LINKÖPINGS UNIVERSITET – IFM/KEMI/JD

TFKE30 – Analytical Chemistry

PROBLEM 1 – Chromatography (6 p)

Below is a GC chromatogram with three peaks. The first peak is methane that travels through the
column without retention. Peaks 2 and 3 are caused by compounds 2 and 3, which thus are retained
by the column to some extent.

(a) Calculate the retention factor k for compounds 1 and 2 using the relevant values given in the
chromatogram below. (2p)
(b) Calculate the separation factor α for compounds 1 and 2. (2p)
(c) Calculate the resolution (Rs) between compounds 1 and 2. Their average peak width at the
baseline is 30 seconds. (2p)

Show equations and your calculations.

Compound 2

Compound 1

tm = 1.00 min

tr = 5.00 min tr = 6.00 min

SVAR:
(a) Retention factor k=(tr-tm)/tm  k1=5-1/1=4 ; k2=6-1/1=5
(b) Separation factor α = k2/k1=5/4=1.25
(c) Resolution Rs=tr2-tr1/Wav=6-5/0.5=2
PROBLEM 2 – gas chromatography (6p)
(a) You develop a GC method and focus on getting resolution (Rs) between the peaks (analytes).
In what two ways can you influence the separation factor α? Explain how the separation
factor is affected in each of the two cases. (2 p)
(b) Explain what chemical tailing is in chromatography. What causes the tailing to occur and
what type of compounds (analytes) suffers from this phenomenon? (2 p)
(c) Which two properties make analytes unsuitable for analysis by GC (1 p)
(d) What is meant with isothermal analysis? What is meant with temperature-programmed
analysis? (1 p)

SVAR:

(a) (i) Change column/stationary phase - the separation factor is affected if the retention factors
k1 and k2 are affected to different degree; α =k2/k1. (ii) Oven temperature – change of
isothermal temperature or temperature program. If the retention factors (k1 and k2) of the
analytes are affected to different degree, the sepraration factor α will also be affected.
(b) Taling : the peaks are tailing when analytes are interacting with free silanol groups in the
stationary phase. Polar compounds that can interact via hydrogen bonding are often tailing
(alcohols, carboxylic acids, amines)
(c) Too high boling points (>approx 400 ºC), thermal instability.
(d) Isothermal analysis: constant oven temperature during entire analysis. Temperature
programming: Temperature starts at low temperature and is increased during the analysis.

PROBLEM 3 – Liquid chromatography (6 p)

( ) and ethyl benzoate ( ).

(a) Propose a suitable stationary phase and a suitable mobile phase. (2 p)

Thereafter you want to develop an isocratic reversed-phase HPLC method that can separate the
same two compounds.

(c) Propose a suitable stationary phase (write also its chemical structure) and a suitable mobile
phase. (2 p)
(d) What will be the elution order of the two analytes? Explain. (1 p)

SVAR:

(a) Stationary phase: Silica. Mobile phase: suitable mixture of two organic solvent (e.g. toluene,
hexane, chloroform).
 (b) Ethyl benzoate is more polar and interacts more strongly with the polar stationary phase and
is therefore retained more strongly. Ethylbenzene therefore elutes before ethyl benzoate.
(c) Stationary phase: Octadecyl silica (C18), Si-O-Si(CH3)2-C18. Mobile phase : mixture of
methanol and water
(d) Ethylbenzene is more nonpolar and interacts more strongly with the nonpolar stationary
phase and is therefore retained more strongly. Ethyl benzoate therefore elutes before
ethylbenzene.

PROBLEM 4 – Gas and liquid chromatography (6 p)

Take a stand on the statements below. Determine if each statement is true or false. The answers
should not be justified or commented on.

Assessment: correct answer +1 p, wrong answer -1 p, no answer 0 p.

The problem as a whole cannot give less than 0 p.

(6 p)
(a) HPLC : In reversed-phase HPLC, it is common to use pure silica as the stationary phase.
(b) GC : The retention factor increases if the film thickness is decreased.
(c) If you double the column length, you will also double the plate number N.
(d) I you double the column length, you will also double the resolution Rs.
(e) HPLC : the plate height decreases with decreased particle size
(f) GC : The peak size of the analyte will increase when changing the split ratio from 20:1 to
50:1.

SVAR:

(a) False
(b) False
(c) True
(d) False
(e) True
(f) False