Column chromatography

 Column chromatography 

Column chromatography is a laboratory technique used to separate, identify, and purify mixtures of compounds based on their chemical properties and affinities.

Principle:

Column chromatography involves passing a mixture through a stationary phase (adsorbent material) packed in a column. The components separate as they interact differently with the stationary phase.

Types:

1. Adsorption Chromatography: Separates based on adsorption to the stationary phase.

2. Ion-Exchange Chromatography: Separates ions based on electrostatic interactions.

3. Size-Exclusion Chromatography: Separates based on molecular size.

4. Affinity Chromatography: Separates based on specific binding interactions.

Steps:

1. Preparation: Choose stationary phase, column, and solvents.

2. Sample Loading: Apply mixture to the column.

3. Elution: Pass solvent through the column.

4. Separation: Components separate based on interactions.

5. Detection: Identify separated components using detectors.

6. Collection: Collect separated fractions.

Stationary Phases

1. Silica Gel

2. Alumina

3. Cellulose

4. Ion-exchange resins

Mobile Phases:

1. Solvents (e.g., hexane, ethyl acetate)

2. Buffer solutions

Applications:

1. Purification of compounds

2. Separation of enantiomers

3. Analysis of biological samples

4. Pharmaceutical industry




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