Optical Methods of Analysis

Dr. Jyoti
Origin of Spectra
 The interaction of radiation with matter gives a
response which can be measured to obtain
information about the analyte.

 The interactions in spectroscopy involve

transitions between different energy levels of
the chemical species.
How the spectra is obtained?
 The analyte is stimulated in some way by applying energy.
 By applying heat
electrical energy
light
chemical reaction
In case of optical spectroscopic methods, the source is the light.
Light has dual nature, both particle (packets of energy, photons) and wave (wavelength and
frequency) nature.

 Prior to applying the stimulus, the analyte is in lowest energy state (ground state).
 The stimulus causes some of the analyte species to undergo transition to a higher energy
(excited state).
 The information about the analyte is obtained by measuring the electromagnetic radiation

emitted as it returns to the ground state and increase of radiant power or energy is
recorded at the detector (Absorption process)
absorbed as a result of excitation and decrease of radiant power or energy is recorded
at the detector (Emission process)
 Information Obtained?
 Information about the identity (qualitative) and
concentration (quantitative) of the analyte.

 The information can be expressed graphically by

plotting a spectrum. For optical information this is a plot
of the emitted radiation as a function of frequency or
wavelength.

𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 (𝑎. 𝑢. )
1
ℎ𝑐
ℎ𝑐
E1 = hν = 𝜆
E2 = hν = 𝜆 1

0 𝜆2 𝜆1
𝑊𝑎𝑣𝑒𝑙𝑒𝑛𝑔𝑡ℎ (𝑛𝑚)
Transitions other than electronic are
also possible:
 In addition to its electronic energy, a molecule may also
Vibrate with a chemical bond.
Rotate around one of its axis.
 Thus the molecule may exhibit various vibrational and
rotational energy states.
 Absorption of a photon can lead to a change or
excitation in one of these states.

 The energy level differences are in the order:

∆Eelectronic > ∆Evibrational > ∆Erotational
Selection rules for electronic spectra:
 Spin Selection Rule, ∆S = 0
The allowed transitions must involve the promotion of
electrons without a change in their spin.

 Orbital Selection Rule or Laporte Rule, ∆ℓ = ±1

The allowed transitions must involve the change in the ℓ.
If the molecule has a center of symmetry, transitions within a
given set of p or d orbitals are forbidden.
Relaxation of the rules can occur through:
 Spin-Orbit Coupling: This give rise to weak spin forbidden
bands.

 Vibronic Coupling: An octahedral complex may have allowed

vibrations where the molecule is asymmetric.

 𝜋-acceptor and 𝜋-donor ligands can mix with the d-orbitals

so that the transitions are no longer purely d-d

[Mn(H2O)6]2+ - spin and laporte forbidden

[Ti(H2O)6]3+ - spin allowed and laporte forbidden
[CoCl4]2- - spin allowed and laporte partially allowed by d-p mixing
MnO4- or [TiCl6]2- - spin and laporte forbidden
Lambert Beer’s Law
 Because of the interactions between the photons and absorbing particles, the
radiant power of the beam decreases from P˳ to P.
 The absorption law tells us quantitatively how the absorption of light depends
upon the concentration of the absorbing molecule and the path length over
which the absorption occurs.
 According to Beer’s law, absorbance is directly proportional to the
concentration of the absorbing species, c and to the path length, b of the
absorbing medium:
𝑃˳
Therefore, A = log 𝑃 = abc
where a is proportionality constant called the absorptivity
c is the concentration in moles per litre
b in cm
The proportionality constant is called molar absorptivity and is therefore written
as
A = εbc
units of ε = Lmol-1cm-1
A = 2 - logT
Plot of Absorbance versus concentration
 A straight line is obtained passing through the origin.
 This relationship is linear for low concentrations.

Absorbance (a.u.)

Wavelength (nm)
Deviations from Beer’s law:
 A non-linear relationship is seen in the following
conditions:
 Real deviations: These are fundamental deviations due to
the limitations of the law itself.
 At high concentrations of the solute, the solute begins
to behave differently due to interactions with the
solvent and other solute molecules and even due to H-
bonding interactions.
 High concentration would result in a shift in the
absorption 𝜆 of the analyte.
 Chemical deviations: This may be due to specific
chemical species present.

 When the concentrations are high, the analyte molecule

can undergo association, dissociation and interact with
the solvent to produce a product which has a different
absorption characteristic and hence the absorption
spectrum of the sample changes.
 Instrument deviations: These occur because of the
method adopted for recording the absorbance.

 If the measurements are not made at 𝜆max there is a lot

of change in the molar absorptivity of the analyte at
other 𝜆 which results in the non-linear relationship.
UV Visible Spectroscopy: Spectrophotometer

 It is an instrument which is composed of two units:

(i) A spectrometer which produces light of definite
wavelength
(ii) A photometer which measures the intensity of the
transmitted or absorbed light.
This serves for the measurement of relative light whether
emitted, transmitted or reflected as a function of wavelength.

A spectrophotometer is composed of
(i) A source for the continuous visible spectrum.
(ii) A device for obtaining monochromatic light (prism, grating,
slit)
(iii) Absorption cells for the sample and blank solution.
(iv) A means of measuring the difference in absorption
between the sample and reference (blank).
Spectrophotometer: Block Diagram
Spectrophotometer

(i) Source used is tungsten/halogen lamp (for visible region)

or hydrogen or deuterium lamp (for UV region).
(ii) Monochromator used can be a prism, grating or a slit. A slit
may be used for picking out the desired wavelength. If the
slit is in a fixed position, rotation of the prism or the
grating is used to obtain the proper wavelength.
(iii) Absorption cell can be a glass cuvette (for visible range) or
quartz (both UV and visible range) cuvette. Cells may be
rectangular or cylindrical.
(iv) Detector’s role is to respond to change in wavelength. For
this photomultiplier tube is used.
Photomultiplier tube is a photoemissive device in which the
absorption of a photon results in the emission of an electron.
There is an electron multiplier which amplify the electrons
generated by a photocathode exposed to a photon flux. Thus it
is useful for the light detection of very weak signals.
Single Beam Spectrophotometer

(i) This is now less commonly used as reproducibility is a

problem sometimes.
(ii) There is only one sample compartment.
(iii) A light beam from a continuous source passes
through a colliminating lens and after that through a
filter. It becomes monochromatic and the beam then
passes through the sample solution and reaches the
detector.
This process is done with the blank first and then the
sample turn by turn.
Double Beam Spectrophotometer
 Double Beam Spectrophotometer
(i) In this there are two sample compartments.
(ii) Both the reference and the analyte can be put simultaneously for
measurement.
(iii) A combination of light source gives the entire spectrum.
(iv) The combined output of these two lamps is focused onto a
diffraction grating which behaves as a monochromator.
(v) A single beam is focused onto a rotating disc where due to the
rotation of the disc, the light falls on the sample and the analyte.
(vi) When the light hits the rotating disc, one of the three things can
happen:
(a) If it hits the transparent section, it will go straight and pass through
the cell containing the sample. After that it hits a mirror from where
it is reflected onto a second rotating disc. This disc is rotating in
such a way that when the light arrives from the first disc it meets
the mirrored section of the second disc. This bounces it onto the
detector.
(b) If it hits the mirrored section, it is bounced down and after being
reflected by a mirror passes through the reference cell. After that it
hits the second rotating disc which is rotating in such a way that it
meets the transparent section and reaches the detector.
(c) If the light meets the black section, it is blocked and for a very short
while and no light passes through the spectrophotometer.
Beer Lambert’s Law:

(i) For each wavelength of light passing through the

spectrophotometer, the intensity of the light passing
through the reference cell is measured.This is I˳
(ii) The intensity of light passing through the sample
cell is also measured which is I.
(iii) If I < I˳

sample has absorbed some light and computer

converts this into absorbance of the sample A.
𝐼˳
A = log
𝐼
A = εcl
The recorder plots absorbance against wavelength.
Flame Photometer
Flame Photometer
 Principle:

The flame transform the solid or liquid into vapor state and
decomposes it to simpler molecules or atoms.
On dispersing the material in a flame, water or solvent is
evaporated (aspirated) and dry salt is left in the flame.
On further heating at a higher temperature, the dry salt is
vaporized and the molecule is dissociated to neutral atoms.
These are excited by the thermal energy of the flame.
When the excited atoms return to the ground state, they
emit radiations.
A particular element emits characteristic spectra of its own
at a particular wavelength.
The line spectra are obtained for atoms whereas the
molecules give band spectra.
For example: Sodium (590 nm)
Potassium (766.5 nm)
Calcium (423 nm)
Flame Photometer
Flame Photometer
Flame Temperature (°C)
Acetylene-Oxygen 3200
Propane-Air 1800
Natural Gas-Air 1700
Acetylene-Nitrous Oxide 2700

The measurement of intensities of spectral lines is dependent

upon:
(1) the amount of salt impregnated in the flame
(2) the amount of salt dissociated
(3) the extent it is ionized
(4) the number of atoms excited
(5) the changes of transition from the excited to ground state
and self absorption
Flame Photometer
The are many factors which contribute to variation in emission
intensity:
(1) Due to the formation of metal hydroxides of alkali metals in
the flame.

Oxyacetylene provides an atmosphere which is more favorable

for the existence of free atoms in those cases where stable
monoxide molecules can be formed.
Interferences in Flame Photometry:
(1) Spectral Interferences: These are encountered while isolating the
desired radiant energy.To remove this, monochromator is used.

(2) Background Emission: This may occur due to certain constituents

present in the flame and can be eliminated by using a blank solution.

(3) Self Absorption: During the emission, some of the energy may be
lost due to self absorption and hence the strength of the spectral line
is weakened. Low conc. can be used.

(4) Ionization: Oxygen gas flame possesses sufficient energy to ionize

the alkali and alkaline earth metals.

(5) Interference due to anions: H2SO4, H3PO4, HNO3 lower the

metallic emission.
The use of a protective chelating agents can reduce these types of
interferences.

(6) Solution properties: Vapor pressure and surface tension influences

droplet size. Use of LiCl can eliminate this effect. Non-ionic
surfactants are also useful in increasing the surface tension of the
solution.
Chemical Interferences in Flame Photometry:
These results from the chemical reactions taking place during the
atomization which in turn change the properties of absorption.

These may occur due to the formation of compounds of low

volatility, dissociation equilibria, ionization in flames.

The anions form compounds of low volatility and reduce the rate of
atomization. PO43- and SO42- reduce atomization of calcium.
High temperature flames can be used.
Use of releasing agents can eliminate this interference.

For example: addition of Sr or La reduces the PO43- content.

protective agents like EDTA eliminate interference of
several ions like Al3+ by process of masking.
Flame Photometry:
Atomic Absorption Spectroscopy AAS:

This includes absorption of light by atoms which are then excited to

a higher level. When they come back to the ground state, emit
radiation which is characteristic of a particular element.

The source used is hollow cathode lamp. It consists of the electrode

of a metal which is to be analysed.
Several combinations are available:
(Ca, Mg, Al) (Fe, Cu, Mn) (Cu, Zn, Pb, Sn) (Cr, Co, Cu, Fe, Mn, Ni)

The atomiser is graphite furnace or high temperature flame.

Metals which can be analysed are Cu, Pb, Zn, Cd, Al, Ti Be, Fe, Co,
Ni

Thus AAS is composed of the following four components:

(1) Atomisation unit
(2) Source of radiation
(3) Optical system
(4) Detector
Interferences in AAS:
(1) Due to the overlap of absorptions of interfering species with the
analytical absorption.

(2) The presence of combustion products give spectral interference

and can be reduced by using a blank.
(3) A source of absorption or scattering from a sample causes an
error in absorbance and hence concentration. This can be avoided
by variation of temperature and fuel to oxidant ratio in the flame.
Thus background correction can be done.
(4) Chemical interferences.
Reference
 Analytical chemistry by S. M. Khopkar