Ion exchange chromatography is a technique for separating ions and molecules based on their affinity for ion exchange resins. These resins consist of an insoluble matrix with ionizable functional groups attached. Separation occurs as molecules exchange ions with the resin based on properties like charge and pH. Ion exchange chromatography has applications in water softening, amino acid separation, and purification of reagents and metals.
Download and play it my friends it contain VIDEO
The technique of ion exchange chromatography is based upon the interaction between charged solute molecules and oppositely charged moieties covalently linked to chromatographic matrix.
The reasons for its widespread success is its applicability, high resolving power, high capacity and simplicity of the technique.
Separation in ion exchange chromatography depends upon the reversible adsorption of charged solute molecules to immobilized ion exchange groups of opposite charge. Most experiments are performed by following : Video For Understanding Play It
Download and play it my friends it contain VIDEO
The technique of ion exchange chromatography is based upon the interaction between charged solute molecules and oppositely charged moieties covalently linked to chromatographic matrix.
The reasons for its widespread success is its applicability, high resolving power, high capacity and simplicity of the technique.
Separation in ion exchange chromatography depends upon the reversible adsorption of charged solute molecules to immobilized ion exchange groups of opposite charge. Most experiments are performed by following : Video For Understanding Play It
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.
ION EXCHANGE CHROMATOGRAPHY
ByM.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
High Performance Liquid Chromatography (HPLC) is a form of column chromatography that pumps a sample mixture or analyte in a solvent (known as the mobile phase) at high pressure through a column with chromatographic packing material (stationary phase).
OHS Laws (National & International) Pro & ConsZainab Arshad
OHS Laws (National & International) are made to protect co-workers, family members, employers, customers, and many others who might be affected by the workplace environment.
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.
ION EXCHANGE CHROMATOGRAPHY
ByM.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
High Performance Liquid Chromatography (HPLC) is a form of column chromatography that pumps a sample mixture or analyte in a solvent (known as the mobile phase) at high pressure through a column with chromatographic packing material (stationary phase).
OHS Laws (National & International) Pro & ConsZainab Arshad
OHS Laws (National & International) are made to protect co-workers, family members, employers, customers, and many others who might be affected by the workplace environment.
Ion exchange chromatography may be defined as a reversible reaction in which free mobile ions of a solids called ion exchange are exchanged for different ions of similar charge present in solution.....................................................................
This presentation was given at the Phycological Society of Southern Africa meetin in 2004 and shows prelimanary investigations into modeling and IMTA seaweed abalone land based recirculting system
Following is my journal documentation during Master's in Biotechnology completed in 2015. I do understand many changes would've occurred in the curriculum since then, but the basics seldom change. Kindly absorb as per your need.
Drug absorption by the human intestine
Models of intestinal absorption of pharmaceutical compounds.
Characteristics of Caco-2 cells
Permeability assessment
Cultivation of Caco-2 cell monolayers
Trans Epithelial Electrical Resistance (TEER) measurement
LY rejection
Caco-2 permeability assay procedure
Apparent permeability, Papp(cm/s) & Efflux Ratio
Ion exchange chromatography, Principles of Ion -exchange chromatographyHigh performance liquid chromatography (HPLC), Isocratic and Gradient Elution , The Ion Exchanger (Matrix)
Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
Ethnobotany and Ethnopharmacology:
Ethnobotany in herbal drug evaluation,
Impact of Ethnobotany in traditional medicine,
New development in herbals,
Bio-prospecting tools for drug discovery,
Role of Ethnopharmacology in drug evaluation,
Reverse Pharmacology.
Students, digital devices and success - Andreas Schleicher - 27 May 2024..pptxEduSkills OECD
Andreas Schleicher presents at the OECD webinar ‘Digital devices in schools: detrimental distraction or secret to success?’ on 27 May 2024. The presentation was based on findings from PISA 2022 results and the webinar helped launch the PISA in Focus ‘Managing screen time: How to protect and equip students against distraction’ https://www.oecd-ilibrary.org/education/managing-screen-time_7c225af4-en and the OECD Education Policy Perspective ‘Students, digital devices and success’ can be found here - https://oe.cd/il/5yV
The Indian economy is classified into different sectors to simplify the analysis and understanding of economic activities. For Class 10, it's essential to grasp the sectors of the Indian economy, understand their characteristics, and recognize their importance. This guide will provide detailed notes on the Sectors of the Indian Economy Class 10, using specific long-tail keywords to enhance comprehension.
For more information, visit-www.vavaclasses.com
This is a presentation by Dada Robert in a Your Skill Boost masterclass organised by the Excellence Foundation for South Sudan (EFSS) on Saturday, the 25th and Sunday, the 26th of May 2024.
He discussed the concept of quality improvement, emphasizing its applicability to various aspects of life, including personal, project, and program improvements. He defined quality as doing the right thing at the right time in the right way to achieve the best possible results and discussed the concept of the "gap" between what we know and what we do, and how this gap represents the areas we need to improve. He explained the scientific approach to quality improvement, which involves systematic performance analysis, testing and learning, and implementing change ideas. He also highlighted the importance of client focus and a team approach to quality improvement.
3. ION EXCHANGE CHROMATOGRAPHY:
It is the
based on their affinity towards the ion
exchangers.
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4. Separation is based on the attraction
between oppositely charged particles.
Net charge exhibited by compound is
dependent on their pKa and pH of the
solution in accordance with HENDERSON
HASSELBACH EQUATION.
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5. ION EXCHANGERS
Special type of polyelectrolyte and consist of
to which are bonded a large
number of electrically charged group.
Classification:
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Ionexchanger
Based on the
ionogenic group
Based on the nature
of the source
Based on the
structure
Madras Medical College
6. CATIONIC
EXCHANGE RESIN
STRONG WEAK
ANIONIC
EXCHANGE RESIN
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STRONG WEAK
R-SO3H R-COOH R-NH4
+R2-NH
Strong ion exchange media : no variation in charge, sample loading capacity is
maintained
Amberlite IR-120
Dowex
QAE-
Sephadex A-25
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Eg.
Clay,Peat,Lignite,Dolomite,
etc
Eg. sephadex
Eg. Acrylic amide,
formaldehyde, silicates
NATURAL
SEMI SYNTHETIC
SYNTHETIC
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PELLICULAR TYPE MACRORETICULAR
RESIN BEADS
MACROPOROUS
30-40µ
Low exchange
capacityVery Low exchange
capacity
5-10µ
Porous highly
efficient
SULPHONATED
RESINS
Low exchange
capacity
9. PROPERTIES
It must be sufficiently cross linked to have only a
negligible solubility.In order to permit diffusion of ions
through the structure at constant and finite rates
Swollen resin must be denser than water
Resin must be chemically stable
Cross linking is of greater importance
Swelling
polar solvents--- swelling
Non-polar solvents----- contraction
Particle size---- decrease higher the rate of ion exchange
50-100mesh/100-200mesh
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10. BASIS FOR SEPARATION
Capacity factor- The number of sites availbale for
the exchange.
capacity = V X N
W
Flow rate of sample - 1ml/min or 30 drops/min
Particle size- 100-200mesh for resins
pH- if exchange capacity of cationic exchanger
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For cation-5 moles/g
For anion-3.5 moles/g
11. Contd…
Distribution coefficient (KD)= Amount of ions in resin
Amount of ions in solution
Separation factor (α) = KD of component 1
KD of component 2
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14. PROCEDURE
Buffer selection and preparation
Column and media preparation
Sample preparation
Sample loading
Elution
Re-equilibration
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15. Eg.
In cation exchange chromatography, using a functional group on the solid
support with a pKa of 1.2, a sample molecule with a pI of 8.2 may be run in a
mobile phase buffer of pH 6.0.
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As a rule, the pH of the mobile phase buffer must be between the
pI (isoelectric point) or pKa (acid dissociation constant) of the
charged molecule and the pKa of the charged group on the solid
support.
USUALLY PREPARED VOLUME – 500ml
16. BUFFER SYSTEM FOR ANIONIC EXCHANGE
RESINS
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17. BUFFER SYSTEM FOR CATIONIC EXCHANGE
RESINS
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18. Equilibrate column with 5–10 column
volumes of start buffer or until the baseline,
eluent pH and conductivity are stable.
Using prepacked columns is highly
recommended to ensure the best
performance and reproducible results. An
evenly packed column ensures that
component peaks are not unnecessarily
broadened as sample passes down the
column so that the best resolution can be
achieved.
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19. Contd…
Allow buffers, media or prepacked columns to reach the same
temperature before use.
Rapid changes in temperature, for example removing packed
columns from a cold room and then applying buffer at room
temperature, can cause air bubbles in the packing and affect
the separation.
Wash away storage solutions and preservatives before using
any IEX medium.
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20. Contd..
Increase the volumes used for column equilibration before
the first run if using buffers containing detergents or a
different counter-ion to the one in which the medium has
been stored.
The volume required for the packed bed is determined by the
amount of sample to be purified and the binding capacity of
the medium.
Pack a column that will have approximately 5-fold excess of
the binding capacity required with a bed height up to 20 cm.
Wet packing
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21. Desalt the sample and transfer to the buffer
Adjust the sample to the chosen starting pH and ionic
strength and apply to the column.
Sample volume should be based on the capacity of ion
exchange resins
For protein samples maximum concentration of about 50-
70mg/ml can be used
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22. Apply samples directly to the column via a chromatography
system, a peristaltic pump or a syringe.
The choice of equipment depends largely on the sample
volume, the size of column, the type of IEX medium and the
requirements for accuracy in gradient elution.
Ensure that the top of the column bed is not disturbed during
sample application
Do not change buffer conditions until all unbound material
has been washed through the column (monitored by UV
absorbance) and until UV and conductivity values have
returned
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23. Achieved by decreasing the affinity of solute. It is done by
using salt of varying concentration
Bound proteins are eluted by controlled changes in ionic
strength or pH. The way in which these changes take place, by
using a linear or step elution, is selected according to the aim
of the separation:
Linear gradient elution
–high resolution separation or analysis
Step elution
–faster separation time, reduced buffer consumption
–group separation
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24. Linear gradient elution:
Begin elution using a linear gradient volume of 10–20
column volumes with an increasing ionic strength up to
0.5 M NaCl (50%B).
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25. Step elution
Elute bound proteins with 5 column volumes of
start buffer + NaCl at chosen ionic strength.
Repeat at higher ionic strengths until the target
protein(s) has been eluted.
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26. Wash with 5 column volumes of 1 M NaCl (100%B) to elute any remaining
ionically-bound material. Include a wash step at the end of every run in order
to remove any molecules that are still bound to the medium. Monitor UV
absorbance so that the wash step can be shortened or prolonged, as
necessary
Re-equilibrate with 5–10 column volumes of start buffer or until eluent pH
and conductivity reach the required values. A re-equilibration step after
washing returns the column to start conditions before applying further
samples. Whenever possible, monitor pH and conductivity to check when
start conditions have been reached. The re-equilibration step can then be
shortened or prolonged as necessary
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29. APPLICATION
• Separation of similar ions from one another
• Removal of interfering radicals
• Softening of hard water
• Complete demineralisation of water
• Separation lanthanides and actinides
• Separation of sugars
• Separation of amino acids
• Preparation of pure reagents
• Hydrometallurgy
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31. REFERENCE
www.harvardapparaturs.com, guide to ion
exchange chromatography
Instrumental analysis of chemical analysis by
Gurudeep R.Chatwal, Sham.K.Anand, Pg. 2.662-
2.672
Instrumental methods of chemical analysis by
B.K.sharma, C-123 -150
http://www.gelifesciences.com/file_source/GELS
/Service%20and%20Support/Documents%20and
%20Downloads/Handbooks/pdfs/Ion%20Exchang
e%20Chromatography.pdf
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