ACH470S TUT 1: INTRODUCTION TO CHROMATOGRAPHY

1. Why is TLC (thin layer chromatography ) useful in chemistry? What type of chromatography is it
classified as? Give a few biochemical applications of chromatography.
2.

3. What are the mobile and stationary phases in paper chromatography ?

4. On what factors does the Rf value of a compound depend ?

5. Give a few biochemical applications of chromatography

6. The following data were obtained for 3 compounds separated on a 20-m capillary column.

Cpd time (min) w (min)

A 8.04 0.15
B 8.26 0.15
C 8.43 0.16

(a) Calculate the number of theoretical plates for each compound and the average number of theoretical plates
for the column

(b) Calculate the average height of a theoretical plate in mm.

(c) Explain why it is possible for each compound to have a different number of theoretical plates.

(d) Using the data from above, calculate the resolution and the selectivity factors for each pair of adjacent
compounds. Discuss how you might improve the resolution between compounds B and C. The retention time
for a non-retained solute is 1.19 min.

7a). Use the partial chromatogram below to determine the resolution between the two solute bands.

7b) The chromatogram above was obtained on a 2-m column with a column dead time of

50 s. Suppose you want to increase the resolution between the two components to 1.5. Without changing the
height of a theoretical plate, what length column do you need? What height of a theoretical plate do you need
to achieve a resolution of 1.5 without increasing the column’s length?

8) Use the chromatogram below obtained using a 2-m column, to determine values or tr, w, t′r′, N, and H.