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Class 12
Class 12
Topic : Chromatography
To separate:
CHEMICALS:
1) Ethanol 4) Nickel(II) chloride
2) Distilled water 5) Potassium Ferrocyanide
3) Ferric(III) chloride 6) Copper(II) chloride
INTRODUCTION
Chromatography is an excellent and most extensively used
technique for separation and purification of compounds. It may
be defined as “the process of selective retardation of one or
more compounds of a liquid or gas uniformly migrating
through a stationary phase”.
Rf Value
3. Development:
The dry strip is positioned in a chromatographic jar in such a
way that its spotted end comes in contact with the mobile
phase. For example, the strip is suspended vertically in such a
way that its spotted end dips about 1 cm in the liquid phase
The mobile phase can be single substance or a mixture of
substance or a mixture of substances. The composition of the
mobile phase (solvent developer) is decided by the nature of
the species to be separated. The selection of an appropriate
mobile phase for a new separation procedure has often been
determined by empirical experiments. However, the following
points are observed while selecting a suitable solvent (mobile
phase):
a) The components should be more soluble in the stationary
phase as compared to mobile phase.
b) The solvent should not react chemically with any of the
component of paper.
c) R f values of any two components mist differ by at least 0.950
in the chosen solvent
d) The composition of solvent doesn’t differ with time.
e) The solvent should not interfere with the detection of the
developed chromatogram
When the solvent has risen to a reasonable distance (say 15cm)
from the original line, the strip is removed from the jar and the
position of the solvent is marked by drawing a small line with a
pencil. This line is called solvent front. The paper strip is then
dried in air is known as chromatogram.
4. Identification:
The components of mixture separate in the form of spots lying
between the original line and the solvent line. Number of
components are usually equal to number of spots appeared.
Visualization of the spots is done from their colors. But if the
sports are not colored, then the strip is prayed with a reagent
called visualization reagent or spraying reagent. This reagent
causes the components to form a derivative which is either
colored or fluoresces when exposed to UV light.
PROCEDURE
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