromatography.
Applications of Chromatography
Pharmaceutical sector
To identify and analyze samples for the presence of trace elements or chemicals.
Separation of compounds based on their molecular weight and element composition.
Detects the unknown compounds and purity of mixture.
In drug development.
Chemical industry
In testing water samples and also checks air quality.
HPLC and GC are very much used for detecting various contaminants such as polychlorinated biphenyl (PCBs) in pesticides and oils.
In various life sciences applications.
In forensic pathology and crime scene testing like analyzing blood and hair samples.
Chromatography : A seperation techniqueSHIVANEE VYAS
Chromatography is a method of seperating mixture of components into individual components through equlibrium distribution between two phases.
Each chromatographic method essentially consists of 2 phases a staionary phase and a mobile phase.
Stationary phase : solid or liquid
Mobile phase : liquid or gas
TLC-Introduction, Principle, Procedure, and Applications.
Paper Chromatography-Introduction, Principle, Procedure, and Applications.
Column Chromatography-Introduction, Principle, Procedure, and Applications.
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Bills have a main role in point of sale procedure. It will help to track sales, handling payments and giving receipts to customers. Bill splitting also has an important role in POS. For example, If some friends come together for dinner and if they want to divide the bill then it is possible by POS bill splitting. This slide will show how to split bills in odoo 17 POS.
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Adsorption Chromatography Assignment.pdf
1. 1
ADSORPTION CHROMATOGRAPHY
Topic Outlines:
✓ Absorption versus Adsorption.
✓ Introduction of Adsorption Chromatography.
✓ Conditions.
✓ Principle of Adsorption Chromatography.
✓ Diagram & Procedure of Adsorption Chromatography.
✓ Experiment (TLC).
✓ Types of Adsorption Chromatography.
✓ Applications of Adsorption Chromatography.
✓ Advantages & Disadvantages of Adsorption Chromatography.
Absorption Versus Adsorption:
These are two different terms:
→ In absorption, one substance penetrates into the bulk of another substance.
→ Adsorption is a surface phenomenon where interaction takes place only on the
surface of one substance.
Introduction of Adsorption Chromatography:
A Russian botanist, Mikhail Tsvet in the year 1901, invented the process called
adsorption chromatography while doing his research on plant pigments.
Adsorption Chromatography:
Adsorption chromatography is the oldest types of chromatography technique. It makes
use of a mobile phase which is either in liquid or gaseous form. The mobile phase is
adsorbed onto the surface of a stationary solid phase. It can be defined as;
“As a technique for the separation of the component in a mixture by adsorption
from a mobile phase into the stationary solid surface.”
Conditions:
Adsorption chromatography depends on the following condition:
❖ The selection of the adsorbent
❖ The choice of the solvent for the sample mixture
❖ The rate of flow of the solvent
❖ The temperature
❖ The selection of the column size
Principle of Adsorption Chromatography:
• Adsorption Chromatography is based on the principle that some solid substances,
which are known as adsorbent, have the power to hold molecules at their surface.
• This holding force is due to weak, non-ionic attractive forces of the van der Waals'
and hydrogen bonding, which only occur at specific adsorption beds.
• As solvent (eluent) is constantly passed over the adsorbent (stationary phase) in
the column. The most commonly used adsorbents are silica, alumina, and carbon.
2. 2
Adsorption Chromatography Diagram:
Adsorption Chromatography Procedure:
Before beginning an adsorption chromatography experiment, we must recognize the
various components essential to perform the process.
o Stationary phase: The stationary phase of adsorption chromatography is
adsorbent.
o Mobile phase: In adsorption chromatography, either a liquid or a gas is used as a
mobile phase.
Apparatus:
▪ Chromatography jar: The glass jar has a lid. It helps to maintain a proper
environment during separation.
▪ Thin-layer chromatography plate: Silicate glass plate with size 20*20 cm,
20*5 cm, 20*10.
▪ Capillary tube: Sample mixture is applied to TLC with the help of this tube.
▪ Mobile phase: Liquid or gas.
▪ Stationary phase: Adsorbents.
3. 3
Experiment (TLC):
o Take a clean and dry chromatographic jar.
o To make sure that the environment in the jar is saturated with solvent vapors, a
paper soaked in the mobile phase is applied to the walls.
o Add the mobile phase to the jar and close it and maintain equilibrium
o Mark the baseline on the adsorbent.
o Apply sample to TLC plate with the help of a capillary tube and allow it to dry.
Put the plates in the jar and close it.
o Wait until the solvent moves from the baseline.
o Take out the TLC plate and dry it.
Types of Adsorption Chromatography:
These are some important types of Adsorption Chromatography.
1. Thin Layer Chromatography (TLC).
2. Paper Chromatography.
3. Column Chromatography.
4. Gas-Solid Chromatography.
1- Thin Layer Chromatography (TLC):
It is an analytical method of separation used for qualitative analysis and
monitoring of the reaction as well as identifying unknown compounds.
In which the mobile phase moves over an adsorbent.
2- Paper Chromatography:
It is a technique that uses paper sheets or strips as the adsorbent being the
stationary phase through which a solution is made to pass is called paper
chromatography.
The solid surface of the paper is the stationary phase and the liquid phase is the
mobile phase.
3- Column Chromatography:
It is a commonly used technique for separating the compounds from the sample
mixture. It is used on small or large scans to separate and purify compounds.
The mixture of samples moves through the stationary phase with the mobile
phase and based on various degrees of adhesion it separates the compounds.
4- Gas-Solid Chromatography:
It is an analytical method for the separation of volatile compounds in which the
mobile phase is gas and the stationary phase is suitable for solid support.
The gas-solid chromatography, work on the principle of separation is adsorption
and generally used for samples that had less solubility in the stationary phase.
Adsorption Chromatography Applications:
Adsorption chromatography have the various applications:
• It is used for the separation of amino acids.
• It is used in the isolation of antibiotics.
4. 4
• It is used in the identification of carbohydrates.
• It is used to determine the concentration of a molecule.
• It is used for the separation and identification of isomers.
• Used to isolate and determine various peptides and proteins.
Advantages of adsorption chromatography:
The advantages of adsorption chromatography are as follows.
▪ Adsorption chromatography is an important tool for separating many molecules
that cannot be distinguished from other methods.
▪ Adsorption chromatography uses a broad range of mobile phases.
▪ The molecules in the complex mixture can be easily separated.
▪ Very few types of equipment are used as opposed to other separation methods.
Disadvantages of adsorption chromatography:
The disadvantages of adsorption chromatography are as follows.
▪ Automation is making it more complicated and expensive.
▪ The major disadvantage of adsorption chromatography is that some solutes have
longer retention times.
▪ It can cause catalytic variations in the sample.
▪ Results obtained by some methods of adsorption chromatography are difficult to
reproduce.
Best of Luck
