Ion exchange chromatography separates ions and polar molecules based on their affinity for an ion exchanger. There are two main types: cation exchange attracts positively charged molecules to a negatively charged resin, while anion exchange attracts negatively charged molecules to a positively charged resin. The stationary phase is an ion exchange resin and the mobile phase is usually a liquid or gas that carries sample components through the resin. The process involves equilibrating the resin, adsorbing the sample so charged molecules bind to the resin, then eluting bound molecules by increasing ionic strength so they detach from the resin in a purified form.

1. FARWA FIAZ
13091514-057
BS 6th (B)

2. TOPIC
“ION EXCHANGE CHROMATOGRAPHY”

3. • INTRODUCTION:
• It is a process that separates ions and polar molecules
based on their affinity to the ion exchanger.
• An ion exchanger is something that is supposed to attract
positive or negative charges.(charged resin in this case)
• This process is used for proteins, small nucleotides, and
amino acids.
• It is mostly used in protein purification and water
analysis.

4. • TYPES
• Following are the two type of ion exchange
chromatography:
1. Cation exchange chromatography
• In this type positively charged molecules are
attracted to a negatively charged ion
exchanger
• Commonly used cation exchanger are sulfate
derivatives, S-resin and
carboxylate derived ions

5. 2. Anion exchange chromatography
In this type negatively charged molecules are
attracted to a positively charged solid
support(ION EXCHANGER)
Commonly used anion exchange resins are
Q-resin and DEAE resin.

6. TWO PHASES
• The stationary phase is a resin on which usually
organic coating provides a charged surface. It is
fixed and immobilized.
• The mobile phase may be a liquid or gas, that
moves the sample components through a region
containing the solid or liquid stationary phase.
Base solutions and salt solutions can be used as
the mobile phase.

7. PRINCIPLE:
1.Equilibration
• At this stage whole surface of charged resin is
equally surrounded by salt molecules or buffer
2.Sample application and Adsorption
• The impure sample for example of protein is
applied from the top of the tube it may contain
+ve,-ve or neutral groups
• Molecules having opposite charge will bound
charged resin. This binding will depend on the
suitable PH.

8. • 3. ELUTION
• By increasing the ionic strength or concentration of
buffer,it will displace or release all the bound protein
charges and unbound impurities and move them down
the tube to elute them out,charged protein molecules
will take longer time to unbound from charged resin so
it can be purified and isolated according to this
property. This process is called elusion.
• 4.Regeneration
• Buffer ions removes protein molecules still bound to
ion exchanger. This ensures that stationary phase is
available for the next run.

9. urple
• At the very first step sample is shown in
purple color
• Then buffer is applied which wash out all the
bound and unbound sample materials
• Liquid in blue color( last test tube) will be
purified desired sample