A basic understanding of chromatography, various types and how it works

1. General Principle and types of
chromatography
By,
Ritwick Ranjan Sarma
M.Sc. Bioscience
Yenepoya Research Centre

2. Contents:
• History
• Introduction
• Principle
• Chromatogram
• Retention Factor
• Types of chromatography
• TLC
• Gas chromatography
• HPLC
• Size exclusion chromatography
• Affinity chromatography
• Ion exchange chromatography
• Applications of chromatography

3. History:
• Tswet, a Russian Botanist
(referred to as the father
of Chromatography) is
credited with the
development of
chromatography in the
year 1903.
• Used this technique to
separate plant pigments.
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4. Chromatography:
• The word “Chromatography”
is derived from two greek
words-Chroma means colour,
and Graphein to write.
• Chromatography is the
collective term for a family
of laboratory techniques for
the separation of mixtures.
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5. Principle of Chromatography:
• Chromatography is
based on the
principle of the
partition of solute
between two
phases/solvents.
• Consist of mobile
phase and stationary
phase.
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6. Chromatogram:
• A graphical representation of detector
response, concentration of analyte in
effluent, or other quantity used as a
measure of effluent concentration.
• The retention time or volume is when a
solute exists the injector and passes
through the column and the detector.
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7. Retention factor:
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8. Types of Chromatography:
• Planar Chromatography
• Column Chromatography
Based on shape of
bed
• Gas chromatography
• Liquid chromatography
Based on physical
state of mobile
phase and stationary
phase
• Ion exchange
• Partition
• Affinity chromatography
Based on
mechanism of
separation

9. Thin Layer Chromatography(TLC)
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10. Gas Chromatography:
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11. HPLC:
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12. Size Exclusion
Chromatography
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13. Affinity Chromatography:
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14. Ion-exchange Chromatography:
• There are two types of Ion exchange
a) Cation exchange
b) Anion exchange
• Column is made up of polymer which is attached
to a charged ion (cation/anion).
• Stationary phase = Solid
• Mobile phase = Liquid

15. Application of chromatography:
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16. References:
• Wilson and Walker Biochemistry, 6th edition.
• https://microbenotes.com/chromatography-
principle-types-and-applications
• https://www.omicsonline.org/special-
issue/chromatography-and-its-applications.php
• www.slideshare.net/chromatography