Infrared Spectroscopy

Dr. Pronit Biswas

Assistant Professor
Department of Forensic Science,
School of Forensic Sciences,
National Forensic Science University (NFSU),
Delhi Campus
MHA Govt. of India, Delhi, India
Background
 Infrared (IR) spectroscopy deals with the infrared region (12800 -10 cm-1)
of electromagnetic spectrum.
 The term infra means beyond, thus infrared means beyond red.
 It covers a range of techniques, the most common being a form of
absorption spectroscopy.
 This spectroscopic technique is used for identifying compounds and
determining the sample composition.
Conti.
 Infrared spectrophotometer is a common laboratory instrument used for this
technique.
 The infrared region in electromagnetic spectrum is divided into the
following three regions named for their relation with the visible spectrum:
1) The far-infrared, approximately 400 - 10 cm-' (1000-30um), lies adjacent to the
microwave region, has low energy and is used for rotational spectroscopy.
2) The mid-infrared, approximately 4000- 400 cm - (30-2.5um) is used to study the
fundamental vibrations and associated rotational –vibrational structure. This region has
wavelengths between 3x10-4 and 3x10-3 cm.
3) The near- infrared, approximately 14000-4000 cm-' (2.5-0.8um) has higher energy and
can excite overtone or harmonic vibrations.
Conti.
Conti.
 Different covalent bonds have different strengths due to the masses of
different atoms at either end of the bond.
 As a result, the bonds vibrate at different frequencies
 The frequency of vibration can be found by detecting when the molecules
absorb electro-magnetic radiation.
 Various types of vibration are possible.
 Examples include... STRETCHING and BENDING
Conti.
 Examples include... STRETCHING and BENDING

SYMMETRIC BENDING ASYMMETRIC

STRETCHING STRETCH
Conti.
Number of Vibrational Modes:
 In order for a vibrational mode in a sample to be “IR-active”. It must be
associated with changes in the dipole moment. A permanent dipole is not
necessary, as the rule requires only a change in dipole moment.
 A molecule can vibrate in many ways, and each way is called a vibrational mode.
For molecules with N-number of atoms.
 Linear molecules have 3N-5 degrees of vibrational modes, whereas, non linear
molecules have 3N-6 degrees of vibrational modes (also called vibrational degree
of freedom) as an examples H2O, a non-linear molecule, will have 3×3-6=3
degree of vibrational freedom or modes.
Conti.
Fundamental Modes of Vibrations in Polyatomic Molecules
Fundamental vibrations are of the following two types:
1) Stretching Vibrations: In this type, the distance between the two atoms increases
or decreases, however, keeping the atoms in the same bond axis. Stretchingvibrations
are of the following two types:
i) Symmetric Stretching Vibrations: In this type, the movement of atoms with
respect to a particular atom in a mole cule is in the same direction.
ii) Asymmetric Stretching Vibrations: In this type, one atom approaches and
the other atom moves apart from the central atom.
Conti.
2. Bending or Deformation Vibrations: In this type, the positions of atoms change
with respect to the original bond axis. The stretching absorptions of a bond appear at
higher frequencies (i.e., higher energy) than the bending absorptions of the same
bond. Bending vibrations are of the following four types:
i) Scissoring: In this type, two atoms approach each other.
ii) Rocking: The movement of atoms is in the same direction.
iii) Wagging: Two atoms move up and down the plane with respect to the central
atom.
iv) Twisting: One atom moves up the plane and the other moves down with
respect to the central atom.
Conti.
No band appears in the IR spectrum of a molecule if no change occurs in its
dipole moment.
Theory
IR-Spectroscopy exploits the fact that molecule absorb frequencies
characteristic of their structure. These absorptions occur at resonance
frequencies i.e., the frequency of the absorbed radiation matches the
vibrational frequency.
The energies are affected by the shape of the molecular potencial energy
surfaces, the masses of the atoms and the associated vibronic coupling.
Principle
Infrared spectroscopy works on the principle that all molecules vibrate and absorb
energy in the infrared region.
Most of the vibrational absorption states correspond to 2.5- 25 um (4000 -400
cm1) wavelength.
The vibrational frequency (v) in
a two atomic system containing
two masses (m1 and m2,) is
related to the force constant (k)
and reduced mass by the
following equation:
Principle (Conti.)
As per the formula, larger the force constant (k) of the bond between two atoms,
higher is the vibrational
frequency.
Organic molecules absorb the IR radiation and convert them into energy of
molecular vibrations. Thus, indicating that a C=C bond will absorb at a higher
frequency than a C - C bond.
Apparatus
Infrared spectrophotometers consists of fallowing components:
1. Light source
2. Monochromators and optical material
3. Sample holder
4. A radiation Detector- which converts radiant energy to a usable signal
5. Instrument for the measurement of response of detector or recorder.
Apparatus. (Conti.)
Radiation
Sampling area Monochromator
source

Detector

Amplifier

Recorder
Apparatus. (Conti.)
The desirable characteristic of these components are:
 Instruments for measuring infrared absorption all require a source of
continuous infrared radiation and a sensitive infrared transducer.
 A source generate a beam of radiation with sufficient power for easy
detection and measurement . In addition, its output power should be stable
for reasonable periods.
Apparatus. (Conti.)

Materials for parts

Apparatus. (Conti.)

Materials for parts

Apparatus. (Conti.)
1:Light source
Sources for different IR regions are:
a. Near IR region: Tungsten filament, Nichrome wire, Rhodium wire.
b. Mid IR region: Nernst glower and Globar source
These two are most commonly used IR sources.
c. Far IR region: Mercury arc lamp and carbon dioxide laser.
Apparatus. (Conti.)
IR sources are hot bodies, which emit continuously through out the IR region and
which approximates a black body radiator in its absorption properties.
The radiation source must emit IR radiation, which must be
1. Intense enough for detection
2. Steady and
3. Must extend over the desired wavelength.
Although these radiations are continuous, only selected frequencies will be
absorbed by samples.
An incandescent solid is used as source of IR radiation. The inert solid is heated
electrically to 1200-1500 o c.
Apparatus. (Conti.)
a. Nernst Glower
Nernst Glower consists of a rod or hollow tube about 2 cm long and 1 mm in
diameter, made by sintering a mixture of oxides of Cerium, Zirconium , Thorium
and Ytterium.
Nernst glower is a non conducting at room temperature and must heated external
means to bring conducting state .
It is maintained at higher temperature by making use of electrical heating and can
be operated in air, since it is not subject to oxidation.
Apparatus. (Conti.)
Disadvantage of Nernst Glower:
1. Mechanical failure
2. Its energy is concentrated in visible and near infrared region of spectrum.
3. The Nernst Glower has a large negative temperature coefficient of electrical
resistance, and it must be heated externally (1000-1800oc) to a dull red heat
before the current is large enough to maintain the desired temperature.
Apparatus. (Conti.)
b. Globar Source:
Globar is a silicon carbide rod usually about 5 cm in length and 0.5 cm in diameter.
It has an advantage of positive co-efficient of resistance. When heated to temperature
between 1300-1700 c, it strongly emits radiation in IR region.
Unlike Nernst Glower, the globar source is self starting and can be controlled
conveniently with a variable transformer .
The disadvantage is its less intense source than nernst glower. It is more satisfactory
because it is worth at wave length longer than 650 cm-1.
However in far infrared region, both these sources loses their effectiveness.
Apparatus. (Conti.)
c. Mercury arc lamp
Beckman devised the quartz mercury lamp for the far IR region, special high-
pressure mercury arc lamps are used.
At the shorter wavelength, the heated quartz envelope emits the radiation whereas
at longer wavelength the mercury plasma provides radiation through the quartz.
d. Tungsten filament lamp
The ordinary tungsten filament lamp is a convenient source for the near-IR region of
4000 to 12800 cm-1.
Apparatus. (Conti.)
e. Rhodium wire
Rhodium wire heater sealed in a cylinder has been used.
f. Nichrome wire
It is used when the required wavelength range and intensity are not too great.
Apparatus. (Conti.)
2. MONOCHROMATOR
The radiation source emits radiations of variable frequencies. But the sample in
the IR spectroscopy absorbs only at certain frequencies. It is, therefore., necessary
to select desired frequencies from the radiation source and reject the others.
The selection has been achieved by means of monochromators .
The prism or grating type monochromators are employed in infrared work.
However prism instruments are favored because of greater range and simplicity.
Apparatus. (Conti.)
It has the fallowing components
1. Entrance slit (to get narrow source)
2. Collimator (to render the light parallel)
3. Grating or Prism (to disperse radiation)
4. Collimator (to reform the image of entrance slit)
5. Exit slit (to fall on sample cell)
Apparatus. (Conti.)
Note : neither quartz nor glass is sufficient transparent to infrared, radiations
above 3.5 microns.
These Can not be used as they absorb satisfactory in IR region. Hence crystals of
certain halogen salts, which transmit infrared freely have been utilized for optical
parts.
Quartz is employed as material for prism construction and is used for near IR
region. It strongly absorbs beyond 4 microns.
Apparatus. (Conti.)
The great bulk of analytical work in IR is done crystalline NaCl as the Prism
material. Beyond 20 microns NaCl absorbs strongly. So cant be used.
Crystalline KBr and Cerium bromide are satisfactory for the far IR region (15-40
microns).
Lithium fluoride provides Prism material in near IR region(1-5 micron).
Apparatus. (Conti.)
The prism disperse the light radiation into its individual wavelength .The
resolution depends on the size and refractive index of the prism. The material that
are transparent to IR is used for the construction for the prism.
There are two types.
1. Refractive type.
2. Reflective type (littrow type).
Apparatus. (Conti.)
1st Refractive type
In the refractive type prism, the light from the source through the entrance slit
falls on a collimator. The parallel radiation from the collimator are dispersed
according to the wavelength. And by using another collimator the image of the
entrance slit are reformed. The required radiation in exit slit can be selected by
rotating the prism or by keeping the prism stationary and moving the exit slit.
Apparatus. (Conti.)
Apparatus. (Conti.)
2nd Reflective type
The principle of working is similar to the refractive type except that, a
reflective surface is present on one side of the prism. Hence the dispersed
radiation gets reflected and can be collected on the same side of the source of
the light.
Apparatus. (Conti.)
Gratings
Are most effective one in converting a polychromatic light to monochromatic
light. As a resolution of +/-0.1 nm could be achieved by using gratings, they
are commonly used in spectrophotometers.
Gratings are of two types.
I. Diffraction grating.
II. Transmission gratings.
Apparatus. (Conti.)
I. Diffraction Grating
Gratings are nothing but rulings made on some material depending upon the
instrument like glass, quartz, plastic and for IR alkyl halides.
The number of rulings per mm is 20 groves or lines per mm in IR spectrophotometer.
these gratings are replica made from master gratings by, coating the original master
grating with a epoxy resin and are removed after setting.
To make the surface reflective, a deposit of aluminum is made on the surface .In
order to minimize to greater amounts of scattered radiation and appearance of
unwanted radiation of other spectral orders, the gratings are blazed to concentrate the
radiation into a single order.
Apparatus. (Conti.)
Apparatus. (Conti.)
II. Transmission grating
It is similar to diffraction grating but refraction takes place instead of
reflection. Refraction produces reinforcement. this occurs when radiation
transmitted through grating reinforces with the partially refracted radiation.
Apparatus. (Conti.)
Sampling procedure in IR
1. Liquids
A drop of a liquid organic compound is placed between a pair of polished
sodium chloride or
potassium bromide plates, referred to as salt plates. When the plates are
squeezed gently, a thin liquid film forms between them. A spectrum determined
by this method is referred to as a neat spectrum since no solvent is used. Salt
plates break easily and are water soluble. Compounds analyzed by this
technique must be free of water. The pair of plates is inserted into a holder that
fits into the spectrometer.
Sampling procedure in IR (Conti.)
2. Solids
There are several methods for determining infrared spectra for solids. One method of choice
has been to mix a finely ground sample with powdered potassium bromide and press the
mixture under high pressure. Under pressure, the potassium bromide melts and seals the
sample into a matrix. The resulting KBr pellet is inserted in the instrument. If a good pellet is
prepared, the spectrum obtained will have no interfering bands since potassium bromide is
transparent down to 400 cm−1
3. Nujol mull thin film methods
Another method, a Nujol mull, involves grinding the compound with mineral oil (Nujol) to
create a suspension of the finely divided sample dispersed in mineral oil. The thick
suspension is placed between two salt plates.
Sampling procedure in IR (Conti.)
4. Attenuated Total Reflectance (ATR)
The two most important sampling techniques are transmission Fourier
transform infrared (FTIR) and attenuated total reflectance FTIR (ATR-FTIR).
Other important FTIR sampling techniques include spectral reflection and
diffuse reflection:
Spectral reflection—liquids, thins films, bulk materials Spectral reflection
sampling can be used for FTIR testing of liquids, thin films, and bulk materials.
The IR light hits the sample at a defined angle and is reflected by the surface.
The reflected light is detected at the same defined angle as the incoming light.
Sampling procedure in IR (Conti.)
5. Sampling of Gases
Gas samples are examined in the IR spectrometer after removal of water
vapour. the simplest gas cell consists of metal bar glass cylinder of 10 cm. long
and closed with a appropriate window. The gaseous sample is passed through a
stop cock via a suitable gas handling apparatus and partial pressure of 5-15 mm
Hg gives a reasonable level of absorption In most cases. The end wall of gas
cell is made of NaCl and for low concentrated gases long path lens are required.
Multi reflection can be used to make effective oath length as long as 40 cm so
that constituent of gas can be determined.
Detectors
Near-infrared: lead sulfide photoconductive
Mid-infrared: Thermopile, Themistor or Pyroelectric
Far –infrared: Golay, pyroelectric.
At the shorter-wavelength end, below about 1.2 microns, the preferred
detection methods are the same as those used for visible and UV radiation. The
detectors used at longer wavelengths can be classified into 2 groups.
Detectors (Conti.)
The detectors used at longer wavelengths can be classified into 2 groups
1. Thermal detectors; in which the infrared radiation produces a heating
effect that alters some physical property of detector, and
2. Photon detectors, which use the quantum effects of the IR radiations to
change the electric properties of a semiconductor.
Detectors (Conti.)
Photon detector
Photon detectors consists of thin layer of lead telluride or lead sulphide
supported on a glass and enclosed in a evacuated glass envelope. Lead sulphide
photoconductive cells are sensitive to infrared radiation upto 3 microns. When
IR radiation is focused on lead sulphide. A sufficiently energetic photon that
strikes an electron in the detector can raise that electron from a non conducting
state into a conducting state, its conductance increases and causes more current
to flow, which is amplified and measured.
Detectors (Conti.)
Photon detector (Conti.)
Advantage of photon detectors
1. Response time is 0.5milli sec.
2. Sensitive,
3. Fast.
Disadvantage
When operated at room temperature, it has a restricted range ( near IR)
Detectors (Conti.)
Thermal detectors
Generally thermal detectors are used in IR spectroscopy. Radiations above 1 micron
are detected by its heating-effects, either by a sensible thermocouple or by resistance
thermistors. When either of these instrument is connected to a galvanometer, a
deflection is noted which is in proportion to the transmitting radiant power.
So thermal detectors give response for all frequencies, the radiant power is low for IR
region, i.e. The detector signal will also be low. In order to locate these low signals, a
preamplifier is fixed to the detector and the radiant beam is modulated with a low
frequency light interrupt 10-26 cps. Thus to detect such signals, thermal detector
must posses a short response time and the absorbed heat must be lost rapidly.
Detectors (Conti.)
Bolometer
A bolometer is based upon the fact that the electrical resistance of a metal changes
with increase of temperature. It is consists of a resistors made up of sintered
oxides of manganese, cobalt and nickel, which have high temperature coefficient
of resistance.
The constructed metal or semiconductor that exhibits a large change in electrical
resistance as a function of temperature. When radiation falls on the bolometer, its
temperature changes and hence the resistance of conductor also changes. The
degree of changes in resistance is a measure of amount of radiation falls on the
bolometer.
Detectors (Conti.)
Golay cell
It uses the expansion of a gas as the measuring device. The unit consists of a small
metal cylinder closed by a rigid blackened metal plate at one end and by a flexible
silvered diaphragm at other end. The chamber is filled with xenon. The radiation
passes through a small IR transmitting window and is absorbed by the blackened
plate. Heat conducted to the gas causes it to expand and deform the flexible
diaphragm (mirror).
Detectors (Conti.)
Golay cell (Conti.)
To amplify distortions of the mirror surface, light from a lamp inside the detector
housing is focused on the mirror, which reflects the light beam on to the phototube.
With the flexible mirror in its rest position, an image off the moiré grid falls on the
other half so that no light passes through. Flexing of the mirror moves the image off
the grid laterally so that varying amount of light can reach the phototube.
The Golay cell has sensitivity similar to that of thermocouple. It is significantly
superior as a detector for the far IR region. Since the angular aperture is 60 degree,
the detector must be used with a system of condensed mirrors to concentrate the
incident radiation.
Detectors (Conti.)
Golay cell (Conti.)
FTIR
Background
Fourier-transform infrared spectroscopy is a technique used for the detection of
the infrared spectrum of absorption and emission of molecules in solid, liquid,
or gaseous phase. The term Fourier-transform has been used because this
process requires a mathematical process termed Fourier transform to convert
the raw data into the actual spectrum.
Principle
• Fourier-transform infrared spectroscopy uses the same principle of dispersive
spectroscopy except that instead of focusing a beam with a single frequency, this
technique uses a beam with multiple frequencies.
• This technique then measures how much of that beam is absorbed by the sample.
• Similarly, a second beam with different frequencies is used for sample processing, and the
data is collected.
• This process is repeated a number of times over a short period of time.
• All the data obtained is then given to a computer that works backward to determine the
absorption at each wavelength.
• The spectrometer has a special structure termed Michelson interferometer which consists
of a certain number of mirrors, one of which moves towards the light source.
• With the movement of the mirror, each wavelength of light is periodically blocked and
transmitted due to wave interference.
• As a result of this, the beam coming out of the interferometer has a different spectrum
each time.
Steps in FTIR
• The sample is placed in the FTIR spectrometer. The sample should be thin enough for the
IR rays to pass through.
• The device directs the IR towards the sample and measures how much and which
frequency of the beam is absorbed by the sample.
• With the help of the reference database, the sample is identified.
Ray diagram
Why FT-IR?
Fourier Transform Infrared (FT-IR) spectrometry was developed in order to overcome the
limitations encountered with dispersive instruments. The main difficulty was the slow
scanning process. A method for measuring all of the infrared frequencies simultaneously,
rather than individually, was needed.
A solution was developed which employed a very simple optical device called an
interferometer. The interferometer produces a unique type of signal which has all of the
infrared frequencies “encoded” into it. The signal can be measured very quickly, usually on
the order of one second or so. Thus, the time element per sample is reduced to a matter of a
few seconds rather than several minutes.
IR spectra interpretation
Applications of IR or FTIR
 FTIR spectroscopy is commonly used for the analysis of organic, polymeric, and
eveninorganic compounds.
 This technique allows the determination of the chemical composition of various samples.
 FTIR has been used with gas chromatography for the detection of substances separated
from the chromatographic technique.
 FTIR can directly detect the polarity of the given site while investigating hydrophobic
membrane environments.
Raman Spectroscopy
Background
 Raman spectroscopy is an analytical technique where scattered light is used to
measure the vibrational energy modes of a sample.
 It is named after the Indian physicist C. V. Raman who, together with his
research partner K. S. Krishnan, was the first to observe Raman scattering in
1928.
 Raman spectroscopy can provide both chemical and structural information, as
well as the identification of substances through their characteristic Raman
‘fingerprint’.
 Raman spectroscopy extracts this information through the detection of Raman
scattering from the sample.
Background (Conti.)
What is Raman Scattering?
Background (Conti.)
Raman Scattering
 When light is scattered by molecule, the oscillating electromagnetic field of a
photon induces a polarization of the molecular electron cloud.
 This leaves the molecule in a higher energy state with the energy of the photon
transferred to the molecule.
 This can be considered as the formation of a very short-lived complex between
the photon and molecule which is commonly called the virtual state of the
molecule.
 The virtual state is not stable and the photon is re-emitted almost immediately, as
scattered light.
Background (Conti.)
Rayleigh Scattering
 In the vast majority of scattering events, the energy of the molecule is unchanged
after its interaction with the photon.
 The energy, and therefore the wavelength, of the scattered photon is equal to that
of the incident photon.
 This is called elastic (energy of scattering particle is conserved) or Rayleigh
scattering and is the dominant process.
Background (Conti.)
Raman Scattering
In a much rarer event (approximately 1 in 10 million photons)Raman scattering
occurs, which is an inelastic scattering process with a transfer of energy between the
molecule and scattered photon.
If the molecule gains energy from the photon during the scattering (excited to a
higher vibrational level) then the scattered photon loses energy and its wavelength
increases which is called Stokes Raman scattering.
Inversely, if the molecule loses energy by relaxing to a lower vibrational level the
scattered photon gains the corresponding energy and its wavelength decreases;
which is called Anti-Stokes Raman scattering.
Background (Conti.)
Raman Scattering (Conti.)
 Quantum mechanically, Stokes and Anti-Stokes are equally likely processes.
 However, with an ensemble of molecules, the majority of molecules will be in the ground
vibrational level (Boltzmann distribution) and Stokes scatter is the statistically more probable
process.
 As a result, the Stokes Raman scatter is always more intense than the anti-Stokes and for this
reason, it is nearly always the Stokes Raman scatter that is measured in Raman spectroscopy.
 The wavelength of the Raman scattered light will depend on the wavelength of the excitation
light.
 This makes the Raman scatter wavelength an impractical number for comparison between
spectra measured using different lasers.
Background (Conti.)
Raman Shift
The Raman scatter position is therefore converted to a Raman shift away from
excitation wavelength.
Principle of Raman spectroscopy
 A Raman spectrum arises from the analysis of scattered light from the sample.
 The largest part of an incident light beam passes through the sample (transmission). A
small part is scattered isotropically, i.e. uniformly in all directions, and possesses the same
wavelength as the incident beam.
 The Raman spectrum arises from the fact that a tiny proportion of light scattered by the
sample will have a different frequency than the incident light.
 As a result of the incident photon, the molecules move to an exciting rotational-
vibrational-electronic state, and the photons are emitted from the sample.
 In order to maintain the energy of the system after the molecule moves to a higher state,
the scattered photons shift to a new state.
 The energy difference between the new state and the original state of the scattered photon
is equal to the difference in energy between the two vibrational states of the molecules.
 The criterion for a peak to appear in the Raman spectrum is a change in polarizability of
the molecule during the transition.
 The depolarization is calculated as the ratio of two intensities with perpendicular and
parallel polarization with respect to the incident beam.
Basic Principals of Raman Spectroscopy:
- light is scattered by the sample at various angles by momentary absorption to virtual state
and reemission

No change in
electronic states

Infinite number
of virtual states
energy absorbed by molecule
from photon of light
not quantized
- some scattered emissions occur at the same energy while others return in a different
state

Rayleigh Scattering
no change in energy
hnin = hnout Raman Scattering
net change in energy
hnin <> hnout

Elastic: collision between photon and molecule results in no change in energy

Inelastic: collision between photon and molecule results in a net change in energy
 Two Types of Raman Scattering

Anti-Stokes: E = hn + ΔE

Stokes: E = hn - ΔE

± ΔE – the energy of the first vibration level of the ground state – IR vibration absorbance

 Raman frequency shift and IR absorption peak frequency are identical

 - Resulting Raman Spectrum

Lower energy higher energy

Probability of Emission Observed Intensity

Raleigh scattering >> Stokes >> anti-Stokes
difference in population of energy levels of vibrational transitions

Intensity of Raman lines are 0.001% intensity of the source

 - need change in polarizability of molecule during vibration
- polarizability related to electron cloud distribution

example:
O = C = O IR inactive
Raman active

O = C = O IR active
Raman inactive

IR & Raman are complimentary. Can be cases where vibration is both IR & Raman active
(eg. SO2 – non-linear molecule)

In general: C
IR tends to emphasize polar functional groups (R-OH, , etc.)
Raman emphasizes aromatic & carbon backbone (C=C, -CH2-, etc.)
- Raman does not “see” many common polar solvents can
use with aqueous samples – advantage over IR

Raman frequency range: 4000 -50 cm-1(Stokes and anti-stokes)

Ray diagram
Instrumentation
1. Light Source: A modern, compact Raman spectrometer consists of several basic
components, including a laser that serves as the excitation source to induce the Raman
scattering. Typically, solid state lasers are used in modern Raman instruments with
popular wavelengths of, 785 nm, 830 nm and 1064 nm.
2. Spectrograph Grater: The 532 nm laser energy is transmitted to and collected from the
sample by fiber optics cables. A notch or edge filter is used to eliminate Rayleigh and
anti-Stokes scattering and the remaining Stokes scattered light is passed on to a
dispersion element, typically a holographic grating.
3. Detector: Charge Coupled Device (CCD) detector captures the light, resulting in the
Raman spectrum. Since Raman scattering yields a weak signal, it is most important that
high-quality, optically well-matched components are used in the Raman spectrometer.
Steps of Raman in spectroscopy
 Mostly aqueous solutions are used the required laser is turned on after selecting the
appropriate wavelength being used.
 The spectrometer is then calibrated by using a reference sample by using appropriate
exposure energy and time.
 The sample is then placed under the microscope while focusing on the layer to be
investigated.
 A range of wavenumbers is scanned by the monochromator, which results in the Raman
spectrum.
 The data is then investigated by appropriate software, and the analysis is made based on
the peaks formed on the spectrum.
- comparison of Raman and IR Spectra
Applications
Raman spectroscopy is employed over infrared spectroscopy as it allows the analysis of the
sample in aqueous conditions as well.
 Investigating carbon bonds in aliphatic and aromatic rings.
 Bonds that are difficult to see in FTIR (i.e., 0-0, S-H, C=S, N=N, C=C etc.)
 Examination of particles in solution is important, e.g. polymorphism
 Lower frequency modes are important (e.g. Inorganic-Oxides)
 Reactions in aqueous media can investigated.
 Reactions in which observation through a reaction window is easier and safer (e.g. high
pressure catalytic reactions, polymerizations)
 Investigating lower frequency lattice modes is of interest
 Investigation of reaction initiation, endpoint, and product stability of biphasic and colloidal
reactions
Similarity in IR and Raman Spectroscopy
 IR and Raman spectroscopy are both techniques used to study molecular
vibrations and provide complementary information about a molecule.
 They both interact with phonons, with IR spectroscopy involving the absorption
of infrared photons and Raman spectroscopy relying on inelastic scattering of
light. Thus, while their spectra can provide similar and overlapping information,
the choice of method often depends on the specific application and the type of
molecular information required.
Difference between IR and Raman Spectroscopy
 Raman spectroscopy reckons on an amendment in polarizability of the molecule,
whereas IR spectroscopy is reliant upon the change in dipole moment.
 Raman spectroscopy calculates relative frequencies in which the sample scatters
radiation. IR spectroscopy evaluates absolute frequencies in which a sample
absorbs radiation.
 Raman spectroscopy is sensitive to homo-nuclear molecular bonds. It is able to
distinguish between single, double, and triple bonds between carbon atoms. IR
spectroscopy has responsiveness with regard to hetero-nuclear functional group
vibrations and polar bonds. An example is OH stretching in water.
Difference between IR and Raman Spectroscopy (Conti.)
 In sample preparation, Raman spectra analysis has no requirement. But IR
spectroscopy has constraints of a thickness of sample, uniformity, and dilution
for avoiding saturation.
 Fluorescence will interfere with the Raman imaging, but it is not an issue in
taking the IR spectrum.
 Both the methods have some limitations when taken separately. But, when used
combinedly, they become a powerful tool in material characterization. Both can
be used with microscopic techniques.
 Raman spectroscopy is a weaker technique, while IR spectroscopy is a
stronger technique.
Difference between IR and Raman Spectroscopy (Conti.)
 In the investigation of crystallization processes, Raman spectroscopy analyses
solid crystal forms in solution, whereas IR spectroscopy is utilized for solution-
phase supersaturation studies.
 Raman spectrometers have laser light sources operating in the near IR or visible
region in the electromagnetic spectrum. IR spectrophotometers possess black
body radiators providing energy in the mid-IR region.
 Raman spectrometers have holographic grating with CCD detectors. IR
spectrometers use interferometers providing modulated radiation, which is
detected by photonic detectors or pyroelectric detectors.
Difference between IR and Raman Spectroscopy (Conti.)
 The Raman spectrum results from the scattering of light by vibrating molecules.
The IR spectrum results from the absorption of light by vibrating molecules.
 Raman activity occurs due to the change of polarizability of a molecule. IR
activity occurs due to changing dipole moments.
 In Raman spectroscopy imaging, monochromatic light of high-intensity laser
beams can be used in IR, UV, or visible regions. In IR spectroscopy, the light
range is limited to IR frequency.
 In Raman spectroscopy instrumentation, scattered light is observed at right
angles to the incident beam. Regarding the IR spectroscopy, the absorption
signal is estimated in the same direction as the incident beam.
Difference between IR and Raman Spectroscopy (Conti.)
 The technique of Raman spectroscopy is non-destructive. Sample measurement
can be done directly in a glass container. In the case of pharmaceutical samples,
they can be measured directly in sachets. IR spectroscopy needs sample
preparation through the KBr pellet for observation of gels, films, and liquids.
 Raman spectrum is a result of Raman spectroscopy, where a laser is used to
analyze the sample due to the inelastic scattering of photons in the sample. IR
spectrum is a result of IR spectroscopy, where IR radiation is utilized to analyze
the sample.
Difference between IR and Raman Spectroscopy (Conti.)
 Vibrational modes are active in Raman spectroscopy if they cause a change in
polarizability. Vibrational modes are active in IR spectroscopy if they cause
changes in dipole moments.
 Raman analysis can use water as a solvent, whereas IR spectroscopy cannot use
water as a solvent.
 Raman spectroscopy is a highly expensive method, while IR spectroscopy is
relatively inexpensive.
 In Raman spectroscopy, laser sources are highly intensive. It facilitates focusing
the beam on a small sample area with small volumes. When the quantity of
samples is limited, this facility serves as a primary advantage.
Difference between IR and Raman Spectroscopy (Conti.)
 A higher degree of amplification of weak Stoke signals is a requisite in the presence
of intense Rayleigh light scattering. This results in a higher cost for the analysis
when compared to IR spectroscopy. However, the higher cost can be easily justified
against the benefits offered by the technique.
 In Raman spectroscopy, bands present below 400 cm-1 can be measured. As
scattering effects are usually weaker, Raman spectroscopy exhibits lower
sensitivity at times. Sometimes, visible light excitation, fluorescence, and
contamination can overlap with the signal.
 In Raman spectroscopy instrumentation, scattered light is observed at right angles to the
incident beam. Regarding the IR spectroscopy, the absorption signal is estimated in the
same direction as the incident beam.
Difference between IR and Raman Spectroscopy (Conti.)
 The Raman spectrum can be recorded at a single exposure, whereas two separate runs
are required with different prisms to cover the whole region of IR.
 Water is used as a solvent for Raman spectroscopy, while water cannot be used as a
solvent for IR spectroscopy as it is opaque to IR
 Raman spectroscopy is very accurate, but not that much sensitive. IR spectroscopy is
both accurate and sensitive.
 In Raman spectroscopy, optical systems are made up of quartz or glass, while in IR
spectroscopy, optical system is composed of special crystals like NaBr and CaF2
 Occasionally, photochemical reactions occur in the regions of Raman lines and create
difficulties for the Raman spectrum. In IR spectroscopy, photochemical reactions do
not occur.
Difference between IR and Raman Spectroscopy (Conti.)
 In Raman spectroscopy, substances to be investigated should be colorless and pure.
 Since Raman lines are weaker in intensity, concentrated solution should be used
in increasing the intensity of Raman lines. In IR spectroscopy, dilute solutions are
preferred for measurement.
 In Raman spectroscopy, vibrational frequencies of larger molecules can be
measured. In the case of IR spectroscopy, vibrational frequencies of larger
molecules cannot be measured.
 Homonuclear diatomic molecules are found to be active in Raman spectroscopy.
In IR spectroscopy, homonuclear diatomic molecules are not active.
Thank you