Definition of thin
Layer Chromatography:
Thin layer chromatography is defined as the method of
separation of the two or more component from the mixture based on the
adsorption or the partition principle.TLC is one of the simplest forms of the chromatography.
Principle of thin layer
chromatography:
The principle of Thin Layer Chromatography is adsorption
Chromatography or the partition chromatography depends on the nature of the
mobile phase and the stationary phase used in the analysis.The analyte molecule which have more affinity towards the stationary phase are travel slower and the molecule which have lower affinity
are travel faster when mobile phase is applied.
Component and
Instrument for Thin layer Chromatography:
In thin layer chromatography the stationary phase coated on
the aluminium or glass plate. Most commonly TLC silica gel is used as the
stationary phase.
The stationary phase is relatively
thin, uniform, layer of dry fine powder of silica which applies on aluminium
glass or plastic material.
TLC Silica Gel, alumina are used as the stationary phase. The
stationary phase material is coated on the aluminium plate or glass plate.
Stationary phase can be categorise as inorganic and organic
stationary phase
Silica gel, Aluminium Silicate, Bentonite are the example of
the inorganic stationary phase.
Cellulose, Activated charcoal, dextran gel polyamide are the
example of the organic stationary phase.
Different solvent are used as the mobile phase. Mobile phase
may be used as the mixture of the solvent
Hexane, Hexane, Chloroform, Acetonitrile, ethyl Acetate,
water
The selection of the stationary phase and the mobile phase is
depending on the nature of the analyte which need to be separated.
TLC chamber:
The TLC chamber is made of glass with
lid or beaker with the lid.
TLC chamber should be made up of
inert material and should be not react with the mobile phase.
TLC chamber should be transparent.
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| Thin Layer Chromatography (TLC) |
Application of thin
Layer chromatography:
Thin layer Chromatography is used to identification of the
component in mixture and find the purity of the analyte and the Identification
of the impurities
In pharmaceutical industry identification of active
pharmaceutical ingredient in tablet or other dosage form, Presence of impurity
into the mixture.
In phytochemical Analysis to find out component into the
plant
In Food Industry also Thin Layer chromatography
In Environmental Analysis also thin layer chromatography to
find out pollutant in air and water
Advantages of thin
layer chromatography:
Easiest, quick and perform in less time
Less expensive, not much instrument and material required
Simple to perform and fast
Disadvantages of Thin
Layer chromatography:
Only non-volatile materials can be analyse by the thin layer
chromatography
Completely manual method human error may cause variation in
result.
Calculation of Rf factor:
Rf value:
Is the ratio of distance traveled by the solute to the distance traveled by the solvent.
Is the ratio of distance traveled by the solute to the distance traveled by the solvent.
Rf value: Distance traveled by sample/ Distance travel by solvent.
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