A

Project Report on

REVIEW ON INFRARED SPECTROSCOPY

Submitted by

INDRAJIT GUCHHAIT

University Regn. No. –014525 OF 2019-20

Roll No. -18901919109

Under the guidance of

MS. SURITA GHOSH
Assistant Professor

In partial fulfilment of the Requirement for the Degree of

Bachelor of Pharmacy

Maulana Abul Kalam Azad University of Technology

West Bengal

Dr. B. C. Roy College of Pharmacy and Allied Health Sciences

Durgapur 713206

Academic Year: 2021 - 2022

DECLARATION
 This is to certify that the Project entitled “REVIEW ON INFRARED
SPECTROSCOPY” in partial fulfilment of the requirement for the award of
Bachelor of Pharmacy submitted to Maulana Abul Kalam Azad University of
Technology, West Bengal, is an authentic record of bona fide work carried out
by me under the guidance ofMS. SURITA GHOSH.

The matter embodied in this Project has not been submitted for the award of any other
degree or diploma to any University / Institution.

Signature

Name of Candidate: INDRAJIT GUCHHAIT

Roll No:18901919109

Date:

Place:
 CERTIFICATE

This is to certify that the Project entitled “REVIEW ON INFRARED

SPECTROSCOPY” by IndrajitGuchhaitin partial fulfilment for the BACHELOR OF
PHARMACY under MAULANA ABUL KALAM AZAD UNIVERSITY OF
TECHNOLOGY is a record of the original research work under our supervision and up to
our satisfaction. To our knowledge, neither his thesis nor any part of it has been submitted
for any other academic award anywhere before. The thesis is forwarded to Controller of
Examination MAULANA ABUL KALAM AZAD UNIVERSITY OF TECHNOLOGY
for assessment.

__________________
Signature of Guide
Name of Guide: MS. SURITA GHOSH
Date:

Place:

_________________________
PROF (DR.) SUBHABRATA RAY
(PRINCIPAL)
Date:

Place:

Signature
Name of External Examiner:

Date:

Place:

iii
 ACKNOWLEDGEMENTS

I would like to express my special thanks of gratitude to my guide ASSISTANT PROF.

MS. SURITA GHOSH ma'am for her valuable guidance, the scientific way of thinking.
Encouragement and support in completing my project work.

I would also like to extend my gratitude to our DIRECTOR DR. SUBRATA

CHAKRABORTY sir and our PRINCIPAL PROF. (DR.) SUBHABRATA RAY sir
who gave me the golden opportunity to do this wonderful project work. They helped me
and provided with their valuable guidance of each and every step.

I would also like to thank my parents and friends who helped me a lot in finishing this
project within the limited time.

iv
 REVIEW ON INFRARED
SPECTROSCOPY

ABSTRACT
The study of Infrared light absorption, which results in vibrational transitions, is known
as Infrared spectroscopy or vibrational spectroscopy. By providing the analytical
knowledge needed to solve problems, relevant IR measurements can be made. We
observe the interaction of functional groups in chemical molecules with Infrared light,
which produces predictable vibrations that offer a "fingerprint" characteristic of chemical
or biological substances present in the sample, using Infrared spectroscopy. It saves the
industry money, time, and labour. It will also provide the analyst with speed,
convenience, precision, and reliability. Infrared spectroscopy is a critical tool since it is
one of the few techniques that can be used to generate metal-complexes. Using Infrared
spectroscopy, we can confirm that a metal compound has formed. Changes in distinctive
bands, as well as an increase or decrease in the number of bands and the atom bond, aid in
the identification of the complex atom. Dispersive instruments, multichannel dispersive
instruments, multiplex instruments, and multichannel/multiplex instruments are among
the IR instruments currently in use. This chapter's guidelines will assist you in making.

v
 Table of Contents
Declaration……………………………………………………………………………... i
Certificate ………………………………………………………………………………ii
Acknowledgements …………………………………………………………………….iii
REVIEW ON INFRARED SPECTROSCOPY..........................................................................................................1
INTRODUCTION..............................................................................................................................................1
PRINCIPLE.......................................................................................................................................................3
CRITERIA FOR A COMPOUND TO ABSORB IR RADIATION...............................................................................6
FUNDAMENTALS OF INFRARED SPECTROSCOPY............................................................................................8
The Intensity of Infrared Radiation...............................................................................................................10
MOLECULAR VIBRATION..............................................................................................................................14
FACTORS EFECTING MOLECULAR VIBRATION..............................................................................................17
Region of Ir Spectroscopy.............................................................................................................................21
INSTRUMENTATION OF IR SPECTROSCOPY..................................................................................................22
APPLICATION OF IR SPECTROSCOPY.............................................................................................................26
Conclusion....................................................................................................................................................27
REFERENCE...................................................................................................................................................28

vi
 INTRODUCTION
Sir William Herschel discovered infrared radiation in 1800. Spectroscopy is the study of
infrared radiation absorption that leads to a vibrational transition. The light we perceive is
only a small part of the electromagnetic spectrum. Ultraviolet is on the high energy side
of the visible spectram, while infrared is on the low energy side. The infrared part is the
most significant for organic chemical analysis. A frequency corresponding to a
wavelength ranging from 2500 to 16,000 nm [1].

It can be used to characterise new materials as well as identify and validate known and
unknown samples. Infrared spectrometer is a method or technique for obtaining infrared
spectrum in IR spectroscopy. A graph of an IR spectrum shows infrared light absorption
on the vertical axis vs. frequency or wavelength on the horizontal axis. In IR spectra, the
notation cm1 stands for reciprocal centimetres, which are the most frequent frequency
units. Micrometres (formerly "microns"), sign m, are used to denote IR wavelengths,
which are reciprocally related to wave numbers. This technology is used in a Fourier
transform infrared (FTIR) spectrometer, which is a standard laboratory gadget. [2]

Because the numbers are more understandable, the frequency scale at the bottom of the
chart is presented in reciprocal centimetres (cm-1) rather than Hz. [3]

The mutual centimetre is the number of wave cycles in one centimetre, where frequency
(Hz) is the number of wave cycles per second and 3*1010 cm is the number of wave
cycles.The infrared spectra of samples in all phases can be obtained (liquid, solid and
gaseous). Liquids are commonly tested by sandwiching a thin film between two polished
salt plates. [4]

For molecular structure research, identification, and quantitative analysis of materials,

infrared spectroscopy is a helpful instrument. The benefit of this technique is that it may
be used to solve a wide range of issues in both the condensed and gaseous states.
Infrared spectroscopy is employed in chemical, environmental, life, materials,
pharmaceutical, and surface sciences, as well as many technical applications, as
mentioned in the book's subsequent chapters. This book's goal is to give readers a step-
by-step guide to the experimental components of this adaptable method. On the
assumption that the readers have no prior knowledge of infrared spectroscopy, this
chapter provides introductory explanations on an infrared absorption spectrum and

1
related basic themes, which readers should learn before reading the next chapters. Visible
light is absorbed by numerous materials, as is widely known, and the absorption of visible
light is related to the colours of materials. Blue materials absorb red-colored radiation,
while red materials absorb blue-colored radiation. The wavelengths of red-colored
radiation are longer than those of blue-colored radiation. A visible absorption spectrum is
a figure that shows quantitatively the absorption of visible light at different wavelengths
from violet to red. The colour of the material from which the spectrum is measured is
closely reflected in the visible absorption spectrum. The wavelengths of infrared light are
longer than the wavelengths of red-colored energy.The wavelengths of visible light with
red colour have the longest wavelengths, which grow from violet to red. Although
infrared radiation is not visible to the naked eye, it is absorbed by practically all
materials. The absorption of infrared radiation against the wavelength of infrared
radiation is plotted quantitatively in an infrared spectrum. Any material, excluding
metals, may typically be observed in the infrared absorption spectrum, regardless of
whether the sample is in the gaseous, liquid, or solid state. Because of this benefit,
infrared spectroscopy is a very helpful tool that is used in a variety of fields. Infrared
spectra are often measured on liquid and solid samples. Only one or two broad bands are
commonly detected in visible absorption spectra of liquids and solids, but infrared
absorption spectra exhibit at least many, often many reasonably sharp absorption bands [5].
The majority of organic molecules have several infrared absorption bands. The distinction
between visible and infrared absorption spectra is owing to the two types of spectra
having different origins. The states of electrons in a molecule are linked to visible
absorption. Infrared absorptions, on the other hand, are caused by the vibrating states of
atoms in a molecule. To put it another way, the visible absorption spectrum is an
electrical spectrum, while the infrared absorption spectrum is a vibrational spectrum.
Vibrational motions of atoms in a molecule are called molecular vibrations. At present,
measurements of infrared spectra are widely performed in materials science, life science,
and surface science. In these fields, the states of targets of research are usually liquids or
solids. This book primarily aims at describing the fundamentals of Infrared spectroscopy
and practical methods of measuring infrared spectra from various samples in the liquid
and solid states.[6]

2
 PRINCIPLE
In every molecule, it is known that atoms or groups of atoms or groups of atoms are
connected by bonds. These bonds are analogous to springs and not ringed in nature.
Because of continuous motion of the molecule they maintain some vibrations with some
frequency,characteristic to every portion of the molecule .This is called the natural
frequency of vibration . When energy in the form of infrared radiation is applied,

Applied infrared frequency = Natural frequency of vibration,

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

Every bond and or portion of a molecule or functional group requires different

frequencies for absorption. Hence a characteristicved for every functional group or part of
the molecule. In other words, IR spectra is nothing but a fingerprint of a molecule. Eg.
Spectra of ethanol is given in upper Fig.
In Pharmaceutical analysis, we use infrared
radiation (mid-IR) of wavelength 25μ to 2.5μ or
wavenumbers from 400cm¹ to 4characteristic
000cm¹. There are other regions like near-IR
(0.8μ to 2.5µ) and far-IR (25μ to 1000μ) which
are not used in Pharmacy.
In IR spectra, we use wave numbers instead of
wavelengths for mentioning the characteristic peak, becauseChemistrylibertexts.com
wave numbers are larger
values and easy to handle than wavelengths which will show only small differences
between functional groups.
Wave number is nothing but the number of waves present per cm which can be calculated
from the wavelength.Type equation here .
1
×104 ¿ wavenumber per cm or cm-1
wavelength∈µ

The premise behind IR spectroscopy is that molecules absorb certain frequencies that are
unique to their structure. All atoms in molecules are in constant vibration with regard to

3
 each other at temperatures above absolute zero. A sample's IR spectrum is recorded by
passing a beam of IR radiation through it. [12]The molecule absorbs the IR radiation when
the frequency of a given vibration is equal to the frequency of the IR radiation directed at
it. The amount of energy absorbed at each frequency can be determined by examining the
transmitted light (or wavelength). Using various sampling accessories, IR spectrometers
can accept a wide range of sample types such as gases, liquids, and solids.

Infrared Spectroscopy is the analysis of infrared light interacting with a molecule .[7]

1. The portion of the infrared region most useful for analysis of organic compounds have
a wavelength range from 2,500 to 16,000 nm, with a corresponding frequency range
from 1.9*1013 to 1.2*1014 Hz.
2. Photon energies associated with this part of the infrared (from 1 to 15 kcal/mole) are
not large enough to excite electrons, but may induce vibrational excitation of covalently
bonded atoms and groups.
3. It is well known that, in addition to the simple rotation of groups around single bonds,
molecules undergo a wide range of vibrational movements that are unique to their
constituent atoms.
4. As a result, practically all organic molecules absorb infrared light that corresponds to
these vibrations in terms of energy.
5. Chemists may get absorption spectra of compounds that are a unique reflection of their
molecular structure using infrared spectrometers, which work in the same way as other
spectrometers.
6. An infrared spectrum is a plot of measured infrared intensity vs wavelength (or
frequency) of light, which is the most basic measurement acquired in infrared
spectroscopy.
• Infrared spectroscopy (IR Spectroscopy) monitors atom vibrations and can be used to
discover functional groups.
• Infrared spectroscopy (IR Spectroscopy) monitors atom vibrations and can be used to
discover functional groups.
Generally, stronger bonds and light atoms will vibrate at a high stretching frequency
(wavenumber).[8]

4
Infrared spectroscopy takes advantage of the fact that molecules rotate or vibrate at
certain frequencies that correspond to discrete energy levels. The form of the molecular
potential energy surfaces, the masses of the atoms, and the related vibronic coupling
dictate these resonant frequencies. A vibrational mode in a molecule must be coupled
with changes in the permanent dipole in order to be IR active. The resonant frequencies
are determined by the normal modes corresponding to the neighbourhood of the
equilibrium molecular in the Born-Oppenheimer and harmonic approximations, i.e. when
the molecular Hamiltonian corresponding to the electronic ground state can be
approximated by a harmonic oscillator in the geometry. [9]

Advantage and Disadvantage of IR Spectroscopy

Spectroscopy is a widely used analytical method for identifying functional groups in
molecules and for studying the interaction of electromagnetic radiation with matter. It's
challenging to work with complicated mixes.

The advantage of IR spectroscopy:

1. Qualitative and quantitative analysis: Infrared spectroscopy has the benefit of

providing qualitative and quantitative chemical information without damaging the
sample.

2. Sample Preparation: Infrared spectroscopy has the benefit of not requiring any
special sample preparation.

3. Sensitive and time-saving technique: Because IR spectroscopy is very sensitive, it

only requires a little amount of sample to scan the sample spectrum, and it only takes a
few seconds to scan the whole IR range.

4. Its versatility: IR spectroscopy can evaluate solid, liquid, gas, and semisolid materials.

5. Easy for interpretation: The Peak intensities, peak positions, peak widths, shapes, and
functional groups provide all helpful information.

Disadvantage:

1.Infrared spectroscopy does not provide information on the relative positions of

functional groups in a molecule.

2.IR spectroscopy cannot be used to determine the molecular weight of a substance.

5
3.Sometimes onerous handling methods and sample cell maintenance are disadvantages.

4.Atoms and monatomic icons do not have Infrared spectra; hence it cannot be
analysed.

5.Using Infrared spectroscopy necessitates highly sensitive and well-tuned equipment.

6.Infrared spectroscopy analysis of samples including aqueous solutions and complex

mixtures is difficult.[10]

CRITERIA FOR A COMPOUND TO ABSORB IR

RADIATION

Absorptions of Infrared Light In order for a molecule to show Infrared absorptions, it

must have a certain property, namely, the molecule's electric dipole moment must change
during vibration. This is the infrared spectroscopy selection rule. An example of an
infrared-active molecule, a heteronuclear diatomic molecule, is shown in the figure. As
the bond stretches and collapses, the dipole moment of such a molecule changes. A
homonuclear diatomic molecule, for example, is an infrared - inactive molecule because
its dipole moment remains zero no matter how long the link is. When allocating infrared
bands, it's crucial to understand chemical symmetry and group theory. [4]

Absorptions in the Infrared spectrum are not endlessly narrow, and numerous variables
contribute to their broadening. The Doppler effect, in which radiation shifts in frequency
as the source moves closer or farther away from the observer, is a factor in gases.
Collisions between molecules also cause band broadening. The finite duration of the
states involved in the transition is another source of line broadening. When the
Schrodinger equation is solved for a system that changes with time in quantum
mechanics, the energy states of the system do not have precisely defined energies,
resulting in lifespan widening.There is a relationship between the lifetime of an excited
state and the bandwidth of the absorption band associated with the transition to the
excited state , and this is a consequence of the Heisenberg Uncertainty Principle . This

6
relationship demonstrates that the shorter the lifetime of a state , then the less well defined
is its energy .`[8]`

1. Correct wavelength of radiation

2. Change in dipole moment

1. Correct wavelength of radiation:

• When the inherent frequency of vibrations of some parts of a molecule is the same as
the frequency of incident radiation, the molecule absorbs radiation.

• A molecule absorbs radiation only when the frequency of the incident radiation is equal
to the natural frequency of vibration of the part of the molecule in question. Following
absorption of the correct wavelength of radiations, the molecule vibrates at a higher
amplitude due to the absorbed IR energy. 8.7x1013 / s (2890 cm 1) is the natural
vibrational frequency of Hcl. Only the radiations with a frequency of 8.7 1013 / s are
absorbed when a hcl sample is exposed to IR radiations, with the rest being transferred.2.
Change in dipole moment:

• A molecule can only absorb IR radiations when its absorption causes a change in its
electric dipole.

• A polar bond is usually IR active whereas non polar bonds in a symmetrical molecule
will absorb weakly or not at all.

• A molecule is said to have an electric dipole when there is a slight positive & a slight
negative charge on its component of atoms.

▪ A molecule can only absorb IR radiation when its absorption causes a change in its
electric dipole .

▪ A polar bond is usually IR - active.

▪ A nonpolar bond in a symmetrical molecule will absorb weakly or not at all.[11]

FUNDAMENTALS OF INFRARED SPECTROSCOPY

7
A Basic knowledge of infrared spectroscopy that should be performed before
performing infrared measurements.

The Ordinate and abscissa Axes of an Infrared Spectrum

The abscissa or x coordinate of a point is its distance from the y-axis when scaled with
the x-axis. The ordinate is the distance between a point and the x-axis when scaled with
the y-axis.

If (x, y) is an ordered pair, then y is the ordinate in this case. An ordered pair is used to
represent a point in the Cartesian plane, and the abscissa is the initial coordinate (x) in
the plane. The abscissa or x coordinate of a point is its distance from the y-axis scaled
with the x-axis.

The abscissa and ordinate are the first and second coordinates of a point in a coordinate
system in mathematics:

abscissa = x-axis (horizontal) coordinate

ordinate= y-axis (vertical) coordinate

In a two-dimensional rectangular Cartesian coordinate system, these are usually the

horizontal and vertical coordinates of a point. The abscissa (horizontal, generally x) and
the ordinate (vertical, usually y) of an ordered pair describe the location of a point in
two-dimensional rectangular space:

A point's abscissa is the signed measure of its projection on the primary axis, whose
absolute value equals the distance between the projection and the axis's origin, and whose
sign is determined by the point's location relative to the origin (before: negative; after:
positive).

The ordinate of a point is the signed measure of its projection on the secondary axis,
whose absolute value is the distance between the projection and the origin of the axis, and
whose sign is given by the location on the projection relative to the origin (before:
negative; after: positive).[12][7]

For a long time, it has been known that vapors, liquids, crystals, powder, glass, and a
variety of other materials absorb infrared light. Although the wavelength range of
infrared radiation is not completely defined, the wavelength regions for near-infrared,

8
mid-infrared, and far-infrared radiation are as follows: 700 nm to 2.5 m for near-
infrared, 2.5–25 m for mid-infrared, and 25 m to 1 mm for far-infrared.[13]

The ordinate axis of an Infrared spectrum is the absorption intensity. In an infrared

spectrum, the wavelength can be used as the abscissa axis. However, it is now common
practise, particularly in the mid-infrared area, to utilise the wavenumber as the abscissa
axis rather than the wavelength. The wavenumber is the reciprocal of the wavelength and
represents the number of light waves per unit length (typically 1 cm).An infrared
spectrum's abscissa axis is usually represented in units of cm1. The abscissa axis of an
infrared spectrum is always labelled "Wavenumber/cm1" in this book. However, if a
near-infrared spectrum is measured as an extension of a visible absorption spectrum, the
wavelength is frequently employed as the abscissa axis in the near-infrared region. The
greater wavenumber (corresponding to the shorter wavelength) is sometimes shown on
the left side of a spectrum, whereas in other circumstances it is placed on the right side. [6]

The wavenumber, which is the number of light waves per centimeter as mentioned above,
corresponds to the frequency divided by the speed of light. Therefore, the wavenumber is
proportional to the energy E of a photon as expressed in the following equation:

E= hc ν ̃ …………….(1) Where h is the

Planck constant, c the speed of light, and 𝜈̃ the wavenumber of infrared radiation. This
proportionality between E and 𝜈

is the reason why the wavenumber is now used as the abscissa axis of an infrared spectrum. In Appendix
infrared radiation correspond to the wavenumber regions of about 14 000–4000 cm−1 for
near-infrared , 4000–400 cm1 for mid-infrared , and 400–10 cm1 for far-
infrared . The wavenumber region of 400–10 cm1 for far-infrared

far-infrared region approximately coincides

with the terahertz frequency region. For this reason, the term terahertz spectroscopy is
recently being increasingly used in place of far-infrared spectroscopy. However, the
term far-infrared spectroscopy is considered a better designation because of its
consistency with other optical spectroscopies.[14]

9
 The Intensity of Infrared Radiation
Because infrared radiation is an electromagnetic wave, electromagnetic theory applies to
it. In this theory, the intensity of an electromagnetic wave irradiating a region is defined
as the average energy of radiation per unit area per unit time. In keeping with
spectroscopic tradition, this quantity is referred to as intensity in this text. It's worth
mentioning, however, that in other fields, the term "irradiance" is increasingly being used
instead of "intensity." This amount is indicated in units of W m2 (= J s1 m2), despite the
fact that its absolute value is rarely stated in infrared spectroscopy unless lasers are
involved.The temporal average of the cube of the amplitude of the electric field E
determines the intensity I. When there's no one around,I is expressed as

I= E0 C0 [E2]t …………………. [2]

where 𝜀0 and c0 denote, respectively, the electric constant and the speed of light in
vacuum,and the symbol ⟨⟩t means time average.[15]

Lambert – Beer’s Law:

Absorption of Infrared radiation occurs when an Infrared beam passes through a sample
layer. As shown in Figure, a collimated Infrared beam of intensity I0 and wavenumber
irradiates a sample of thickness l at right angles to its surface. If the sample is transparent
to the infrared beam, it can pass through it without losing its intensity. The Infrared
laser's reflection at the sample's surface is not taken into account here. If the sample
absorbs the wavenumber Infrared

radiation, the intensity of the Infrared radiation decreases as the beam travels through i

-dI=𝛼Idl ……………….[3]

10
where I is the intensity of the beam entering the thin layer dl and is a proportionality
constant describing the amount of absorption (called the absorption coefficient). Equation
(2) is integrated to produce the following equation.

ln I = 𝛼l + a ………….[4]

where the integration constant a must be equal to ln I 0 (I0 is the intensity at l = 0).
Then,the following two equations are derived.

ln(l0 ¿ l ¿=¿ 𝛼l ……………..[5]

l=l0 exp(−αl ¿ ……………….[6]

It should be remembered that each of I0, I, and 𝛼 is a function of 𝜈̃ and, strictly speaking,
should be expressed, respectively, as I0(𝜈̃), I(𝜈̃), and 𝛼(𝜈̃). Equation (5) means that the
absorption intensity expressed as ln(I0
I) is proportional to the thickness of the sample. This relation is known as Lambert’s lawThe value l

0/I) is known as absorbance (abbreviated as A) or optical density (OD).

Transmittance is the percentage scale expression of the number I/I 0 (the term
"transmission factor" is also approved by IUPAC [1]). It may also be used to represent the
strength of absorption. The ordinate axis of an infrared spectrum is the absorption
intensity represented in either absorbance or transmittance. If the sample is a solution, the
absorption coefficient in Equations (3)– (6) should be substituted by cs, where cs is the
concentration of a solute in the solution and a function of is a proportionality constant.
This means that the absorbance is proportional to both the solution thickness (also known
as the cell pathlength) and the solute concentration. This relationship is well-known as
Lambert–Beer’s law, holds for most dilute solutions, but deviation from this law occurs
in various cases , particularly in concentrated solutions .

11
Complex Refractive Index

The relation in Equation (6) can also be derived in the following way. The electric field E
of an electromagnetic wave (or an infrared beam) traveling in the x direction in vacuum
may be expressed as

x
E=A exp2πi( −vt ¿ … … … … … .[7 ]
λ

Where A is a vector describing the light wave's amplitude, frequency, time, and
wavelength, and t is the time. The right-hand side of Equation (7) (and a nearly related
form of A exp 2i(t x)) is addressed in greater detail in Appendix B. If the light wave
travels through a medium other than vacuum, the complex refractive index n must be
included, which is defined as A is a vector indicating the amplitude of the light wave, the
frequency of the light wave, t time, and the wavelength of the light wave. In Appendix B
the meaning of the right-hand side of Equation (7) (and a closely related form of A exp
2πi(𝜈 t x 𝜆)) is discussed in more detail. If the light wave travels in a medium other
than vacuum, it is necessary to introduce the complex refractive index ̂n defined as

̂ n = n + ik ………..[8]

Where n is the (real) refractive index, and k, which is called the absorption index or the
imaginary refractive index, is a constant to express the absorption of light by the medium.
The wavelength 𝜆 in the medium is related to the wavelength 𝜆0 in vacuum by the
equation 𝜆 = 𝜆0 n. By substituting 𝜆 0n for 𝜆 in Equation (7), the following equation is
derived

−2 πkx nx
E=Aexp( )exp2πi( −¿vt) ……[9]
λ0 λ0

As is clear from Equation (9), the amplitude of the light wave decreases in proportion to the factor exp (2kx
𝜆0). Since the

12
intensity is proportional to the square of the amplitude of the electric field, the ratio of the
intensity I at x = l to I0 at x = 0 is given by the following equation.

i [|A|exp exp (−2 πkl¿ 0 ) ] 2

=
I0 ¿ A∨2

−4 πkl
= exp( ) ……………..[10]
λ0

Equation (10) is similar to Equation (10) if 4k0 in Equation (10) is equated with (6). The
complex refractive index is defined as n = n ik in certain literature. In this scenario, E = A
exp 2i(t x) should be used as the right side of Equation (7). The complicated refractive
index is a crucial figure. It demonstrates that refraction and absorption are inextricably
linked, and that they should always be viewed as a pair. The goal of experimental
spectroscopy is to determine the real and imaginary components of the complicated
refractive index of any material quantitatively. This goal, however, is difficult to achieve
since it is impossible to entirely separate absorption from reflection (and refraction) in the
laboratory. A cell is commonly used to test the transmission infrared spectrum of a liquid
or solution, for example. Reflection occurs at the cell window surfaces in this example,
making determining I0 problematic. If the sample is a dilute solution, the problem can be
handled by measuring the intensity of infrared light transmitted from a cell containing just
solvent. The intensity obtained in this manner may be used to calculate I0. There is no
simple, dependable approach that can be used with concentrated solutions, liquids, solid
films, and other materials. However, in most practical applications, qualitative absorption
measurements provide useful information, therefore the quantitative calculation of the
complicated refractive index is typically avoided. [15]

MOLECULAR VIBRATION
When a molecule absorbs energy, E, corresponding to the vibration's frequency, v,
according to the relation E = h, where h is Planck's constant, the vibration is activated.
When a molecule in its ground state absorbs one of these quanta of energy, a fundamental

13
vibration is elicited. When a large number of quanta are absorbed, the first and maybe
higher overtones are elicited.

To a first approximation, the motion of a normal vibration can be described as a simple

harmonic motion. In this approximation, the vibrational energy is a quadratic function
(parabola) in terms of atomic displacements, and the first overtone has double the
frequency of the fundamental. Vibrations are actually anharmonic, with the first overtone
being slightly less than twice the fundamental frequency. Excitation of the higher
overtones takes less and less extra energy when the molecule's potential energy
approaches that of a Morse potential, or more accurately, a Morse/Long-range potential,
and eventually leads to separation.

A molecule's vibrational states can be investigated in a variety of methods. Infrared

spectroscopy is the most direct method, because vibrational transitions often need an
amount of energy that corresponds to the infrared portion of the spectrum. Raman
spectroscopy, which employs visible light to measure vibration frequencies, can also be
used to directly measure vibration frequencies. The two methodologies are compatible,
and comparing them can provide useful structural information, such as in the case of
centrosymmetric molecules and the law of mutual exclusion. In the ultraviolet-visible
region, vibrational excitation can coexist with electronic excitation. The combined
excitation is known as a vibronic transition, giving vibrational fine structure to electronic
transitions, particularly for molecules in the gas state. Simultaneous excitation of a
vibration and rotations gives rise to vibration-rotation spectra.[16]

• A molecule may not be looked upon as a rigid assemblage of atoms.Rather it may be

regarded as a sort of flexible system consisting of balls of varying masses representing the
atoms of a molecule and springs of varying strengths representing the chemical bonds of a
molecule.

• The vibrations for molecules are of two types , namely : Stretching , and Bending.[15]

•The sole bond in a simple diatomic molecule is one that can stretch. Each atom in a
polyatomic molecule has three degrees of freedom in three directions that are
perpendicular to one another. As a result, an n-atom molecule possesses 3n degrees of
freedom. Two degrees of freedom explain rotation and three degrees of freedom describe
translation for a linear molecule, leaving 3n -5 for fundamental vibrations. While three

14
degrees of freedom indicate rotation and three degrees of freedom describe translation for
a non-linear molecule, the remaining 3n -6 are the number of basic vibrations. [17] More
complicated molecules may have more than one link, resulting in a variety of vibrations.
Stretching and bending are the two basic kinds of vibrations.

1. Stretching vibration(bond length altered)

The bond length is increased or decreased at regular intervals in this sort of

vibration. Stretching vibrations can be divided into two categories. Stretching that
is both symmetrical and asymmetrical. A molecule's bond is subjected to a variety
of vibrations and rotations. As a result, the atom is no longer stationary and is
constantly fluctuating. Stretching and bending modes define vibrational motions.
For diatomic or triatomic molecules, these movements are easily characterized.
This is not the case for big molecules because they will be subjected to many
vibrational movements and interactions. Stretching vibrations occur when the
interatomic distance between two atoms changes continuously along the axis of
the link between them. A bending vibration arises when the angle between two
bonds changes. Wagging, twisting, rocking, and scissoring are the four forms of
bending vibrations.

a.Symmetric Vibration

When atoms move in opposite directions when bonds shrink or stretch, this is known as a
symmetric stretch. In relation to the central atom, this is symmetric. As a result, there is
no change in the dipole moment, therefore this movement cannot be due to infrared
radiation absorption. [18]

b.Assymetric Vibration

When some atoms travel in the same direction while others move in the opposite
direction, this is known as an asymmetric stretch. There is no change in dipole moment
because this is not symmetric with regard to the core atom. The absorption of infrared
radiation causes an asymmetric stretch. When various atoms travel downward and upward
away from the core atom, a bend forms. This motion generates an electronegativity

15
imbalance and polarity shift. The absorption of infrared radiation can cause this motion.
The bending motion is seen in the image below, together with the other water vibrational
modes.[18]

Pharmatutor.org

2.Bonding Vibration(Bond angle altered)

• They entail the movement of atoms connected to a central atom.

• A change in the angle between two bonds characterises these.

• Deformation vibration is another name for it.

• It uses less energy, allowing it to appear at a longer wavelength.

• It happens less frequently: 1400-666cm-1.

• It's also categorised as:

a.In plane bending-

* In these types of vibrations, there is a change in bond angle.

* This type of bending takes place within the same plane.

i. Scissoring:

• Two atoms approach each other i.e. they move back & forth.

• Bond angles are decreased.

• Also known as symmetrical bending.

ii. Rocking:

• Movement of atoms takes place in the same direction i.e. they swing back & forth along
the central atom.

b) Out plane bending-

16
Outside of the molecule's plane, this kind of bending happens.

i. Wagging:

• Two atoms oscillate back and forth, moving to one side of the plane.

• They are able to move up and down the plane.

ii. Twisting:

• One atom moves above the plane & other move below the plane i.e. they rotate around
the bond which joins the central atom of molecule.[19]

FACTORS EFECTING MOLECULAR VIBRATION

1.Coupled Vibration

2.Fermi resonance

3.Electronic effect

4.hydrogen bonding

1.Coupled vibration

Coupled vibrations are those that occur at different frequencies with a greater wave
number.

Lower wavenumber—symmetrical

17
Asymmetrical—Wave number is higher

The following are the prerequisites for a successful coupling interaction:

I When the individual absorption frequencies of the coupled groups are approximately
identical, the interaction is strongest. (ii) When two groups share a common atom, strong
coupling between stretching vibrations occurs.

(iii) When groups are separated by one or more carbon atoms and the vibrations are
mutually perpendicular, coupling is insignificant.

(iv) When there is a common bond, such as between the bending vibrations of adjacent C-
H bonds in aromatic rings, coupling between bending vibrations can occur.

(v) If the stretching bond forms one side of the changing angle, then coupling between
stretching and bending vibrations can take place.[13]

2.Fermi resonance

A large amplitude vibration induced by a little oscillation is known as resonance. Two

new vibration modes occur when two fundamental vibration modes are coupled together,
with frequencies higher and lower than those measured in the absence of contact. Fermi
Interactions between basic vibrations and overtones or composite tone vibrations are
referred to as resonance.

If two different vibrational levels, belonging to the same species, have nearly the same
energy.

• There may be a reciprocal energy perturbation.

• One frequency shifts to a lower frequency, while the other shifts to a higher frequency.

• The strength of the corresponding bands increases significantly.

• A molecule's energy is transferred from the fundamental vibrational level to an overtone

or combination tone in this process. • Resonance separates the two levels and blends their
characters, resulting in each level having a fundamental and an overtone or combination
tone character.[20]

18
Hydrogen Bonding

proton's s - orbital can effectively overlap the acceptor group's P or orbital

• The stronger the hydrogen bond is, the longer the 0 - H bond is, the lower the vibration
frequency is, and the absorption band is broader and more intense.

• Hydrogen bonding affects the N-H stretching frequencies of amines in the same way
that it affects the hydroxyl group, but the frequency shifts are smaller for amines than for
hydroxyl compounds.

• Amines' hydrogen bonding is weaker than that of hydroxy compounds because nitrogen
is less electronegative than oxygen.[21]

• Intermolecular hydrogen bonds give rise to broad bands, while intramolecular hydrogen
bonds give sharp and well defined bands .

• Dilution distinguishes between intermolecular and intramolecular hydrogen bonding.

• Intramolecular hydrogen bonding is unaffected by dilution, and therefore the absorption

band is unchanged, whereas intermolecular bonds are broken by dilution, resulting in a
drop in the bonded 0 - H absorption.

• The strength of hydrogen bonding is also affected by :

• Ring strain

• Molecular geometry

• Relative acidity and basicity of the proton donor and acceptor groups.[22][18]

Electronic Effect

When the substituents in the vicinity of a group are modified, the absorption frequencies
for that group change.

It includes:

• Inductive effect.

19
• Mesomeric effect

• Field effect

• Inductive Effect

•The addition of an alkyl group creates the + I effect, which causes the bond to stretch or
weaken.

• As a result, the force constant is reduced, and the absorption wavenumber drops.
Compare the wave numbers of the following substances' v (C = 0) absorptions:

•Formaldehyde (HCHO).

•Acetaldehyde (CH3CHO).

• Acetone (CH3COCH3).

• When an electronegative atom or group is introduced, the -I effect occurs, causing the
bond order to increase.

•As a result, the force constant rises and the absorption wavenumber rises. [20]

Mesomeric effect

It causes lengthening or the weakening of a bond leading to the lowering of absorption

frequency.

Because the electron pair on the nitrogen atom in amide is less electronegative than the
electron pair on the oxygen atom, the electron pair on the nitrogen atom in amide is more
labile and participates in conjugation more.

20
The C = 0 absorption frequency in amides is substantially lower than in esters due to the
higher degree of conjugation. [23]

Field effect

•The lone pair of electrons on two atoms interact and affect the vibrational frequencies of
both groups in ortho substituted compounds through space interactions. This phenomenon
is known as the field effect. [20]

Region of Ir Spectroscopy
The wavelength range of the IR sector of the electromagnetic spectrum is 2 -15 m. The IR
region is traditionally separated into three regions: near IR, mid IR, and distant IR. The
majority of the IR utilized comes from the mid-IR area.

InfraRed Near Spectroscopy:

Because of vibrational overtones and combination bands, absorption bands in the near
infrared (NIR) range (750-2500 nm) are feeble. When two molecular vibrations are
excited at the same time, combination bands form. For each succeeding overtone, the
strength of the overtone bands decreases by one order of overtone. An overtone
absorption occurs when a molecule is stimulated from its ground vibrational state to a
higher vibrational state with a vibrational quantum number v larger than or equal to 2.
From v = 0 to v = 2, the first overtone appears. When v = 0 changes to v = 3, the second
overtone occurs. Near IR absorption produces weak transitions, which are referred to as
forbidden transitions. However, these transitions are important when non-destructive
observations are needed, such as on a solid sample. Although near infrared spectra have
low absorption, they have a high signal to noise ratio because of powerful radiation
sources, and NIR may penetrate undiluted substances and employ longer route lengths,
making it particularly useful for quick analysis. measurement of more representative
samples.[23]

Far InfraRed Spectroscopy:

Due to stretching and bending vibrations of bonds between metal atoms and ligands, the
far IR region is particularly relevant for inorganic research. These vibrations are typically
recorded at frequencies lower than 650 cm-1. When a persistent dipole moment is

21
present, pure rotational absorption of gasses is observed in the far IR region. H2O, O3,
and HCl are some examples. [24]

INSTRUMENTATION OF IR SPECTROSCOPY
The main parts of the IR spectrometer are as follows:

1.Radiation source

2.Sample cells and sampling of substances

3.Monochromators

4.Detectors

5.Recorder

A. IR radiation sources
IR instruments require a source of radiant energy that emits IR radiation that is
stable, powerful enough to be detected, and spans the appropriate wavelength
range. [24]

Various sources of IR radiations are as follows.

1.Nernst glower

2.Incandescent lamp

3.Mercury arc

22
4.Tungsten lamp

5.Glober source

6.Nichrome wire
B
. Sample cells and sampling of substances

IR spectroscopy has been used for the characterization of solid, liquid, or gas samples.

i. Solid - Solid samples are prepared using a variety of processes, including the pressed
pellet technique, solid run in solution, solid films, mull technique, and so on.

ii. Liquid — A liquid sample cell built of alkali halides may hold samples. Aqueous
solvents are ineffective because they dissolve alkali halides. Only organic solvents, such
as chloroform, are permitted.

iii. Gases– Gas sampling is comparable to liquid samples.

C. Monochromators

• Various types of monochromators are prism, gratings and filters.

• Prisms are made of Potassium bromide, Sodium chloride or Caesium iodide.

• Filters are made up of Lithium Fluoride and Diffraction gratings are made up of alkali
halides.

D . Detectors

• Detectors are used to measure the intensity of unabsorbed infrared radiation.

• Detectors like thermocouples, Bolometers, thermistors, Golay cells, and pyro-electric

detectors are used.

E . Recorders

• Recorders are used to record the IR Spectrum [24]

Radiation source

Incandescent lamp

● Nichrome wire

23
Nerst Glower

● Composed of zirconium, yttrium, thorium

● Diameter 2 mm, length 30 mm Oxide ●

● Provide radiation at 7100 cm - ¹ ● Temperature range = 1000-1800 ° C

● Disadvantage: emits IR over a wide wavelength.

Globar cell

● Intered silicon carbide

● Diameter 4 mm , length 50 mm

● Heated at 1300-1700 ° C

● Provide radiation at 5200 cm –

Mercury arc

● Made up of quartz.

MonoChromator

Prism

● NaCl (IR)

● Lithium /Ca fluoride (region of stretching)

Grating

● Made with aluminum, not attacked by moisture.

● Use Over considerable wavelength.

Detectors

Thermocouple

● Two dissimilar metal Bismuth and Antimony.

24
● Due to the temperature difference between two points, a potential difference is created,
which leads to the flow of electricity.

● Respone time is 60 m / sec.

Bolometer

● Made of 1arm of wheatstone bridge IR falls on conductor,

● temp changes, resistance changes ---- determined by current.

For 10 ° C in temp - resistance by C %.

Thermistors

● Made of a fused mixture of metal oxides.

As temp increases, electrical resistance 10 ° C 1 temp - resistance 5 % .

● Response time is 80 m / sec .

Golay cells

IR falls on metal plate heat gas expand deform diaphragm.

Light from lamp is made to fall on diaphragm reflect on photocell (any deviation Is determined)

● Response time is 20 m / sec.[25]

Semiconductor

● LeadSulphide / lead telluride . Insensitive to longer wavelengths.

As IR falls it is displaced conductivity changes.

● Other materials : Indium antimonide , germanium doped with copper or mercury used
as a detector .

Germanium detectors cooled

25]
in lig helium sensitive to longer wavelength.[

Pyroelectric

25
● When a dielectric is placed in an E.F., it gets polarised based on the dielectric constant;
when the field is withdrawn, the polarisation normally vanishes, except in ferroelectric
compounds, which maintain considerable residual polarisation. Their residual polarisation
can be temperature sensitive, which is known as pyroelectricity.

● E.g. :Triglycerine sulphate ( TGS ) cooled in liq N2 since response deteriorates above
450 and lost at 490 ( Curie point ) .[26]

APPLICATION OF IR SPECTROSCOPY
1. Drugs in Formulations Determination

2. The cis-trans isomer is determined.

3. Separate and describe the primary, secondary, and tertiary amine salts.

4. Investigation of complicated forms

5. In a study of quantitative reaction sequences.

6. When it comes to identifying functional groupings.

7. Explain the differences between two types of hydrogen bonds.

Characterization of proteins

9.Analysis of nanoscale semiconductors and

10.Exploration of space.

11.Gaseous, liquid, or solid sample analysis

12.Compound identification.

Quantitative analysis is the thirteenth step.[27]

Conclusion
Infrared spectroscopy (IR) identifies the components of a sample (liquid, solid or gas)
Infrared (IR) spectrometers determine how IR radiation interacts with substances. The
FTIR spectrometer determines the frequencies at which the samples absorb radiation, as
well as the intensity of those absorptions. A spectrum depicts the intensity and frequency
of sample absorption in a two-dimensional graphic. The quantity of light absorbed by a
sample is measured in absorbance, whereas the amount of light that flows through it is
measured in percent transmittance. This approach can provide vital information on what

26
makes up an unknown sample and how much of each component is present in that
sample. It has a wide range of uses, including research and development of new products.

REFERENCE
1. R. Escobedo, R. Miranda, and J. Martínez, “Separation of organic and inorganic
compounds for special applications,” International Journal of Molecular
Sciences, vol. 17, pp. 1–26, 2016.
2. K. Arora and A. Parma, “Simulation of IR spectra of some organic compounds-A
review,” IOSR Journal of Applied Chemistry, vol. 6, no. 1, pp. 10–24, 2013
3. R. M. Silverstein, G. C. Bassler, and T. C. Morrill, Spectrometric Identification
of Organic Compounds, 4th ed. (New York: Wiley, 1981), 166
4. G. Bellisola and C. Sorio, “Infrared spectroscopy and microscopy in cancer
research and diagnosis,” American Journal of Cancer Research, vol. 2, no. 1, pp.
1–21, 2012.
5. K. Hashimoto, V. R. Badarla, A. Kawai, and T. Ideguchi, “Complementary
vibrational spectroscopy,” Nature Communications, vol. 10, no. 1, pp. 1–6, 2019.
6. F. Long, Z. Chen, K. Han, L. Zhang, and W. Zhuang, “Differentiation between
enamines and tautomerizable imines oxidation reaction mechanism using
electron-vibration-vibration two-dimensional infrared spectroscopy,” Molecules,
vol. 24, no. 5, pp. 1–15, 2019.
7. H. Chen, C. Lu, J. Liu et al., “Synchrotron radiation X-ray powder diffraction
techniques applied in hydrogen storage materials-a review,” Progress in Natural
Science: Materials Internationa, vol. 77, pp. 66–73, 2017.

27
8. A. L. Le Sueur, R. E. Horness, and M. C. Thielges, “Applications of two-
dimensional infrared spectroscopy,” The Analyst, vol. 140, no. 13, pp. 4336–
4349, 2015.
9. M. R. Siddiqui, Z. A. Al Othman, and N. Rahman, “Analytical techniques in
pharmaceutical analysis: a review,” Arabian Journal of Chemistry, vol. 10, pp.
S1409–S1421, 2017.
10. P.L. Theophanis, J.M.Blackwell, Fourier transform infrared spectroscopic
characterization for detecting soil minerals and toxicological effects of selected
minerals in soils,” Acta Chimica and Pharmaceutica Indica, vol. 2, pp. 242–253,
2012.
11. Trishna B, Impact of the Revisit of Thermal Infrared Observations on
Evapotranspiration Uncertainty—soil mineralogy, pp 73 - 77 I pub 2019.
12. Dyer, jasen, ‘On the word Abscissa’ (2009)http://numberwarrior.wordpress.com/
pub, accessed may 2022.
13. F. Meirer and B. M. Weckhuysen, “Spatial and temporal exploration of
heterogeneous catalysts with synchrotron radiation,” Nature Reviews Materials,
vol. 3, no. 9, pp. 324–340, 2018
14. D. F. Barbin, A. L. d. S. M. Felicio, D.-W. Sun, S. L. Nixdorf, and E. Y. Hirooka,
“Application of infrared spectral techniques on quality and compositional
attributes of coffee: an overview,” Food Research International, vol. 61, pp. 23–
32, 2014.
15. J. Coates, “Interpretation of infrared spectra, a practical approach,”
in Encyclopedia of Analytical Chemistry, pp. 117–124, John Wiley & Sons,
Hoboken, NJ, USA, 2000.
16. L. Harper, J. Powell, and E. M. Pijl, “An overview of forensic drug testing
methods and their suitability for harm reduction point-of-care services,” Harm
Reduction Journal, vol. 14, no. 1, pp. 1–13, 2017.
17. S. M. Oliveira Junior, M. F. Diniz, R. C. L. Dutra, M. Massi, and C. Otani,
“Applications of FT-IR techniques and goniometry on characterization of carbon
fiber surfaces,” Journal of Aerospace Technology and Management, vol. 8, pp.
26–32, 2016

28
18. K. H. Michaelian, M. D. Frogley, G. Cinque, and L. Quaroni, “Infrared spectra of
micro-structured samples with MicroPhotoacoustic spectroscopy and synchrotron
radiation,” The Analyst, vol. 145, no. 4, pp. 1483–1490, 2020.
19. S. Simona, C. Carla, O. Giulia, and G. Elisabetta, “Infrared spectroscopy as a
new tool for studying single living cells: is there a niche?” Biomedical
Spectroscopy and Imaging, vol. 6, pp. 85–99, 2017.
20. M. Pilling and P. Gardner, “Fundamental developments in infrared spectroscopic
imaging for biomedical applications,” Chemical Society Reviews, vol. 45, no. 7,
pp. 1935–1957, 2016.
21. D. T. D. Childs, R. A. Hogg, D. G. Revin, I. U. Rehman, J. W. Cockburn, and S.
J. Matcher, “Sensitivity advantage of QCL tunable-laser mid-infrared
spectroscopy over FTIR spectroscopy,” Applied Spectroscopy Reviews, vol. 50,
no. 10, pp. 822–839, 2015.
22. E. Stavitski, R. J. Smith, M. W. Bourassa, A. S. Acerbo, G. L. Carr, and L. M.
Miller, “Dynamic full-field infrared imaging with multiple synchrotron
beams,” Analytical Chemistry, vol. 85, no. 7, pp. 3599–3605, 2013.
23. H.-Y. N. Holman, H. A. Bechtel, Z. Hao, and M. C. Martin, “Synchrotron IR
spectromicroscopy: chemistry of living cells,” Analytical Chemistry, vol. 82, no.
21, pp. 8757–8765, 2010.
24. Gurudeep R. Chatwal, Sham K. Anand,Instrumental Methods of analysis, pg
no :2.55 – 2.59.
25. V. Balan, C. Mhai, F. Cojocaru, C. Uritu, B. Dodi, and I. Gardikiotis,
“Vibrational spectroscopy fingerprinting in medicine: from molecular to clinical
practice,” Materials, vol. 12, no. 18, pp. 1–40, 2019.
26. https://www.vedantu.com/chemistry/infrared-spectroscopy,accessd 18th
may,2022.
27. T. P. Wrobel and R. Bhargava, “Infrared spectroscopic imaging advances as an
analytical technology for biomedical sciences,” Analytical Chemistry, vol. 90, no.
3, pp. 1444–1463, 2018.
28. L. M. Miller and P. Dumas, “From structure to cellular mechanism with infrared
microspectroscopy,” Current Opinion in Structural Biology, vol. 20, no. 5, pp.
649–656, 2010.

29
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