This document provides an overview of column chromatography, including its history, definition, principle, types, experimental technique, requirements, applications, advantages, and disadvantages. Column chromatography was developed in 1901 by Russian botanist Mikhail Tsvet as a method to separate plant pigments by passing an organic solution through an adsorptive material in a glass column, resulting in discrete colored bands. It involves using a column packed with a stationary phase and flowing a liquid mobile phase through to separate components of a mixture based on differential adsorption between the phases.

1. PRESENTED BY
RD

2.  Introduction
 History
 Definition of column chromatography
 Principle
 Classification of column chromatography
 Experimental technique of column chromatography
 Requirements
 Application
 Advantages
 Disadvantages
 References

3.  DEFINITION:-
 Chromatography is a physical method of separation in which the
components to be separated are distributed between two phases :
 Stationary phase
 Mobile phase
o The stationary phase may be a solid or liquid supported on a solid or gel.
• May be packed in a column Eg.-Column Chromatography.
• Spread as a layer on glass/aluminium plate Eg. – TLC, HPTLC.
• Distributed as a liquid film Eg.- GLC
o The mobile phase may be a liquid /solvent/mixture of solvent or gases.

4.  The first chromatography column was developed by the
Russian botanist Mikhail Tsvet in 1901.
 Who washed an organic solution of plant pigments
through a vertical glass column packed with an
adsorptive material.
 He discovered that the pigments separated into a series
of discrete colored bands on the column, divided by
regions entirely free of color.

5.  Column chromatography is a separation technique in
which components of mixture is separated by using a
glass column packed with stationary phase and the
liquid mobile phase flowing continuously through the
column.

6.  Column chromatography is separated into two categories, depending on
how the solvent flows down the column.
 Gravity column chromatography:- If the solvent is allowed to flow down
the column by gravity, it is called gravity column chromatography.
 Flash chromatography:-If the solvent is forced down the column by
positive air pressure, it is called flash column chromatography, currently
used in organic chemistry research laboratories

7.  A solid stationary phase and a liquid mobile phase is used and the principle
of separation is adsorption.
 The mixture to be separated is dissolved in a suitable solvent and allowed
to pass through a tube containing the adsorbent
 The component which have greater absorbing power is adsorb in the upper
part of the column.
 The next component is adsorb in lower portion of the column which have
less adsorbing power than the first component.
 The process is continued. As a result the materials are partially separated
and adsorbed in the various part of the column. The type of interaction
between the stationary phase and solute is reversible in nature.

9.  Column
 Adsorbents
 Packing the column
 Wet packing
 Dry packing
 Solvent used
 Application of the sample
 Identification and separation of compound
 Recovery of components
 Detection of compound

10.  Separation of mixture of compound.
 Removal of impurities or purification process.
 Isolation of active constituents.
 Isolation of metabolites from biological fluids.
 Estimation of drug in formulation and crude drug extract.

11.  Any type of mixture can be separated by column chromatography.
 Any quantity of mixture can also be.
 Wider choice of Mobile Phase

12.  Time consuming method.
 More amount of solvent are required which are expensive.
 Use of maximum adsorbent.
 Drying and bubble presence are possible.

13.  Kar Ashutosh, “pharmaceutical Analysis’’,CBS
Publishers & Distributors pvt.ltd , Volume 2nd , page
no 161-181.
 Indian Pharmacopoeia , ministry of health sciences
government of India 2014 edition , volume 1st , page
no 163-166.