SUGGESTED ANSWER FOR TUTORIAL 1

QUESTION 1

a) Very long columns are not often used in chromatographic separations in order to
achieve good separations because using infinitely long columns would allow more
separation process (plates) to occur, they would also lead to unacceptably long
separations. Longer separations are also more susceptible to broadening as a
result of longitudinal diffusion. As a result, overall separation quality is
diminished.

b) Two ways to increase efficiency in a chromatographic separation using packed column:

i) Use smaller packing material; This diminishes band broadening due to

multipath.
ii) Decrease the thickness of the stationary phase coating; This diminished
band broadening due to mass transfer terms.

(Q1 JAN 2012)

QUESTION 2

a) i) NA = 2568

NB = 1044

Higher N = more efficient

ii) RA = 1.65

RB = 1.37

RA = 1.65 Baseline resolution between two peaks (as shown in the figure)
requires an Rs >1.5. Good separation for column A.

RB = 1.35. Resolution is more than 1.0, therefore the separation is considered

adequate for column B.

However, Column A better than column B

(Q1a JUN 2012)

b) i)

ii)

iii)

(Q1b JUN 2012)

QUESTION 3

Flow rate need to be optimized in order to improve the efficiency of a column:

For B/v, high flow rate will decrease the term and thus reduces band broadening

But for Cv, low flow rate is required to give the molecules enough time to reach
equilibrium.

Thus, an optimum flow rate is high enough to reduce diffusion and low enough to give
enough time for achieving equilibrium.

(Q1 JAN 2013)

QUESTION 4

(a)

Rs1 = 2.11 ; Rs2 = 0.78

Rs1 = 2.11. Peak X and Y were well separated as Rs > 1.5.

Rs2 = 0.78. Resolution is less than 1.0, therefore overlapping of peak Y and Z.
(b) i) 30 cms-1. Optimum flow rate is at minimum H.

ii) Increase in mass transfer. When the flow rate is high (above the optimum
value), the analyte has a strong affinity for the stationary phase and the
analyte in the mobile phase will move ahead of the analyte in the stationary
phase (without reaching equilibrium).
(Q1a & b JUN 2013)

QUESTION 5

In a chromatographic analysis, the effect of the following on the plate height of a column.

i) Increasing the flow rate

H increases. At high flow rate, term Cv increases. Analytes in mobile phase will
move faster than those in the stationary phase (without reaching equilibrium).

ii) Reducing the particle size of the packing

H decreases. Smaller particle size of packing will reduce differences in path of
molecules (reduce Eddy diffusion). Smaller particle size of packing will give
higher surface area, thus easier to reach equilibrium, reduce Cv.
(Q1a DEC 2013)
QUESTION 6

a) Navg = (