Partition Chromatography

Partition Chromatography Definition

Partition Chromatography technique is defined as the separation of components between two liquid phases viz original solvent and the film of solvent used in column. This separation theory was introduced in the year 1940s which was published by Richard Laurence Millington Synge and Archer Martin. It is also known as Liquid-liquid chromatography (LLC). Or if gas is the mobile phase it is called Gas-liquid chromatography (GLC)

Partition Chromatography Principle

The separation of the components from the sample mixture is carried out by the process of partition of the components between 2 phases. Both the phases are in liquid form. In this process the immiscible solid surface coated with the liquid surface on the stationary phase is in the mobile phase. The liquid surface is immobilized by stationary phase which results in making it stationary phase. The mobile phase moves from the stationary phase and components get separated. The separation depends on different partition coefficient.

Partition Chromatography Diagram

Diagram of paper chromatography

Partition Chromatography Procedure

Below we have explained the procedure to conduct Paper Chromatography Experiment for easy understanding

Apparatus required – chromatography jar, liquid impregnated paper (stationary phase), capillary tube (to apply sample mixture), mobile phase (example chloroform, methanol,acetone,ethanol).

  1. Take a clean and dry chromatography jar.
  2. To make sure that the environment of the jar is saturated with solvent vapors, a paper impregnated in mobile phase is set to the walls.
  3. Add mobile phase to the jar. Around 0.5 cm to 1 cm from the bottom of the jar.
  4. Close the jar.
  5. Allow to attain equilibrium.
  6. Mark the baseline on adsorbent.
  7. Apply sample to the paper with the help of capillary tube.
  8. Air dry the sample spot.
  9. Place the paper in the jar and close it.
  10. Allow the system to stand till the solvent moves to some distance from the baseline.
  11. Take out the paper and dry it.
  12. If the sample components are separated showing colors then dry it in ordinary light. If it is a colorless component then dry it in UV lamp.
  13. Store the chromatogram
  14. Calculate Rf value.

Partition Chromatography Applications

There are various applications of Paper Chromatography. Some of the applications are mentioned below:

  • To separate and identify amino acids.
  • To separate and identify tannins.
  • To separate and identify alkaloids.
  • To separate and identify carbohydrates.
  • To separate and identify glycosides.

Types of Partition Chromatography:

  1. Liquid-liquid Chromatography – It is a chromatography technique where a sheet of blotting paper, is used instead of adsorption column. The components are separated based on their differential migratory velocities. On separating they are stained to make the chromatogram visible.
  2. Gas liquid Chromatography – A chromatography technique in which the separation of the mixture is done by an inert gas along a tube. The tube is filled with finely divided inert solid. The solid is coated with a nonvolatile oil. The migration of each component occurs at a rate determined by its solubility in oil as well as its vapor pressure.

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