Unit-5

Instrumental methods
and applications
 Electromagnetic spectrum
The electromagnetic spectrum is the full range of
electromagnetic radiation, organized by frequency or wavelength. The
spectrum is divided into separate bands, with different names for the
electromagnetic waves within each band. From low to high frequency these
are: radio waves, microwaves, infrared, visible light, ultraviolet, X-rays,
and gamma rays. The electromagnetic waves in each of these bands have
different characteristics, such as how they are produced, how they interact
with matter, and their practical applications.
 SPECTROSCOPY
• spectroscopy is one of the most powerful tool
available for the study of atomic and molecular
structure and is used in the analysis of a most of
the samples.
• Spectroscopy deals with the study of interaction
of electromagnetic radiation with the matter.
• During the interaction, the energy is absorbed
(or) emitted by matter.
• The measurement of this radiation & frequency
are made using spectroscopy.
 Types of spectroscopy
The study of spectroscopy can be carried out
under the following types.
1.Atomic spectroscopy
2.Molecular spectroscopy
1.Atomic spectroscopy:- It deals with the interaction of
the electromagnetic radiation with atoms. During which
the atoms absorb radiation and gets excited from the
ground state electronic level to another.
2.Molecular spectroscopy:-It deals with interaction of
electromagnetic radiation with molecules. This results
in transition between rotational, vibrational and
electronic energy levels.
 Differences between atomic & molecular
spectroscopy
ATOMIC SPECTROSCOPY MOLECULAR SPECTROSCOPY

1.It occurs from the interaction 1.It occurs from the interaction
of atoms at electromagnetic of molecules of electromagnetic
radiation. radiation.

2.Atomic spectra is a line 2.Molecular spectra is a

spectra. complicated spectra.

3.It is due to electronic transition 3.It is due to vibrational,

in an element. rotational and electronic
transition in a molecule.
Spectrum types:-
1.Absorption spectrum:-
• Consider a molecule having only two energy levels E 1
and E2 as shown in figure.
E2

E1
• When a beam of electromagnetic radiation is allowed to
fall on the molecule in the ground state, the molecule
absorbs photon of energy and undergoes a transition
from lower energy level to higher energy level.
• The measurement of this decrease in the intensity of the
radiation is the basis of absorption spectroscopy. The
spectrum thus obtained is called absorption spectrum.
2.Emission spectrum:-
• If the molecule comes down from excited state
to the ground state with the emission of
photons of energy (hʋ), the spectrum is called
emission spectrum.
E2

E1
 Absorption of radiation
When electromagnetic radiation is passed through
a matter, the following changes occur.
• As the photons of electromagnetic radiations are
absorbed by the matter, electronic transition, vibrational
changes (or) rotational changes may occur.
• After absorption, molecules get excited from the
ground state to excited state.
• Then they liberate energy quickly in the form of heat
(or) re-emit electromagnetic radiation.
• But in some cases, the portion of electromagnetic
radiation, which passes into the matter, instead of being
absorbed may be scattered or reflecting or re-emitted.
• When the electromagnetic radiation is absorbed or scattered,
it may undergo changes in polarisation or orientation.
• In some cases the molecules absorbs radiation and get
excited.
a) Fluorescence:- If the excited molecules re-emits the
radiation almost instantaneously (with in 10 -8 sec) it is
called fluorescence.
b) Phosphorescence:- If the excited molecules re-emits the
radiation after some time (slowly) it is called
phosphorescence.
Factors affecting absorbance:-
1.The nature of the absorbing molecules.
2.The concentration of the molecules.
3.The length of the path of the radiation through the matter.
Limitations of Beer’s – Lambert’s law:-
• Beer-Lambert’s law is not obeyed if the radiation
used is not monochromatic.
• It is applicable only for dilute solutions.
• The temperature of the system should not be
allowed to very to a large extent.
• It is not applied to suspensions.
• Deviation may occur, if the solution contains
impurities.
• Deviation also occurs if the solution undergoes
polymerisation (or) dissociation.
 UV-VISIBLE SPECTROSCOPY
• Instrumentation
• Principle
• Types of transitions involved in organic
molecules
• Applications
 UV-Visible instrumentation:-
Light --> Mono chromater --> Sample holder-->
Detector--> Recorder.
Components of instrument:
1.Light source. a)Deuterium lamp (UV 200-400nm)
b)Tungsten halogen lamp (Visible
400-700nm)
2.Mono chromater. a) Filters b) Prisms c) Gratings
3.Sample holder- Cuvette
4.Detector
5.Recorder
 PRINCIPLE
Absorption spectroscopy:-
• UV-Visible spectra arises from the transition of valence electrons
with in a molecule or ion from a lower electronic energy level
(Eo) to higher electronic energy level (E1).
• This transition occurs due to the absorption of UV (Wave length
1-400nm)or Visible (Wave length 400-700nm) region of the
electronic spectrum by a molecule or ion.
• The actual amount of energy required depends on the difference
in energy b/w the Eo and E1 of the electrons.
ΔE = E1-Eo = hʋ
• Electronic transition depends on the electronic structure of the
absorbing medium.
• The absorption of UV-Visible radiation in organic molecule is
mainly due to presence of certain functional groups.
• The two important groups responsible for absorption and position of
absorption in UV-Visible spectra are
1.Chromophores
2.Auxochromes.
1.Chromophores:-
• The presence of one or more unsaturated linkages (π-electrons) in a
compound is responsible for the colour of the compound, these
linkages are referred to as chromophores.
• Chromophores undergo π-π* transitions in the short wave length
regions of UV radiation.
Ex:- Ethylene, Acetylene, Cyanide group, Ketone group etc.
2.Auxochromes:- It refers to an atom or a group of atoms which does
not give rise to absorption band on its own, but when conjugate to
chromophore will cause a red shift.
Ex:- OH group, NH2 group, Cl, Br , I , etc.,
Illustration: In Chloro ethylene (CH2= CHCl)
 Possible electronic transitions
Types of transitions involved in organic molecules:-
The energy level diagram for a molecule is shown in the
figure. The energy values for different transitions are in the
following order.
n-π < π-π* < n-σ* < < σ-σ*
Fig: UV-Vissible spectrum of unsaturated Ketone
 Applications
• Predicting relationship b/w different groups.
• Qualitative analysis: UV absorption spectroscopy is
used for characterizing and identification of aromatic
compounds.
• Detecting of impurities in sample.
• Quantitative analysis of compounds.
• To determine the molecular weight of the compound.
• Study of tautomeric equilibrium
• Studying kinetics of chemical reactions.
• Determination of calcium in blood serum.
INFRARED SPECTROSCOPY

 Introduction
 Principle
 Molecular vibration
 Instrumentation
 Application
 Advantages
 Disadvantages
 IR SPECTROSCOPY
 The study of absorption of infrared radiation, which
causes vibrational transition in the molecule is known as
Vibrational spectroscopy.

Mainly used in structure elucidation to determine the

functional groups.
 PRINCIPLE
Molecules are made up of atoms linked by chemical bonds. The
movement of atoms and the chemical bonds is like spring and balls
(vibration).

This characteristic vibration are called Natural frequency of

vibration.
Principle ...
 When energy in the form of infrared radiation is applied then

it causes the vibration between the atoms of the molecules.

Applied infrared frequency = Natural frequency of vibration,

Absorption of IR radiation takes place, a peak is observed.

Different functional groups absorb characteristic

frequencies of IR radiation, gives the characteristic
peakvalue.

IR spectrum of a chemical substance is a finger print of a

 Molecular vibrations
Stretching vibrations Bending vibrations

Symmetrical Asymmetrical In plane Out plane

scissoring
scissoring rocking wagging twisting
1. Stretching vibrations:

 Vibration or oscillation along the line of bond

 Change in bond length

 a) Symmetrical stretching:

 Two bonds increase or decrease in length

simultaneously.

H
 b) Asymmetrical stretching

 one bond length is increased and other is

decreased.

H
 2. Bending vibrations

• Vibration or oscillation not along the line of bond

• Deformations

• Bond angle is altered

 A) In plane bending

i) Scissoring:
 2 atoms approach each other
 Bond angles are decrease

H
 ii) Rocking:
 Movement of atoms take place in the same direction.

H
 B) Out plane bending

i .Wagging:
Two atoms move to one side of the plane. They move
up and down the plane.

H H

C
C

H
H

ii. Twisting:
One atom moves above the plane and another atom
moves below the plane.
INSTRUMENTATION:

2 types - IR frequencies are handled.

1) dispersive type -

IR is separated into individual frequencies by

dispersion, using a grating monochromator.

2) interferometric type (FTIR) -

IR frequencies are allowed to interact to produce an
interference pattern and this pattern is then analyzed, to
determine individual frequencies and their intensities.
The main parts of IR spectrometer are as follows:

1. Radiation source
2. Sample cells and sampling of substances
3. Monochromators
4. Detectors
1.IR source
 Nernst Glower -Composed of rare earth
oxides (Zirconia, Yttria & Thoria )
 Globar - silicon carbide

 W filament lamp

 Special high pressure Hg arc lamp

 CO2 laser
 2. SAMPLE CELL

• Made up of alkali halides like NaCl or KBr .

•Aqueous solvents cannot be used -they dissolve alkali
halides.
• Only organic solvents like chloroform is used.
•IR spectroscopy has been used for the characterization of
solid, liquid, gas samples
 3. Monochromators

 convert polychromatic light into monochromatic light.

 They are of 3 types.

a) metal halide prisms
b) NaCl prisms
c) gratings
 4. DETECTOR

 convert the radiation into electrical signal.

 Two Types Of Detectors
1. Photon detector
2. Thermal detector
•Biological and biomedical fields like detection of water in

biological membranes.

• Analysis of Aircraft exhausts.

• Measurement of toxic gas in fuels.

• Combustion.

• Gas analysis.
 Applications
• Compositional analysis of organic, inorganic and
polymers.

• Detection of Impurities.

•
 ADVANTAGES:

 Easy to use

 Analysis time typically < 10 minutes

 Inexpensive, found in most labs.

 DISADVANTAGES:

Sensitive to the absorption of water .If there is significant

moisture in the sample the penetration distance of the light
decreases.