MS3014 tutorial
UV-Vis spectroscopy
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Recap: UV-Vis Spectroscopy
1. Principles of UV-VIS Spectroscopy
o Electronic transitions in molecules
(organic molecules & transition metals)
o Ligand field theory
2. Instrumentation
o Qualitative analysis
o Quantitative analysis
3. Applications of UV-Vis: Examples
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Cobalt (II) chloride, also known as Drierite™, is a color indicator used in
de-humidifying agents commonly found in the laboratory. This compound
changes from deep blue to light pink in the presence of water:
Co[CoCl4] + 12 H2O ↔ 2 Co(H2O)6Cl2
(deep blue, tetrahedral CoCl42-) (light pink, octahedral [Co(H2O)6]2+)
Use your knowledge on ligand field splitting to explain the color change.
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Co[CoCl4] + 12 H2O ↔ 2 Co(H2O)6Cl2
(deep blue, tetrahedral CoCl42-) (light pink, octahedral [Co(H2O)6]2+)
Co2+ has a d7 configuration
• The energy gap between the eg and
t2g orbitals is greater in [Co(H20)6]2+
than in CoCl42-
• Hence more energy is required to high-spin complex low spin complex
make a d-d transition in the case of ~ 600 nm ~ 450 – 560nm
[Co(H20)6]2+ (orange) (green)
• Higher energy = absorb light of
LOWER wavelength (likely green) to
appear as pink (complementary
color).
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1. Which of these molecules would appear as colored under visible light?
A B C D
Benzene Naphthalene Anthracene Tetracene
255 nm 315 nm 357 nm 471 nm
colorless white Off-white to yellow orange
As the number of fused rings increases, the long wavelength
absorption shifts to higher values.
Absorption of light in visible range will result in the compound
having a color that is complementary to the wavelength of
light absorbed.
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2. Which of the following transitional metal complexes are colorless?
A. [FeCl6]4- C. [Co(CN6)]3-
Fe(26): [Ar]4s23d6 Co(27): [Ar]4s23d7
Fe2+ : [Ar]4s03d6 Co3+ : [Ar]4s03d6
B. [Zn(OH)6]4- D. [Cr(H2O)6]2+
Zn(30): [Ar]4s23d10 Cr(24): [Ar]4s13d5
Zn2+ : [Ar]4s03d10 Cr2+ : [Ar]4s03d4
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3. You come across two unlabelled colored solutions, one orange and one green. Both
solutions are known to be made up of a hypothetical transitional metal octahedral
complex; however, one contains chloride ions while the other has ammonia ligands.
The green solution is now mixed with excess hydrogen cyanide. What color would
the new mixture likely be? Explain.
Since we know that NH3 is a stronger ligand than Cl-, the solution containing NH3 will
absorb light with a lower wavelength (due to higher E gap).
Hence the green solution contains Cl- ions.
Now the green solution is added with excess HCN (strong
ligand). This increases the E gap, and now shift the light
absorption wavelength to lower than 600 - 640nm. Since the
complex with NH3 absorbs blue, the complex containing the CN-
ligand will likely absorb violet and appear yellow.
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4. Erythrosine (MW = 836) is an organoiodine compound, used primarily as a food
colorant. The figure below shows the UV-VIS spectrum of erythrosine aqueous
solution in a 0.1 cm long cell, where the maximum absorbance is at 524 nm. The
molar absorption coefficient is 83,000 M-1cm-1.
The table below shows the absorbances of some erythrosine solutions with known
concentrations. By constructing a calibration curve, determine the concentration of an
unknown sample of erythrosine with an absorbance of 0.58 measured at 524 nm.
Concentration of
Abs524nm
erythrosine (mM)
0.1 0.16
0.2 0.37
0.4 0.75
0.5 0.97
0.8 1.22
1.0 1.21
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5. Erythrosine (MW = 836) is an organoiodine compound, used primarily as a food
colorant. The figure below shows the UV-VIS spectrum of erythrosine aqueous
solution in a 0.1 cm long cell, where the maximum absorbance is at 524 nm. The
molar absorption coefficient is 83,000 M-1cm-1.
In aqueous solution, erythrosine loses protons
and gains negative charges. Delocalization of
electrons causes this molecule to absorb UV-VIS.
Erythrosine is also known
as Red Dye No. 3
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The table below shows the absorbances of some erythrosine solutions with known
concentrations. By constructing a calibration curve, determine the concentration of an
unknown sample of erythrosine with an absorbance of 0.58 measured at 524 nm.
Calibration curve
Concentration of
Abs524nm 1.6
erythrosine (mM)
1.4
0.1 0.16
1.2
Absorbance
0.2 0.37 1
y = 1.4576x
0.4 0.75 0.8
R² = 0.9661
0.6
0.5 0.97 0.4
0.8 1.22 0.2
1.0 1.21 0
0 0.2 0.4 0.6 0.8 1 1.2
Concentration of erythrosine (mM)
Abs524nm = 1.4576 [C]
[C] = 0.58/1.4576 = 0.400 mM
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The table below shows the absorbances of some erythrosine solutions with known
concentrations. By constructing a calibration curve, determine the concentration of an
unknown sample of erythrosine with an absorbance of 0.58 measured at 524 nm.
Calibration curve
Concentration of
Abs524nm 1.2
erythrosine (mM)
0.1 0.16 1
Absorbance
0.2 0.37 0.8
0.4 0.75 0.6 y = 1.9022x
0.5 0.97 0.4 R² = 0.9991
0.8 1.22 0.2
1.0 1.21 0
0 0.1 0.2 0.3 0.4 0.5 0.6
Concentration of erythrosine (mM)
Abs524nm = 1.9022 [C]
[C] = 0.58/1.9022 = 0.305 mM
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5. Which of the following situations would you NOT recommend UV-VIS
spectroscopy?
A. To identify species with known absorption maximas.
B. To determine the extent of a chemical reaction by measuring the absorbance
intensity of a product at a known wavelength.
C. To determine the types of functional groups present in a sample.
D. To quantify concentrations of DNA in water.
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Lecture learning outcomes
You should be able to:
1. describe how molecules (organic molecules & transition metals) undergo
electronic transitions due to UV-VIS absorption;
2. derive the type of complexes (and their colors) using ligand field theory;
3. calculate concentrations of absorbing species using Beer-Lambert law
(A=ecL); and
4. identify applications for UV-Vis spectroscopy (and understand the limitations
of this technique) 13