Unit IV.

IR SPECTROSCOPY
 IR SPECTROSCOPY
Definition:
It is the spectroscopy which deals with the infrared
region(700 nm to 1000 μm) of the electromagnetic
spectrum with a longer wave length and lower frequency
than visible light.
Principle
IR spectra is produced by the absorption of energy by a
molecule in the infrared region and the transitions occur
between vibrational levels. Hence IR spectroscopy is
also known as vibrational spectroscopy
 IR spectrum
It is divided into three regions.
Near IR – 12500 to 4000 cm-1
IR – 4000 to 670 cm-1
Far IR – 670 to 50 cm-1
The most useful IR region lies between 4000 to
670 cm-1
 Theory of IR absorption

IR radiation does not have sufficient energy to induce

electronic transitions like UV-Visible spectroscopy. It
causes only vibrational and rotational changes. For a
molecule to absorb IR radiation, two conditions must
be satisfied.
 There must be change in the net dipole moment of
the molecule during the vibration.
 The energy of the IR radiation must match th
energy difference between two vibrational levels.
 Theory of IR absorption
The bonds of a molecule experience various types of
vibrations. The atoms are not stationary and fluctuate
continuously. Vibrational motions are defined by
stretching and bending modes. There are two types of
vibrations.
• Stretching Vibration – Symmetric and Asymmetric
• Bending Vibration –
(a) Inplane bending – Rocking, Scissoring,
(b) Outplane bending – Wagging and Twisting.
Stretching Vibration
Stretching Vibration
Bending Vibration
Bending Vibration
Bending Vibration
Modes of Vibrations
Modes of Vibrations
 Instrumentation

Fig. Block diagram of IR spectrophotometer

 Instrumentation
The basic components of an infrared
spectrophotometer are as follows.
1. Source
The most common sources used are the Nernst glower
and the globar. The Nernst glower is a tube made up
of zirconium, yttrium and thorium. The globar is a
cylindrical rod made up of Si-C. Both need to be
heated to 10000C to 18000C to emit IR radiation.
2. Sample cells and Sampling techniques
The sample cells are made up of NaCl and it is
transparent to IR light. Gaseous samples are taken
in a 10 cm long cell. Liquid samples are placed
between two discs of NaCl. Solid samples are
made into a mull by grinding with Nujol (mineral
oil) or a pellet by grinding with KBr pellets.
Monochromator
The monochromator separates polychromatic radiation
into individual wavelengths. Eg., NaCl, LiF, CaF 2, etc.
Detectors
They convert the light signal into electrical signal.
Photovoltaic cells, photoconductive cells, bolometers,
thermocouples, etc are used as detectors.
Amplifier / Recorder
The electrical signal is amplified and converted to
percentage transmittance as a function of wave
number and recorded.
 Typical apparatus
A beam of infrared light is produced and split into
two separate beams.
One is passed through the sample, the other passed
through a reference which is often the substance the
sample is dissolved in.
The beams are both reflected back towards a
detector, however first they pass through a splitter
which quickly alternates which of the two beams
enters the detector.
The two signals are then compared and a printout is
obtained
 IR spectroscopy Application

1. Identification of functional group and

structure elucidation
2. Identification of substances
3. Studying the progress of the reaction
4. Detection of impurities
5. Quantitative analysis
Theory of IR absorption
 Alkanes
The spectra of simple alkanes are characterized by
absorptions due to C–H stretching and bending (the C–C
stretching and bending bands are either too weak or of too
low a frequency to be detected in IR spectroscopy). In
simple alkanes, which have very few bands, each band in
the spectrum can be assigned.
C–H stretch from 3000–2850 cm-1
C–H bend or scissoring from 1470-1450 cm-1
C–H rock, methyl from 1370-1350 cm-1
C–H rock, methyl, seen only in long chain alkanes,
from 725-720 cm-1
 Alkanes
The IR spectrum of octane is shown below. Note
the strong bands in the 3000-2850 cm -1 region
due to C-H stretch. The C-H scissoring (1470),
methyl rock (1383), and long-chain methyl rock
(728) are noted on this spectrum. Since most
organic compounds have these features, these C-
H vibrations are usually not noted when
interpreting a routine IR spectrum.
 Vibrational Spectroscopy
A molecule composed of n-atoms has 3n
degrees of freedom, six of which are
translations and rotations of the molecule
itself.
This leaves 3n-6 degrees of vibrational
freedom (3n-5 if the molecule is linear).
Vibrational modes are often given descriptive
names, such as stretching, bending, scissoring,
rocking and twisting.
 Vibrational Spectroscopy
 The four-atom molecule of formaldehyde, the
gas phase spectrum of which is shown below,
provides an example of these terms.
 If a ball & stick model of formaldehyde is not
displayed to the right of the spectrum, press
the view ball & stick model button at the
lower right.
 We expect six fundamental vibrations (12
minus 6), and these have been assigned to the
spectrum absorptions.
Gas Phase Infrared Spectrum of
Formaldehyde, H2C=O
 Vibrational Spectroscopy

The exact frequency at which a given vibration occurs is

determined by the strengths of the bonds involved and the
mass of the component atoms. For a more detailed discussion
of these factors. In practice, infrared spectra do not normally
display separate absorption signals for each of the 3n-6
fundamental vibrational modes of a molecule. The number of
observed absorptions may be increased by additive and
subtractive interactions leading to combination tones and
overtones of the fundamental vibrations, in much the same
way that sound vibrations from a musical instrument interact.
 Vibrational Spectroscopy

• Furthermore, the number of observed absorptions

may be decreased by molecular symmetry,
spectrometer limitations, and spectroscopic
selection rules. One selection rule that influences
the intensity of infrared absorptions, is that a
change in dipole moment should occur for a
vibration to absorb infrared energy. Absorption
bands associated with C=O bond stretching are
usually very strong because a large change in the
dipole takes place in that mode.
 Some General Trends
I. Stretching frequencies are higher than
corresponding bending frequencies. (It is easier to
bend a bond than to stretch or compress it.)
II. Bonds to hydrogen have higher stretching
frequencies than those to heavier atoms.
III. Triple bonds have higher stretching frequencies than
corresponding double bonds, which in turn have
higher frequencies than single bonds. (Except for
bonds to hydrogen).
Spectrophotometric and Fluorometric Methods for the
Determination of Fe(III) Ions in Water and
Pharmaceutical Samples
Chemical sensors based on mesoporous silica nanotubes
(MSNTs) for the quick detection of Fe(III) ions have been
developed.
The nanotubes’ surface was chemically modified with
phenolic groups by reaction of the silanol from the silica
nanotubes surface with 3-aminopropyltriethoxysilane
followed by reaction with 3- formylsalicylic acid (3-fsa) or
5-formylsalicylic acid (5-fsa) to produce the novel
nanosensors.
The color of the resultant 3-fsa-MSNT and 5-fsa-MSNT
sensors changes once meeting a very low concentration of
Fe(III) ions.
 Continuation
Color changes can be seen by the naked eye and tracked
with a smartphone or fluorometric or spectrophotometric
techniques.
Many experimental studies have been conducted to find
out the optimum conditions for colorimetric and
fluorometric determining of the Fe(III) ions by the two
novel sensors. The response time, for the two sensors, that
is necessary to achieve a steady spectroscopic signal was
less than 15 s.
The suggested methods were validated in terms of the
lowest limit of detection (LOD), the lowest limit of
quantification (LOQ), linearity, and precision according to
International Conference on Harmonization (ICH)
guidelines.
 Continuation

The lowest limit of detection that was obtained

from the spectrophotometric technique was 18
ppb for Fe(III) ions.
In addition, the results showed that the two
sensors can be used eight times after recycling
using 0.1 M EDTA as eluent with high
efficiency (90%). As a result, the two sensors
were successfully used to determine Fe(III) in a
variety of real samples (tap water, river water,
seawater, and pharmaceutical samples) with
great sensitivity and selectivity.
 The absorption spectra and color change of different Fe(III) ion
concentrations with the 3-fsa-MSNT sensor at pH 4 (A) and with
the 5-fsa-MSNT sensor at pH 2 (B).
Calibration plots of the 3-fsa-MSNT sensor (A) and 5-fsa-
MSNT sensor (B) with different Fe(III) concentrations were
measured at absorbances of 378 and 495 nm, respectively
 Chromatographic methods
 Chromatography is a technique employed for separation
of the components of mixture by continues distribution
of the component between two phases.
 One phase moves (mobile phase) over the other phase
(stationary phase) in a continuous manner.
 When the stationary phase is a solid support of
adsorptive nature and mobile phase is liquid or gaseous
phase it is called Adsorption chromatography.
 When the stationary phase is liquid in nature and the
mobile phase is liquid or gaseous it is called Partition
Chromatography.
 Theory of Chromatography

Two theoretical approaches have been developed to describe

the processes involved in the passage of solutes through a
chromatographic system.
1. The plate theory:
According to martin and synge, a chromatographic system
consists of discrete layers of theoretical plates. At each of
these, equilibration of the solute between the mobile and
stationary phases occurs. The movement of solute is
considered as a series of stepwise transfers from plate to plate.
 Theory of Chromatography

2. The rate theory:

This theory considers the dynamics of the solute
particles as it passes through the void space between
the stationary phase particles in the system as a well
its kinetic as it is transferes to and from the stationary
phase.
 Phases of Chromatography
Normal Phase Mode:
In normal phase mode the stationary phase is polar and the
mobile phase is non polar in nature. In this technique, non
polar compounds travel faster and are eluted first. This is
because of the lower affinity between the non polar
compounds and the stationary phase. Polar compounds are
retained for longer times because of their higher affinity
with the stationary phase. These compounds, therefore take
more time to elute. Normal phase mode of separation is
therefore, not generally used for pharmaceutical
applications because most of the drug molecules are polar
in nature and hence take longer time to elute.
 Reversed Phase Mode
It is the most popular mode for analytical and preparative separations
of compound of interest in chemical, biological, pharmaceutical, food
and biomedical sciences. In this mode, the stationary phase is non
polar hydrophobic packing with octyl or octa decyl functional group
bonded to silica gel and the mobile phase is polar solvent. The polar
compound gets eluted first in this mode and non polar compounds are
retained for longer time. As most of the drugs and pharmaceuticals are
polar in nature, they are retained for longer times and hence elute
faster. The different columns used are Octa Decyl Silane (ODS) or C 18,
C8, C4 ( in the order of ncreasing polarity of the stationary phase). An
aqueous mobile phase allows the use of secondary solute chemical
equilibrium (such as ionization control, ion pairing and complexation)
to control retention and selectivity.