UV-Vis Spectroscopy

• We know that:
• Substances are colored because of the
absorption and the emission of light in the
visible part of the Electromagnetic (EM)
spectrum.
• What is the origin of these transitions?
• Chromophores
• These describe a group of atoms on the molecule
which absorb light in the UV and/or Visible
region
Chromophores
How does absorption occur?
• Transitions from the HOMO to the LUMO
Effect of conjugation
• Conjugation refers to alternating double and single
bonds or non-bonding systems
• The higher the level of conjugation in the molecule
the wavelength of absorption and the closer it gets to
the visible part of the EM spectrum.
• The gap between HOMO and LUMO narrows as
conjugation increases.
• Examples: Methane has a λmax at 125nm
• Ethene-165nm, butadiene-217nm, hexatriene-
258nm
Can you identify the chromophores
and conjugation in each?
Identify the chromophores in the Food
dyes?
Absorption Spectra
Why are we interested in the
(λ)max?

Leads to
errors
Beer Lambert’s Law
• For dilute solutions the concentration of the
analyte is directly proportional to Absorbance.
• A=εcl
• A= Absorbance, c= concentration, l=path length,
ε=molar absorptivity/extinction coefficient (constant
for a particular molecular)
Calibration Curve
• This is done with a series of standards
containing the analyte at varying concentrations.
• Question
• A water source believed to be contaminated with
potassium permanganate is collected. 10ml of
the sample is pipetted into a 250ml volumetric
flask. This solution was used to fill a cuvette and
analysed at 500nm. The absorbance was found
to be 0.45 Determine the concentration of
potassium permanganate in the water source.
• What is the concentration of the unknown?
Problem with Concentration
• Serial Dilution