1. Substances A and B have retention times of 16.40 and 17.63 min.

respectively, on a
30.0-cm column. An unretained species passes through the column in 1.30 min. The
peak widths (at base) for A and B are 1.11 and 1.21 min. respectively. Calculate (a)
the column resolution. (b) the average number of plates in the column, (c) the plate
height, (d) the length of column required to achieve a resolution of 1.5. (e) the time
required to elute substance B on the column that gives an R, value of 1.5, and (f) the
plate height required for a resolution of 1.5 on the original 30-cm column and in the
original time.
2. Two solutes were eluted at 350 and 372 s on a 2-m column with a column dead time
of 50 s. The sum of the baseline widths for two solutes was found as 45 seconds.
Determine the resolution between two solute bands. How long a column is needed to
achieve a resolution of 1.5? What height of a theoretical plate is needed to achieve a
resolution of 1.5 without increasing the length of the column?
3. It is desired to just resolve two gas-chromatographic peaks with retention times of 85
and 100 s, respectively, using a column that has an H value of 1.5 cm/plate under the
operating conditions. What length columns is required? Assume the two peaks have
the same base width.

4. Trihalomethanes, CHCl3, CHBr3, CHCl2Br, and CHClBr2 in drinking water were

eluted using a packed column with a nonpolar stationary phase. Predict the order in
which these four trihalomethanes will elute.

5. A mixture of n-heptane, tetrahydrofuran, 2-butanone, and n-propanol were eluted in

using a polar stationary phase such as Carbowax and a nonpolar stationary phase such
as polydimethyl siloxane. Predict the order of elution.

6. What is the expected order of elution of the following compounds from an alumina
column by a liquid mobile phase: n-butanol, 1-butylchloride, n-hexanoic acid, n-
hexane, n-hexene.
7. Predict the elution order of the following compounds from CaO column: n-butanol,
methanol, ethanol.
8. List the following compounds in their expected order of elution from LSC column:
benzoic acid, benzene, chlorobenzene, phenol.
9. What volume of mobile phase is required for the separation of component A from a
mixture on a stationary phase of 4.8 ml with a partition coefficient of 9.6? The
retention volume of the component A is 58.4 ml.
10. A substance was found to have a retention time of 59 sec. the peak width at the
baseline was found to be 8.3 sec. Calculate the number of theoretical plates and plate
height.
11. It takes 2 min 30 sec for a solute to be eluted from a chromatographic column to its
maximum concentration. Calculate its retention volume if the flow rate is 24 ml min -1.
12. A GC column exhibits 2400 theoretical plates. If the retention times for benzene and
toluene are 12 and 24 min, respectively, will baseline resolution will be achieved?
13. Retention times for several compounds separated by GC are given below:

Substance tr (s)
Air 75
n-pentane 190
n-hexane 330
1-pentene 302
a) Calculate the relative retention of 1-pentene with respect to n-pentane.
b) Calculate the relative retention of n-haxane with respect to n-pentane.
14. A non-retained substance is eluted from an HPLC column in 0.6 min at a flow rate of
1.8 ml min-1. What is the void volume in the column? If morphine is eluted in 4.7 min,
what is its capacity factor?
15. A chromatographic peak had a retention time of 72 sec. The base width obtained from
intersection of the base line with extrapolated sides of the peak was 6 sec. If the
column was 90 cm in length, what was the H?
16. A chromatographic separation of a two component sample on a 50-cm column gave
the retention times for the solutes A and B are 2.5 and 3.1 min and base widths of the
two chromatographic peaks being 0.24 and 0.3 mm, respectively. Calculate:

a) the number of theoretical plates

b) plate height and

c) resolution of the two peaks. 4

17. A substance C, which is not retained in the stationary phase, exits from a
chromatography column 5 min after injection. Another substance D, which is retained
exits in 20 min. The flow rate of the mobile phase is 250 ml min-1.

a) How long does D spend in the mobile phase? b) How long does D spend in the
stationary phase? c) What is the value of k1 for D? d) What is the value of KD for C?

e) What is the retention volume, Vr for D? f) What is Vm?

18. The following data are obtained from a GC analysis: t r = 10 mm, W= 0.30 mm, t air=
0.50 mm, Vs= 4 ml and F=40ml min-1. Calculate: a) k1 b) Vm c) Vr and d) N.
For a chromatographic column, the values of A, B and C in Van-Deemter equation are 0.06
cm, 0.57 cm2 s-1 and 0.13 S. Calculate the optimum velocity of the carrier gas.

A 30-m long open tubular capillary column is used for GC having an optimum velocity at 12
cm s-1 and exhibits 1.6 ×105 plates at this optimum velocity. Determine B and C parameters
for this column.

For a gas chromatographic column, the values for A, B and C in the Van-Deemter equation
were 0.15 cm, 0.36 cm2 sec-1 and 4.3×10-2 S, respectively. Calculate the best flow rate and the
minimum plate height.
The acetaminophen content of tablet is determined by HPLC using C 8 column and 38:62
MeOH and water mobile phase. The UV detection is set at 254 nm. A 10 µL standard
solution containing 12 ppm acetaminophen is eluted out with response from the detector as
0.324 arbitrary units. Ten tablets are ground into fine powder and their extract prepared
into 500 mL. A 1 mL of extract is diluted to 25 mL with the mobile phase and a 10 µL is
injected into the column. The resulted absorption is 0.232 arbitrary units. What weight of
acetaminophen is present in each tablet?

Ion-exchange chromatography is employed to analyse waste water for total cation concentration. A
50 mL of waste water sample, has total alkalinity of 234 ppm CaCO 3, is passed through cation
exchange column containing H+ form of resin (Res-SO3-)H+ and the effluent is titrated with 23.4 mL of
0.025 mol L-1 NaOH to phenolphthalein end-point. Calculate the total cation concentration of water
sample and report in ppm CaCO3.

A sample containing three amino acids namely, glycine, alanine and methionine, is spotted on the
marked line on TLC plate and placed in TLC chamber containing 45:25:30 (v/v) mixture of CHCl 3,
C6H5CH2OH and CH3COOH as mobile phase. The solvent is travelled by a distance of 16 cm from
sample introduction line of TLC plate. The calculated retardation factors for glycine, alanine and
methionine are 0.24, 0.32 and 0.46, respectively. Find out the distance in cm migrated by each
amino acid on TLC plate from the origin of the sample spot. Calculate the retention factors for each
compound.

Use the following data of TLC separation and calculate the values of R f and k for solutes A, B
and C.
TLC Stationary Phase: Bonded Silica
Mobile Phase: 45:25:30 (v/v) Mixture of CHCl3, C6H5CH2OH and CH3COOH
Species Migrated Distance Migrated from sample spot, cm
Solute A 9.12
Solute B 6.34
Solute C 3.11
Mobile Phase 14.00