Work of Chromatography – Basic Principle of Chromatography

Chromatography is a technique which is used to separate component of the mixture. Chromatography derives from a Greek word. Chrome mean Color and Graphy mean writing. Khromatos mean color writing.

First Mikhail Tsvet worked on plant pigments like Chlorophyll, Xanthophyll, and carotenoids in 1900.  He separated plants Pigments. Many types of Chromatography techniques are used named are. Biochemical techniques may be Preparative or Analytical.

Preparative Procedure is most commonly used to a purification of a large number of biological samples that future used to study and characterization. The analytical procedure is applied to purification of Biological samples. However, they may be used to elute any physical, chemical and biological characterization of biomolecules. There are many techniques of chromatography listed below. These techniques are studied in chemistry, biology as well as biochemistry.

Types of Chromatography

  • Paper Chromatography

  • Thin layer Chromatography

  • Gas Chromatography

  • HPLC

  • Affinity Chromatography

  • Ion exchange chromatography

On what basis Chromatography separate substance/ Work of chromatography?

Chromatography technique worked on basic principle. Two phases are involved. One is called the stationary phase and 2nd is called the mobile phase. Stationary phase. As the name indicated stationary mean station, it may be solid, gas, or liquid.  Mobile phase. Mobile phase may be gas or liquid. It’s mean mobility.

A sample to be examined is called solute. The substance is separated due to their relative affinity. Solute distribute itself between stationary phase as well as mobile phase. The distribution of components of a mixture between the stationary phase and the mobile phase is governed by the distribution coefficient. The component of a mixture with small k value remains in stationary phase as the moving phase flow over it. The component with a greater value of K remains largely dissolved in the mobile phase.

For Example, Particles having a great affinity to remain with mobile phase travel fast. On the other hand, particles having low affinity with mobile phase remain below and stick with the stationary phase. Like as you have seen many stones present on the Seashore. These stones are of different sizes. Small stones will move with the flow of water because these stones have an affinity to move with water. On the other hand, larger and heavy stone which are stick with land remain as such could not move with water because these have not affinity to flow with water.

What is Adsorption chromatography?

Chromatography in which the stationary phase is solid called adsorption chromatography.  In which substance leave the mobile phase to become adsorbed on the stationary phase.

What is partition chromatography?

Chromatography on which stationary phase is liquid called parathion chromatography. In this type, the substances being separated are distributed are throughout both the stationary and mobile phases.

Uses of Chromatography

Chromatography is used for many purposes. This is use full in organic synthesis for separation, isolation, and purification of the products. They are equally important in quantitative and qualitative analyses and for determination of the purity of a substance.

Different chromatography techniques are used for separate components. As chromatography used in industry to purify samples. Mostly HPLC and gas chromatography used in industry. Drug industry chromatography used are very important to purify drug. Paper chromatography used to separate color pigments. Ion exchange chromatography uses to separate large or small ion.

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