Infra Red (IR) spectroscopy

Dr Minaketan Sahoo
Dept. of Pharma. Analysis
IPT Salipur
 Infrared Spectroscopy
Despite the Typical Graphical Display of Molecular Structures, Molecules are
Highly Flexible and Undergo Multiple Modes Of Motion Over a Range of Time-
Frames

Motions involve rotations, translations, and changes in bond lengths, bond angles, dihedral
angles, ring flips, methyl bond rotations.
 Electromagnetic Radiation
In the late 1800’s James C. Maxwell postulated that instead of visible light being composed
of small tiny divisible particles of matter, as is described by Newton in his Corpuscular
Theory of Light, it was comprised of electromagnetic waves.

Y
He further stated that this form of energy was
in the form of waves that contained two
fields, electrical and magnetic.

These two components having equal

wavelength and frequency are X
perpendicular to each other.
5
Electromagnetic Radiation

Wave nature of light

Electromagnetic Radiation
Electromagnetic Radiation
 Wide Range of Types of Electromagnetic Radiation in nature.
1. Only a small fraction (350-780 nM is visible light).
2. The complete variety of electromagnetic radiation is used throughout spectroscopy.
3. Different energies allow monitoring of different types of interactions with matter.

E=hn = hc/l
Electromagnetic Radiation
Electromagnetic Radiation

 The absorption of IR radiations can be expressed either in terms of wavelength (λ)

or in wave number (ū).
 Mostly IR spectra of organic compounds are plotted as percentage transmittance
against wave number.
Wave number = 1/ wavelength in cm
e.g. Wavelength ( λ) = 2.5 μ= 2.5 x 10-4cm
Wave number (ū) = 1 / λ = 1 / 2.5 x 10-4cm = 4000 cm-1
if, λ= 15μ = 15x10-4 cm
ū = 1 / λ = 1 / 15x10-4 cm = 667 cm
 H
Wagging
C
H
iii.) Types of Molecular Vibrations

Bond Stretching Bond Bending

symmetric In-plane rocking

asymmetric In-plane scissoring

Out-of-plane wagging

Out-of-plane twisting
 symmetric asymmetric In-plane scissoring

Out-of-plane twisting In-plane rocking Out-of-plane wagging

Another Illustration of Molecular Vibrations
C) Instrumentation
1.) Basic Design
- normal IR instrument similar to UV-vis
- main differences are light source & detector
C) Instrumentation
 These are either single beam or double beam.
 In single beam spectrometer the radiations emitted from the source are
passed through a cell containing the sample and through the prism which
disperses the light.
 Single beam spectrophotometers are simple, sensitive, accurate, versatile
and used to study fine details. But these instruments have two
disadvantages.
(1)When the spectra of the solution is to be recorded the absorption bonds
due to solvent are also obtained, thus, making the interpretation of the
bonds and identification of the compounds more difficult.
(2)The base line i.e. the line obtained without the use of the sample in the
light path; slopes because the intensity of the source changes
continuously with the wavelength.
C) Instrumentation

Components of Double beam IR spectrophotometer:-

(1)Radiation source.
(2)Detector.
(3)Other components
a) Sampling area.
b) Monochromator and optical material.
(4)Instrument for the measurement of response of the
recorder (Amplifier).
i.) Light Source:
- must produce IR radiation
- can’t use glass since absorbs IR radiation
- several possible types

a) Nernst Glower

Zr, Ce, Th

- rare earth metal oxides (Zr, Ce, Th) heated electrically

- apply current to cylinder, has resistance to current flow
generates heat (1200o – 2200o C).
- causes light production similar to blackbody radiation
- range of use ~ 670 – 10,000cm-1
- need good current control or overheats and damaged

b) Globar
- similar to Nernst Glower but uses silicon carbide rod instead of
rare earth oxides
- similar usable range
c) Incandescent Wire Source

- tightly wound nichrome or rodium wire that is electrically heated

- same principal as Nernst Glower
- lower intensity then Nernst Glower or Globar, but longer lifetime

d) CO2 Laser
- CO2 laser gas mixture consists of 70% He, 15% CO2, and 15% N2
- a voltage is placed across the gas, exciting N 2 to lowest vibrational levels.
- the excited N2 populate the asymmetric vibrational states in the CO 2 through
collisions.
- infrared output of the laser is the result of transitions between rotational states of
the CO2 molecule of the first asymmetric vibrational mode to rotational states of
both the first symmetric stretch mode and the second bending mode
- gives off band of ~ 100 cm-1’s in range of 900-1100 cm-1
- small range but can choose which band used & many compounds have IR
absorbance in this region
- much more intense than Blackbody sources

e) Others
- mercury arc (l > 50 mm) (far IR)
- tungsten lamp (4000 -12,800cm-1) (near IR)
ii.) Detectors:
- two main types in common IR instruments

a) Thermal Detectors
1.) Thermocouple
- two pieces of dissimilar metals fused together at the ends
- when heated, metals heat at different rates
- potential difference is created between two metals that varies with their
difference in temperature
- usually made with blackened surface (to improve heat absorption)
- placed in evacuated tube with window transparent to IR (not glass or
quartz)
- IR “hits” and heats one of the two wires.
- can use several thermocouples to increase sensitivity.

metal1 metal2
hn

- +
IR transparent
material (NaCl)
V
 2.) Bolometer
- strips of metal (Pt, Ni) or semiconductor that has a large change in
resistance to current with temperature.
- as light is absorbed by blackened surface, resistance increases and
current decreases
- very sensitive i

hn

A
b) Photoconducting Detectors
- thin film of semiconductor (ex. PbS) on a nonconducting glass surface
and sealed in a vacuum.
- absorption of light by semiconductor moves from non-conducting to
conducting state
- decrease in resistance  increase in current hn
vacuum
- range: 10,000 -333 cm-1 at room temperature

semiconductor

glass
Transparent
to IR
c) Pyroelectric Detectors
- pyroelectric (ceramic, lithium tantalate) material get polarized (separation
of (+) and (-) charges) in presence of electric field.
- temperature dependent polarization
- measure degree of polarization related to temperature of crystal
- fast response, good for FTIR
 iii.) Other Components
a.) Sample Cell
- must be made of IR transparent material (KBr pellets or NaCl)

Liguid Sample Holder NaCl plates

Sampling area:- (Sampling Technique):-

Various techniques can be employed for placing the sample in the
path of the IR beam depending upon the state of sample i.e.
solid, liquid or Gas.
(1)Sampling area:- (Sampling Technique):-
Solid samples: - Solid samples may be used in the form of solution, powder, glassy
film or pellet.
(i) Solids run in solution: - It the solid is soluble in some suitable solvent, its solution can be
made and run in one of the cells for liquids. Carbon disulphide is transparent below 1330cm -
1
. These solvents can be used for solution.
(ii) Solid films: - Amorphous solid samples may be cast into films from solution on an IR
transmitting window or solid melted between salt plates and allowed to cool into a film. A
solid film can also be deposited over the alkali metal halide (NaCl or KBr) disc by putting a
drop of the solution of the sample (sample dissolved in a volatile solvent) on the disc and
evaporating the solvent. Polymers, fats and waxy materials show excellent spectra in this
way.
(iii)Mulling Techniques: - Mulls are prepared by thoroughly grinding 1mg of a solid in a smooth
agate mortar. The powdered sample is mixed with little mulling reagents, such as, (1) Nujol
(a mixture of liquid paraffinic hydrocarbons with high molecular weights), (2)
Hexachlorobutadiene, (3) Chlorofluoro-carbon oil. (4) Per-fluoro kerosene. The powdered
sample is mulled to form a paste which is then transferred to flat plates of NaCl. The use of
mulling reagent makes possible a scan, essentially free of interfering bonds, over the 4000-
667 cm-1regions.
 Sampling area:- (Sampling Technique):-
Solid samples: -
iv. Pressed Pellet Techniques:-
This technique depends on the fact that dry, powdered alkali halide such as
KBr, KI, CsBr; can be pressed under high pressure in vacuum to form
transparent discs. The resulting transparent discs are inserted into a special
holder of the spectrophotometer. The substance under investigation should
be absolutely dry as water absorbs strongly at about 3710cm-1 and also near
1630 cm-1. The sample (solid substance) is ground with KBr and is made into a
disc after drying and then pressing it under elevated temperature at high
pressure. Also a blank disc is prepared with pure KBr which may be placed in
the path of the reference beam. It is often advisable to carry out grinding
under IR lamp to avoid condensation of atmospheric moisture. Grinding is
usually done in agate mortar and pestle. Discs obtained from poorly ground
mixture scatter more light than they disperse. The particle size of the ground
mixture should be less than 2mµ to avoid scattering. A complete spectrum
can be scanned by mixing 1-2% of the solid sample with KBr and then grinding
it to the desired particle size. KBr is transparent to the IR region (2.5µ-15µ).
Sampling area:- (Sampling Technique):-

Liquid samples: -
The simplest technique consists of sampling a liquid in a thin
film (0.1 to 0.3mm) squeezed between two NaCl plates i.e.
plates made of the material transparent to IR light. Non-
corrosive and too viscous liquids are measured in sealed tubes
to prevent undue evaporation. The cells (which may be cavity
cell, sandwich cell, demountable cells) are usually rinsed, filled
and emptied with the aid of a hypodermic syringe (without
needle). The cap of the cell is plugged with Teflon. CCl 4 and CS2
are used as solvents for dissolve limited number of compounds.
These are volatile, toxic and transparent in 4000-1350 cm-1 for
CCl4 and 1330-625 cm-1 for CS2. A drop of the low melting
substance can be placed on the NaCl plate and another NaCl
plate is placed on it. The pair of NaCl plates enclosing the liquid
film is then placed in the path of the sample beam.
Sampling area:- (Sampling Technique):-

Gases samples:-
The vapour or gas is introduced into a special gas
cell, usually about 10cm long. The walls of the cell
are made of NaCl, which allows the cell to be
placed directly in the path of the sample beam.
Very few organic compounds can be examined as
gases. The low frequency rotational changes in the
gaseous phase often split the high frequency
vibrational bonds. Only few thermally stable metal
carbonyls, Cyclopentadienyl compounds and
certain acetyl acetone complexes can be examined
in the gas phase.
iii.) Other Components
b.) monochromator
- reflective grating is common
- can’t use glass prism, since absorbs IR
iv.) Overall Instrument Design

- Need chopper to discriminate source

light
from background IR radiation
- Monochromator after sample cell
- Not done in UV-Vis since letting in all

hn to sample may cause

photdegradation (too much energy)
- IR lower energy
- Advantage that allows
monochromator to be used to
screen
out more background IR light
- Problems:
- Source weak , need long scans
- Detector response slow – rounded
peaks
v.) Fourier Transfer IR (FTIR) – alternative to Normal IR
- Based on Michelson Interferometer

Principle:
1) light from source is split by central mirror into 2 beams of equal intensity
2) beams go to two other mirrors, reflected by central mirror, recombine and pass
through sample to detector
3) two side mirrors. One fixed and other movable
a) move second mirror, light in two-paths travel different distances before
recombined
b) constructive & destructive interference
c) as mirror is moved, get a change in signal
Advantages of FTIR compared to Normal IR:
1) much faster, seconds vs. minutes
2) use signal averaging to increase signal-to-noise (S/N)

increase S / N  number scans

3) higher inherent S/N – no slits, less optical equipment, higher light intensity
4) high resolution (<0.1 cm-1)

Disadvantages of FTIR compared to Normal IR:

1) single-beam, requires collecting blank
2) can’t use thermal detectors – too slow

In normal IR, scan through frequency range. In

FTIR collect all frequencies at once.
D) Application of IR

1.) Qualitative Analysis (Compound Identification)

- main application
- Use of IR, with NMR and MS, in late 1950’s revolutionized organic
chemistry
► decreased the time to confirm compound identification 10-
1000 fold
i.) General Scheme
1) examine what functional groups are present by looking at group
frequency region
- 3600 cm-1 to 1200 cm-1
ii.) Group Frequency Region
- approximate frequency of many functional groups (C=O,C=C,C-H,O-H) can be
calculated from atomic masses & force constants
- positions changes a little with neighboring atoms, but often in same general region
- serves as a good initial guide to compound identity, but not positive proof.
Abbreviated Table of Group Frequencies for Organic Groups
Bond Type of Compound Frequency Range, cm-1 Intensity
C-H Alkanes 2850-2970 Strong
C-H Alkenes H 3010-3095 Medium
C C 675-995 strong

C-H Alkynes C C H 3300 Strong

C-H Aromatic rings 3010-3100 Medium

690-900 strong
0-H Monomeric alcohols, phenols 3590-3650 Variable
Hydrogen-bonded alchohols, phenols 3200-3600 Variable, sometimes broad
Monomeric carboxylic acids 3500-3650 Medium
Hydrogen-bonded carboxylic acids 2500-2700 broad

N-H Amines, amides 3300-3500 medium

C=C Alkenes 1610-1680 Variable
C=C Aromatic rings 1500-1600 Variable
C C Alkynes 2100-2260 Variable
C-N Amines, amides 1180-1360 Strong
C N Nitriles 2210-2280 Strong
C-O Alcohols, ethers,carboxylic acids, esters 1050-1300 Strong
C=O Aldehydes, ketones, carboxylic acids, esters 1690-1760 Strong
NO2 Nitro compounds 1500-1570 Strong
1300-1370
iii.) Fingerprint Region (1200-700 cm-1)
- region of most single bond signals
- many have similar frequencies, so affect each other & give pattern characteristics of
overall skeletal structure of a compound
- exact interpretation of this region of spectra seldom possible because of complexity
- complexity  uniqueness Fingerprint Region
iv.) Computer Searches
- many modern instruments have reference IR spectra on file (~100,000 compounds)
- matches based on location of strongest band, then 2 nd strongest band, etc

overall skeletal structure of a compound

- exact interpretation of this region of spectra seldom possible because of complexity
- complexity  uniqueness

Bio-Rad SearchIT database

of ~200,000 IR spectra
2.) Quantitative Analysis
- not as good as UV/Vis in terms of accuracy and precision
► more complex spectra
► narrower bands (Beer’s Law deviation)
► limitations of IR instruments (lower light throughput, weaker detectors)
► high background IR
► difficult to match reference and sample cells
► changes in e (A=ebc) common
- potential advantage is good selectivity, since so many compounds have different IR

spectra
► one common application is determination of air contaminants.
Contaminants Concn, ppm Found, ppm Relative error, %
Carbon Monoxide 50 49.1 1.8

Methylethyl ketone 100 98.3 1.7

Methyl alcohol 100 99.0 1.0

Ethylene oxide 50 49.9 0.2

chloroform 100 99.5 0.5

Example 9: The spectrum is for a substance with an empirical formula of C 3H5N. What is the
compound?

Nitrile or No aromatics
Aliphatic
alkyne group One or more
hydrogens
alkane groups

H2C C N

H3C
Applications of IR:-
1) Identification of organic compounds:-
The identity of an organic compound can be established from its finger print
region. The identity of an organic compound is confirmed if its finger print
region exactly matches with the known spectrum of that compound. The
compounds containing the same functional groups may have similar
absorptions above 1500cm-1 but they differ considerably in finger print
region.
2) Structure determination:-
IR helps to establish the structure of an unknown compound. All major
functional groups absorb at their characteristic wave numbers. From the
data available due to absorption frequencies the probable structure can be
predicted.
3) Qualitative analysis of functional groups:-
The presence or the absence of absorption bonds help in predicting the
presence of certain functional groups in the compound. The presence of
oxygen reveals that the group may be –0H, C=0, C00R, C00H, anhydride, etc.
Applications of IR:-
4) Identification of drug substance: - IR spectrum of sample and standard
compound to identify a substance. If the spectra are same, then the identity
of the sample can be confirmed.
5) Identifying the impurities in a drug sample: - Impurities have different
chemical nature when compared to the pure drug. Hence these impurities
give rise to additional peaks that of the pure drug.
6) Study of polymers.
7) Ratio of cis-trans isomers in a mixture of compounds.
8) Quantitative analysis: - The quality of a substance can be determined either
in pure form or as a mixture of 2 or more components. In this, the peak which
is characteristic for the drug is chosen and a comparison of the area of peaks
for standard and sample is done. This method is called as Baseline technique
and is thus used to determine the quantity of a substance.
9) Analysis of Petroleum Hydrocarbons, Oil, and Grease Contents.
10)Qualitative Analysis of Multilayered Polymeric Films using FTIR
Microspectroscopy.