Ion Exchange Chromatography

Presented by,

Mahendra JadhaoPhD Research Scholar,Sungkyunkwan University,Rep. of Korea.

Introduction

• Separation or Purification technique based on electrostatic interaction of charged surface

Types of Ion Exchange Chromatography

• Cation Exchange Chromatography

• Anion Exchange Chromatography

Principle of Ion Exchange Chromatography

Reversible electrostatic interaction of charged species with separation matrix

Mechanism of Separation in IEC

• pH Based Binding

pI

pH = pI – No net charge

pH > pI – Net negative charge

pH < pI – Net positive charge

Cation exchange chromatography

• Positively charged molecules are attracted to a negatively charged solid support. Commonly used cation exchange resins are S-resin, sulfate derivatives; and CM resins, carboxylate derived ions

  

Anion exchange chromatography

• Negatively charged molecules is attracted to a positively charged solid support. Commonly used anion exchange resins are Q-resin, a Quaternary amine; and DEAE resin, DiEthylAminoEthane

Ion Exchange Resins• Resins are amorphous particles of organic materials (e.g. polystyrene, cellulose)• Used for separation of small molecules.

Classification

Working Principle

• The basic process of chromatography using ion exchange can be represented in following steps:

1. Equilibrium

2. Sample application

3. Elution

4. Regeneration

Factors affecting Ion Exchange Chromatography

1. Nature and properties of Resins a. Cross linking and Swelling: if more cross-linking, they are more rigid but swelling is less Less swelling- separation of ions of different size is difficult b. Nature of Exchange ions : valency of ions , concentration of solution

2. pH of Mobile phase ( Buffer)3. Mobile phase modifiers

Applications• Softening of hard water• To separate protein mixtures• Organic separation: additives in food and drug

samples• Demineralization/ deionization of water• Purification of solutions to be free from ionic

impurities• Concentration of ionic solutions.• Separation of inorganic ions.

Thank you…