SPECTROSCOPY

SPECTROSCOPY
 Spectroscopy is defined as the area of science
concerned with the absorption, emission, and
scattering of electromagnetic radiation by atoms and
molecules, which may be in the gas, liquid, or solid
phase.
 Spectroscopy is the study of the interaction of
electromagnetic radiation with the matter.
 During the interaction the energy is absorbed or
emitted by the matter.
 ELECTROMAGNETIC RADIATIONS

 Electromagnetic radiation—light—is a form of energy

whose behavior is described by the properties of both
waves and particles.
 It has wave properties like refraction, reflection etc.
 particle properties like absorption ,emission etc.
 Electromagnetic (EM) radiation is a form of energy
propagated through free space or through a material
medium in the form of electromagnetic waves.
ELECTROMAGNETIC RADIATIONS
 WAVE PROPERTIES OF
ELECTROMAGNETIC RADIATIONS
 SPECTRUM
 A spectrum is defined as the characteristic wavelengths

of electromagnetic radiation (or a portion thereof) that is

emitted or absorbed by an object or substance, atom, or
molecule.
Example
 Examples of a spectrum include the rainbow, the
emission colors from the Sun, and the infrared
absorption wavelengths from a molecule.
 ELCRTOMAGNETIC
SPECTRUM
 The electromagnetic (EM) spectrum is the range of all
types of EM radiation.
 Electromagnetic spectrum, the entire distribution of
electromagnetic radiation according to frequency or
wavelength.
 All electromagnetic waves travel at the speed of light in a
vacuum.
 ELCRTOMAGNETIC
SPECTRUM

 Gamma rays: These are part of the electromagnetic

spectrum which includes visible as well as invisible
radiations. Gamma rays have the highest frequency
which is greater than 10 Hz and the wavelengths are
short which is below 100pm.
 ELCRTOMAGNETIC
SPECTRUM
 X-rays: These are parts of the electromagnetic spectrum
and their wavelengths are in the range 0.01nm to 10nm
and their frequency is in the range 30 PHz to 30 Ehz. X-
rays find applications in the healthcare industry.

 Ultraviolet (UV) rays: These are the part of the

electromagnetic spectrum with a wavelength between
10nm to 400nm. Their frequency is between 800 THz to
30 PHz. When there is a continuous exposure of the skin
to UV rays, they will result in skin and eye diseases.
 ELCRTOMAGNETIC
SPECTRUM
 Visible light: These are part of the electromagnetic
spectrum which lies between UV rays and infrared rays.
The wavelengths of visible light are 400nm to 700nm and
the frequency range is between 400 THz to 800 THz.

 Infrared radiation: These are the part of the

electromagnetic spectrum and their wavelength range is
between 710nm to 1mm. Infrared radiation is also referred
to as thermal waves as they have heat-inducing property.
 ELCRTOMAGNETIC
SPECTRUM
 Microwaves: These are part of the electromagnetic
spectrum and are known as microwave radiation. The
frequency range between 300 MHz to 300 GHz. Their
wavelength range is between 1nm to 30cm. In the
electromagnetic spectrum series, microwaves are
between infrared radiation and radio waves.
 Radio waves: These are part of the electromagnetic
spectrum and have the longest wavelengths and huge
band of frequencies. The frequency range of radio waves
is 3 kHz and 300 GHz. Radio waves find applications in
radio communication, broadcasting etc.
 ELCRTOMAGNETIC
SPECTRUM
Wave length Frequency in
S. No. Type of radiation Applications
in meters Hz

Medicine for killing

1 Gamma rays 10-11 1019
cancer cells

Used to view
2 X ray 10-9 1017 inside of the body
and object
Absorbed by the
3 UV 10-7 1015 skin ,used in fluorescent
tubes
Making things able
4 Visible 10-6 1014
to see
Transmit heat from
5 IR 10-5 1013 sun ,fires ,radiator
s
Used in cooking
6 Micro waves 10 -3
10 11
radar ,telephone
and other signals
radio
7 Radio waves (low energy) 10 2
10 6
communication,
broadcasting etc
 SPECTROSCOPY
 Spectroscopy is a general term used for the instrumental
processes by which information about molecular structure
is obtained through careful analysis of the absorption,
scattering, or emission of electromagnetic radiation by
compounds.
 Spectroscopy is the study of the interaction of
electromagnetic radiation with the matter.
 During the interaction the energy is absorbed or emitted
by the matter.
 It is used to study the atomic and molecular structure of
the substance.
 TYPES OF SPECTROSCOPY
Spectroscopy is two types
1. Atomic spectroscopy.
2. Molecular spectroscopy.

1) Atomic spectroscopy
 Atomic spectroscopy deals with the interaction of the
electromagnetic radiation with atoms.

 During which the atoms absorb radiation and gets

excited from the ground state electronic energy level
to another.

2) Molecular spectroscopy
 Molecular spectroscopy deals with the interaction of the
electromagnetic radiation with molecules.

 This results in transition between rotational, vibrational

and electronic energy levels.
 UV SPECTROSCOPY

o The wave length range of UV radiation starts 10 nm to

400 nm.
o The ultra violet region is sub divided into two spectral
regions
 THEORY OF ULTRA VIOLET SPECTRA

 Ultraviolet absorption spectra arise from transition of

electron with in a molecule from a lower level to a higher
level.
 A molecule absorb ultraviolet radiation of frequency (𝜗),
the electron in that molecule undergo transition from
lower to higher energy level
 LAWS OF ABSORPTION & EXPRESSION
 There are two laws which govern the absorption of light by
the molecules.
 Lambert’s law
 Beer’s law

LAMBERT’S LAW
 When a beam of monochromatic light is passed through a
solution of an absorbing substance, the rate of decrease
of intensity of radiation (‘dI’) with thickness of the
absorbing solution (‘dx’) is directly proportional to the
intensity of incident radiation (I).
 Mathematically, the law is expressed as,
 LAWS OF ABSORPTION & EXPRESSION

Where, I = Intensity of radiation.

x = Thickness of the medium.
k = Proportionality constant or absorption
coefficient.
Let I0 be the intensity of radiation before entering the
absorbing medium (x=0)
Then I is the intensity of radiation after passing through any
thickness, say x of the medium can be calculated as:
On integrating the above expression between the limits, I = Io
at x = 0 and I = I at x = x
We get,
LAWS OF ABSORPTION & EXPRESSION
LAWS OF ABSORPTION & EXPRESSION

The intensity of the radiation absorbed, Iabs is given by

I abs I 0  I

I abs I 0  I 0 e  kx

I abs I 0 1  e  kx

 LAWS OF ABSORPTION & EXPRESSION

BEER’S LAW:
When a beam of monochromatic light is passed
through a solution of an absorbing substance, the rate of
decrease of intensity of radiation (‘dI’ ) with thickness of
absorbing solution (‘dx’) is directly proportional to the
intensity of incident radiation (I), as well as concentration
of the solution (c).
Mathematically, it is expressed as
dI l
 k IC
dx
 LAWS OF ABSORPTION & EXPRESSION

Where, C = Concentration of the solution

kl = Molar absorption coefficient
Suppose I0 be the intensity of the radiation before
entering the absorbing solution(when x=0).
Then the intensity of radiation, I after passing
through the thickness x of the medium can be calculated as:
On integrating the above expression between
the limits, I = Io at x = 0 and I = I at x = x
I x x
We get, dI
  Cdx
l
 k
I0
I x 0
LAWS OF ABSORPTION & EXPRESSION

I  k l cx
e
I0
 k l cx
I I 0 e

On combining the two laws the Beer-Lambert law can

be formulated as

I
log  ECl  A
I0
 LAWS OF ABSORPTION & EXPRESSION

This equation is known as Beer – Lambert’s Law.

Where, I0 = Intensity of incident light

I = Intensity of transmitted light
c = Concentration of solution
l = Path length of the sample
Ɛ = Molar extinction coefficient or molar absorbtivity
A = Absorbance.
 APPLICATION &LIMITATIONS
APPLICATION OF BEER-LAMBERTS LAW
By using Beer-Lamberts law, the concentration of unknown
solution can be determined.
LIMITATIONS OF BEER – LAMBERT’S LAW
1. This law is not obeyed if the radiation used in polychromatic.
2. This law is not obeyed when fluorescent compounds are
present.
3. It is not applied to suspensions.
4. It is applicable only for dilute solutions.
5. Deviation may occur, if the solution contains impurities.
6. Deviations may also occur if the solution undergoes association
or dissociation.
7. This law is not obeyed when different forms of the absorbing
molecules are in equilibrium as in keto-enol tautomers.
 FRANCK CONDON PRINCIPLE

 According to this principle, “An electronic transition takes

place so rapidly that a vibrating molecule does not

change its internuclear distance appreciably during the

transition.”

 Since the electrons move so much faster than the nuclei

that during the electronic transition, the nuclei do not

change their positions.

 Hence an electronic transition may be represented by a

vertical line on the plot of potential energy versus

 FRANCK CONDON PRINCIPLE

 Classically, the Franck–Condon principle is the

approximation that an electronic transition is most likely
to occur without changes in the positions of the nuclei in
the molecular entity and its environment. The resulting
state is called a Franck–Condon state.
 The Franck-Condon Principle Electronic transitions are
“vertical”, that is the nuclei don’t move while the
electron(s) are being excited.
FRANCK CONDON PRINCIPLE
 UV INSTRUMENTATION
 Principle: Visible and UV spectra arises from the
transition of valency electrons within a molecule or
ion from a lower electronic energy level (E0) to higher

electronic energy level (E1).

 This transition occurs due to the absorption of UV or
visible radiation 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-E0 = hυ.
 UV INSTRUMENTATION
The various components of UV-visible

spectrometer are as follows.

1. Radiation source

2. Monochromator

3. Cells

4. Detectors

5. Recorder
INSTRUMENTATION
 INSTRUMENTATION
Radiation source: The most commonly used radiation
sources are hydrogen or deuterium lamps.
Monochromator: It 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.
Cells: The cells (containing sample and reference for
analysis) must transmit the light of the wavelength
used. The most commonly used cells are made of quartz
or fused silica.
Detectors: It converts the radiation, falling on which,
into current. Commonly used detectors are Barrier layer
cell, photomultiplier tube, photocell.
Recorder: The signal from the detector is recorded by
recorder pen.
 INSTRUMENTATION
Working:
The radiation from the source is allowed to pass
through the Monochromator which selects a narrow range of
wavelength. The beam of radiation coming out of the
Monochromator is split into two equal beams. One-half of the
beam is passed through the sample solution and the another
half is passed through the solvent (reference). The
instrument compares the intensity of the two beams. If the
compound absorbs light at a particular wavelength then the
intensity of the sample beam (I) will be less than that of the
reference beam (I0). The instrument gives output graph which

is a plot of wavelength vs absorbance. This graph is known as

absorption spectrum.
INFRARED SPECTROSCOPY
 INFRARED SPECTROSCOPY
• IR spectroscopy provides useful information about

the structure of molecule quickly over the other

methods.

•When infrared light is passed through a sample of an

organic compound, some of the frequencies are

absorbed while other frequencies are transmitted

through the sample.

•If we plot the percent absorbance or transmittance

against frequency, the result is an infrared spectrum.

INFRARED SPECTROSCOPY
PRINCIPLE
• Infrared spectra 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 :
IR = 650 – 4000 cm-1;
Far IR = lower than 650 cm-1;
Near IR = higher than 4000 cm-1
 FT IR SPECTROMETER

FTIR stands for Fourier Transform Infrared.

FTIR spectrometers consist of
• IR source
• Interferometer
• Sample cell or chamber
• Detector
• Laser.
FT IR SPECTROMETER
 FT IR SPECTROMETER
IR source: IR radiation is emitted from a glowing

black body source.

Common IR sources are: Silicon carbide rods

which are resistively heated and commonly known as

a Globar.

Interferometer: The interferometer consists of a

beam splitter, a fixed mirror, and a moving mirror.

 FT IR SPECTROMETER
Sample cells and sampling of substances: The

material containing the sample must be transparent to

IR radiation.

For, solid sample – KBr pellet technique is used.

Liquid sample – NaCl cell (or) Nujol is used (solvent

can also be used).

 FT IR SPECTROMETER
Detector:

There are two classes of infrared detectors;

1.thermal detector

2.photonic detector

Thermal detectors use the IR radiation as heat

quantum mechanical (photonic) detectors use the

IR radiation as light which results in a more sensitive

detector.
 FT IR SPECTROMETER

Laser: Many instruments employ a Helium-Neon

laser as an internal wavelength calibration standard.

 FT IR SPECTROMETER

FTIR working:
Most interferometers consist of a beam splitter
which splits the incident infrared beam into two
separate optical beams.
One beam is reflected from a fixed mirror, while
the other beam is reflected from a mirror that is
constantly moving in the instrument.
The two beams are reflected from their respective
mirrors and recombine at the beam splitter.
 FT IR SPECTROMETER

FTIR working:
The path length of the beam that is reflected from
the fixed mirror remains constant, while the path
length of the beam that is reflected from the moving
mirror is constantly changing as the mirror moves.
The signal that exits the interferometer is the
result of these two beams interfering with each other,
and is called an interferogram.
 FT IR SPECTROMETER
FTIR working:

The interferogram is unique in that every data

point, which is a function of the moving mirror

position, has information about every infrared

frequency emitted from the source.

This allows for all frequencies to be measured

simultaneously.
 FT IR SPECTROMETER

FTIR working:

The interferogram is converted to a more familiar

IR spectrum (wavenumber vs. % transmittance) using

the well-known mathematical technique called Fourier

transformation.

The transformation of the interferogram is carried

out by the instrument software.

 FT IR SPECTROMETER
FTIR working:

IR spectra are presented on a relative scale (%T),

therefore, a background spectrum must be measured.

A background spectrum is taken with no sample in

the beam and is then subtracted from the sample

spectrum to remove artifacts generated by the

instrument or air (i.e. water, carbon dioxide, etc.).

 FT IR SPECTROMETER

FTIR Advantages:

FTIR instruments have several advantages over

dispersive IR instruments including:

•Speed

•Sensitivity

•Simplicity

•Internal calibration