CHAPTER 3

UV/VISIBLE ABSORPTION SPECTROSCOPY

Major application;

Identification and determination of inorganic and organic

species.

In the uv/vis region, many types of organic and inorganic

compounds absorb radiation directly. Others can be
converted to absorbing species by means chemical reaction.

3.1 . Absorbing Species

 The absorbing of UV or visible radiation by an atomic or

molecular species, M can be considered to be a two-step
process.

a. Involves electronic exitation

M + hv M*
 - The product of the reaction between M and the
photon hv is an electronically excited species
symbolized by M*

- The lifetime of the excitation, 10-8 – 10-9

(b) Involves relaxation in the conversion of the

excitation energy to heat

M* M + heat

- relaxation may occur by decomposition of M* to

form new species; such as a process is called a
photochemical reaction.
 The absorption of UV and Vis radiation generally results
from excitation of bonding electrons. Therefore, the
wavelengths of absorption peaks can be correlated with
the types of bonds in the species under study.

 More important, the application of UV and Vis absorption

spectroscopy to the quantitative determination of
compounds containing absorbing groups.

 Three types of electronic transitions:

a. π, δ and n electrons
b. d and f electrons
c. charge transfer electrons
3.2 Absorbing species containing π,σ and n electrons

 Absorbing species containing π,σ and n electrons include

organic molecules and several inorganic anions.

 All organic compounds are capable of absorbing

electromagnetic radiation because all contain valence
electrons that can be excited to higher energy levels.

 The electronic spectra of organic molecules containing

chromophores are usually complex because the
superposition of vibrational transitions on the electronic
transition leads to an intricate combination of overlapping
lines; the result is a broad band of absorption that often
appears to be continuous.
 The complex nature of the spectra makes detailed theoretical
analysis difficult or impossible.

3.3 Types of Absorbing Electrons

The electrons that contribute to absorption by an organic

molecule are:

i. Those that participate directly in bond formation

between atoms and thus associate with more than one
atom.

ii. Nonbonding or unshared outer electrons that are

largely localized about such atoms as oxygen, the
halogen, sulphur and nitrogen.
 When two atomic orbitals combine, either a low-energy
bonding molecular orbital or a high-energy antibonding
molecular orbital results. The electrons of a molecule
occupy the former in the ground state.

Electrons in a molecule can be classified into four different

types:

1. Closed-shell electrons;

That are not involved in bonding. These have high an

excitation energies and do not contribute to absorption in
the vis and UV regions.

2. Covalent single-bond electrons (σ or sigma electrons)

These also posses too high an excitation energy to

contribute to absorption of Vis or UV radiation.
 E.g: single-valence bonds in saturated hydrocarbons
–CH2-CH2-

3. Paired nonbonding outershell electrons (n electrons)

E.g. N, O, S and halogens. These are less tightly held than σ

electrons and can be excited by Vis or UV radiation

4. Electons in π (pi) orbitals

E.g. In double or triple bonds. These are the most readily

excited and are responsible for a majority of electronic spectra
in the UV and Vis regions.
 Many organic compounds contain nonbonding (n) electrons.
An example showing three types of electrons in formaldehyde
is shown in figure below:

Types of molecular orbitals in formaldehyde

 The energies for various types of molecular orbitals differ
significantly. The energy level of a nonbonding electron lies
between the energy level of the bonding and the antibonding
σ and π orbitals.

Electronic molecular energy levels.

Electronic transitions of energy levels can be brought about
by the absorption of radiation.

Four types of transitions are possible:

σ σ* - vacuum UV (λ<185nm)
n σ* - longer than 185nm
π π* - Near UV Such functional groups capable to
n π* – Near / Vis UV absorb uv-called chromophore

σ σ* transition

 An electron in a bonding σ orbital of a molecule is

excited to the corresponding antibonding orbital by the
absorption of radiation.

 Large energy required to induced σ σ* transition.

 Methane – contain only single C-H bonds and thus undergo
only σ σ* transitions ( absorption max at
125 nm ).

 Ethane - has an absorption peak at 135 nm

- less energy is required because strength of the C-C
bond is less than that of C-H bond.

 Absorption maxima due to σ σ* transition are never

observed in the ordinarily accessible UV region.

n σ* transitions

 Occur to compounds containing atoms with unshared electron

pairs (nonbonding electrons).
 Require less energy than the σ σ* transitions.

 Radiation region : 150 to 250 nm

n π* and π π* transitions

 Occur to most organic compounds because the energies

required for these processes bring the absorption peaks into
an experimentally convenient spectral region – 200 to 700 nm.

 Both transitions require the presence of an unsaturated

functional group to provide the π orbitals.

 Molar absorptivities, ε value:

n π*: 10 to 100 Lcm-1mol-1

π π*: 1000 to 10000 Lcm-1mol-1
 The effect of the solvent on the wavelength for the peak
(with increasing polarity of the solvent):

n π* - generally shifted to shorter wavelengths

(a hypsochromic or blue shift).

π π* - shifted to longer wavelengths

(a bathochromic or red shift).

 Not all molecules contain σ, π and n molecular orbitals.

The molecule contain σ, π and n orbitals
How λmax results from electron transitions?

Example 1:
Example 2:
3.4 Organic Chromophores

 The absorbing groups in a molecule are called chromophores.

A molecule containing a chromophore is called chromogen.

 An auxochrome does not itself absorb radiation, but if present

in a molecule, it can enhance the absorption by shift the
wavelength of absorption when attached to the chromophore.
Examples are –OH, -NH2 and halogens. These posses
unshared electron (n) that can interact with the π electrons in
the chromophore (n-π conjugation).

 Hypsochromism – a decrease in molar absorptivity

 Hypersochromism – an increase in molar absorptivity

 Aromatic compounds are good absorbers of UV radiation.

 UV spectra of aromatic hydrocarbons are characterized by
three sets of bands that originate from π π* transitions:

- Strong absorption peak at 184 nm (εmax ~ 60,000)

- Weak absorption peak at 204 nm (εmax ~ 7900)

- Weak absorption peak at 256 nm (εmax ~ 200)

3.5 Inorganic Anions

 A number of inorganic anions exhibit UV absorption peaks

that are a consequence on n π* transitions. Examples:

- Nitrate – 313 nm
- Nitrite – 360 and 280 nm
- Carbonate – 217 nm
 Nitrate Nitrite

Azide
Carbonate
- Azide – 230 nm
- Trithiocarbonate – 500 nm

3.6 Application of Absorption Measurement to Qualitative

Analysis

Solvent

 In choosing a solvent, consideration must be given not only to

its transparency, but also its possible effects upon the
absorbing system.

 Polar solvent such as water, alcohols, esters and ketones

tend to obliterate spectral fine structure arising from
vibrational effects.

 Common solvents for the UV spectrophotometry include

water, 95 % ethanol, cyclohexane and 1,4-dioxane.
 Visible region: any colorless solvent

Detection of Functional Group

 Even though it may not provide the perfect identification of

an organic compound, an absorption spectrum in the UV
and the visible regions is nevertheless useful for detecting
the presence of certain functional groups that act as
chromophore.

- Carbonyl group: 280 – 290 nm

- Aromatic ring: 260 nm
3.7 Calibration of Instrumental Methods

 Calibration – a process that relates the measured analytical

signal to the concentration of analyte.

 The three common calibration methods include:

► calibration curve
► standard addition methods
► the internal standard method

Calibration curve
Calibration curve is a regression model used to predict the
unknown concentrations of analytes of interest based on the
response of the instrument to the known standards.
Standard addition methods

 Several standards containing exactly known concentration

of the analyte are introduced into the instrument with the
present of a blank - spiking the sample

 Ideally, the blank contains all of the components of the

original sample except for the analyte

 The resulting data are then plotted to give a graph of

corrected instrument response versus analyte concentration.

The internal standard method

 An internal standard is a substance that is added in a

constant amount to all samples (analyte), blank and
calibration standard in analysis.
 From calibration methods, the concentration of
unknown can be determined by comparing the
unknown signal to a set of standard samples of known
concentration signal.
Calibration curve
 Is often used in the determination of trace elements in metals
by emission spectroscopy.

3.8 Instrumentation

Source;

 Deuterium and hydrogen lamps

 Tungsten filament lamps
 Xenon arc lamps

Types of spectrometers;

 single-beam
 double-beam
Single-beam spectrophotometers

 Use a single light beam in order to irradiate the cell.

 the light enters the sample and a phototranducer monitor

the transmitted radiation as it emerges from the sample.

 Disadvantage: fluctuations, voltage fluctuations,

mechanical component's instability
 Overcome: Most modern spectrophotometers store baseline
spectrum to normalized spectrum.

Double-beam spectrophotometers
 Employ two light beams of equal intensity with two photo-
multipliers to record and subtract the baseline from spectra.

 One light beam irradiates the cell containing the analyte. The
other irradiates a cuvette containing solvent.

 The two outputs are amplified and determined electronically by

the readout device.

 Most modern UV and visible recording instruments are

double-beam.