Column Chromatography Principle

Definition –

Column chromatography is a separation technique that is used to isolate compounds from complex mixtures. Either small or large-scale column chromatography can be carried out to separate and purify the analytes. Column chromatography has two phases viz a mobile phase and a stationary phase.

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Column Chromatography Principle

Column chromatography process involves the uniform percolation of a liquid solute through a column packed with finely divided material. It involves adsorption, partition or ion exchange phenomenon. The adsorption column chromatography is based on the retention of solute by surface adsorption. This technique is useful in the separation of nonpolar substances and constituents of low volatility.

The sample mixture is set on the top of the column and is made to absorb on the top of the stationary phase. Eventually, the mobile phase which is applied to the column is used to remove the mixture via the stationary phase. Separation of compounds in column chromatography by utilizing the molecule’s polarity. The rate at which molecules move in the column during the separation process depends on their polarity. As the mobile phase comes out of the column it is collected in small fractions in the test tube. Then it is allowed to isolate and purify. Later, the removal of salt is done by a rotary evaporator to obtain the separated compound. Column chromatography is the most widely used technique for the purification of compounds due to its versatility. Also, it is a very convenient process.

diagram of column chromatography 1

Advantages And Disadvantages Of Column Chromatography

Advantages Of Column Chromatography

  • All different kinds of complex mixtures can be separated by column chromatography.
  • The mobile phase has a wide range.
  • There is no limit for quantity as any amount of mixture can be separated by this technique.
  • It is a robust method.
  • The separated analytes can be reused.
  • This process can be automated.

Disadvantages Of Column Chromatography

  • It is a time-consuming process for the separation of compounds.
  • It is expensive as higher quantities of solvents are required.
  • The automated process becomes complicated and therefore costly.
  • It has a low separation power.

Applications of Column chromatography

  1. This technique is used for the separation of diastereomers.
  2. It is used to isolate racemates.
  3. It is used to separate geometrical isomers.
  4. Tautomeric mixtures can be separated from this process
  5. To purify homogeneity of various coloured compounds.


  • Column chromatography is a chromatography technique used to separate chemical substances of a mixture into individual compounds.
  • This separation method consists of two phases viz: contiguous stationary phase and mobile phase.
  • The column is prepared by mixing the silica and a suitable solvent. It is later poured into a column made of glass.
  • The mobile phase (suitable solvent) is moved along with a compound mixture through the Column based on the polarity.

For the detailed experiments on Column chromatography Procedure and Theory

You may visit: Column Chromatography

Other important links:

Chromatography and its types Thin layer chromatography
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