UNIVERSITY OF LIMPOPO

TURFLOOP CAMPUS

FACULTY OF SCIENCE & AGRICULTURE

SCHOOL OF PHYSICAL & MINERAL SCIENCES

DEGREE AND DIPLOMA TEST MEMORANDUMS

MODULE: CHEM322 DATE : 10th AUGUST 2013

(ANALYTICAL CHEMISTRY 3)
TIME: 2.5 HOURS (150 MINUTES) TEST : 1
MARKS : 100

INTERNAL EXAMINERS : MODIBANE K. D

: MULAUDZI V. L

THIS PAPER CONSISTS OF …6…PAGES INCLUDING COVER

PAGE

INSTRUCTIONS:
1. Answer only two questions from each section
2. Write neatly and legibly
3. Use allocated marks to gauge the amount of information to give as answer
TEST 1 10 AUGUST 2013

SECTION A (UV, IR & Fluorescence Spectroscopies)

QUESTION 1 (25 marks)

1.1.

 The UV-Vis spectrometry is a spectroscopic technique involving the use of light in

the visible, near ultra-violet and near infrared regions to cause electronic transitions in
the target analyte.
 A light source of a fixed wavelength is shone through an analyte and its absorption
(or transmission) intensity measured against a background using a detector.
 The wavelength is then varied slightly using perhaps a diffractometer, and the process
repeated until the absorption ratio for a spectrum of wavelengths is obtained.
 UV-Visible spectrometry is instrumental in characterizing materials to help determine
their absorption bands, band gap energy as well as their particle sizes.
 It may also be used to characterize an unknown sample since the spectrum is
effectively a chemical “fingerprint” for a given molecule.
 To determine unknown concentration as a quantitative analysis
1.2.

 A monochromator is a dispersive instrument with an entrance slit and an exit slit.

 It is designed to isolate a single band of wavelengths.

 A spectrograph has an entrance slit, but no exit slit. It is designed to image an entire
spectrum at its focal plane.

 Spectrographs are used with multichannel detectors like CCD arrays and diode
arrays.

 A spectrophotometer is an instrument with a monochromator or spectrograph

designed to obtain the ratio of two beam intensities.

 To calculate absorbances and transmittances in absorption spectroscopy.

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1.3.

 When analyte absorb energy, their electrons are excited or promoted from their
valence band to a conduction band resulted with absorption spectrum.
 In an excited state, the atoms can rotate and vibrate with respect to each other.
 These vibrations and rotations also have discrete energy levels, which can be
considered as being packed on top of each electronic level.
 The excited electrons, now at the lowest vibronic level of valence band, may then lose
energy through the emission of light to conduction band.

1.4.

 A = bc

 c = A/b

 = 0.612/(210 L/mol/cm x 1.0 cm)

 = 2.91 x 10-3 mol/L

1.5.

 One common method is to crush the sample with an oily mulling agent
(usually Nujol) in a marble or agate mortar, with a pestle.

 The second method is to grind a quantity of the sample with a specially purified salt
(usually potassium bromide) finely (to remove scattering effects from large crystals).

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 A third technique is the "cast film" technique, which is used mainly for polymeric
materials.

 The final method is to use microtomy to cut a thin film from a solid sample.

QUESTION 2 (25 marks)

2.1.

 A,

 Because PdPc has heavy metal effect

 which enhance population of the triplet state through inter system crossing leading to
less molecule involve in the loss of energy from lowest vibronic level of singlet
excited state to ground state.

2.2.

 The Beer-Lambert law (or Beer's law) is the linear relationship between absorbance
and concentration of an absorbing species.

 The general Beer-Lambert law is usually written as:

A = abc

where A is the measured absorbance, a is a wavelength-dependent absorptivity

coefficient, b is the path length, and c is the analyte concentration.

When working in concentration units of molarity, the Beer-Lambert law is written as:
A = bc

where  is the wavelength-dependent molar absorptivity coefficient with units of M-

1
cm-1.

 Real limitations: Non-linearities due to intermolecular interactions, Self aggregation

effects and electrolyte effects and Dynamic dissociation or association of analyte

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 Instrumental: Polychromatic radiation

 Different molar absorptivities at different wavelength leads to non-linearities in

Beer’s Law

 Stray radiation, Mismatched cells, Non-zero intercept in calibration curve

2.3. [6 marks]

 (a) molar extinction coefficient=slope=1.003 L/mol/cm and

 (b) the unknown concentration of C= 0.2 mol/L

2.4.

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TEST 1 10 AUGUST 2013

 A single-beam spectrophotometer employs one beam of radiation that irradiates one

cell.

 To obtain the absorbance, the reference cell is replaced with the sample cell
containing the analyte.

 Single-beam instruments have the advantages of simplicity and lower cost.

 With a double-beam instrument, the reference cell and sample cell are irradiated
simultaneously or nearly so.

 These instruments have the advantages that fluctuations in source intensity are
cancelled as is drift in electronic components.

 The double-beam instrument is readily adapted for spectral scanning.

2.5.

 The infrared spectrum of a sample is recorded by passing a beam of infrared light

through the sample.

 When the frequency of the IR is the same as the vibrational frequency of a bond,
absorption occurs. Examination of the transmitted light reveals how much energy was
absorbed at each frequency (or wavelength).

 This can be achieved by scanning the wavelength range using a monochromator.

Alternatively, the whole wavelength range is measured at once using a Fourier
transform instrument and then a transmittance or absorbance spectrum is generated
using a dedicated procedure.

 Analysis of the position, shape and intensity of peaks in this spectrum reveals details

about the molecular structure of the sample.

QUESTION 3 (25 marks)

3.1.

 It is non-destructive, solids, liquids and gases can be analysed

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TEST 1 10 AUGUST 2013

 It is possible to easily identify and distinguish between many organic compounds and
inorganic compounds

Precise measurement method that requires no external calibration

FTIR measurements can be made within seconds

 Optimal sensitivity – detectors are more sensitive and the optical throughput is higher.
FTIR can identify small concentrations of contaminants.

 Mechanical simplicity – The mirror in the interferometer is the only moving part in
the FTIR instrument, therefore making mechanical breakdown minimal.

Simultaneous analysis of multiple gaseous compounds

3.2. -C-H stretching [7 marks]

M1 for carbon=12.0 x10-3 kg/mol /(6.0 x 1023 atom/mol) x 1 atom=2.0 x 10-26 kg

M2 for H2=1.0 x10-3 kg/mol /(6.0 x 1023 atom/mol) x 1 atom=1.67 x 10-27 kg

Reduced mass ()= (M1 x M2)/(M1+M2)= (2.0 x 10-26 kg x 1.67 x 10-27 kg)/( (2.0 x 10-26 kg

+ 1.67 x 10-27 kg)= 1.54 x 10-27 kg

= 5.305 x 10-10(5.0 x102 N/m / 1.54 x 10-27 kg)0.5

= 3.07 x 105 m-1

= 3.07 x 103 cm-1

Therefore: absorption frequency=wavenumber x speed of light= 9.21 x 1013 Hz

3.3.

 These humps show that the molecule is absorbing radiation over a band of
wavelengths.
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TEST 1 10 AUGUST 2013

 One reason for this band, rather than line absorption is that an electronic level
transition is usually accompanied by a simultaneous change between the more
numerous vibrational and rotational levels.

 Thus, a photon with a little too much or too little energy to be accepted by the
molecule for a 'pure' electronic transition.

 That can be utilized for a transition between one of the vibrational levels associated
with the lower electronic state to one of the vibrational levels of a higher electronic
state.

3.4.

 Fluorescence spectroscopy is analytical tool that deals the behaviour of the analyte

when exposure to the light by measuring fluorescence and excitation spectra using

two monochromators and detector.

 In a fluorescence emission spectrum, the excitation wavelength is held constant and

the emission intensity is measured as a function of the emission wavelength.

 In an excitation spectrum, the emission is measured at one wavelength while the

excitation wavelengths are scanned.

 The excitation spectrum closely resembles an absorption spectrum since the emission

intensity is usually proportional to the absorbance of the molecule.

 Factors: temperature, concentration, pH, central metal of the coodination sphere,

molecule its self, solvents

3.5. [5 marks]

A=bc

bc=-log(0.212)/(2.00 cm x 3.78 x 10-3 M)= 0.673 / 7.56 x 10-3= 89.02 L/mol/cm

Therefore c= -log(0.212 x 3)/(1.00 cm x 89.02 L/mol/cm)= 0.197/89.02= 2.21 x 10-3 M

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TEST 1 10 AUGUST 2013

SECTION B (AA, ICP & MS)

QUESTION 4 (25 marks)

4.1.

 Nebulaization

 sample is aspirated into the nebulizer where it is converted into an aerosol which is

then pushed into the plasma

 Laser ablation

 a laser is focused on the sample and creates a plume of ablated material which can be

swept into the plasma

 Electrothermal/intorch vaporization-both uses a hot surface to vaporize sample for

introduction into the plasma

4.2.

 In AAS, an external light source, such as a hollow cathode lamp, must be used to
excite the atom of interest.

 Given that HC lamps provide narrow bandwidth excitation, and given that the
absorption spectra for elements vary.

 Multielement analysis by AA requires multiple excitation sources.

 In Plasma AES, the energy of the plasma is sufficient to excite the atoms in the
sample,

 So the requirement for a separate excitation source is removed.

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TEST 1 10 AUGUST 2013

4.3.

 A Quadrupole is a mass analyzer that uses an electric field to separate ions.

 The Quadrupole consists of 4 parallel rods/ poles, where adjacent rods have opposite
voltage polarity applied to them.

 The electric force on the ions causes the ions to oscillate/orbit in the area between the
4 rods, where the radius of the orbit is held constant.

 The ion moves in a very complex motion that is directly proportional to the mass of
the ion, voltage on the quadrupole, and the radio frequency.

4.4.

 An HCL usually consists of a glass tube containing a cathode, an anode, and a

buffer gas (usually a noble gas) and the cathode is made of element of interest

 A large voltage across the anode and cathode will cause the buffer gas to ionize,
creating a plasma which causes excitations of atoms.

 As these excited atoms decay to lower states, they will emit photons, which can then
be detected and a spectrum can be determined.

 An HCL can also be used to tune light sources to a specific atomic transition by
making use of the optogalvanic effect, which is a result of direct or
indirect photoionization.

4.5.

 Its unequalled sensitivity,

 Detection limits, speed and diversity of its applications.
 A mass spectrum will usually be presented as a vertical bar graph and each bar
represents an ion having a specific mass-to-charge ratio (m/z).
 The length of the bar indicates the relative abundance of the ion.
 The most intense ion is assigned an abundance of 100, and it is referred to as the base
peak.

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TEST 1 10 AUGUST 2013

 The ions formed in a mass spectrometer have a single charge, so the m/z value is
equivalent to mass itself.
 The highest-mass ion in a spectrum is normally considered to be the molecular ion
and lower-mass ions are fragments from the molecular ion, assuming the sample is a
single pure compound.

QUESTION 5 (25 marks)

5.1.

 The radiation from the deuterium lamp is passed through the flame alternately with
HCL beam.

 The D2 lamp is mostly absorbed by the background while,

 The HCL radiation is absorbed by the atoms.

 By comparing the radiant power of the two beams,

 the atomic absorption can be corrected for any background absorption

5.2. [7 marks]

 MS has a small sample of compound is ionized, usually to cations by loss of an

electron (Ion Source).

 The ions are sorted and separated according to their mass and charge (Mass
Analyzer).

 The separated ions are then detected and tallied, the results are displayed on a chart
(Detector).

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TEST 1 10 AUGUST 2013

5.3.

 Full line of UV filtering safety glasses

 to offer protection against harmful UV rays emitted from Cu or Pb.

 heat resistant gloves

 as appropriate when setting up the AAS and

 when running samples due to high flame temperature in AAS

5.4.

 To provide a relatively quiet flame, and

 To provide a long path length

 To enhance sensitivity for AA and

 Reproducibility

5.5.

 The higher resolution of the double focusing spectrometer

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 Which allows the ions of interest to be better separated from background ions than
with a relative low resolution quadrupole spectrometer

 The higher signal to background ratio of the double focusing instrument,

 leads to lower detection limits than with the quadrupole.

QUESTION 6 (25 marks)

6.1.

 The sample solution is converted into mist-form using a nebulizer, and then

introduced into the flame.

 It is atomized by the temperature of the flame.

 The flames vary in temperature

 It uses Ar-H2, Air-H2, Air-acetylene & ONO-acetylene

 Measurement time: A few dozen seconds

6.2.

 ICPMS is a type of mass spectrometry which is capable of detecting metals and

several non-metals at concentrations as low as one part in 1012 (part per trillion).

 This is achieved by ionizing the sample with inductively coupled plasma and then
using a mass spectrometer to separate and quantify those ions.

 ICP-MS has greater speed, precision, and sensitivity.

 Metal speciation

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TEST 1 10 AUGUST 2013

 Quantification of proteins and biomolecules

 Elemental analysis

6.3. [8 marks]

Setting up two equations in the two unknowns: y=mx+c

0.599=0.600m+c…………………………………(1)

0.396=0.400m+c…………………………………(2)

Subtraction: (1)-(2) and solve for m

0.203=0.200m

m=1.015………………………………………..(3)

Substitute (3) into (1)

0.599=0.600(1.015)+c

c=-0.01

The unknown Pb is then: y=mx+c

0.502=1.015x+c

x=(0.502+0.01)/(1.015)=0.504ppm

x=0.504ppm=0.504 mg/L=(0.504 x 10-3 g/L)/(207.2 g/mol)= 2.43 x 10-6 mol/L

6.4.

 The temperature setting of an ICP is generally very high, this will effectively favour
the formation of a higher percentage of atoms and
 ions will easily form at this higher temperature.

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 Sample residence time is prolonged rendering the process to complete isolation and
differentiation.
 The percentage of electron is high, constant and
 the probability of interferences stemming from ionization interferences is
consequently diminished.
 The stable and inert chemical environment for formation of ions and atoms is ideal for
this process without undue interferences and erratic consequences.

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