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Chromatography Worksheet

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10 views3 pages

Chromatography Worksheet

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rheasa04
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CHEM 222- LABORATOTY METHODS II

Unit I – Chromatography. Worksheet #1. Prepared by Dr. C. D. Barker.


BARBADOS COMMUNITY COLLEGE
DIVISION OF SCIENCE
CHEM 222 – LABORATORY METHODS II

UNIT I – CHROMATOGRAPHY

1. Two different brands of purple food colouring are analysed by paper chromatography.
Both samples contained a blue pigment. In the first sample, the solvent travelled 8.25 cm
while the blue pigment travelled 5.68 cm. In the second sample, the solvent travelled
7.86 cm while the blue pigment travelled 5.41 cm.
(a) Sketch the chromatogram, indicating the relevant distances and indicating an
approximate point for the red dye.
(b) Calculate the retention factor of the blue pigment in each sample.
(c) Is it a different blue pigment present in the two brands of the purple colouring? Justify
your answer.

2. Explain the principles upon which chromatographic methods of separation are based.

3. Distinguish between the stationary phase and the mobile phase in chromatography.

4. Give examples of the stationary and mobile phases for each of the following:
a. Thin layer chromatography
b. Column chromatography
c. Gas-liquid chromatography

5. Explain the following terms:


a. retention factor c. solvent front
b. retention time d. visualising agent

6. Define the following terms:


a. Chromatography
b. Stationary phase
c. Mobile phase

1
CHEM 222- LABORATOTY METHODS II
Unit I – Chromatography. Worksheet #1. Prepared by Dr. C. D. Barker.

7. Explain the following terms:


a. Gradient elution
b. Isocratic elution

8. State the two different types of chromatography.

9. State the uses of chromatography in criminal investigations.

10. Explain how chromatography can be used to separate a mixture of dyes.

11. What is the main basis for separation in electrophoresis? What factor also influences
separation in electrophoresis?

12. Explain how electrophoresis may be used to separate a mixture of amino acids.

13. Explain how (i) TLC (ii) ultraviolet/visible (UV/Vis) analysis may be used along with
HPLC in the separation of a mixture of amino acids.

14. Figure 1 shows the separation of a mixture of amino acids, obtained in the hydrolysis of a
dipeptide, on a TLC plate.

Solvent front
R 13.5 cm

12.5 cm
Q
2.5 cm

Mixture of amino acids

Figure 1: The separation of a mixture of amino acids

a. Name two materials commonly used as the stationary phase in TLC.


b. Explain how the presence of the colourless components can be detected.
c. Calculate the Rf values of each component.
d. Suggest two reasons, other than the distance travelled by the solvent, for the
difference in the Rf values of the components.

2
CHEM 222- LABORATOTY METHODS II
Unit I – Chromatography. Worksheet #1. Prepared by Dr. C. D. Barker.

15. The methods of TLC and column chromatography are both similar and different. Assess
the methods of these two types of chromatography in relation to their uses.

16. What is the main difference between column chromatography and HPLC?

17. Name two materials commonly used as the stationary phase in column chromatography
and HPLC.

18. Explain the differences between HPLC and GLC, and relate these to their uses.

19. What is the basis of separation in GLC?

20. Draw a diagram of a GLC trace, and use it to explain how peak areas and percentage
compositions may be calculated in GLC.

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