Iub Pha404 Autumn 2022 Chapter-2 SH Full

Infrared Spectroscopy
Department of Pharmacy
• Infrared spectroscopy is a type of

absorption spectroscopy that deals with
the infrared region of the electromagnetic
spectrum.
• Infrared radiation which extends from 0.8

to 200 μ affects only the vibrational and
rotational energy levels.
 The infrared region is subdivided into-
1. Near infrared region (NIR): extends from 0.8

to 2.5μ. It is concerned with low energy
electronic transitions as well as vibrational
and rotational changes.
2. Infrared region (IR): extends from 2.5 to 16μ
(frequency 4000 to 625cm-1). It affects only
the vibrational and rotational energy levels.
3. Far infrared region (FIR): extends from 16
to 200μ. It affects only the rotational energy
level of the molecule.
• The absorption of infrared radiation can be expressed either in

terms of wavelength (λ) or wave number (v).
• Mostly infrared spectra of organic compounds are plotted as

percentage of transmittance against wave number.
• Relation between wave length and wave number is:
wave number = 1/wavelength in centimeter

(1 μ = 10 -4 cm)
0.8 2.5 15 200 μ
Near IR Middle IR Far IR

12500 4000 667 50 cm -1
Theory of infrared absorption
 The energy of a molecule consists of translational,

rotational, vibrational and electronic energy .
E = Eelectronic + Evibrational + Erotational + Etranslational
 Translation energy of a molecule is associated

with the movement of the molecule as a whole, for
example in a gas. Rotational energy is related to
the rotation of the molecule, whereas vibrational
energy is associated with the vibration of atoms
within the molecule. Finally, electronic energy is
related to the energy of the molecule's electrons.
Theory of infrared absorption
 IR radiation does not have enough energy to induce

electronic transitions as seen with UV. Absorption of IR is
restricted to compounds with small energy differences in the
possible vibrational and rotational states.
 For a molecule to absorb IR, the vibrations or rotations within

a molecule must cause a net change in the dipole moment of
the molecule. The alternating electrical field of the radiation
interacts with fluctuations in the dipole moment of the
molecule. If the frequency of the radiation matches the
vibrational frequency of the molecule then radiation will be
absorbed, causing a change in the amplitude of molecular
vibration.
Molecular rotations
• Rotational transitions are of little use to us. Rotational

levels are quantized and absorption of IR by gases
yields line spectra. However, in liquids or solids, these
lines broaden into a continuum due to molecular
collisions and other interactions.
Molecular rotations
• The positions of atoms in a molecule are not

fixed; they are subject to a number of different
vibrations. Vibrations fall into the two main
categories of stretching and bending.
Molecular rotations
Stretching
• In this type of vibration, the distance

between the two atoms increases or
decreases but the atoms remain in the
same bond axis.
Stretching
 Stretching vibrations are two types-
 Symmetric stretching: In this type, the

movement of the atoms with respect a particular
atom in a molecule is in the same plane.
 Asymmetric stretching: In this type, one of the

atom moves toward the central atom while the
other moves away from it.
Stretching
Bending
• In this type of vibration, the position of the

atoms changes with respect to the original bond
axis. Bending vibrations are four types-
Bending
 Rocking: Movement of the atoms takes place in

the same direction.
 Scissoring: Two atoms move toward and away

from each other.
 Wagging: two atoms move up and below the plane

with respect to the central atom.
 Twisting: One of the atoms moves up the plane,

while the other moves down the plane with respect
to the central atom.
Bending
Calculation of stretching frequencies using
Hooke’s law:
Hooke’s law:
Problem-1: Calculate the stretching frequencies for C=C

bond. The force constant of a C=C bond is 10×105
dynes/cm.
Hooke’s law:
Problem 2: The force constant of C-H bond is 5×105

dynes/cm. Calculate the stretching frequency.
Instrumentation
 Two types of infrared spectrophotometer are in common use

in the laboratory.
1. Dispersive infrared spectrophotometer
2. Fourier transform infrared spectrophotometer (FTIR)

Dispersive infrared spectrophotometer
Dispersive infrared instruments are sometimes called

grating or scanning spectrometers. ... This grating is
similar to a prism. It separates the wavelengths of light in
the spectral range and directs each wavelength individually
through a slit to the detector.
 Dispersive IR spectrophotometer consists of the following
parts.
 Radiation source
 Sample compartment
 Monochromator
 Detector
 Recorder
Radiation source
• An inert solid is electrically heated to a temperature in
the range 1500-2000 0C. The heated material will then
emit infrared radiation.
Sample compartment
 After preparation of the sample (solid, liquid or gas), it
is inserted into a holder and then placed in the sample
compartment in the path of the IR beam. For solutions,
a reference cell is needed that contain solvent only.
After passing through the sample and reference, the
beams are chopped by a mirror which focuses each
beam alternately into the entrance slit of the
monochromator.
 Glass and plastics absorb strongly throughout the IR

region of the spectrum. Thus cells are constructed by
ionic substances like-
 Sodium chloride
 Potassium bromide
3. Monochromator: A monochromator
selects specific frequency from other
extraneous radiation using either a prism or a
diffraction grating.
4. Detector: The detector senses the

difference and determines which frequencies
have been absorb by the sample and which
frequencies remain unaffected and send a
signal which is amplified by an amplifier.
6. Recorder
The amplified detector signal is recorded as % of
transmitted light vs. frequency on a chart.
Fourier Transform Infrared Spectroscopy (FT-IR)
• Fourier transform infrared spectroscopy (FTIR) is a

technique which is used to obtain infrared spectrum of
absorption, emission, and photoconductivity of solid, liquid,
and gas.
• In FTIR, instead of recording the amount of energy absorbed

when the frequency of the IR radiation is varied by a
monochromator, the IR radiation is guided through an
interferometer. A diagram of a Michelson interferometer is
shown below:
The Michelson interferometer consists of a source, beamsplitter,

two mirrors, a laser and a detector.
The energy goes from the source to the beamsplitter which splits
the beam into two parts. One part is transmitted to a moving
mirror; one part is reflected to a fixed mirror. The moving mirror
moves back and forth at a constant velocity. This velocity is
timed according to the very precise laser wavelength in the
system which also acts as an internal wavelength calibration.
The two beams are reflected from the mirrors and recombined at
the beamsplitter. The beam from the moving mirror has traveled
a different distance than the beam from the fixed mirror.
When the beams are combined an interference

constructively and some destructively occurs. This
interference pattern is called an interferogram.
This interferogram then goes from the beamsplitter to

the sample, where some energy is absorbed and some
is transmitted. The transmitted portion reaches the
detector. The detector reads information about every
wavelength in the infrared range simultaneously.
To obtain the infrared spectrum, the detector signal is sent

to the computer, and an algorithm called a Fourier
transform is performed on the interferogram to convert it
into a single beam spectrum. A reference or “background”
single beam is also collected without a sample and the
sample single beam is ratio-ed to the background single
beam to produce a transmittance or
“%T” spectrum. This transmittance spectrum can be
converted to absorbance by taking the negative log10 of
the data points.
The x-axis of the FT-IR spectrum is typically displayed in

“wavenumbers”, or cm-1. This unit is a product of the
Fourier transform algorithm operating on the
interferogram and is the reciprocal of the actual
wavelength of light measured in centimeters at a point in
the infrared spectrum.
Preparation of sample
• IR spectra may be obtained from all phases: solid,

liquid and gaseous.
• Vapor or gaseous phase: The vapor or gas is
introduced into a special cell usually about 10cm
long which is then placed directly in the path of one
of the infrared beams. The end walls of the cell are
usually made of sodium chloride which is
transparent to infrared from 4000 to 650 cm-1.
• Liquids: The liquids are usually examined as a

thin film sandwiched between two polished salt
plates made of NaCl or KBr. The pair of plates is
inserted in a holder which is then placed in the
path of the IR beam. It is the simplest of all
procedures. KBr is transparent throughout the
4000 to 400 cm-1.
Solids
• Three methods are commonly used for preparing a solid

sample for measurement of IR absorption.
• a) KBr disc: About 1mg of the sample is finely ground

with about 300mg of KBr in a mortar. The mixture is
pressed into a disk using a special mould and hydraulic
press. Under pressure KBr melts and seals the
compound into a matrix and form a KBr disk. The KBr
disk can be inserted into a holder which is then placed in
the spectrophotometer.
• b) As Nujol mull (in the form of suspension): About

1mg of the sample is finely ground in a mortar with a
drop of nujol mull. The mull is then placed between flat
plates of NaCl.
• c) In solution: The compound is dissolved to give a 1-

5% solution in CCl4 or CHCl3. The solution is introduced
into a special NaCl cell. A second cell of equal thickness
but containing pure solvent is placed in the path of the
other beam of the spectrometer in order to cancel the
solvent absorption. For aqueous solutions, special
calcium fluoride cells are used
Advantages FT-IR Over Dispersible IR
• There are three major advantages in the

performance of an FT-IR spectrometer over a
dispersive infrared spectrometer.
• These advantages have been the reason for

the switch to the more modern FT-IR technique
in the last decade by infrared spectroscopists.
1. Multiplex Advantage
• An interferometer in an FT-IR instrument does not separate
energy into individual frequencies for measurement of the infrared
spectrum. Each point in the interferogram contains information
from each wavelength of light being measured. Every stroke of
the moving mirror in the interferometer equals one scan of the
entire infrared spectrum, and individual scans can be combined to
give better representation of the actual absorbance of the sample.
• In contrast, every wavelength across the spectrum must be
measured individually in a dispersive spectrometer. This is a slow
process, and typically only one measurement scan of the sample
is made in a dispersive instrument.
• The FT-IR advantage is that many scans can be completed and
combined on an FT-IR in a shorter time than one scan on a
dispersive instrument. The multiplex advantage results in faster
data collection of an FT-IR spectrum.
2. Throughput Advantage
• An FT-IR instrument does not use a slit to limit the individual
frequency reaching the sample and detector as a dispersive
instrument does.
• There are also fewer mirror surfaces in an FT-IR
spectrometer, so there are less reflection losses than in a
dispersive spectrometer.
• Overall, more energy reaches the sample and hence the
detector in an FT-IR spectrometer than in a dispersive
spectrometer. This means that the signal-to-noise ratio of an
infrared spectrum measured on an FT-IR is higher than the
signal-to-noise ratio attained on a dispersive instrument
which increase the sensitivity of small peaks that result in
sharp and more distinguisable peak in FTIR.
3. Precision Advantage
• An FT-IR spectrometer requires the use of a laser to control the
velocity of the moving mirror and to time the collection of data points
throughout the mirror stroke length for each scan.
• This laser is also available as a source of wavelength calibration within
the instrument. The laser wavelength is a constant value, and the x-
axis data points of the FT-IR spectrum are automatically referenced to
this known value to maintain internal precision and accuracy of the
wavelength positions.
• Spectra collected with an FT-IR spectrometer can be compared with
confidence whether they were collected five minutes or five years
apart. This capability is not available on a dispersive infrared system.
External calibration standards are required to control the accuracy of a
dispersive instrument, making spectra less comparable due to
instrumental unknowns during and between scans.
• Accuracy and precision in infrared spectra are much higher when
collected on an FT-IR.
Factors Influencing the Vibrational Frequencies
1. Electronic effect:
• Changes in the absorption frequencies of a particular group take
place when the substituent in the neibghour group of that
particular group is changed. The frequency shifts are due to
electronic effects which changes the force constant or the bond
strength resulting in the change of absorption frequency from the
normal value.
• Example: The abosrption of carbonyl group in the folloing
compounds are different:
Formaldehyde (HCHO) 1750 cm-1
Acetaldehyde (CH3CHO) 1745 cm-1
Acetone (CH3COCH3) 1715 cm-1
Chloroacetone (ClCH2COCH3) 1725 cm-1
Dichloroacetone (ClCH2COCH2Cl) 1740 cm-1
HOW THE FACTORS AFFECT C=O

STRETCHING VIBRATIONS
acid carboxylic
chloride ester aldehyde ketone acid amide
B A C
O O O O O O
R C R C R C R C R C R C
Cl OR' H R OH NH2
1800 1735 1725 1715 1710 1690
D
anhydride
O O
BASE
R C O C R VALUE
1810 and 1760

( two peaks ) A E-donating C Resonance
B E-withdrawing D H-bonding
2. Coupled Vibrations and Fermi resonance:

• We expect one stretching absorption frequency for an
isolated C-H bond but in case of methylene (-CH2-) group,
two absorptions occur which correspond to symmetric and
asymmetric vibration as follows:
• In such case asymmetric vibrations always occur at

higher wave number compared to symmetric vibrations.
These are called coupled vibrations because these
vibrations always occur at different frequencies than that
occurred for an isolated C-H stretching.
• Sometimes it happens that two different vibrational levels

have nearly the same energy. If such vibrations belong to
the same species then a mutual perturbation of energy
may occur, resulting in the shift of one towards lower
frequency and the other towards higher frequency. This
phenomenon is called Fermi resonance.
• Example:
- Fermi resonance is observed by aldehydes in which C-H
stretching absorption usually appears at a doublet (2820
and 2720 cm-1) due to interaction between C-H stretching
(fundamental) and the C-H bending.
- CO2 shows two bands at 1285.5 and 1388.3 cm-1.
- Acid anhydride shows two C=O stretching absorption
bands at 1810 and 1760 cm-1.
3. Hydrogen Bonding:
• Hydrogen bonding brings about remarkable downward frequency shifts.
Stronger the H-bonding greater is the absorption shifts towards lower
wave number than the normal value.
• Two types of H-bond (intramolecular and intermolecular) can be readily
distinguished by IR spectrum.
- Intermolecular H-bond: Gives broad bands, absorption is
concentration dependent and the absorption difference between free
and associated molecules is more.
- Intramolecular H-bond: Gives sharp and well defined bands,
absorption is concentration independent and the absorption difference
between free and associated molecules is less.
• In aliphatic alcohols, a sharp bands appear at 3650 cm-1 in dilute
solution due to free O-H group while a broad band is noticed at 3350
cm-1due to H-bonded O-H group.
4. Bond Angle:
Small rings also have a shift in vibrational frequency to

higher energy. Therefore 5,4, or 3 membered rings have
the carbonyl stretching frequency shifted.
Angle strain in these rings causes the carbonyl group to

have more electron density, Therefore a “stiffer” bond.
QUESTION??
What is the effect of concentration of salicylic acid or

benzoic acid on the absorption bands of carbonyl groups
at different concentration?
COOH COOH
OH
What would be the effect of increase temperature on the

infrared spectrum of a hydrogen-bonded compound?
Why FT-IR spectroscopy?
 Non-destructive technique
 Good precision
 No external calibration
 High speed
 Signal-Noise ratio
 Mechanically simple
What information can FT-IR provide?
 Identify unknown material
 Determine quality or consistency of sample
 Determine amount of components in mixture

IR Spectrum
The spectrum tells us;
• The infrared spectrum for a molecule is a graphical

display
• The spectrum has two regions:
 The fingerprint region 600-1200 cm-1

 Functional group region 1200-3600 cm-1
IR Spectrum
IR Spectrum
2-Methylpropanoic Acid
O-H
C-O C
C-H C=O CH3 CH OH
CH3
Application of IR spectroscopy
 1. Identification of functional group and

structure elucidation
 Entire IR region is divided into group frequency

region and fingerprint region. Range of group
frequency is 4000-1350 cm-1 while that of finger
print region is 1350-400 cm-1.
 In group frequency region, the peaks corresponding
to different functional groups can be observed.
According to corresponding peaks, functional group
can be determined.
2. Identification of substances
• Each atom of the molecule is connected by bond and

each bond requires different IR region so
characteristic peaks are observed. It contains a very
complicated series of absorptions. These are mainly
due to all manner of bending vibrations within the
molecule. This region of IR spectrum is called as
finger print region of the molecule. Identification of a
compound can be revealed by comparing its finger
print region with the spectra of authentic sample with
identical conditions.
• 3. Detection of impurities
• 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.
• 4. Quantitative analysis
• The quantity of the substance can be determined in
pure form. Most simple quantitative infrared
methods of analysis use the intensities of the C=O,
N–H or O–H groups.

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Infrared Spectroscopy

• Infrared spectroscopy is a type of

• Infrared radiation which extends from 0.8

 The infrared region is subdivided into-

1. Near infrared region (NIR): extends from 0.8

• The absorption of infrared radiation can be expressed either in

• Mostly infrared spectra of organic compounds are plotted as

• Relation between wave length and wave number is:

wave number = 1/wavelength in centimeter

0.8 2.5 15 200 μ

Near IR Middle IR Far IR

 The energy of a molecule consists of translational,

E = Eelectronic + Evibrational + Erotational + Etranslational

 Translation energy of a molecule is associated

 IR radiation does not have enough energy to induce

 For a molecule to absorb IR, the vibrations or rotations within

• Rotational transitions are of little use to us. Rotational

• The positions of atoms in a molecule are not

• In this type of vibration, the distance

 Stretching vibrations are two types-

 Symmetric stretching: In this type, the

 Asymmetric stretching: In this type, one of the

• In this type of vibration, the position of the

 Rocking: Movement of the atoms takes place in

 Scissoring: Two atoms move toward and away

 Wagging: two atoms move up and below the plane

 Twisting: One of the atoms moves up the plane,

Problem-1: Calculate the stretching frequencies for C=C

Problem 2: The force constant of C-H bond is 5×105

 Two types of infrared spectrophotometer are in common use

1. Dispersive infrared spectrophotometer

2. Fourier transform infrared spectrophotometer (FTIR)

Dispersive infrared instruments are sometimes called

 Glass and plastics absorb strongly throughout the IR

4. Detector: The detector senses the

• Fourier transform infrared spectroscopy (FTIR) is a

• In FTIR, instead of recording the amount of energy absorbed

The Michelson interferometer consists of a source, beamsplitter,

When the beams are combined an interference

This interferogram then goes from the beamsplitter to

To obtain the infrared spectrum, the detector signal is sent

The x-axis of the FT-IR spectrum is typically displayed in

• IR spectra may be obtained from all phases: solid,

• Liquids: The liquids are usually examined as a

• Three methods are commonly used for preparing a solid

• a) KBr disc: About 1mg of the sample is finely ground

• b) As Nujol mull (in the form of suspension): About

• c) In solution: The compound is dissolved to give a 1-

• There are three major advantages in the

• These advantages have been the reason for

HOW THE FACTORS AFFECT C=O

1810 and 1760

2. Coupled Vibrations and Fermi resonance:

• In such case asymmetric vibrations always occur at

• Sometimes it happens that two different vibrational levels

Small rings also have a shift in vibrational frequency to

Angle strain in these rings causes the carbonyl group to

What is the effect of concentration of salicylic acid or

What would be the effect of increase temperature on the

 Identify unknown material

 Determine quality or consistency of sample

 Determine amount of components in mixture