Paper chromatography is a type of chromatography that separates mixtures by running them upwards on a sheet of paper. It was first introduced in 1865. The paper acts as the stationary phase while the solvent acts as the mobile phase. Components of a mixture travel at different rates depending on how strongly they interact with the stationary and mobile phases. This allows the components to separate into distinct spots. The Rf value can be calculated to characterize separation. To perform it, analytes are spotted on a paper strip and developed in a solvent, then Rf values are determined. Separation occurs due to differences in how analytes partition between the stationary cellulose fibers and mobile solvent.

1. PAPER CHROMATOGRAPHY
BY
Dr. Suman Pattanayak
Associate Professor
Department of Pharma Analysis & QA.
Vijaya Institute of Pharmaceutical Sciences for Women
IV B. Pharm/ I Sem
Pharmaceutical Analysis

2. Chromatography
It is a method of separating components of a mixture by differential
movement through a two-phase system: the mobile phase and the secondary
phase. This movement is effected by the flow of a liquid or a gas ( mobile phase )
which percolates through an adsorbent ( secondary phase ).

3. What is Paper
Chromatography?

4. Paper Chromatography
Paper chromatography was first introduced by
the German scientist, Christian Friedrich Schonbein in
1865.
It is a type of a planar chromatography. It is the
simplest and widely used type of chromatography
procedures which runs on a specialized paper.

5. Two principles of Paper Chromatography:
Paper Partition Chromatography
Paper Adsorption Chromatography
Paper impregnated with silica or alumina acts as the
adsorbent ( stationary phase) and solvent as the mobile
phase.
The moisture or water present in the pores of the cellulose fibers
present in the filter paper acts as the stationary phase and
another solvent as the mobile phase.
IN GENERAL, PAPER CHROMATOGRAPHY = PAPER PARTITION CHROMATOGRAPHY

6. The principle of Separation
The principle of separation is mainly partition rather
than adsorption.

7. Paper Chromatography has different types or modes:
Ascending chromatography: As the
name indicates, the chromatogram
ascends. Here the development of
paper occurs due the solvent
movement or travel in upward
direction on the paper.
Descending chromatography: Here the development of paper occurs due to
solvent travel downwards on the paper.
Ascending- descending mode: Here solvent first travels upwards
and then down wards on the paper.

8. Radial mode: Here the solvent travels from center(mid point) towards
periphery of Circular chromatography paper.
Two dimensional chromatography: Here the
chromatogram development occurs in two
directions at right angles.

9. Rf ( Retention Factor )Value
distance traveled by component from application point
Rf = -----------------------------------------------------------------------------------
distance traveled by solvent from application point

10. How to Perform Paper
Chromatography?

11. 1. Preparing the Paper Strips
• Cut the filter paper into
5 x4 measurement.
• Draw a line 0.5 cm
above the bottom edge
of the strip with the
pencil.
• Label each strip with its
corresponding solution.
•
• Place a spot from each
pen on your starting
line.

12. 2. Developing the Chromatograms
• Place the strips in the beakers.
• Make sure the solution does not
come above your start line.
• Keep the beakers covered.
• Let strips develop until the
ascending solution front is about
2 cm from the top of the strip.
• Remove the strips and let them
dry.

13. Paper Chromatography result I- Blue II- Yellow III- Green
3. Calculate the Rf value and make
conclusions.

14. How does Paper
Chromatography work?

15. Property of Water

16. Property of the Paper

17. Attraction of the water molecules to the molecules of the glucose in
the paper

18. Water molecules thoroughly scattered throughout
the paper

19. Property of dye:
Smaller- rise up faster than the bigger dye molecule

20. Dye molecules with more amount of charge rise up
faster than the other one that did not

21. Dipole-dipole interaction of water molecules ( polar ) and dye
molecules ( polar )

22. Prepared by:
Group 5