CHEMISTRY

UNIT 2 MODULE 2
Introduction to Spectroscopy
1. explain the nature of electromagnetic radiation
2. state the approximate wavelength ranges of the X-ray, UV/VIS, IR and radiofrequency regions of the
EM spectrum
3. appreciate the relative energies and dangers associated with exposure to high energy wavelengths
4. perform calculations using the equation E = h = hc /
5. understand that energy levels in atoms and molecules are quantised
UV-VIS Spectroscopy
1. explain the origin of absorption in UV/VIS spectroscopy
2. explain why some species will absorb light in the uv/vis region but some others will not
3. describe the basic steps involved in analysing samples by uv/vis spectroscopy (also mention use of
complexing agents to form coloured compounds, as well as sensitivity and detection limits)
4. state the Beer-Lambert Law and be able to use the equation to calculate the concentration of a given
species in solution (implies the use of standards and calibration curves)
5. list examples of the use of uv/vis spectroscopy in the quantitation of substance (refer to iron tablets,
glucose and urea in blood, cyanide in water)
Introduction to spectroscopy
Electromagnetic radiation can be described in terms of a stream of photons, which are massless particles each
travelling in a wave-like pattern and moving at the speed of light. Each photon contains a certain amount (or
bundle) of energy, and all electromagnetic radiation consists of these photons.

The electromagnetic (EM) spectrum is just a name that scientists give a bunch of types of radiation when they
want to talk about them as a group. Radiation is energy that travels and spreads out as it goes-- visible light
visible light that comes from a lamp in your house and radio waves that come from a radio station are two types
of electromagnetic radiation.

Other examples of EM radiation are microwaves, infrared and ultraviolet light, X-rays and gamma-rays. Hotter,
more energetic objects and events create higher energy radiation than cool objects. Only extremely hot objects or
particles moving at very high velocities can create high-energy radiation like X-rays and gamma-rays.
EM waves are typically described by any three of the following physical properties: the frequency,
wavelength, , and photon energy, E.

, and

Frequencies range from about a million billion Hertz (gamma rays) down to a few Hertz (radio waves).
Wavelength is inversely proportional to the wave frequency, so gamma rays have very short wavelengths that
are fractions of the size of atoms, whereas radio wavelengths can be as long as a several thousand kilometers.
Photon energy is directly proportional to the wave frequency, so gamma rays have the highest energy around a
mega electron volt and radio waves have very low energy around femto electron volts (femto = 10 15).

These relations are illustrated by the following equation E = h

or

and its variations below:or

Where: c = 3 x 108 m s-1 (speed of light in vacuum) &

h = 6.626 1034 Js-1 (Planck's constant).

Questions
a. State the relationships between wavelength ( ), frequency ( ) and energy (E) of a wave.
b. A compound absorbs light of frequency 927.8MHz. Calculate the wavelength of the light
absorbed.
High energy EM radiation e.g. ultraviolet rays, X-rays and gamma rays are very dangerous to living tissue.
These energetic rays have to ability to disrupt and destroy DNA in tissue which can result in mutations or even
death. All three types of rays would result in electronic transitions and even ionisation.

In atoms, energy levels do NOT form a continuum. In fact, these energy levels are discrete or quantised.
For example, energy levels may be at 5J, 10 J, 20 J but never 5.1J, 5.2 J, 10.3 J etc. There are no in
between energy levels or values.

UV-VIS Spectroscopy
The absorption of ultraviolet or visible radiation corresponds to the excitation of outer electrons and thus
electronic transitions. When an atom or molecule absorbs energy, electrons are promoted from their ground state
(lowest energy level) to an excited state. Absorption of ultraviolet and visible radiation in organic molecules is
restricted to certain functional groups (chromophores) that contain valence electrons of low excitation energy.
Chromophores generally contain either or lone pair(s) of electrons
Electronic transitions generally seen are n* and * transitions. In essence molecules containing lone
pairs of electrons OR electrons would absorb uv-vis radiation.
All other transitions either do NOT occur or are TOO SMALL to be considered. Below is a diagram that shows
the various electronic transitions.

Question:
Underline the molecules in the list below which would absorb uv-vis radiation. O2, C6H6, NH3, CH4, C2H4, HCl
Sample preparation
UV-VIS samples are usually liquids or solutions. They are placed in a cell or cuvette which is a rectangular
block, with two sides frosted and the other two sides clear and one end open and the other closed. The frosted
sides can be touched but not the clear sides. Cells are made of fused synthetic silica.
Solutions or liquids are placed in the cell in increasing concentrations. A blank (which contains
everything else but the analyte in question) is first used to use as a reference point.
Sometimes, complexing agents must be added to provide colour to the analyte which allows it to be
analysed via uv-vis more easily. For example iron present in aqueous samples can be
determined spectrophotometrically by complexation with a suitable complexing agent. The absorbance
of the metal-ligand complex is usually measured in the visible region and is related to metal ion
concentration. Colorimetric determination of iron can be done using several known complexing agents.
Among the routinely used is 1,10-phenanthroline (phen) which reacts with Fe2+ to form an orangered complex. Also dimethylglyoximeis is a complexing agent using to determine the amount of
nickel present.
The procedure depends on the construction of a calibration curve from standard Fe 2+, followed by
measurement of the unknown Fe2+ concentration
Standards
from
and
thecalibration
curve. curves are
used for UV-VIS spectroscopy
coupled with
Using absorbance to determine the concentration
of Beer-Lambert
an analyte Law as

concentration of the analyte MUST

show a linear signal response for the
Law to be of any worth.
NB

Io = incident light,
I = transmitted light

The Beer-Lambert Law

= molar absorbtivity
= cell length in cm (usually 1 cm in length)
c = molar concentration of solution
A = absorbance
Spectrophotometer
The basic parts of a spectrophotometer are a light source, a holder for the sample, a diffraction grating or
monochromator to separate the different wavelengths of light, and a detector. The radiation source is often a
Tungsten filament (300-2500 nm), a deuterium arc lamp which is continuous over the ultraviolet region (190400 nm), and more recently light emitting diodes (LED) and Xenon Arc Lamps for the visible wavelengths. The
detector is typically a photodiode. Photodiodes are used with monochromators, which filter the light so that only
light of a single wavelength reaches the detector.
The dual-beam design greatly simplifies this process by simultaneously measuring P and Po of the sample and
reference cells, respectively. Most spectrometers use a mirrored rotating chopper wheel to alternately direct the
light beam through the sample and reference cells. The detection electronics or software program can then
manipulate the P and Po values as the wavelength scans to produce the spectrum of absorbance or transmittance
as a function of wavelength.
Schematic of a dual-beam uv-vis spectrophotometer

Applications of UV-VIS spectroscopy

UV/VIS spectroscopy is routinely used in the quantitative determination of solutions of transition metal ions and
highly conjugated organic compounds, of iron in iron tablets, glucose and urea in blood and cyanide in water
Solutions of transition metal ions which are too pale to give a reasonable absorbance can be complexed with
various ligands to form more intense colours (i.e., absorb visible light) because d electrons within the metal
atoms can be excited from one electronic state to another. The colour of metal ion solutions is strongly affected
by the presence of other species, such as certain anions or ligands. For instance, the colour of a dilute solution of
copper sulphate is a very light blue; adding ammonia intensifies the colour and changes the wavelength of
maximum absorption (max)
Question: