Principle

VISIBLE AND ULTRAVIOLET (UV) SPECTROSCOPY

A
Visible and Ultraviolet (UV) spectra arise from the transition of valence electrons within a
molecule or ion from a lower electronic energy level (ground state E0) to higher electronic energy
level (excited state E1).
This transition occurs due to the absorption of UV (wavelength 100-400 nm) or visible (wave
length 400-750 nm) region of the electronic spectrum by a molecule (or) ion.
The actual amount of energy required depends on the difference in energy between the
ground state and the excited state of the electrons.
E1 − Eo = hν.

INSTRUMENTATION:

1. Radiations source
In visible – UV spectrometers, the most commonly used radiation sources are hydrogen (or)
deuterium lamps.

2. Monochromators

The monochromator is used to disperse the radiation according to the wavelength. The
essential elements of a monochromator are an entrance slit, a dispersing element and an exit slit. The
dispersing element may be a prism or grating (or) a filter.

3. Cells (sample cell and reference cell)

The cells, containing samples or reference for analysis, should fulfil the following conditions
 They must be uniform in construction.
 The material of construction should be inert to solvents.
 They must transmit the light of the wavelength used.

4. Detectors
There are three common types of detectors used in visible UV spectrophotometers. They are Barrier
layer cell, Photomultiplier tube, Photocell.
The detector converts the radiation, falling on which, into current. The current is directly proportional
to the concentration of the solution

5. Recording system

The signal from the detector is finally received by the recording system. The recording is
done by recorder pen.

WORKING OF VISIBLE AND UV SPECTROPHOTOMETER:

 The radiation from the source is allowed to pass through the monochromator unit.
 The monochromator allows a narrow range of wavelength to pass through an exit slit.
 The beam of radiation coming out of the monochromator is split into two equal beams.
 One-half of the beams (the sample beam) is directed to pass through a transparent cell
containing a solution of the compound to be analysed.

Dr. X. JOSEPH RAJ, Assistant Professor, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology
  The another half (the reference beam) is directed to pass through an identical cell that
contains only the solvent. The instrument is designed in such a way that it can compare the
inten sities of the two beams.
 If the compound absorbs light at a particular wavelength, then intensity of the sample beam
(I) will be less than that of the reference beam (Io).The instrument gives output graph, which
is a plot of wave length Vs absorbance of the light. This graph is known as an absorption
spectrum.

Applications of UV-Visible:
 Predicting relationship between different groups
 Qualitative analysis
 Detection of impurities
 Quantitative analysis
 Determination of molecular weight

B
Dr. X. JOSEPH RAJ, Assistant Professor, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology
 COLORIMETRY
Colorimetry is concerned with the visible region (400-750 nm) of the spectrum. The instrument, used
for measuring absorption of radiant energy in the visible region from the substances is called
colorimeter.
Principle:
This method is convenient for the coloured substances or coloured solutions. The intensity of
colour can be easily measured by using a photo electric colorimeter, from which the concentration of
coloured solution can be obtained by using Beer-Lambert’s law.
If the substance is colourless, then a suitable complexing agent is added to the solution so that
a coloured complex is obtained, which can absorb the light.
Example
For the estimation of cuprous ions, complexing agent, ammonium hydroxide, is added to get blue
coloured solution.
Instrumentation:
Components:
1. Radiation sources
The wavelength range of visible light lies between 400-750 nm. In this region, a tungsten-
filament lamp is most widely used.

2. Filter (or) monochromator

It is a instrument, which allows the light of the required wavelength to pass through, but
absorbs the light of other wavelengths.

3. Slits
(a) Entrance slit: It provides a narrow source of the light.
(b) Exit slit: It selects a narrow band of dispersed spectrum for observation by the detector.

4. Cell
The cell, holding the test sample (usually a solution), should be transparent. For visible region
the cell is made of colour-corrected fused glass.

5. Detector
It is used for measuring the radiant energy transmitted through the sample. Photosensitive
devices are used to detect radiations. These detectors produce current, which is directly proportional
to the intensity of the incident radiation.

6. Meter
It is used to measure directly the fraction of light absorbed.

WORKING OF COLORIMETER:
 In a colorimeter, a narrow beam of light is passed from radiation source through the test
solution (cell) towards a sensitive detector (photocell).
 Usually colorimeter is provided with the arrangement of filter and slits, which select the light
of required wave length.

Dr. X. JOSEPH RAJ, Assistant Professor, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology
  The detector (photocell) generates the current, which is proportional to the amount of light
transmitted by the solution.
 The amount of light transmitted depends on the depth of colour of the test solution. Thus, the
current from the Photocell will be more when the light transmitted is more. This is possible
only if the coloured solution is most dilute.

Current α Light transmitted α 1/Concentration

 The transmitted light is allowed to send through a meter, which is calibrated to show not the
fraction of light transmitted but the fraction of light absorbed. The light absorbed is
proportional to the concentration of the test solution.

Applications of colorimetry:

 Molar compositions of complexes can be determined.

 The instability constants of metal complexes are also determined.
 Dissociation constants (Pk) of an indicator can be determined.
 Structure of inorganic compounds, complexes (cis & trans isomer) can be determined.
 Molecular weight of a compound can also be determined using colorimetric measurements.

Dr. X. JOSEPH RAJ, Assistant Professor, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology
C
 FLAME PHOTOMETRY (or) FLAME EMISSION SPECTROSCOPY

Flame photometry is a method in which, the intensity of the emitted light is measured, when a
atomised metal is introduced into a flame. The wavelength of the colour tells us what the element
is, and the intensity of the colour tells us how much of the element is present.

Theory (or) Principle:

When a metallic salt solution is introduced into a flame, the following processes will occur.
(i) The solvent is evaporated leaving behind the solid salt particle.
(ii) The salt is vapourised into the gaseous state and dissociated into atoms.
(iii) Some of the atoms from the ground state are excited to higher energy state by absorbing
thermal energy from the flame.
The excited atoms, which are unstable, quickly emit photons of different wave lengths and
return to the lower energy state. Then the emitted radiation is passed through the filter, which permits
the characteristic wavelength of the metal under examination. It is then passed into the detector, and
finally into the recorder.

Instrumentation:

The various components of the flame photometer are described as follows.

1. Burner: The flame must possess the following characteristics.
 It should evaporate the solvent from the sample solution.
 It should decompose the solid into atoms.
 It should excite the atoms and cause them to emit radiant energy.
2. Mirror: The radiation from the flame is emitted in all directions in space. In order to increase the
amount of radiation reaching the detector, a convex mirror is used which is set behind the burner.
3. Slits
Entrance slits: It is kept between the flame and monochromator. It permits only the radiation coming
from the flame and mirror.
Exit slit: It is kept between the monochromator and detector. It prevents the entry of interfering lines.

4. Monochromator (or) Prism (or) Grating (or) Filter

It allows the light of the required wave length to pass through, but absorbs the light of other
wavelengths.
5. Detector
The radiation coming out from the filter is allowed to fall on the detector, which measures the
intensity of the radiation falling on it. Photo multiplier (or) photocell is used as detector, which
converts the radiation into an electrical current.

Dr. X. JOSEPH RAJ, Assistant Professor, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology
 6. Amplifier & Recorder
The current coming out from the detector is weak, so it is amplified and recorded.
Working of Flame photometer:
 Air, at a given pressure, is passed into an atomiser. The suction so-produced draws some
solution of the sample into the atomiser.
 Air + sample solution is then mixed with fuel gas in the mixing chamber. The Air + sample
solution + fuel gas mixture is then burnt in the burner.
 The radiation, emitted by burner flame, is passed successively through the lens, filter,
detector, amplifier and finally into a recorder.

Applications of flame photometry

1. Qualitative Analysis
 The elements of group I & II. (K, Na, Li, Ca, Mg, etc) can be detected visually from the
colour of the flame. Eg; Na Brick red colour at 422nm and K red colour at 766nm.
 Non-radiating elements such as carbon, hydrogen and halides cannot be detected using this
method.
2. Quantitative analysis
 The amount of the elements in group I & II (alkali & alkaline-earth metals) can be determined
from the sample.
 Certain transition elements, such as Cu, Fe & Mn can also be determined using flame
photometry.
3. Other applications
 The measurement of these elements is very useful in medicine, agriculture and plant science.
 Flame photometry is extensively used in the analysis of biological fluids and tissues
 In soil analysis the elements like, Na, K, Al, Ca, Fe, etc., are determined.
 Industrial and natural waters, petroleum products, cement, glass, and metallurgical products
can also be analysed by this method.
Limitations of flame photometry:
 It cannot be used for the determination of all metal atoms and inert gases.
 Only liquid samples must be used.

ATOMIC ABSOPTION SPECTROSCOPY:

D
Dr. X. JOSEPH RAJ, Assistant Professor, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology
 Atomic absorption spectroscopy is based on the atomization of the sample followed by absorption of
characteristic radiation by the ground state gaseous atoms.
When the light of the required wavelength is allowed to pass through a flame, having atoms of the
metallic species, part of that light will be absorbed and the absorption will be proportional to the
concentration of the atom in the flame.
Thus, in atomic absorption spectroscopy, the amount of light absorbed is determined.

Instrumentation:

Various components

1. Radiation source

The radiation source should emit, stable, intense, characteristic radiation of the
element to be determined. The hollow cathode lamp, which consists of a glass tube containing noble
gases like a argon (anode) and hollow cathode, made of the analyte metal, is generally used.

2. Chopper

A rotating wheel is interposed between the hollow cathode lamp and the flame. It breaks the
steady light, from the lamp, into an pulsating light (because the recorder will record only the
pulsating (alternating) current).

3. Burner (or) Flame

The flame is used for converting the liquid sample into the gaseous state. It converts the
molecule into atomic vapour. Two types of burners are used

1. Total consumption burner.

2 Premixed burner.

4. Nebulisation of the liquid sample

Before the liquid sample enters the burner, it is first of all converted into small droplets. This
method of formation of small droplets from the liquid sample is called nebulisation.

5. Monochromators

The monochromators select a given absorbing line from the spectral lines emitted from the
hollow cathode. The most common monochromators are

(i) Prisms.

(ii) Gratings.

6. Detectors

The photomultiplier tube is a most suitable detector. When the photon strikes the
photomultiplier tube, an electric current (emf) is produced.

7. Amplifier

Dr. X. JOSEPH RAJ, Assistant Professor, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology
 The electric current, from the photomultiplier detector, is fed into the amplifier, which
amplifies the electric current many times.

8. Read-Out Device (or) Recorder

The signal coming out from the amplifier is recorded using chart recorder (or) digital read-out
devices. The characteristic radiation, obtained from the hollow cathode lamp, is passed through a
flame into which the sample is aspirated. The metallic compounds are decomposed into atoms of the
element to be measured.

The atoms absorb a fraction of radiation in the flame. The unabsorbed radiation from the
flame is allowed to pass through a mono chromator. From the monochromator the unabsorbed
radiation is led into the detector. From the detector, the output is amplified and measured on a
recorder.

IR SPECTROSCOPY
E
Dr. X. JOSEPH RAJ, Assistant Professor, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology
 Principle:
An IR spectrum is produced by the absorption of energy by a molecule in the infrared region
and the transitions occur between vibrational levels. So, IR spectroscopy is also known as vibrational
spectroscopy.
Range of Infrared Radiation
The range in the electromagnetic spectrum extending from 12500 to 50 cm− 1 (0.8 to 200 μ)
is commonly referred to as the infrared. This region is further divided into three sub regions.

Molecular Vibrations and Origin of IR Spectrum:

Since atoms in a molecule are continuously vibrating, molecules are also vibrating. There are two
kinds of fundamental vibrations in the molecule.
Stretching vibrations: During stretching the distance between two atoms decreases or increases, but
bond angle remains unaltered.
Bending (or) deformation vibrations:During bending bond angle increases and decreases but bond
distance remains unaltered.
When IR light of the same frequency is incident on the molecule, energy is absorbed resulting in
increase of amplitude of vibration.
When the molecule returns from the excited state to the original ground state, the absorbed energy is
released as heat.
Thus every compound shows characteristic absorption bands in the IR region of the spectrum.
Different functional groups produce easily recognisable band at definite positions in the IR spectral
range (12500 to 50 cm− 1).
Finger Print Region: The vibrational spectral (IR spectra) region at 1400 − 700 cm− 1 gives very
rich and intense absorption bands. This region is termed as fingerprint region. The region 4000 −
1430 cm− 1 is known as Group frequency region.

USES OF FINGERPRINT REGION:

1. IR spectra are often characterized as molecular finger prints, which detect the presence of
functional groups.
2. Fingerprint region is also used to identify and characterize the molecule just as a fingerprint can be
used to identify a person.

IR SPECTROSCOPY INSTRUMENTATION:
1. Radiation source
The main source of IR radiation is,
1) Nichrome wire
2) Nernst glower (filament containing oxides of Zr, Th, Ce)
When they are heated electrically at 1200 to 2000°C, they glow and produce IR radiation.
2. Monochromator

Dr. X. JOSEPH RAJ, Assistant Professor, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology
 It allows the light of the required wave length to pass through, but absorbs the light of other
wave length.
3. Sample Cell
The cell, holding the test sample, must be transparent to IR radiation.
4. Detector
IR detectors generally convert thermal radiant energy into electrical energy. There are so
many detectors, of which the followings are important.
(a) Photoconductivity cell.
(b) Thermocouple.
(c) Pyroelectric detectors.
5. Recorder
The recorder records the signal coming out from the detector.

WORKING OF IR SPECTROPHOTOMETER:
The radiation emitted by the source is split into two identical beams having equal intensity.
One of the beams passes through the sample and the other through the reference sample.
When the sample cell contains the sample, the half-beam travelling through it becomes less intense.
When the two half beams (one coming from the reference and the other from the sample) recombine,
they produce an oscillating signal, which is measured by the detector. The signal from the detector is
passed to the recording unit and recorded.

Applications of IR spectroscopy
1. Identity of the compound can be established
2. Detection of functional groups:
3. Testing the purity of a sample.
4. Study of progress of a chemical reaction
5. Determination of shape (or) Symmetry of a molecule

Dr. X. JOSEPH RAJ, Assistant Professor, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology

Dr. X. JOSEPH RAJ, Assistant Professor, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology