CONFIDENTIAL

AS/OCT2010/CHM510
UNIVERSITI TEKNOLOGI MARA
FINAL EXAMINATION
COURSE
COURSE CODE
EXAMINATION
TIME
ANALYTICAL SEPARATION METHODS
CHM510
OCTOBER 2010
3 HOURS
INSTRUCTIONS TO CANDIDATES
1. This question paper consists of six (6) questions.
Answer ALL questions in the Answer Booklet. Start each answer on a new page. 2.
3. Do not bring any material into the examination room unless permission is given by the
invigilator.
Please check to make sure that this examination pack consists of:
i) the Question Paper
ii) an Answer Booklet - provided by the Faculty
DO NOT TURN THIS PAGE UNTIL YOU ARE TOLD TO DO SO
This examination paper consists of 5 printed pages
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AS/OCT 2010/CHM510
QUESTION 1
A separation of three compounds on a column (25 mm x 4.6 mm x 10 um) produced the
following data:
Peak
Unretained
1
2
3
Calculate:
a) The average number of plates from the above data.
Retention time (min)
1.2
3.4
3.8
4.7
Peak width (min)
-
0.11
0.13
0.17
b) The average plate height for the column.
QUESTION 2
(3 marks)
(3 marks)
Gas chromatography (GC), liquid chromatography (LC), and supercritical fluid
chromatography (SFC) are three types of separation methods with different mobile phases.
a) Compare and discuss how each mobile phase affects
i) the B term of van Deemter in each of the above separation method.
(6 marks)
ii) the types of sample that are suitable for each of the above separation method.
(6 marks)
b) For each of the above separation methods, discuss one main parameter (exclude
flow rate) that can be manipulated to improve the separation of analytes.
(6 marks)
QUESTION 3
a) You are required to analyse relatively polar analytes in an aqueous sample using
solid phase microextraction (SPME) followed by gas chromatography (GC) with
electron capture detector (ECD). For each of the following pairs, choose the more
appropriate parameter and justify your choice.
i) Poly(phenylmethyl) siloxane (50% phenyl) column or polyethylene glycol column.
(3 marks)
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ii) Split injection mode or splitless injection mode.
(3 marks)
iii) Isothermal temperature or temperature programming.
(3 marks)
iv) Nitrogen or helium mobile phase
(3 marks)
b) Suggest with reason(s), a specific approach/condition for the gas chromatographic
separation of the following samples:
i) A food sample containing non-volatile additive compounds.
(3 marks)
ii) A sample mixture consists of compounds with boiling point in the range of 60C
to 250C.
(3 marks)
iii) An industrial soil sample containing thermally labile contaminants.
(3 marks)
iv) A gaseous sample with very volatile analytes.
(3 marks)
QUESTION 4
For each of the following, give a reason for the mentioned problem and suggest an approach
to overcome the problem.
a) In a normal phase separation of 5 compounds using isocratic mixture of
hexane:acetone (80:20), the retention time for the last two peaks were too long.
(5 marks)
b) For the separation of organic acids using ion exchange chromatography, the
compounds were separated using carboxylic acid -COO" H
+
resin as the stationary
phase at pH 4. The compounds were not separated due to overlapping of peaks.
(5 marks)
c) You are given the task to separate anions in a water sample. In your laboratory, the
liquid chromatograph available can only run partition chromatography. When the
separation of anions in the sample was carried out using reverse phase
chromatography with methanol:water (50:50) as the mobile phase, no peak was
observed.
(5 marks)
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AS/OCT2010/CHM510
d) When a Bio-gel 150 with a molecular mass range of 22,000 - 250,000 was used for
size exclusion chromatography packing, the first three compounds were separated
well. However, the last two compounds were eluted as a single peak.
(5 marks)
QUESTION 5
Analyte species (A: polar neutral, B: non-polar neutral,
20
X
+
,
25
X
+
,
20
Y",
25
Y") in an aqueous
sample were separated by capillary electrophoresis (CE) in 15 mM borate buffer (pH 5.8).
The CE instrument was set up with the detection end at cathode.
a) Describe the order of elution for the above separation.
(5 marks)
b) Explain what would happen to the separation if:
i) the pH was decreased to 4.0
(2 marks)
ii) the pH was decreased to 2.0 (assume that the charges of the components do
not change).
(3 marks)
iii) the pH was 5.8 and treated with ethyl ammonium bromide.
(3 marks)
iv) the pH was 5.8 and added with high concentration of anionic surfactant (above
critical micellar concentration/CMC level).
(3 marks)
QUESTION 6
a) For each of the following analysis, discuss the reason for the underlined parameter/
approach used.
i) In the solid phase extraction (SPE) process for the extraction of chlorphyrifos
using C18 sorbent, column conditioning was performed before sample
application.
(4 marks)
ii) Fragrance analytes from plant sample were extracted using headspace-solid
phase microextraction (HS-SPME) followed by desorption process.
(4 marks)
iii) Pressurized liquid extraction (PLE) is used to extract solutes from solid and
semi solid samples by utilizing high temperature and pressure.
(2 marks)
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b) For each of the following, give a reason to the mentioned problem and suggest an
approach to overcome the problem without changing the extraction method used.
i) Alkanes can be extracted from urban air particulates using supercritical fluid
extraction (SFE) with C0
2
at 75 bar (45C), whereas the polyaromatic
hydrocarbons (PAHs) remain unextracted.
(3 marks)
ii) Non-polar contaminants from soil sample cannot be extracted using hexane
with microwave extraction method (MAE).
(3 marks)
END OF QUESTION PAPER
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