SPECTROSCOPY

THEORY & APPLICATIONS

Complimentary Colours
It is possible to predict which
wavelengths are likely to be absorbed
by a coloured substance.

When white light passes through or

is reflected by a coloured substance,
a characteristic portion of the mixed
wavelengths is absorbed. The
remaining light will then assume the
complementary colour to the
wavelength(s) absorbed.

This relationship is demonstrated by

the colour wheel shown on the right.
Complementary colours are
diametrically opposite each other.
The test tube in the figure contains an orange
solution. The solar spectra is white light. When
sunlight shines through an orange solution, the
violet, blue and green wavelengths are absorbed.
The other colours pass through. The transmitted
light is the light we see, and it looks orange.
Diffraction refers to various phenomena
that occur when a wave encounters the
corners of an obstacle or an opening
resulting in bending of wave.
 UV-Vis Spectroscopy
Region Wavelength (nm)  extensively used in teaching, research
Far ultraviolet 10–200
and analytical laboratories for the
quantitative analysis of all molecules
Near 200–380 that absorb UV and visible
ultroviolet electromagnetic radiations
Visible 380–780  stimulates molecular vibrations and
Near infrared 780–3000 electronic transitions

Middle infrared 3000–30,000

 analyses compounds using the
electromagnetic radiation spectrum
Far infrared 30,000–300,000 from 10 nm to 700 nm
 measures absorption or transmittance
Microwave 300,000–
1,000,000,000  identifies inorganic and organic
species
Health issues
UV exposure
UV radiation has an impact
on human health, both
beneficial and harmful. Too
much exposure can be
harmful while moderate
exposure has beneficial
effects.
Harmful Effects
Excessive UV exposure is harmful to the eye, skin, and immune system.
UVA radiation causes little or no immediate reaction, but at UVB band (315--280 nm),
more damage to eyes and skin occur(lighter skin is more sensitive).
UVC band (265 nm to 275 nm) is the most damaging to the eyes and skin.
UVB overexposure can not only cause sunburn, but is also responsible for skin cancer.
Beneficial effects
If one can moderate their UV exposure, the three primary health benefits are the
production of vitamin D, improvement in mood, and increased energy.
 Internal Energy of Molecule

Etotal=Etrans+Eelec+Evib+Erot+Enucl

Eelec: electronic transitions (UV, X-ray)

Evib: vibrational transitions (Infrared)
Erot: rotational transitions (Microwave)
Enucl: nucleus spin (nuclear magnetic resonance)
or (MRI: magnetic resonance imaging)
The molecules normally have many excited
Molecular Energy Levels
modes of vibration and rotation within the
electronic transitions at room temperature.
The vibrations cannot be completely “frozen
out” even at very low temperatures.
Consequently, a collection of molecules
generally has its members in
many states of vibrational and rotational
excitation. The energy levels for these states
are quite closely spaced, corresponding to
energy differences considerably smaller
than those of electronic levels. The
rotational and vibrational levels are thus
“superimposed” on the electronic levels.
A molecule may therefore undergo electronic
and vibrational–rotational excitation
simultaneously.
Molecular Energy Levels
 Electronic Transitions
Absorption of UV-Vis radiation is associated with
excitation of electrons, in both atoms and
molecules, from lower to higher energy levels.
Energy levels of matter are quantised, only
light with a precise amount of energy can cause
transition from one energy level to another.
Light with a different energy would not be absorbed
and would not change the level.
Electronic Transition Examples
Transitions Wavelength
Both π to σ* and σ to σ* transitions require a great
deal of energy and therefore occur in the far UV
region or weakly in the region 180-240nm.
Consequently, saturated groups (single bonds) do not
exhibit strong absorption in the ordinary UV region.
Transitions of the n to π* and π to π* occur in
molecules with unsaturated centres; they require less
energy and occur at longer wavelengths than
transitions to σ* antibonding orbitals.

σ→σ∗ <200 nm C–C, C–H

n→σ∗ 160–260 H2O, CH3OH, CH3Cl

π→π∗ 200–500 C=C, C=O, C=N, C≡C

n→π∗ 250–600 C=O, C=N, N=N, N=O

 Electronic Transitions
All molecules will undergo electronic excitation following absorption of light, but saturated
molecules require very high-energy radiation (in the vacuum ultraviolet, <200 nm)

The larger the gap between the energy levels,

the greater the energy required to promote
the electron to the higher energy level,
resulting in light of higher frequency, and
therefore shorter wavelength, being
absorbed.

Therefore in order to absorb light in the region

from 200 to 800 nm (where most UV-Vis
spectra are measured), the molecule must
contain either π bonds or atoms with non-
bonding orbitals. A non-bonding orbital is a
lone pair on, say, oxygen, nitrogen or a halogen.
π→π∗ transitions are more likely to occur
(HOMO to LUMO)while n→π∗ transitions are
less likely to take place