BY

P.Sravanthi
M.pharmacy 1styear-pharmaceutical analysis
TRINITY college of pharmaceutical sciences
INFRARED SPECTROPHOTOMETER
Classification of analytical
techniques

 1.Separation techniques →Chromatography

 2.Spectrophotometric → SPECTROSCOPY
 3. Electro analytical → Potentiometry,conductometry
 4. Titrimetric analysis→ Titrations
 SPECTROSCOPY
 Spectroscopy is the branch of science deals with the
study of interaction of electromagnetic radiation with
matter.
 Electromagnetic radiation is a type of energy that is
transmitted through space at enormous velocities.
 EMR→ANALYTE→SPECTROPHOTOGRAPH
↓
concentration should be lower
 IR SPECTROSCOPY
INTRODUCTION
 Infrared spectroscopy (IR) measures the bond
vibration frequencies in a molecule and is used to
determine the functional groups.
 The infrared region of the spectrum encompasses
radiation with wave numbers ranging from about
12,500 to 50cm-1 (or) wave lengths from 0.8 to 200µ.
 Infrared region lies between visible and microwave
region.
 The infrared region constitutes 3 parts

 a) The near IR (0.8 -2.5µm) (12,500-4000cm-1)

 b) The middle IR (2.5 -15µm) (4000-667cm-1)
 c) The far IR (15-200µm) (667-50cm-1)

 most of the analytical applications are confined to the

middle IR region because absorption of organic molecules
are high in this region.
 Wave number is mostly used measure in IR absorption
because wave numbers are larger values & easy to handle
than wave length which are measured in µm.
 E = hν = hc/λ = hcν¯
 It gives sufficient information about the structure of a
compound.
 PRINCIPLE OF IR SPECTROSCOPY
 The absorption of infra red radiation » excitation of
molecule.
 Vibrational transitions which are accompanied by a
change in dipole moment of the molecule are called
infrared active transitions. Because they absorb the ir
region.
 Atoms or groups of atoms are connected by bonds.
These are non rigid in nature.
 Due to continuous motion of the molecule they
maintain vibration with frequency.
 Applied frequency = natural frequency of vibration
 MOLECULAR VIBRATIONS
 There are 2 types of vibrations.
1) Stretching vibrations
2) Bending vibrations
• 1)Stretching vibrations: in this bond length is
altered.
• They are of 2 types
• a) symmetrical stretching: 2 bonds increase or
decrease in length.
b) Asymmetrical stretching: in this one bond length is
increased and other is decreased.

2)Bending vibrations:
• These are also called as deformations.
• In this bond angle is altered.
• These are of 2 types
• a) in plane bending→ scissoring, rocking
• b) out plane bending→ wagging, twisting
 Scissoring:
 This is an in plane bending.
 In this bond angles are decreased.2 atoms approach
each other.

Rocking:
• In this movement of atoms takes place in same
direction.
 Wagging:
 It is an out of plane bending.
 In this 2 atoms move to one side of the plane. They
move up and down the plane.

Twisting:
• In this one atom moves above the plane and the other
atom moves below the plane.
 NUMBER OF VIBRATIONAL MODES

 A molecule can vibrate in many ways, and each way is

called a vibrational mode.
 If a molecule contains ‘N’ atoms, total number of
vibrational modes
 For linear molecule it is (3N-5)
 For non linear molecule it is (3N-6)

 Eg: H2O, a non-linear molecule, will have 3 × 3 – 6 = 3

degrees of vibrational freedom, or modes.
 VIBRATIONAL FREQUENCY
 occurs when atoms in a molecule are in periodic motion
while the molecule as a whole has constant translational
and rotational motion.
 The frequency of the periodic motion is known as a
vibration frequency.
 The value of stretching vibrational frequency of a bond can
be calculated by the application of hooke’s law.
ν/c = ν¯ = 1/2пc[k/m1m2/m1+m2]1/2
= 1/2пc√k/µ
Where, µ→reduced mass
m1&m2 →masses of the atoms
k →force constant
c →velocity of radiation
 Factors influencing vibrational
frequencies
 Calculated value of frequency of absorption for a
particular bond is never exactly equal to its
experimental value.
 There are many factors which are responsible for
vibrational shifts
1) Vibrational coupling:
• it is observed in compounds containing –CH2 &
-CH3.
EG. Carboxylic acid anhydrides
amides
aldehydes
 2)Hydrogen bonding:
 Hydrogen bonding brings about remarkable
downward frequency shifts.
 Stronger the hydrogen bonding, greater is the
absorption shift towards lower wave length than the
normal value.
 There is 2 types of hydrogen bonding
a) inter molecular→broad bands
b) intra molecular → sharp bands
• hydrogen bonding in O-H and N-H compounds
deserve special attention.
• Eg: alcohols&phenols
enols & chelates
 3)Electronic effects:
 In this the frequency shifts are due to electronic effects
which include conjugation, mesomeric effect,
inductive effect.
 a) conjugation: conjugation lowers the absorption
frequency of C=O stretching whether the conjugation
is due to α,β- unsaturation or due to an aromatic ring.
 b) mesomeric effect: a molecule can be represented by
2or more structures that differ only in the arrangement
of electrons.
 c) inductive effect: depends upon the intrinsic
tendency of a substituent to either release or withdraw
electrons.
 INSTRUMENTATION
 There are 2 types of infrared spectrophotometer,
characterized by the manner in which the ir
frequencies are handled.
 1) dispersive type
 2) interferometric type
 In dispersive type the infrared light is
separated into individual frequencies by dispersion,
using a grating monochromator.
 In interferometric type the ir frequencies are
allowed to interact to produce an interference pattern
and this pattern is then analyzed, to determine
individual frequencies and their intensities.
 The main parts of an IR spectrometer are as follows
 1) IR radiation source
 2) monochromators
 3) sample cells
 4) detectors
 Infrared sources:
 require a source of IR radiant energy.
 The rod is electrically heated to 1500K to produce IR
radiations.
 a) Nernst glower or Nernst filament: they are made up of
sintered mixtures of oxides of Zr, Th, Ce, Y, Er, etc.
 Used in near IR region.
 Intensity of radiation produced is more intense.
 Used for detection of carbohydrates & proteins.
 b) globar source: made up of silicon carbide.
They are self starters.
used in middle IR region.
used to detect simple functional groups.
 C) incandescent lamp: it is made up of nichrome wire.
used in near IR instruments.
Has a low spectral emissivity.
It is heated up to 1100K.
Used to detect complex organic molecules
 d) mercury arc: high pressures are used.
used in far IR region.
intensity of radiation is greater.
used to detect inorganic complexes.
 e) tungsten lamp: they heated up to 3500K
used in mid IR region.
intensity of radiation is mild.
used to detection of organic functionalgrps

MONOCHROMATORS
• They convert polychromatic light into mono chromatic
light.
• They must be constructed of materials which transmit
the IR.
• They are of 3 types.
• a) metal halide prisms
• b) NaCl prisms
• c) gratings
 a) metal halide prisms:
 prisms which are made up of KBr are used in the
wavelength region from 12-25µm.
 LiF prisms are used in the wavelength region from
0.2-6µm.
 CeBr prisms used in wavelength region from 15-38µm.
 b) NaCl prisms:
 Used in the whole wave length region from 4000-
650cm-1.
 they have to be protected above 20•c because of
hygroscopic nature.
 c) gratings:
• They offer better resolution at low frequency than prisms.
 sample cells

 Sample cells made up of alkali halides like NaCl or

KBr are used.
 Aqueous solvents cannot be used as they dissolve alkali
halides.
 Only organic solvents like chloroform is used.
 IR spectroscopy has been used for the characterization
of solid, liquid, gas samples.
 detectors
• They convert the radiation into electrical signal.
• There are mainly 5 types of detectors used in IR.

 Thermo couple detectors:

• made up of bismuth& antimony coated
by metal oxides. If wires of 2 dissimilar
metals joined head to tail, then a difference
in temperature between head & tail causes
a current to flow in the wire. This current
proportional to the intensity of radiation.
• also called as thermopile
detectors.
• material should be thermally
active. used in dispersive instruments.
• Response time is 30 seconds.
• They give responses for all frequencies.
 Thermister detectors
 These are made up of sintered oxides
of Mn, Co, Ni.
• The material sh0uld be thermally
sensitive.
• The response time is 4 seconds.

pyro electric detectors

• These are made up of TGS, LiTaO3.
• They are used in FTIR instruments.
• It involves multiple scanning.
• Principle involved is electric polarization.
 Golay detectors
 The material used is CO2.
 The material should be
inert in nature.
• used in non dispersive IR
instruments.
• Response time is 0.01 sec.
• Principle involved is
expansion of gases.
 Photo conductive detectors
 Photoelectric detectors such as the mercury cadmium
telluride detector comprise a film of semiconducting
material deposited on a glass surface, sealed in an
evacuated envelope.
 Absorption of IR promotes nonconducting valence
electrons to a higher, conducting state.
 These detectors have better response characteristics
than pyroelectric detectors and are used in FT-IR
instruments - particularly in GC - FT-IR.
 DISPERSIVE INSTRUMENTS
 These are often double-beam recording instruments,
employing diffraction gratings for dispersion of
radiation.
 These 2 beams are reflected to a chopper which
consists of rotating mirror.
 It sends individual frequencies to the detector
thermopile.
 Detector will receive alternately an intense beam & a
weak beam.
 This alternate current flows from detector to amplifier.
INTERFEROMETRIC INSTRUMENTS
THE MICHELSON INTERFEROMETER:
 It is used to produce a new signal of a much lower
frequency which contains the same information as the
original IR signal.
 The output from the interferometer is an interferogram.
 Radiation leaves the source and is split.
 Half is reflected to a stationary mirror and then back to the
splitter.
 The other half of the radiation from the source passes
through the splitter and is reflected back by a movable
mirror. Therefore, the path length of this beam is variable.
The two reflected beams recombine at the splitter, and they
interfere .
 interference alternates between constructive and
destructive phases.
 The accuracy of this measurement system means that the
IR frequency scale is accurate and precise.
 FOURIER TRANSFORM IR
SPECTROMETER
 In the FT-IR instrument, the sample is placed between the
output of the interferometer and the detector. The sample
absorbs radiation of particular wavelengths.
 An interferogram of a reference is needed to obtain the spectrum
of the sample.
 After an interferogram has been collected, a computer performs a
Fast Fourier Transform, which results in a frequency domain
trace (i.e intensity vs wavenumber).
 The detector used in an FT-IR instrument must respond quickly
because intensity changes are rapid .
 Pyroelectric detectors or liquid nitrogen cooled photon detectors
must be used. Thermal detectors are too slow.
 To achieve a good signal to noise ratio, many interferograms are
obtained and then averaged. This can be done in less time than it
would take a dispersive instrument to record one scan.
 Advantages of Fourier transform
IR over dispersive IR
 Improved frequency resolution
 Improved frequency reproducibility (older dispersive
instruments must be recalibrated for each session of
use)
 Faster operation
 Computer based (allowing storage of spectra and
facilities for processing spectra)
 Easily adapted for remote use (such as diverting the
beam to pass through an external cell and detector, as
in GC - FT-IR)