APPLIED
CHEMISTRY
CHROMATOGRAPHY
Presented by
Shumaila Munawar
M.Phil Chemistry
Introduction
Chromatography
Technique used to separate different components of a mixture
Chromo means color
Graphy means to write or
representation of something on
paper
Phases in Chromatography
There are 2 phases in chromatography
Mobile phase
Stationary phase
Moving phase that moves along the column
That remain fixed and mobile phase moves through it and
components get separated
e,g glass metal or plastic sheet
Strip of paper is stationary phase
Mobile phase
Principle
 Physical method of separation that distributes
components between 2 phases.
 Components move with different affinities
 Component with lesser affinity towards SP move
faster the one with greater affinity move slower.
Classification ofChromatography
On basis of chromatographic bed shape
On basis of solute interaction to the
stationary phase
paperThin layerion Exchangesize Exclusion
partition
2 Dimensional 3 Dimensional
Column
Adsorption
PC Chromatography
There are 2 types of PC chromatography
Paper adsorption Chromatography
paper impregnated with silica act as a stationary phase and
solvent act as mobile phase.
Paper Partition Chromatography
Water present in pores of cellulose fibers present in filter paper
act as a stationary phase and solvent act as mobile phase.
Christian Friedreck
Schonbein in 1865
Material required
Mobile phase solvent
Sheet of paper used as SP
Chromatographic tank
Lead pencil
Visualizing agents
Methodology
 Cut a strip of filter paper
 A horizontal line is drawn
near one end about 1.5cm
from the bottom edge of
paper
 The sample need to be
separated placed as a
small drop with help of
capillary tube
Filter paper spot of sample eg ink.
 Place the paper in chromatographic tank which contain suitable
solvent as MP.
 Solvent level must be lower than the drop.
 Cover the Chromatographic tank
 The solvent rises up the paper taking each component of
sample with it.
 When the solvent rises near the end of the paper then
the paper should be taken out from sealed container and
air dried.The paper with separated bands of components
are then observed under UV-light
Chromatogram
The pattern of separated
substances obtained by
chromatography
Spraying agents
If the components are not
colored themselves sprayed
with visualizing agenst which
reacts with component and
convert them into visible
products.
Rf value
Filter paper

Paper Chromatography

  • 1.
  • 2.
    Introduction Chromatography Technique used toseparate different components of a mixture Chromo means color Graphy means to write or representation of something on paper
  • 4.
    Phases in Chromatography Thereare 2 phases in chromatography Mobile phase Stationary phase Moving phase that moves along the column That remain fixed and mobile phase moves through it and components get separated e,g glass metal or plastic sheet
  • 5.
    Strip of paperis stationary phase Mobile phase
  • 6.
    Principle  Physical methodof separation that distributes components between 2 phases.  Components move with different affinities  Component with lesser affinity towards SP move faster the one with greater affinity move slower.
  • 7.
    Classification ofChromatography On basisof chromatographic bed shape On basis of solute interaction to the stationary phase paperThin layerion Exchangesize Exclusion partition 2 Dimensional 3 Dimensional Column Adsorption
  • 8.
    PC Chromatography There are2 types of PC chromatography Paper adsorption Chromatography paper impregnated with silica act as a stationary phase and solvent act as mobile phase. Paper Partition Chromatography Water present in pores of cellulose fibers present in filter paper act as a stationary phase and solvent act as mobile phase. Christian Friedreck Schonbein in 1865
  • 9.
    Material required Mobile phasesolvent Sheet of paper used as SP Chromatographic tank Lead pencil Visualizing agents
  • 10.
    Methodology  Cut astrip of filter paper  A horizontal line is drawn near one end about 1.5cm from the bottom edge of paper  The sample need to be separated placed as a small drop with help of capillary tube Filter paper spot of sample eg ink.
  • 11.
     Place thepaper in chromatographic tank which contain suitable solvent as MP.  Solvent level must be lower than the drop.  Cover the Chromatographic tank
  • 12.
     The solventrises up the paper taking each component of sample with it.  When the solvent rises near the end of the paper then the paper should be taken out from sealed container and air dried.The paper with separated bands of components are then observed under UV-light
  • 13.
    Chromatogram The pattern ofseparated substances obtained by chromatography Spraying agents If the components are not colored themselves sprayed with visualizing agenst which reacts with component and convert them into visible products.
  • 14.