Page 71 - Practical book PC406-MNU-2025

MANSOURA NATIONAL UNIVERSIY
PHARM D- CLINICAL PHARMACY LEVEL II INSTRUNENTAL ANALYSIS (PC 406)

A) Paper chromatography

Paper chromatography:
PC is an analytical technique for separation, purification and identification of mixtures.

Separation of a dye into its components

 Sample: green dye.

 Mobile phase: methanol: water (75:25).

 Stationary phase: water bound to cellulose.

 Separation principle: the mobile phase will move through the paper fibers by

capillary attraction.

 The distance that the solvent will carry each component a long the paper during a

given time interval is determined by:
1. The degree of solubility of the components of the dye in the solvent

2. The degree of attraction of these components to the wet cellulose
molecules in the paper fibers.

Note:
The component that's more soluble in the mobile phase, or that has a low affinity
for cellulose (stationary phase), will be carried the greatest distance from the base

line.

P Pr ro oc ce ed du ur re es s: :

1) Cut a filter paper as 4-cm × 10-cm rectangle.

2) Draw a line at 2-cm from the base of the filter paper, this line is called "base line".

3) Apply a small spot of the sample in the middle

of the base line using a capillary tube.

4) Transfer 10-mL of the mobile phase into a beaker.

5) The paper is then dipped into the mobile phase
taking care that the spot is above the surface of the

mobile phase.
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