PRESENTED BY:-

HIRA SALEEM

BS CHEMISTRY.SEM-4

COURSE CODE:CHE-402

DEPARTMENT OF CHEMISTRY
 CHROMATOGRAPHY
Introduction of chromatography
Classification of chromatography
Types of chromatography
Paper chromatography
Principle, Procedure, Types, Application, Advantages & disadvantages

Thin layer chromatography

Principle, Procedure, Application, Advantages & disadvantages
Comparison of Paper chromatography & Thin layer chromatography
Introduction of Chromatography
Definition:- Chromatography is a technique employed
For separation of the components of mixture by continuous
Distribution of the components between two phases.(mobile phase
and stationary phase).

Chromatography is relatively a new technique which was first

Invented by Michael Tswett a Russian botanist in 1906.
 Classification of Chromatography
CHROMATOGRAPHY

Adsorption Partition
Competition between solid and liquid Competition between liquid and liquid

Competition between Competition between Competition between Competition

solid & liquid solid & gas liquid &liquid between liquid &
gas
 Types of Chromatography
Thin layer Chromatography
Paper Chromatography
Column Chromatography
Gas Chromatography
High Performance liquid Chromatography
Procedure of Paper
Chromatography
 In this technique a drop of the test
solution is applied as a small spot on a
filter paper and the spot is dried.
 The paper is kept in a close chamber and
the edge of the filter paper is dipped into
a solvent called a developing solvent.
 As soon as filter paper gets the liquid
through its capillary axis and when it
reaches the spot of the test solution.
 The movement of substances relative to
the solvent is expressed in terms of Rf
values.
For Example: Migration parameters.
Advantages & Disadvantages of Paper
Chromatography:
Advantages:
o Paper chromatography requires very less quantitative material.
o Paper chromatography is cheaper compare to other chromatography methods.

Disadvantages:
o Large quantity of sample cannot be applied on paper chromatography.
In quantitative analysis paper chromatography is not effective.
o Less accurate compare to HPLC or HPTLC.
Advantages & Disadvantages of Thin layer
Chromatography:
Advantages:
o The components are separated in very little time as the components will elute
out very quickly.
o The solvents for the TLC plate can be changes.

Disadvantages:
o In this method the plate length is limited and hence separation takes place only
up to certain length.
o The separation takes place in an open system.
Comparison of Paper Chromatography &
Thin layer Chromatography:
Paper Chromatography
• The principle of separation is
partition.
• Less amount of substance is
required.
• More time consuming (2-3
hours)
• Separation is less sharp.
Thin layer Chromatography
• The principle of separation is
adsorption.
• More amount of substance is
required.
• Less time consuming (15-45
minutes).
• Separation is more sharp.
 REFRENCES
• BIOCHEMISTRY
C.B.Power
G.R.Chatwal
• www.slideshare.net
• m.authorstream.com
• https:/en.m.Wikipedia.org
THANK YOU