elegant chromatographic technique

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4. Introduction..
Chromatography:
Chroma - color
Graphein - to write
Chromatography was first developed by
the Russian botanist Mikhail Tswett in 1903
as he produced a colorful separation of plant pigments
through a column of calcium carbonate
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5. Cont...
 Mobile phase : A solvent that flows through the
supporting medium
 Stationary phase : A layer or coating on the
supporting medium that interacts with the analyte
 Supporting medium : A solid surface on which the
stationary phase is bound or coated.
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6. Cont...
 HPTLC is a well known and versatile separation method
which shows a lot of advantages and options in
comparison to other separation techniques.
 The method is fast and inexpensive and does not need
time consuming pretreatments.
 Traditional Thin Layer Chromatography & its modern
instrumental quantitative analysis version HPTLC are
very popular for many reasons such as
 visual chromatogram,
 simplicity,
 multiple sample handling,
 low running and maintenance costs, disposable layer etc.
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7. Cont...
Features:
Simultaneous processing of sample and standard -
better analytical precision and accuracy, less need for
Internal Standard
Several analysts work simultaneously
Lower analysis time and less cost per analysis
 Low maintenance cost
Simple sample preparation - handle samples of
divergent nature
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8. Cont...
 No prior treatment for solvents like filtration and
degassing
 Low mobile phase consumption per sample
No interference from previous analysis - fresh
stationary and mobile phases for each analysis - no
contamination
 Visual detection possible - open system
 Non UV absorbing compounds detected by post-
chromatographic derivatization
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9. Principle..
 HPTLC have similar approach and employ the same
physical principles of TLC (adsorption chromatography)
i.e. the principle of separation is adsorption.
 The mobile phase solvent flows through because of
capillary action. The components move according to their
affinities towards the adsorbent. The component with
more affinity towards the stationary phase travels slower.
The component with lesser affinity towards the stationary
phase travels faster. Thus the components are separated
on a chromatographic plate.
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10. Steps involved..
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11. Instrumentation..
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Application..

13. Comparisons..
 Advantages of HPTLC over TLC Visual
chromatogram
 Simplicity
 Multiple sample handling
 Low running and maintenance costs and disposable
layer etc
 Detection by TLC scanner
 Spotting by applicator
 Highly précised
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14. Cont..
 Advantages of HPTLC over HPLC
Sample and standard, both can be used at a time.
 Evaluation time is less.
 Use of internal standard is not necessary.
 Simultaneously 2 or 3 persons can work but not in case
of HPLC.
 Solvent degassing and removal of impurities is not
necessary.
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15. Cont..
HPCPC ( High Performance Centrifugal Partition Chromatography):
HPCPC, is anew technique of Liquid Chromatography, LC,
which is support free and which finds its place among the several Liquid
Chromatographic methods as shown on the following chart .
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17. References:
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