UV –Spectroscopy - Absorbance

Introduction

Many molecules absorb ultraviolet or visible light. The absorbance of a solution

increases as attenuation of the beam increases. Absorbance is directly proportional
to the path length, b, and the concentration, c, of the absorbing species. Beer's Law
states that

A = ebc, where e is a constant of proportionality, called the absorbtivity.

Different molecules absorb radiation of different wavelengths. An absorption

spectrum will show a number of absorption bands corresponding to structural
groups within the molecule. For example, the absorption that is observed in the UV
region for the carbonyl group in acetone is of the same wavelength as the
absorption from the carbonyl group in diethyl ketone.

Electronic transitions

The absorption of UV or visible radiation corresponds to the excitation of outer

electrons. There are three types of electronic transition which can be considered;

1. Transitions involving p, s, and n electrons

2. Transitions involving charge-transfer electrons

3. Transitions involving d and f electrons (not covered in this Unit)

When an atom or molecule absorbs energy, electrons are promoted from their
ground state to an excited state. In a molecule, 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.
Absorbing species containing p, s, and n electrons

Absorption of ultraviolet and visible radiation in organic molecules is restricted to

certain functional groups (chromophores) that contain valence electrons of low
excitation energy. The spectrum of a molecule containing these chromophores is
complex. This is because the superposition of rotational and vibrational transitions
on the electronic transitions gives a combination of overlapping lines. This appears
as a continuous absorption band.

Possible electronic transitions of p, s, and n electrons are;

s ® s* Transitions

An electron in a bonding s orbital is excited to the corresponding antibonding

orbital. The energy required is large. For example, methane (which has only C-H
bonds, and can only undergo s ® s* transitions) shows an absorbance maximum at
125 nm. Absorption maxima due to s ® s* transitions are not seen in typical UV-Vis.
spectra (200 - 700 nm)

n ® s* Transitions

Saturated compounds containing atoms with lone pairs (non-bonding electrons) are
capable of n ® s* transitions. These transitions usually need less energy than s ® s *
transitions. They can be initiated by light whose wavelength is in the range 150 -
250 nm. The number of organic functional groups with n ® s* peaks in the UV region
is small.

n ® p* and p ® p* Transitions

Most absorption spectroscopy of organic compounds is based on transitions of n or

p electrons to the p* excited state. This is because the absorption peaks for these
transitions fall in an experimentally convenient region of the spectrum (200 - 700
nm). These transitions need an unsaturated group in the molecule to provide the p
electrons.
Molar absorbtivities from n ® p* transitions are relatively low, and range from 10
to100 L mol-1 cm-1 . p ® p* transitions normally give molar absorbtivities between
1000 and 10,000 L mol-1 cm-1 .

The solvent in which the absorbing species is dissolved also has an effect on the
spectrum of the species. Peaks resulting from n ® p* transitions are shifted to
shorter wavelengths (blue shift) with increasing solvent polarity. This arises from
increased solvation of the lone pair, which lowers the energy of the n orbital. Often
(but not always), the reverse (i.e. red shift) is seen for p ® p* transitions. This is
caused by attractive polarisation forces between the solvent and the absorber,
which lower the energy levels of both the excited and unexcited states. This effect
is greater for the excited state, and so the energy difference between the excited
and unexcited states is slightly reduced - resulting in a small red shift. This effect
also influences n ® p* transitions but is overshadowed by the blue shift resulting
from solvation of lone pairs.

Charge - Transfer Absorption

Many inorganic species show charge-transfer absorption and are called charge-
transfer complexes. For a complex to demonstrate charge-transfer behaviour, one
of its components must have electron donating properties and another component
must be able to accept electrons. Absorption of radiation then involves the transfer
of an electron from the donor to an orbital associated with the acceptor.

Molar absorbtivities from charge-transfer absorption are large (greater that 10,000
L mol-1 cm-1).

Luminescence
Introduction

Luminescence is the emission of light by a substance. It occurs when an electron

returns to the electronic ground state from an excited state and loses it's excess
energy as a photon.

Luminescence spectroscopy is a collective name given to three related

spectroscopic techniques. They are;

• Molecular fluorescence spectroscopy

• Molecular phosphorescence spectroscopy

• Chemi-luminescence spectroscopy
 Fluorescence and phosphorescence (photoluminescence)

The electronic states of most organic molecules can be divided into singlet states
and triplet states;

Singlet state: All electrons in the molecule are

spin-paired
Triplet state: One set of electron spins is
unpaired

Fluorescence
Absorption of UV radiation by a molecule excites it from a vibrational level in the
electronic ground state to one of the many vibrational levels in the electronic
excited state. This excited state is usually the first excited singlet state. A molecule
in a high vibrational level of the excited state will quickly fall to the lowest
vibrational level of this state by losing energy to other molecules through collision.
The molecule will also partition the excess energy to other possible modes of
vibration and rotation. Fluorescence occurs when the molecule returns to the
electronic ground state, from the excited singlet state, by emission of a photon. If a
molecule which absorbs UV radiation does not fluoresce it means that it must have
lost its energy some other way. These processes are called radiationless transfer of
energy. Have a look at this diagram:

Possible physical process following absorption of a photon by a molecule

Intra-molecular redistribution of energy between possible electronic and vibrational

states
The molecule returns to the electronic ground state.The excess energy is converted
to vibrational energy (internal conversion), and so the molecule is placed in an
extremely high vibrational level of the electronic ground state. This excess
vibrational energy is lost by collision with other molecules (vibrational relaxation).
The conversion of electronic energy to vibrational energy is helped if the molecule is
"loose and floppy", because it can reorient itself in ways which aid the internal
transfer of energy.

A combination of intra- and inter-molecular energy redistribution

The spin of an excited electron can be reversed, leaving the molecule in an excited
triplet state; this is called intersystem crossing. The triplet state is of a lower
electronic energy than the excited singlet state. The probability of this happening is
increased if the vibrational levels of these two states overlap. For example, the
lowest singlet vibrational level can overlap one of the higher vibrational levels of the
triplet state. A molecule in a high vibrational level of the excited triplet state can
lose energy in collision with solvent molecules, leaving it at the lowest vibrational
level of the triplet state. It can then undergo a second intersystem crossing to a
high vibrational level of the electronic ground state. Finally, the molecule returns to
the lowest vibrational level of the electronic ground state by vibrational relaxation.

Phosphorescence
A molecule in the excited triplet state may not always use intersystem crossing to
return to the ground state. It could lose energy by emission of a photon. A
triplet/singlet transition is much less probable than a singlet/singlet transition. The
lifetime of the excited triplet state can be up to 10 seconds, in comparison with 10-5
s to 10-8 s average lifetime of an excited singlet state. Emission from triplet/singlet
transitions can continue after initial irradiation. Internal conversion and other
radiationless transfers of energy compete so successfully with phosphorescence
that it is usually seen only at low temperatures or in highly viscous media.

Chemi-luminescence

Chemi-luminescence occurs when a chemical reaction produces an electronically

excited species which emits a photon in order to reach the ground state. These sort
of reactions can be encountered in biological systems; the effect is then known as
bioluminescence. The number of chemical reactions which produce chemi-
luminescence is small. However, some of the compounds which do react to produce
this phenomenon are environmentally significant.

A good example of chemiluminescence is the determination of nitric oxide:

NO + O3 ® NO2* + O2

NO2* ® NO2 + hv (l = 600 - 2800 nm)

The following graph shows the spectral distribution of radiation emitted by the
above reaction: