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Chromatography

The document discusses different types of chromatography techniques including column chromatography, planar chromatography, paper chromatography, thin layer chromatography, and gas chromatography. It explains the principles and key steps of column chromatography, planar chromatography such as thin layer chromatography, and paper chromatography.

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DENISE COLE
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15 views16 pages

Chromatography

The document discusses different types of chromatography techniques including column chromatography, planar chromatography, paper chromatography, thin layer chromatography, and gas chromatography. It explains the principles and key steps of column chromatography, planar chromatography such as thin layer chromatography, and paper chromatography.

Uploaded by

DENISE COLE
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd
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Analytical Chemistry: CHE3724 Module

Dr RC Chokwe
Florida Science Campus
Eureka building
Chemistry Department
K-M-032
[email protected]
+27 11-670-9393
Types of chromatography

Chromatography

Adsorption Partition
chromatography chromatography

CC, GC & HPLC


Paper
TLC
chromatography
Mobile phases and stationary phases
Supporting medium for stationary

Supporting medium

Planar Column

TLC, Paper CC, HPLC, GC


chromatography, HPTLC
Column chromatography

• This is a solid liquid technique where the stationary


phase is a solid and the mobile phase is a liquid
Principle of column chromatography
• When a mixture of components dissolved in the mobile
phase is introduced into the column, the components
move through the stationary phase with different rates
depending on their relative affinity for the stationary
phase.

• The compound with lesser affinity for the stationary


phase moves faster through the stationary phase and is
eluted out of the column first.

• The one with greater affinity for the stationary phase


moves slower and is eluted last
Planar chromatography

• The stationary phase is supported on a flat plate


or is a paper and the mobile phase moves
through the stationary phase by capillary action
or gravity.
Principle of paper chromatography
• The principle is based on partition chromatography
where the components of a mixture are partitioned
between liquid phases.

• One phase is the water, which is held in the pores of the


filter paper used, and the other is the mobile phase which
moves over the paper.

• The components of the mixture are separated based on


the differences in their affinity towards water( SP) and the
mobile phase during the movement of the mobile phase
under capillary action of pores in the paper
Principle of TLC
• The separation is based on the affinity of the compounds
towards the stationary and mobile phases.

• The components of the mixture move up the stationary


phase due to capillary action under the influence of the
mobile phase whereby compounds with a lesser affinity
for the stationary phase move up the plate faster and
those with greater affinity move slower leading to the
separation of the components of the mixture.

• Once separated the compounds are identified using a


suitable detection technique
Four steps of TLC experiment

• Sample application- each sample is spotted


on the plate using capillary
• Development- separation process
• Visualization- viewed under UV light if not
visible
• Interpretation of the results- Calculating and
comparing the different retardation factors
TLC sample application
Development of the TLC plate
Development of the TLC plate
Visualization of TLC Results
Interpretation of TLC Results
TLC results

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