TUTORIAL 7: BASIC CHROMATOGRAPHY

1. Briefly describe the following chromatographic terms:

i. Stationary phase
ii. Partition chromatography
iii. Adsorption chromatography
iv. Solid support (in liquid-liquid chromatography)
v. Thin layer chromatography

2. State two ways to visualize colourless spot on thin layer chromatography (TLC)
plate.

3. A student removed his TLC plate from the developing chamber after the solvent
height reaches ¾ of his plate. However, he could not seen any spot on the plate.
Suggest any three techniques to visualize the spots.

4. In a paper chromatography experiment, a student draws the straight line using a pen
before spotting the sample. Give a reason whether the procedure is correct or
wrong.

5. The diagram below is an illustration of the chromatogram obtained from thin layer
chromatography (TLC) of three known compounds (A, B and C) and an unknown
mixture, X. The mixture X contains two of the known compounds.

i. Calculate the Rf value of A.

ii. Identify the components of X.
iii. Give the type of force that moves the mobile phase.

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 iv. Name another type of chromatography where the mobile phase is driven by
the same force as in TLC.
v. Give an example of the stationary phase used in TLC.

6. One of the methods employs in chromatography is Thin Layer Chromatography

(TLC). State whether the separation in TLC involves adsorption or partition.

7. Draw a TLC slide using the information below:

The solvent front is 6.90 cm; and the Rf values for spots X, Y and Z on the TLC
slide are 0.82, 0.28 and 0.63, respectively.

8. In an experiment using thin layer chromatography (TLC), a colourless sample was

spotted on a TLC plate and developed in a chromatographic tank. Three spots, P, Q
and R were observed on the TLC plate and the distance from the original spot were
5.5, 3.2, and 9.3 cm, respectively. The distance of the solvent front from the original
spot was 12.0 cm. Illustrate the TLC plate and show how the Rf values of P and Q
can be determined.

9. A sample of brown dye from a lollipop is placed at the origin on a strip of a thin
layer chromatography plate. The solvent front moves 9.0 cm from the origin. A blue
component of the dye moves 7.5 cm and a red component moves 5.2 cm. Determine
the Rf values of the two components.

10. A mixture of two dyes – yellow and blue was separated using a paper
chromatography. The yellow dye has a higher Rf value than the blue dye.

i. Sketch the diagrams of the paper chromatogram before and after the
development of the chromatography.
ii. What is the stationary phase in this chromatography?

11. TLC is used in a separation of mixture of two components, X and Y. X is a higher

than Y. By using a polar solvent, explain the principles of separation using a TLC.
Identify the components that are left behind at baseline and the component at the
solvent front.

12. State two examples of the applications of

i. chromatography.
ii. planar chromatography.

13. Give the difference between paper chromatography and TLC in term of their
stationary phase.

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14. Column chromatography and planar chromatography are the two basic types of
chromatographic methods. Explain the differences between the two
chromatographic methods.

15. Identify the stationary phase and mobile phase in thin layer chromatography.

16. If silica gel is used as stationary phase in column chromatography, it is considered

as polar stationary phase. Mixture of compounds, A and B were separated using this
column. Compound A is less polar than compound B. Predict which compound will
come out first from the column. Give your reasons.

17. A column chromatography was set up using water coated on solid support as the
stationary phase and an organic solvent as the mobile phase. Indicate how two
compounds of different polarity can be separated through this column.