Column chromatography

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.