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Partition Chromatography

We know that mixture is the combination of more than one substance in a certain ratio. There are various techniques to separate the constituents of a mixture such as distillation, crystallization, separating funnel, chromatography etc. Out of all these techniques, chromatography is most reliable technique as it gives 99% pure substance.

This technique is usually used to separate constituent from mixture to get high purity. In this technique the substance which has to be analysed is poured into a vertical glass tube. This vertical tube contains an adsorbent. Various components of the mixture adsorbed at different rates of speed over absorbent. The absorption depends on the degree of attraction between absorbent and various components of the mixture. It results the formation of bands of color at different levels in column.

So we can say that chromatography consists of two phases, mobile and stationary phase. The mobile phase is usually a fluid which carries the sample components whereas the stationary phase is a solid material like Kaolin, alumina, silica, and activated charcoal which takes up the particles passing through it.  Chromatography is mainly used in clinical laboratory to detect and identify in body fluids. On the basis of stationary phase, the nature of the adsorptive force, the nature of the mobile phase etc. chromatography can be classified in different types.

For example; adsorption chromatography contains an absorbent as a stationary phase whereas affinity chromatography is based on a highly specific biologic interaction. The column chromatography contains a column in which various solutes of a solution are travelled and separate individual components on the stationary phase.

The column chromatography is used to separate biomolecules like vitamins, steroids, hormones etc.  It also determines the amounts of constituents of mixture. In the exclusion chromatography, the stationary phase is a gel with closely controlled pore size. In this method, molecules are separated on the basis of molecular size and shape.

Gas chromatography is used an inert gas as the mobile phase to separate the components of a mixture. Paper chromatography contains a sheet of blotting paper or filter paper in the adsorption column. The partition chromatography is a process of separation that utilizes the partition of the solutes between two liquid phases.

 

Difference Between Adsorption and Partition Chromatography

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Adsorption and Partition chromatographic techniques are based on different principles however both of them are efficient analytical techniques for the separation of components over a stationary phase with mobile phase on the components. In partition chromatography, separation on the stationary phase occurs by partition whereas adsorption chromatography is based on adsorption. Both of these techniques are based on the nature of component and samples.

However partition chromatography is higher or well-developed techniques used for separation of component. Adsorption chromatography is based on the relative differences in adsorption of constituents of given sample. Because of differences in their affinity towards stationary phase, the components of the mixture adsorb with different rates. Adsorption stands for the physical bonding between the mobile and stationary phase. Polarity of compounds determines the adsorption tendency of them. This technique can be used only for solid-liquid or solid-gas chromatography.

On the other hand, partition chromatography involves the separation of components by the distribution of them between two liquid phases. Components get separated due to differences in partition coefficients.

Partition Chromatography Definition

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The chromatographic technique that involves the separation of components between two liquid phases; original solvent and film of solvent used in column is called as partition chromatography. In this technique the molecules get preferential separated between two liquid phases. One of the liquid phases is stationary phase whereas another is mobile phase in liquid state.

The components of mixture get dispersed into both phases preferentially. Remember polar molecules get partitioned into polar phase and non-polar molecules get attracted towards non-polar phase only. This method can be applied to Liquid-liquid, liquid-gas chromatography only not on solid-gas type as solid phase cannot move.

Partition Chromatography Technique

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Partition chromatography is a type of liquid chromatography which is used to separate components of a given sample. The separation of components is based on the interactions of the particles of sample with the mobile and stationary phases.

In partition chromatography, both phases are in liquid state. 

Partition Chromatography

Partition Chromatography Principle

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The principle of partition chromatography is that the separation of components of given sample occurs due to partition of components between two liquid phases. In this process, the stationary phase is coated with a liquid surface. The solid surface must be immiscible in the mobile phase. Due to partition of component between both phases reduces some components compare to others.

It results the separation of components. Here the stationary phase immobilizes the liquid surface and makes it stationary phase. The mobile phase passes over the stationary phase and separate out. The separation depends on the relative solubility in the stationary liquid layer. Because of different partition coefficient, different component of sample are separated.

Partition Chromatography Procedure

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In the partition chromatography, the separation of components of the mixture occurs due to distribution of components between two liquid phases. The separation of components occurs on the basis of partition coefficients of constituents of mixture. The flow of components or mobile phase separates all components over stationary phase.

The polar phase attracts polar components of sample and non-polar components are attracted by non-polar phase of the system. Partition chromatography can be done with different phases like Liquid-liquid and liquid-gas chromatography.

Partition Chromatography Applications

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Partition chromatography is one of best chromatographic method to get high performance and pure components. Partition Chromatography is used for final purification natural extracts, synthetic mixtures and biological matrices. It is also used for fractionization of complex crude extracts.
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