Infrared Spectroscopy

 Introduction
 Principle
 Theory
 Instrumentation
 FT-IR
 Application

By
[Link] [Link]
S.B. College of Pharmacy

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Introduction to IR Spectroscopy”

 Infrared Spectroscopy is an absorption spectroscopy

 Infrared Spectroscopy is the analysis of infrared light
interacting with a molecule.
 Infrared spectroscopy (IR spectroscopy) is the
spectroscopy, that deals with the infrared region of the
electromagnetic spectrum which is light with a longer
wavelength and lower frequency than visible light.
 All molecules are made up of two or more atoms.
 The atoms are connected by bonds in a molecules
 These bonds are not rigid structures they are analog to a
spring and they are continuously in motion.

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 The total energy of the molecule constitutes three types of
energy that are
Electronic Energy,
Vibrational Energy and
Rotational Energy.
Total Energy of the Molecules=Electronic Energy +
Vibrational Energy + Rotational Energy

Structure of a Molecule

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 INFRARED REGIONS RANGE

INFRARED REGIONS RANGE

800nm(0.8µm) To 1000000nm(1000µm)
Near IR Mid IR Far IR
0.8-2.5 µm 2.5-25 µm 25-1000 (µm)
Near-infrared Mid-infrared Far-infrared region
region region (667-100 cm-1)
(12,500-4000 cm-1) (4000-400cm-1)
Mid-IR region (4000-400cm-1)
Functional Group frequency region Finger Print region
4000Cm-1 to 1500 Cm-1 1500 Cm-1to 400Cm-1

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Principle of IR Spectroscopy:-
 All the atoms in molecules are in continuous vibration to
each other. When Mid-IR radiation falls on a molecule,
the frequency of a specific bond vibration is equal to the
frequency of the IR radiation, the molecule absorbs the
radiation.
 Infrared spectroscopy uses the fact that molecules absorb
frequencies that are characteristic of their structure.
 These absorptions occur at resonant frequencies, i.e. the
frequency of the absorbed radiation matches the
vibrational frequency.
 The energy or frequency absorbed serves to increase
the amplitude of the vibrational motions of the bonds
in the molecules.
 Infrared light interacts only with those vibrations whose
dipole moment (µ) periodically changes due to the
oscillation of the atom. If the oscillating electromagnetic

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 field of the incident photon couples with the dipole
oscillating at the same frequency, it is absorbed.
Criteria for IR Absorption
 A molecule must undergo a net change in dipole
moment.
 The frequency of IR radiation should exactly match the
natural vibrational frequency of the molecule.
Natural vibrational frequency is the frequency at which
a bond vibrates.
Absorption of IR frequency takes place when
Applied Frequency of = Natural vibrational frequency
IR Radiation of a bond
Dipole moment: [Link]

Compounds that don’t absorb IR Radiations

 Symmetrically substituted alkenes and alkynes

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  Many types of C-C Bonds
 Symmetric diatomic molecules
Ex. O2 (O-O), H2(H-H), Cl2(Cl-Cl), N2(N-N)
Wave Number: It is the number of fully completed waves
within a particular distance.
IR spectroscopy is expressed in wave number. (ie) number
of fully completed waves per centimeter (CM -1) as it is easy
to express in hole numbers but nm and micro meter has
decimals.
The unit of IR is Cm-1
1
Wave Length in μm = X 10000
Wave No. of wave cm-1

1
Wave Number (cm-1) = X 10000
Wave Length in μm
The IR radiation used in the IR spectroscopy is Mid-IR
Region only between 2.5-25 µm (4000-400Cm-1).

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Different Types Vibrations of Bonds

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 Two different types of vibration that take place in the bond
between two atoms of the molecules. They are
1. Stretching Vibrations
2. Bending vibrations
1. Stretching Vibrations: These vibrations are by a change in
the bond length between two bonds.
There are two types of Stretching Vibrations.
a) Symmetric Stretching Vibrations
b)Asymmetric Stretching Vibrations
a) Symmetric Stretching Vibrations: Stretching Vibrations in
which Inter atomic bond distance between 2 atoms
increases/decreases simultaneously.

Ex
b) Asymmetric Stretching Vibrations: Stretching Vibrations
in which Inter atomic bond distance between 2 atoms
increases/decreases alternatively.

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 Ex

Bending vibrations: It is a characterized vibration between

neighboring atoms by a change in the bond angle between
two bonds.
It requires less energy so appears at a longer wavelength.
Bending vibration is divided into two types:
1. In-Plane Bending
2. Out of Plane Bending
1. In-Plane Bending: Vibrations in which a change in bond
angle takes place within the same plane.
In Plan Bending vibration is divided into two types:
a) Scissoring Vibrations
b)Rocking Vibrations

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a) Scissoring Vibrations:
When 2 atoms move away or close to each other and bond
angle changes in the same plane.

OR EX
b)Rocking Vibrations
When 2 atoms move away or close to each other and bond
angle does not changes in the same plan.
The bond angle is maintained but both bonds move within
the same plane.

Or Ex
2. Out of Plane Bending
This type of bending takes place outside of the plan of the
molecule.
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 Out of Plan Bending vibration is divided into two types:
a) Wagging Vibrations
b)Twisting Vibrations
a) Wagging Vibrations: Out of Plan Bending vibration in
which both atoms move to one side of the plane.

Ex
b)Twisting Vibrations: Out of Plan Bending vibration in
which one atom is above the plane and the other is below
the plane.

Ex
Sample handling techniques in IR spectroscopy:

 In IR samples of different phases should be treated

differently
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 The common point to the sampling of different phases is
that the material used to dilute the sample must be
transparent to IR radiations.

 So the diluents used are NaCl or KBr.

Solid sample:

 Solid sample is done by pressed pellet technique in this

small amount of finely powdered solid is mixed and diluted
with 100 parts of powdered KBr salt.

 KBr will eliminate the problem of additional bands because

it won’t absorb IR radiation light in the IR radiation
wavelength region of 2.5- 15 µm. and a complete spectrum
of the sample is obtained.

 This mixture is pressed under a high pressure of 10000-

15000 pounds/[Link] in a die form a small pellet.

 Alternatively, the sample can be mixed with Nujol (mineral

oil) and a film of liquid (Nujol oil) is applied liquid sample
cell.

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 Pellet High Pressure Press

Liquid Samples: The liquid samples are mixed with nujol

oil and transferred to a sample shell made up of NaCl or

KBr.

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Instrumental Analysis

Light Source: The light should give sufficient, consistent,

and continuous IR radiation.
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ex. Nernst Glower, globar source, Incandescent wire source,
CO2 Laser

Nernst Glower:

Nernst Glower

 A cylinder made up of an alloy of Zirconium, Cerium,

and Thorium with resistance is heated electrically to a
temperature of about 1200-2200°C by a flow of current.

 This high temperature and flow of current cause the

production of light similar to blackbody radiation.

 The light radiation produced ranges from 670-

10,000Cm-1.

 It required good current control if it gets over heated

and gets damaged.

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Globar Source:

It is similar to the Nernst glower but the cylinder is made up

of silicon carbide rode instead of earth oxides and has a
similar radiation range.

CO2 Laser:

 Carbon dioxide laser has a gas mixture containing 70%

helium, 15% CO2, and 15% N2.

 When a voltage is passed across the gas it excites N2 to

lowest vibration levels.

 The excited N2 populates the asymmetric vibrational

state in the CO2 through collisions.

 IR output of the laser is produced when a transition

between the rotational state of the CO 2 molecule of the
first vibrational mode to the rotational states of both the
first symmetric stretch mode and the second bending
mode.

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Advantage: It has more intensity than the black body
source.

It has less source range but most of the compound has IR

absorption in this range only.

Mirrors and Monochromator: All mirrors and

monochromators are made up of KBr or NaCl as glass and
quartz absorb IR radiations. Monochromators used are
grating types like UV spectroscopy. The monochromator
most commonly used is reflective grating. |*In IR the
monochromator is present after the sample cell because the
energy of IR radiation is very weak so it will not enhance
photodegradation.

The chopper mirror is needed to discriminate source light

from the background IR radiation.

Sample cell: The sample cell is made up of IR transparent

material potassium bromide or sodium chloride.

In IR glass and quartz cannot be used as glass and quartz

absorb IR radiations.
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This sample cell is used only for liquid samples and water
shall not be used in IR spectroscopy as water dissolves the
KBr shell and also water gives IR spectrum.

Detectors: The detectors used are Thermocouple,

Bolometer, Photoconducting Detectors, and Pyroelectric
Detectors.

Thermocouple:

It has two pieces of dissimilar metal fused at the ends and

placed in an evacuated tube with a window transparent to IR
(KBr and not Glass or quartz as glass and quartz absorb IR
radiations).

When the IR radiation falls on the metals get heated up at

different rates.
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Due to the different temperatures in the two metals, a
potential difference is created between the metals.

The metal surface is blackened to improve heat absorption.

IR hits and heats one of the two wires.

Many thermocouples are used to increase the sensitivity.

Bolometer

It has a strip of metal like platinum or nickel or a semi-

conductor that has a large change in resistance to the current
with temperature.

As IR radiations fall in the black metal’s surface the metal

gets heated, resistances vary and current flowing through the
metal changes which shall be detected.

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Photo conducting Detectors:

It has a thin film of semiconductors (ex Pbs) on a non-

conducting glass surface and is sealed in a vacuum.

When IR radiations fall in the semiconductor it gets heated

and moves from non-conducting to conducting state.

An increases in temperature reduces the resistance and

increase in conductivity.

It has a range of 10,000-333 Cm-1at room temperature.

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Pyroelectric Detectors:

It is made up of pyroelectric materials like ceramic and

lithium tantalite which get polarized (separation of (+) and
(-) charges) in the presence of an electric field.

The polarization depends on the temperature produced when

the IR radiation falls in the pyroelectric material.

They measure the degree of polarization related to the

temperature of the crystals.

It has a very fast response and it is very good for FT-IR.

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Number of vibrations in the molecules

The number of vibrations in nonlinear molecules shall be

found with the formula.

No. of vibration =3N-6

N= No. molecules

The number of vibrations in linear molecules shall be found

with the formula.

No. of vibration =3N-5

N= No. molecules

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 Fourier-Transform Infra-Red (FT-IR)

Fourier-transform infrared (FT-IR) spectroscopy is based on

the idea of the interference of radiation between two beams
to yield an interferogram. The latter is a signal produced as
a function of the change in pathlength between the two
beams. The two domains of distance and frequency are
inter-convertible by the mathematical method of Fourier
transformation. The basic components of an FT-IR
spectrometer are a light source, Michelson Interferometers,
sample holder, detector, interface, and computer. The
radiation emerging from the source is passed through an
interferometer to the sample before reaching a detector. The
light from the source is split by a central beam splitter into
two beams of equal intensity. These two split beams are
refluxed by the fixed mirror and moving mirror. The
reflected beam recombines and passes through the sample to
the detector.
In the moving mirror, light travel different distances than
the fixed mirror before recombining and causing
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 constructive and destructive interference to get different
wavelength.
The beam splitter is made up of potassium bromide or
caesium iodide for the mid or near-infrared regions.
Instrumentation of FT-IR
Michelson
Compute
Source Interferomete Sample Detector Amplifier Interface
r
r

Michelson Interferometer

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 Qualitative Analysis

a)Identification of Unknown compounds by matching IR

Spectra:

 The IR spectrum is a fingerprint of compound and it is very

unique for every the compound and it doesn’t change for a
particular compound.

 The IR spectrum of known shall be matched with the IR

spectrum of Reference compounds.

 The IR spectrum of known shall be matched with the IR

spectrum of Reference compounds in standard books.

b)Confirmation of raw material and API in Dosage forms

by matching IR Spectra:

 Similarly, the raw material and active pharmaceutical

ingredients in Dosage shall also be confirmed by spectral
overlay and matched with IR spectrum of Reference
compounds in standard books.

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 IR spectrum of Sample

IR spectrum of Reference Standard

Overlaid IR Spectrum of Test sample & Reference STD

c) Identification of functional group and structure

elucidation

The entire IR region, the region used in IR spectroscopy is

the mid IR region of 4000-400Cm-1 which is divided into
functional group frequency region and fingerprint
region. The range of group frequency is 4000-1500 cm -
1
while that of the fingerprint region is 1500-400 cm-1.

In the group frequency region, the peaks corresponding to

different functional groups can be observed. According to
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 corresponding peaks, the functional group can be
determined.

Each atom of the molecule is connected by a bond and each

bond requires a different IR region so characteristic peaks
are observed. This region of the IR spectrum is called as
fingerprint region of the molecule. It can be determined by
characteristic peaks.

d)Identification of unknown substances

IR spectroscopy is used to establish whether a given sample

of an organic substance is identical to another or not. This is
because a large number of absorption bands is observed in
the IR spectra of organic molecules and the probability that
any two compounds will produce identical spectra is almost
zero. So if two compounds have identical IR spectra then
both of them must be samples of the same substances.

IR spectra of two enantiomeric compounds are identical. So

IR spectroscopy fails to distinguish between enantiomers.

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For example, an IR spectrum of benzaldehyde is observed
as follows.

C-H stretching of aromatic ring - 3080 cm-1

C-H stretching of aldehyde - 2860cm-1 & 2775cm-1
C=O stretching of an aromatic - 1700 cm-1
aldehyde
C=C stretching of an aromatic - 1595 cm-1
ring
C-H bending - 745 cm-1and 685 cm-1
No other compound than benzaldehyde produces the same
IR spectra as shown below.

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e) Studying the progress of the reaction

Progress of chemical reaction can be determined by

examining the small portion of the reaction mixture
withdrawn from time to time. The rate of disappearance of a
characteristic absorption band of the reactant group and/or
the rate of appearance of the characteristic absorption band
of the product group due to the formation of the product is
observed.

f) Detection of impurities

The IR spectrum of the test sample to be determined is

compared with the standard compound. If any additional
peaks are observed in the IR spectrum, then it is due to
impurities present in the compound.

g)Quantitative analysis

The quantity of the substance can be determined either in

pure form or as a mixture of two or more compounds. In
this, the characteristic peak corresponding to the drug
substance is chosen and the log I0/It of peaks for standard
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and test samples are compared. This is called base line
technique by selecting a characteristic peak of the
compound to determine the quantity of the substance.

But quantification in IR spectroscopy is not as good as

UV/Visible in terms of accuracy and precision

 More complex spectra

 Narrow linearity and range ( Beer’s law deviation)

 Limitations of IR instrument (low light through put,

weak detectors)

 High background IR

 Difficult to match the reference and sample cells

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 IR-Spectrum

X-Axis-wave number from 4000-400cm-1

Y-Axis-% Transmittance

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