FACULTY OF SCIENCE, TECHNOLOGY

&
ENVIRONMENT

School of Biological & Chemical Sciences (SBCS)

CH414
INSTRUMENTAL ANALYSIS
FINAL EXAMINATION – SEMESTER II, 2015

Time Allowed: 3 hours plus 10 minutes reading

100 marks (50% of final grade)

INSTRUCTIONS

1. There are five questions. Answer ALL the questions.

2. Each question is worth 20 marks.
3. Answer each question on a new page of your answer booklet.
4. The minimum final exam mark required to pass this course is 40/100.
5. A table of physical constants and a copy of the Periodic Table is attached at
the end of the examination paper.
6. You can use non-programmable calculator.

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Question 1
a. What is an attenuated reflectance IR (AT-IR) spectroscopy and how does it differ from
transmission IR spectroscopy? You may use illustration in your answer.

5 marks

b. Aspartame is a methyl ester of the didpeptide aspartyl-phenylalanine. It is a well-known low

calorie sweetener. It is crystallized as aspartame hemihydrate, which has a stable
conformation at room temperature. If heated, however, aspartame hemihydrate undergoes
dehydration followed by cyclization.

Figure 1. The pathway of water evaporation, dehydration and intramolecular cyclization

of aspartame hemihydrate

With FTIR spectroscopy, it is possible to monitor the gradual changes occurring as

aspartame hemihydrate is heated. The portions of FTIR spectra shown below show these
changes (50, 63, 75, 110, 153, 165 and 174oC).

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 i. Using the FTIR spectra explain what does the peak positions and height mean
with respect to the mechanism seen in Figure 1
5 marks

ii. If the sample of aspartame hemihydrate has spent several weeks in the heat at
200oC, describe how you would best determine the amount of the original
compound left relative to the starting amount, in %.
5 marks

c. The chemical and physical properties of enantiomers are identical but they differ in a
property called optical activity, in which a sample rotates the plane of polarized light beam
passing through. Use illustrations to explain how the polarimeter works.

5 marks
QUESTION 2

a. Compare and contrast between TG(thermogravimetric analysis) and DTA(differential

Thermal Analysis) in terms of:

i. Their thermograms
ii. Quantity measured
iii. Instrument design
iv. Nature of the sample and the reference
5 marks

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b. Below is a typical DSC thermogram obtained for a sample.

i. Explain the origins of the different peaks.

ii. If the Ho (100% crystalline) is 93 J/g, calculate the percentage crystallinity of the
sample at room temperature (i.e., before the sample was subjected to heating in
the DSC scan).

5 marks

c. There are two types of sample injector port used in Gas Chromatography: split and split-less
injector.
i. Explain the difference between the injector ports.
ii. Briefly explain the advantages and disadvantages of the two injector ports.
5 marks

d. Some of the major problems encountered in the operation of Gas Chromatography are:
i. Negative peaks
ii. Loss of resolution
iii. Split Peaks
iv. Baseline drift

Choose any two problems listed above and for each discuss the cause of the problem and
briefly state how the problem could be resolved.
5 marks

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QUESTION 3
a. What is the difference between gradient elution and isocratic elution method? Explain
the advantages of gradient elution over isocratic elution method.
4 marks

b. Size Exclusion Chromatography is a very useful form of liquid chromatography to

separate proteins. Explain how this chromatography can help separate proteins of
different molecular weight.
3 marks

c. During an analysis on HPLC, a student tried to separate natural products in a crude

mixture that was isolated from a plant using methanol as the extracting fluid. The polar
compounds from the plant would be extracted.

The following instrument conditions were chosen by the student:

 C18 reverse phase column
 UV detector
 Flow rate of 0.5mL/minute
 Sample run of 5 minutes
 Isocratic mobile phase (hexane)

i. List all the possible errors the student has made in terms of choice of column,
detector, mobile phase and choice of elution method.
4 marks

ii. Explain the changes the student needs to make to see a proper chromatogram.

3 marks

iii. Explain the difference between normal phase and reverse phase column

2 marks

d. Shown below is the chromatogram from the HPLC separation of a series of organic
compounds. Note: Econosil is a trademark name of the column.

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 Assign each compound to a peak on the chromatogram given below and explain your
answer.

Benzoic acid Phenol Ethyl benzoate Ethyl benzene

4 marks

QUESTION 4

a. Select TWO detectors used in GC and explain briefly the advantages and its application of
such detectors.
5 marks

b. The DEPT spectra of 1-isopropyl-4-methylcyclohexa-1,3-diene is given below. Analyze the

spectral data and assign all the peaks of the structure given. What 1H-NMR information does
DEPT give?

5 marks
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c. During the total synthesis of cholesterol, an intermediate product was isolated that shows
strong absorption in the UV region at 291 nm. Using Woodward’s rule calculate UV-visible
absorption band positions for the following three compounds and explain which is the
correct structure of the product.
6 marks

HO

O O
O
OH OH
(I) (II) (III)

d. The mass spectra of Ethyl 2-hydroxybenzoate (xx) contain daughter ion peaks at m/z 93,
121 and 138. Suggest structures for any two of the daughter ions and give mechanisms for
their formation.
4 marks

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QUESTION 5
The unknown organic compound B, has a molecular formula of C11H14O3. The IR, Mass
spectra, UV, 1H NMR, 13C NMR, 1H-1H COSY and 1H-13C HMQC spectral data is
provided.
Determine the double bond equivalence of the molecule and outline all necessary
information about the structure of the compound.
Write down a plausible structure for unknown compound B and explain how you arrived at
the structure.
20 marks

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Appendix

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