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Thin Layer Chromatography Principle

Thin Layer Chromatography Definition:-

A separation technique used to isolate non-volatile mixtures. TLC is performed with a sheet of plastic, aluminum foil, or glass coated with a thin layer of adsorbent material, such as silica gel, alumina, and cellulose is called Thin-layer chromatography.

Thin Layer Chromatography Principle:-

Thin-layer Chromatography (TLC) has two phases viz mobile phase and stationary phase. The stationary phase is prepared by coating a thin layer of alumina or silica on metal, plastic or glass. The compound mixture moves along with the mobile phase via the stationary phase and separation occurs as per the degree of adhesion.

The separation process depends on the relative affinity of the compounds towards the mobile phase and stationary phase. The compounds which are under the influence of the mobile phase move on the surface of the plate (stationary phase). During this movement, the compounds with lower affinity to stationary phase travel fast while the others move slow. Therefore, the separation is achieved. Once separation occurs, the individual components are visualized at their respective level of travel on the plate.

Advantages And Disadvantages Of Thin Layer Chromatography

Advantages Of Thin Layer Chromatography –

  • An easy method of separation of the components.
  • In this technique, fewer types of equipment are used. The separation is done in a very short time as the components elute rapidly.
  • All components of UV light is achievable to visualize.
  • The non-volatile compounds can be separated by this method.
  • Microlitre quantity of sample can also be separated in TLC
  • The components of complex mixtures easily separate and recover.

Disadvantages Of Thin Layer Chromatography –

  • The results obtained from the experiment are difficult to reproduce.
  • Applicable for soluble mixture components only.
  • Qualitative analysis, not quantitative analysis.
  • Not an automatic process.
  • The humidity and temperature can affect the results as Thin layer chromatography works in an open system.
  • Separation process takes place only to a certain length as plate length is limited.

Applications of thin layer chromatography (TLC)

  1. To purify the given samples.
  2. Compound identification. Compounds such as alcohols, alkaloids, antibiotics, acids, proteins, amines, etc.
  3. To know the level of purity of the sample.
  4. To isolate the component from complex mixtures.


  • TLC (Thin layer chromatography) is a chromatography technique used to separate chemical substances of a mixture into individual compounds.
  • This technique comprises two phases viz contiguous stationary phase and mobile phase.
  • The stationary phase is prepared by mixing calcium sulfate, silica gel, and water.
  • The mobile phase is moved along with the mixture of compounds via the TLC plate (stationary phase).

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