Introduction To Infrared Spectrs

Introduction to Infrared
Spectroscopy
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Contents Page
1 Introduction to IR Spectroscopy 2
2 Electromagnetic Spectrum 3
3 Electromagnetic Radiation and Spectroscopy 4-5
4 Infrared Regions 6-7
5 Molecular Vibrations 8-9
6 Calculation of Molecular Vibrations 10-13
7 Infrared Active Modes 14
8 Absorption Considerations 15-16
9 The IR Spectrum 17
10 Infrared Absorption Signals 18
11 Dispersive IR Instruments 19-20
12 FTIR Instruments 21-23
13 FTIR Operation 24-25
14 Sample Preparation 26-31
14.1 As a Liquid 26-27
14.2 As a Solution 28-29
14.3 As a Nujol Mull 30
14.4 As a KBr Disc 31
15 Attenuated Total Reflectance (ATR) 32-33
16 Applications of IR Spectroscopy 34
17 References 35
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1. Introduction to IR Spectroscopy
Spectroscopy can be defined as the interaction between matter and light. Infrared spectroscopy is a
very powerful technique which uses electromagnetic radiation in the infrared region for the
determination and identification of molecular structure as well as having various quantitative
applications within analytical chemistry (Figure 1).
We do not aim to provide a mechano-quantic description of light and its interaction with atoms, as
this is out of the scope of this module. However, it is important to note that atoms can absorb
energy from electromagnetic radiation; this absorbed energy alters the state of the atoms within the
molecule. These changes are usually manifest in alterations to the frequency and amplitude of
molecular vibrations, which may be measured and plotted to produce an infrared spectrum. 1-4
Infrared spectrometers use optical devices for dispersing and focusing electromagnetic radiation of
IR frequency which is passed through the sample and any changes in absorbance measured against a
reference beam.
There are three well defined IR regions (near, mid and far). The boundaries between them are not
clearly defined and debate still persists, but broadly they are defined as:
 Near infrared (12820-4000 cm-1): poor in specific absorptions, consists of overtones and
combination bands resulting from vibrations in the mid-infrared region of the spectrum.
 Mid-infrared (4000-400 cm-1): provides structural information for most organic molecules.
 Far Infrared (400-33 cm-1): has been less investigated than the other two regions; however,
it has been used with inorganic molecules.
The low energies, typically encountered within the infrared region, are not sufficient to cause
electronic transitions; however, they are large enough to cause changes in the frequency and
amplitude of molecular vibrations.
Figure 1: The electromagnetic spectrum and the infrared region.
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2. Electromagnetic Spectrum
The electromagnetic spectrum is the range of all possible frequencies of electromagnetic radiation,
each of which can be considered as a wave or particle travelling at the speed of light, often referred
to as a photon. These waves differ from each other in length and frequency.
Frequency ν - the number of wave cycles that pass through a point in one second. Measured in
Hertz (Hz).
Wavelength λ - The length of one complete wave cycle (cm).
Frequency and wavelength are inversely related (Equation 1):
Where:
c = speed of light 3 x 1010 cm/sec
The energy of a photon (E in Joules) is related to wavelength and frequency as follows (Equation 2):
Where:
h = Planck’s constant 6.6 x 10-34 Joules-sec
Energy is directly proportional to frequency; therefore, high energy radiation will have a high
frequency.
Energy is inversely proportional to wavelength, hence, short wavelengths are high energy and vice
versa (Figure 2).
Figure 2: Electromagnetic spectrum.
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3. Electromagnetic Radiation and Spectroscopy
The frequency and wavelength of electromagnetic radiation varies over many orders of
magnitude. The electromagnetic spectrum is divided according to the type of atomic or molecular
transition that gives rise to the absorption or emission of photons; UV, IR, microwave, radio wave
etc. (Table 1).
Absorption spectroscopy relies on the absorption of energy from a photon which subsequently
promotes the analyte from a lower-energy state to a higher-energy, or excited, state. As the energy
of the photon changes the type of transition that the analyte undergoes will change. For example in
IR spectroscopy, the absorption of relatively low IR radiation results in the vibration of chemical
bonds within the analyte; a process which requires a fairly low energy input. Whereas, higher
energy photons, such as those found in the UV-visible region of the electromagnetic spectrum, will
promote valence electrons to move from their ground state to excited state energy levels within the
atoms of an analyte; a process that requires a much greater energy input.
Type of Energy Transfer Region of the Spectroscopic Technique

Electromagnetic
Spectrum
Absorption γ-ray Mossbauer
X-ray X-ray absorption
UV-Vis UV-Vis
Atomic absorption
Infrared Infrared (IR)
Raman
Microwave Microwave
Electron spin resonance
(EPR)
Radio waves Nuclear magnetic
resonance (NMR)
Emission (thermal UV-Vis Atomic emission
excitation)
Photoluminescence X-ray X-ray fluorescence
UV-Vis Fluorescence
Phosphorescence
Atomic fluorescence
Table 1: Electromagnetic spectrum region, type of energy transfer, and the associated spectroscopic
technique.
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Figure 3: Relationship between frequency and wavelength.
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4. Infrared Regions
Infrared spectroscopy can be rationalized as the spectroscopy that deals with electromagnetic
radiation of infrared frequency. As previously explained, there are three well defined infrared
regions; each of them has the potential to provide different information:
(Figure 4)
 Far-Infrared (400-33 cm-1): vibrations of molecules containing heavy atoms, molecular

skeleton vibrations and crystal lattice vibrations
 Mid-Infrared (4000-400 cm-1): useful for organic analysis
 Near Infrared (12820-4000 cm-1): overtones; very useful for quantitative analysis
Infrared spectroscopy is one of the most useful and widely used methods to perform structural
analysis.
Given that the molecule under investigation is infrared active, (i.e. it absorbs Infrared radiation),
then different types of structural information can be obtained.
Information achievable with Infrared spectroscopy includes:
1. The type of atoms within the molecule.
2. The type of bonds between atoms.
3. The molecular structure. More often than not, infrared spectroscopy is insufficient to determine
the complete structure and additional techniques (such as NMR, mass spectroscopy, etc.) are used
to solve the puzzle.
Both structures have the molecular formula C2H4O
4. From a quantitative point of view, infrared spectroscopy has a very well gained reputation for its
power, flexibility, and reliability.
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Figure 4: Infrared spectroscopy regions (oversimplified).
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5. Molecular Vibrations
The absorption of light will increase both amplitude and frequency of molecular vibrations. 4-5 When
the radiant energy matches the energy of a specific molecular vibration, absorption occurs.
Molecules with a permanent dipole moment, such as water, HCl, and NO, are infrared active.
The HCl molecule possesses a permanent dipole moment, so it is infrared active.
The O2 molecule does not possess a permanent dipole moment, so it is not infrared active.
In the case of alkenes (C=C) and alkynes (C≡C) if the bond is symmetrically substituted no band will
be seen in the IR spectrum, however, if the bond is asymmetrically substituted a stretching
frequency corresponding to the alkene or alkyne bond will be present (Table 2).
Oscillator Wavenumber (cm-1)

C-H 3320-2700
-C=C- 1690-1590
C=O 1870-1590
C-O 1300-1050
C≡C 2250-2150
C-Cl 800-600
Table 2: Wavenumbers for selected diatomic oscillators.
In order to understand molecular vibrations, a bond can be treated as a simple harmonic oscillator
composed of two masses (atoms) joined by a spring. Figure 6 depicts a diatomic molecule with two
generic atoms (of masses m1 and m2) connected by a spring.
Figure 5: Representation of a polyatomic molecule.
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Figure 6: Representation of a diatomic molecule. If masses m1 and m2 are equal, no change in the
dipole moment will occur as the molecule vibrates.
The classical vibrational frequency for a diatomic molecule (with force constant k and masses m1 and
m2) has been derived from Hooke’s Law (Equation 3):
√ √
Where:
µ = reduced mass =
In terms of the wavenumber ( ̅ ) (Equation 4):
̅ √ √
Where:
c = speed of light = 3 x 1010 cm/sec
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6. Calculation of Molecular Vibrations
The stretching frequency ̅ (cm-1) of C-H can be calculated as follows:
k = 5x105 dyne/cm
m1 = C = 12/6.02x1023 = 1.99x10-23 g
m2 = H = 1/6.02x1023 = 0.167x10-23 g
c = 3x1010 cm/sec
̅ √
̅ √
The observed values for C‒H bonds are in the region 3320-2700 cm-1, which differ from the
calculated values as the calculation does not take into account the environment of the C‒H group
within the molecule.
Force constants (k):
Single bond = 5x105 dyne/cm

Double bond = 10x105 dyne/cm
Triple bond = 15x105 dyne/cm
Remember higher wavenumber = higher frequency
The vibrational frequency of a bond will increase as 1) the strength of the bond increases and/or 2)
the reduced mass (μ) decreases.
For example compare the calculated frequencies for the following bonds:
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1) Increase in bond strength from single to double to triple bond (Figure 7).
C‒N bond
k = 5x105 dyne/cm ̅ √
m1 = C = 12/6.02x1023 = 1.99x10-23 g
m2 = N = 14/6.02x1023 = 2.33x10-23 g
̅ √
c = 3x1010 cm/sec
C=N bond
k = 10x105 dyne/cm ̅ √
m1 = C = 12/6.02x1023 = 1.99x10-23 g
m2 = N = 14/6.02x1023 = 2.33x10-23 g
̅ √
c = 3x1010 cm/sec
C≡N bond
k = 15x105 dyne/cm ̅ √
m1 = C = 12/6.02x1023 = 1.99x10-23 g
m2 = N = 14/6.02x1023 = 2.33x10-23 g
̅ √
c = 3x1010 cm/sec
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Figure 7: Comparison of wavelength, stretching frequency, and wavenumber of bonds with different
strengths (i.e. single, double, and triple bonds).
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2) Decrease in reduced mass (μ) (Figure 8).
C‒S bond
̅ √
k = 5x105 dyne/cm
m1 = C = 12/6.02x1023 = 1.99x10-23 g
m2 = S = 32/6.02x1023 = 5.32x10-23 g
̅ √
c = 3x1010 cm/sec
C‒H bond
̅ √
k = 5x105 dyne/cm
m1 = C = 12/6.02x1023 = 1.99x10-23 g
m2 = H = 1/6.02x1023 = 0.167x10-23 g
̅ √
c = 3x1010 cm/sec
Figure 8: Comparison of wavelength, stretching frequency, and wavenumber of bonds with different
reduced masses (µ).
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7. Infrared Active Modes
A molecule that is infrared active must undergo a change in its dipole moment when vibrating. The
simplest modes of vibration that are infrared active are stretching and bending modes (Figure 9).3-5
Figure 9: Modes of vibration (oversimplified).
For simplicity, we are going to illustrate the vibration modes in a linear molecule (acetylene in this
case).
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The symmetric C≡C stretching will not alter the acetylene’s dipole moment and it is not infrared
active.
However, substitution of either of the H atoms in acetylene produces an asymmetric alkyne bond
which will be IR active.
Homonuclear diatomic molecules such as Cl2, H2, N2, etc. will exhibit no infrared active modes as no
change in their dipole moment is experienced during vibration.
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8. Absorption Considerations
The amount of infrared radiation absorbed by the sample, at any given wavelength, can be
measured in different ways. The use of ‘transmittance’ is common place in infrared spectroscopy. 2-
4, 5-7
Figure 10 illustrates a beam of infrared radiation of power I0, directed at a sample solution.
Figure 10: Absorption of infrared radiation of radiant power I0. The radiation leaving the sample has
radiant power I.
The transmittance can be calculated as follows (Equation 5):
Where:
I0 = intensity of the incident radiation (light)
I = intensity of the radiation (light) leaving the sample
Absorbance is given by Equation 6:
( )
From the previous expression it follows that (Equation 7):
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Absorbance is related to concentration via the Beer-Lambert law (Equation 8):
Where:
ε = molar absorptivity (Lmol-1cm-1)
l = path length (cm)
c = concentration (molL-1)
ε is sometimes referred to as the extinction coefficient.
Infrared (IR) radiation of all wavelengths is transmitted from the source. Some of the wavelengths of
IR radiation will be absorbed by the sample and some of them will pass through (they are
transmitted). The IR radiation which is transmitted is measured by the detector resulting in a unique
IR spectrum for the sample of interest. This spectrum represents the IR absorption and transmission
of that molecule. No two unique molecules will produce the same IR spectrum, resulting in IR
spectroscopy being a very useful tool for molecular characterization and quantification.
Figure 11: Sample absorption of IR radiation.
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9. The IR Spectrum
Historically, infrared spectra have been represented as percent of transmittance versus either the
wavenumber or the wavelength. The use of wavenumbers, is standard, with the use of wavelength
(expressed in nm or µm) having fallen out of favor.4-6, 8-10
In terms of wavenumbers the infrared region spans from 33 to 12820 cm -1. However, most infrared
analyses are carried out in the mid-infrared region (400 to 4,000 cm-1).
By convention, the wavenumbers are plotted in decreasing order from left to right.
A typical IR spectrum is illustrated in Figure 12.
Figure 12: Typical IR spectrum.
The wavenumber ̅ can be found by using the following expression (Equation 9):
The wavenumber is usually expressed as cm-1.
Example: Calculate the wavenumber (in cm-1) corresponding to infrared radiation of wavelength
2,500 nm.
Note: 1 nm = 1 x 10-7 cm
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10. Infrared Absorption Signals
Selected IR frequencies.11
The vibrational frequencies of certain functional groups will correspond to the absorption of certain
wavelengths of IR energy. Absorption of these wavelengths by a sample can therefore act as a
diagnostic or fingerprint, to indicate the possible presence of these groups within the analyte
molecule. Some typical frequencies are shown in the table above.
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11. Dispersive IR Instruments
Most IR spectrometers can be categorized into two classes: dispersive and Fourier Transform
instruments.2-4
The basic design of a dispersive single beam instrument includes a source of infrared radiation, a
monochromator, and the detector (Figure 13).
After interacting with the sample (or the blank), infrared radiation is dispersed by a monochromator
into its individual frequency components and information on which frequencies were absorbed can
be obtained using a photodiode array detector.
Figure 13: Basic concept of a single beam IR instrument.
Sources and detectors for infrared radiation have limited stability; with light intensity and detector
sensitivity changing over time, or with fluctuations in temperature etc. The blank (reference or
background) and sample measurements should be made one after the other to ensure they are
made under the same analytical conditions. This limitation is minimized by the use of double beam
instruments which are capable of measuring the sample and reference simultaneously.
Double beam instruments use ‘choppers’ to control the path of the radiation, alternating between
the sample and the reference (Figure 14). These instruments use the known speed of rotation of the
beam chopper to compare and resolve the information reaching the detector.
The use of an opaque surface provides the means for adjusting the 0% transmittance response of
the detector.
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Figure 14: The double beam IR instrument.
Finally, it is easier to correct for absorption of infrared radiation by carbon dioxide and water
(present within the instrument background) with double beam instruments than with their single
beam counterparts.
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12. FTIR Instruments
FTIR stands for Fourier Transform Infrared. FTIR spectrometers consist of an IR source,
interferometer, sample cell or chamber, detector and a laser. 7,9,11 A schematic of an FTIR instrument
is shown below (Figure 15).
IR source
IR radiation is emitted from a glowing black body source. IR radiation passes through an aperture
which controls the amount of radiation that reaches the sample, and therefore, the detector.
Common IR sources are:
1. Silicon carbide rods which are resistively heated and commonly known as a Globar. An
electric current is passed through the rod which becomes very hot (1300 K) and emits large
amounts of IR radiation. Previously, cooling with water was required to avoid damaging
electrical components; however, advances in metal alloys have led to the production of
Globars that do not require cooling by water.
2. Nichrome and Kanthanl wire coils were once popular IR sources and did not require cooling
as they ran at lower temperatures than Globars, however, this also resulted in lower
amounts of IR radiation being emitted.
3. Nernst Glowers are manufactured from a mixture of refractory oxides and are capable of
reaching hotter temperatures than a Globar; however, they are not capable of producing IR
radiation above 2000 cm-1.
Interferometer
The first interferometer was invented by Albert Abraham Michelson, who received a Nobel Prize for
his work in 1907. Without this essential piece of optical equipment the modern day FTIR system
would not exist. The interferometer consists of a beam splitter, a fixed mirror, and a moving mirror.
Beam Splitter
The beam splitter is made of a special material which transmits half of the incident radiation and
reflects the other half. IR radiation from the source strikes the beam splitter and is separated into
two beams. One beam is transmitted through the beam splitter to the fixed mirror while the other
beam is reflected from the beam splitter to the moving mirror. Both mirrors reflect the radiation
back to the beam splitter where the two beams interfere to produce an interferogram.
Moving Mirror
The moving mirror is a flat highly reflective surface mounted on air bearings that allow for high
speed movement of the mirror (movements are made once every millisecond). The moving mirror
only moves a few millimeters away from the beam splitter.
Fixed Mirror
The fixed mirror is a flat highly reflective surface.
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Figure 15: Operational Schematic of a Thermo Nicolet FTIR Instrument. Image reproduced with
permission from Thermo Fisher Scientific (Madison, WI, USA).
Laser
Many instruments employ a Helium-Neon laser as an internal wavelength calibration standard. It is

imperative that the position of the moving mirror is known at any given moment. The moving mirror
moves back and forth at a precise constant velocity that is timed using a very accurate laser
wavelength.
The intensity of the laser beam is measured at two points in the interferometer. As the mirror
moves the intensity at these two points will rise and fall due to the enhancement and cancellation of
the HeNe beam paths, producing a sine wave of intensity vs. mirror position. The number of
“fringes” in the sine wave allows the instrument to know exactly how far the mirror has moved, and
the relative phase of the sine wave tells the instrument in which direction the mirror is moving.
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Detector
There are two classes of infrared detectors; thermal and photonic detectors. Thermal detectors use
the IR radiation as heat; whereas, quantum mechanical (photonic) detectors use the IR radiation as
light which results in a more sensitive detector.
Thermal detectors: detect changes in temperature of an absorbing material (lithium tantalate

(LiTaO3), lead selenide (PbSe), germanium etc.). Many temperature dependent phenomena can be
followed to measure the effects of the incident IR radiation. Bolometers and microbolometers use
changes in resistance, while thermocouple and thermopiles use the thermoelectric effect. Golay
cells monitor thermal expansion.
Photonic Detector: exhibit faster response times and higher sensitivity in comparison to their
thermal counterparts, therefore, they are much more prolific in FTIR instruments. The materials
used in these detectors are semiconductors with narrow band gaps. The incident IR radiation causes
electronic excitations between the ground and first excited states, which in photoconductive
detectors result in a change in resistivity which is monitored.
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13. FTIR Operation
Prior to the development of FTIR spectrometry, the limitation within IR was the slow scanning
process. FTIR allows for all the infrared frequencies to be scanned simultaneously, allowing for data
to be collected in a matter of seconds rather than several minutes. This is achieved through the use
of an optical device called an interferometer which produces a signal which is made up of all of the
infrared frequencies.
Most interferometers consist of a beam splitter which splits the incident infrared beam into two
separate optical beams. One beam is reflected from a fixed mirror, while the other beam is reflected
from a mirror that is constantly moving in the instrument. The moving mirror typically moves by
only a few millimeters from the beam splitter (Figure 16).
Figure 16: FTIR based on the Michelson interferometer. By changing the position of the moving
mirror, a different optical path is established and different information is obtained.
The two beams are reflected from their respective mirrors and recombine at the beam splitter. The
path length of the beam that is reflected from the fixed mirror remains constant, while the path
length of the beam that is reflected from the moving mirror is constantly changing as the mirror
moves. The signal that exits the interferometer is the result of these two beams interfering with
each other, and is called an interferogram (Figure 17).
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Figure 17: Interferogram.
The interferogram is unique in that every data point, which is a function of the moving mirror
position, has information about every infrared frequency emitted from the source. This allows for all
frequencies to be measured simultaneously.
The interferogram is converted to a more familiar IR spectrum (wavenumber vs. % transmittance)

using the well-known mathematical technique called Fourier transformation. The transformation of
the interferogram is carried out by the instrument software.
IR spectra are presented on a relative scale (%T), therefore, a background spectrum must be
measured. A background spectrum is taken with no sample in the beam and is then subtracted from
the sample spectrum to remove artifacts generated by the instrument or air (i.e. water, carbon
dioxide, etc.).
FTIR Advantages
FTIR instruments have several advantages over dispersive IR instruments including:
Speed
All IR frequencies are measured simultaneously, resulting in measurements being taken in seconds
rather than minutes. This is often referred to as the Felgett Advantage.
Sensitivity
The detectors utilized in FTIR instruments are highly sensitive which results in lower signal to noise
ratios. This is known as the Jacquinot Advantage.
Simplicity
The only moving part in an FTIR instrument is the mirror in the interferometer; therefore, there is
very little need for mechanical maintenance.
Internal calibration
The internal laser is used to self-calibrate the moving mirror in the FTIR instrument negating any
need for timely or complicated external calibration. This is denoted as the Connes Advantage.
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14. Sample Preparation
Proper sample preparation is required to obtain meaningful spectra with sharp peaks, which have
good intensity and resolution. Ideally the largest peaks should be attributable to the compound
being analyzed opposed to the background or sample matrix (water, CO2, solvent etc.) and should
ideally have an intensity of 2-5 %T for the strongest peaks in the spectrum (Figure 18). A
transmission of 5 % is equivalent to an absorbance (A) = 1.3 (i.e. the amount of light that is absorbed
by the sample), which is the upper detection limit for most detectors.
The equation above is worth remembering as it allows the absorbance of a sample to be calculated
from the percentage transmittance data.
Peaks that are of higher intensity will be cut off and the sample will need to be prepared again.
Compounds can be analyzed in the vapor phase, as pure liquids, in solution, and as solids.
Figure 18: Classification of IR signals.
15.1. As a Liquid
A drop of the liquid is squeezed between two sodium chloride (NaCl) plates, which are transparent in
the 4000-625 cm-1 region (Figure 19).
The plates are then placed in a holder and a spectrum is taken.
If the peaks in the spectrum are too intense the liquid can be wiped from one plate, then the
spectrum taken again.
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Figure 19: Sodium chloride (NaCl) plates in various conditions.
NaCl plates are very fragile and sensitive to water. Samples should never be dissolved in water and
placed on a NaCl plate as it will fog up or dissolve. The plates should be held by the edges to avoid
moisture from fingers damaging them. After a sample has been run, ethanol can be used to clean
the plates. Moisture in the air can also damage NaCl plates; therefore, they should be stored in a
desiccator. Cloudy or damaged plates (pitted, fingerprints etc.) will result in poor spectra with broad
bands and spectra with less than optimum transmission (Figure 20). Cloudy plates can be restored
by polishing.
Liquids can also be placed directly on an Attenuated Total Reflectance (ATR) plate which will be
discussed later.
Figure 20: Representative spectra obtained with sodium chloride (NaCl) plates in various conditions.
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15.2. As a Solution
Samples can be dissolved in an appropriate solvent to give a solution. The spectrum is then taken by
placing a drop on a NaCl plate or by using a sodium chloride solution cell (Figure 21). Solvents
should be free of water to avoid damaging the sodium chloride cell surfaces. A reference spectrum
of the blank solvent should be obtained and subtracted from the sample spectrum.
When solvents absorb ~80% of the incident light, spectra cannot be obtained because insufficient
light will be transmitted and detected. The regions in which common solvents absorb too strongly to
give meaningful spectral information from a sample are shown in the table below.
If aqueous solvents must be used for solubility, special calcium fluoride cells can be used.
Figure 21: Sodium chloride solution cell. Image reproduced with permission from International
Crystal Laboratories (Garfield, NJ, USA).
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Transmission Characteristics of Common Solvents.
Transmission below 80%, with a 0.10 mm cell path are shown as darkened areas.
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As a Nujol Mull
IR spectra of solid samples can be obtained using a Nujol mull. Nujol is a mineral oil which itself has
an IR spectrum (Figure 22).
Figure 22: IR spectrum of pure Nujol.
A small amount of sample is ground using a small agate mortar and pestle and a drop of Nujol
(Figure 23). The mull is then pressed between two NaCl plates and the spectrum obtained. The mull
should appear transparent and free of bubbles when properly prepared. If the peaks in the
spectrum are too strong one plate can be wiped clean and the spectrum re-run.
Figure 23: Agate mortar and pestle. Image reproduced with permission from Cole-Parmer (Hanwell,
London, UK).
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As a KBr Disc
A solid sample can be ground with 10-100 times its mass of pure potassium bromide (KBr). Solid
samples should be finely ground before adding the KBr. This is then pressed into a disc using a
special mold and a hydraulic press (Figure 24). The use of KBr eliminates any bands that may
obscure analyte signals when using a Nujol mull. A band at 3450 cm -1 will often be present and is
attributable to the OH group from traces of water. Water can be minimized by drying the KBr in an
oven. Excessive grinding of the hygroscopic KBr can increase the water content.
Solid state spectra can differ greatly from solution state spectra due to intermolecular interactions
between functional groups, i.e. hydrogen bonding. Conversely, solid state spectra will often exhibit
a greater number of resolved bands which can aid in compound identification.
Material Wavelength Range (µm) Wavelength Range (cm-1) Refractive Index at 2 µm

NaCl 0.25-17 40,000-590 1.52
KBr 0.25-25 40,000-400 1.53
KCl 0.30-20 33,000-500 1.5
Table 3: Material used for obtaining solid state IR spectra. Note these materials can also be used to
produce plates and solution cells for obtaining spectra with liquids and mulls.
Reasons for Cloudy Discs
 KBr mixture not properly ground

 Sample was not dry
 Sample:KBr ratio too high
 Disc too thick
 Sample has a low melting point
Figure 24: KBr Press. Image reproduced with permission from Specac (Orpington, Kent, UK).
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15. Attenuated Total Reflectance (ATR)
As has been discussed previously, IR spectra can be obtained from samples as liquids, solids, or
mulls; however, the primary drawback is the sample preparation that is required to obtain good
quality spectra. IR instruments which utilize an attenuated total reflectance (ATR) stage negate the
necessity for complex and timely sample preparation resulting in good quality, reproducible
spectra.
With traditional means of IR spectroscopy the IR radiation is passed through the sample and the
resulting radiation which is transmitted is measured. Attenuated total reflectance measure the
changes which occur in a totally internally reflected IR beam when it is in contact with a sample
(Figure 25).
Figure 25: Evanescent wave.
The infrared beam enters the crystal which is made of an optically dense material (i.e. it has a high
refractive index) at a particular angle of incidence, the IR beam is internally reflected (usually
between five and ten times), this internal reflectance results in the production of an evanescent
wave which can extend beyond the crystal surface and into the sample itself. The wave will usually
penetrate into the sample with a depth of 0.5-2 µm. The depth to which the wave penetrates is
dependent on the angle of the incident IR beam and the refractive index of the crystal material and
sample itself.
When a samples absorbs the infrared radiation there is a change in the evanescent wave; in other
words the wave is attenuated. The attenuated energy from each of the evanescent waves is then
transferred back to the IR beam which exits the crystal and is measured by the detector to produce
an IR spectrum.
Typical ATR Crystal Materials
In ATR instruments the crystal is an optically dense material which has a refractive index that is
greater than the sample. Common ATR crystal materials are listed in Table 4. The most common are
zinc selenide (ZnSe) and Germanium (Ge).
Zinc selenide is applicable to the analysis of liquids and non-abrasive pastes. It has a working pH
range of 5 – 9. Germanium is more robust with a working pH range of 1 – 14 and can be used to
analyze weak acids and alkalis. For a greater initial cost, instruments which utilize diamond as the
ATR crystal material exhibit greater durability and robustness, with the crystal having to be replaced
less in comparison to ZnSe and Ge.
Materials such as ZeSe and Ge can scratch easily; therefore, care must be taken when cleaning the
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crystal surface. It is recommended that crystal surfaces are cleaned with lint free tissues soaked
with solvents such as water, methanol or isopropanol.
Material Wavelength Range (cm-1) Refractive Index

ZnSe 20,000-500 2.43
ZnS 22,000-750 2.25
Ge 5,000-600 4.01
Si 10,000-100 3.42
Diamond 45,000-10 2.40
Table 4: Attenuated total reflectance (ATR) crystal materials.
ATR Instrument
In traditional ATR instruments the sample was clamped against the vertical face of the crystal. This
design has now been replaced by horizontal ATR stages where the upper surface of the crystal is
exposed (Figure 26). ATR accessory kits can be purchased which can be used to modify existing IR
instruments.
Similarly to FT-IR instruments a background spectrum must be collected; this is taken from the clean
ATR crystal. The background spectrum which is obtained can be a useful indication of the cleanliness
of the ATR crystal; a line at 100% T should be obtained with no spectral features. In order for total
internal reflectance to occur there must be good contact between the sample and crystal
surface. Liquid samples can be placed directly onto the ATR crystal; the whole crystal must be
covered. Similarly pastes or other viscous substances can be spread onto the crystal. In the case of
solids, these are more readily analyzed on single reflection ATR instruments which are often made of
diamond. High quality spectra can be obtained directly from powder samples placed on the ATR
crystal. The amount of sample should entirely cover the crystal and does not need to be more than
a few millimeters thick. In order to ensure that there is good contact with the crystal surface the
instrument pressure arm is positioned over the sample and tightened; it may be necessary to apply
greater pressure when analyzing high density polymers or coatings on metal surfaces, however, the
user manual should always be consulted for optimum operating parameters.
The major advantages of ATR instruments are the lack of sample preparation, the ability to obtain
high quality reproducible spectra, and due to their ease of use, the variation between users is
minimized.
Figure 26: ATR Instrument. Image reproduced with permission from Specac (Orpington, Kent, UK).
16. Applications of IR Spectroscopy
34
IR spectroscopy has primarily been used for structural elucidation and identification of unknowns (by
comparison with a spectrum of a standard).
Modern advances have seen the development of 2D IR techniques which have been applied to a
myriad of different applications including isotope labelling studies of biological species, 12 the
investigation of proteins, peptides, and hydrogen bond dynamics,13 and also the study of
nanocrystalline thin films.14
Other areas of note where IR spectroscopy is being utilized are in stem cell studies, 15 materials
science,16 catalysis,17 and reaction kinetics.18 Which demonstrates the applicability and flexibility of
this analytical technique.
35
17. References
1. Satinder Ahuja and Neil Jespersen “Modern Instrumental Analysis (Comprehensive Analytical
Chemistry)” Volume 47. Chapters 1 and 5. First Edition. The Netherlands 2006.
2. David Harvey. “Modern Analytical Chemistry” First edition. Chapter 10. McGraw-
Hill. United States of America 2000.
3. F.W. Fifield and D. Kealey. “Principles and Practice of Analytical Chemistry” Chapters 7 and
9. Fifth Edition. Blackwell Science Ltd. UK 2000.
4. G. H. Jeffrey, J. Bassett, J. Mendham and R. C. Denney. “Vogel’s Textbook of Quantitative
Chemical Analysis” Chapter 19. Fifth Edition. UK 1999.
5. Donald L. Pavia, Gary M. Lampman and George S. Kriz. “Introduction to Spectroscopy. A
Guide for Students of Organic Chemistry” Chapter 2. Thompson Learning. United States of
America 2001.
6. Francis Rouessac and Annick Rouessac. “Chemical Analysis. Modern Instrumentation
Methods and Techniques” Chapter 10. Second Edition. John Wiley & Sons Ltd. England
2007.
7. R. R. Willey. “Fourier Transform Infrared Spectrophotometer for Transmittance and Diffuse
Reflectance Measurements” Applied Spectroscopy. Volume 30, Number 6, Pp 593-601.
1976.
8. John Coates. “Interpretation of Infrared Spectra, A Practical Approach”. Encyclopedia of
Analytical Chemistry. Pp 10815 – 10837. John Wiley & Sons Ltd, Chichester, 2000.
9. Peter J. Larkin. “Infrared and Raman Spectroscopy. Principles and Spectral Interpretation”
Chapters 1 to 6. Radarweg 29, PO Box 211, 1000 AE Amsterdam, The Netherlands. Elsevier
2011.
10. Brian C. Smith. “Fundamentals of Fourier Transform Infrared Spectroscopy” Chapters 1 to
3. CRC Press. United States of America. Taylor and Francis Group. 2011.
11. John A. Dean. “Lange’s Handbook of Chemistry” Section 7.5. Fifteenth Edition. United
States of America. McGraw Hill. 1999.
12. Sang-Hee Shim, Ruchi Gupta, Yun L. Ling, David B Strafeld, Daniel P. Raleigh, and Martin T.
Zanni. “Two-dimensional IR Spectroscopy and Isotope Labelling Defines the Pathway of
Amyloid Formation with Residue Specification” PNAS 106 (2009) 6614-6619.
13. Yung Sam Kim and Robin M. Hochstrasser. “Applications of 2D IR Spectroscopy to Peptides,
Proteins, and Hydrogen-Bond Dynamics” J. Phys. Chem. B 113 (2009) 8231-8251.
14. Wei Xiong, Jennifer E. Laaser, Peerasak Paoprasert, Ryan A. Franking, Robert J. Hamers,
Padma Gopalan and Martin T. Zanni. “Transient 2D IR Spectroscopy of Charge Injection in
Dye-Sensitized Nanocrystalline Thin Films” J. Am. Chem. Soc. 131 (2009), 18040-18041.
15. Diletta Amia, Tui Nerib, Antonino Natalelloa, Paolo Mereghettic, Silvia Maria Dogliaa, Mario
Zanonib, Maurizio Zuccottid, Silvia Garagnab, and Carlo Alberto Redib. “Embryonic Stem Cell
Differentiation Studied by FT-IR Spectroscopy” Biochimica et Biophysica Acta 1783 (2008)
98-106.
16. Petru Pascuta, Lidia Pop, Simona Rada, Maria Bosca, and Eugen Culea. “The Local Structure
of Bismuth Borate Glasses Doped with Europium Ions Evidenced by FT-IR Spectroscopy”
Journal of Materials Science: Materials in Electronics 19 (2008) 424-428.
17. Carlo Lamberti, Adriano Zecchina, Elena Groppob, and Silvia Bordiga. “Probing the Surfaces
of Heterogeneous Catalysts by in situ IR Spectroscopy” Chem. Soc. Rev. 39 (2010) 4951-
5001.
18. Michael Schleeger, Christoph Wagner, Michiel J. Vellekoop, Bernhard Lendl, and Joachim
Heberle. “Time-Resolved Flow-Flash FT-IR Difference Spectroscopy: The Kinetics of CO
Photodissociation from Myoglobin Revisited” Analytical and Bioanalytical Chemistry, 394
(2009) 1869-1877.

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Introduction to Infrared

Figure 1: The electromagnetic spectrum and the infrared region.

Wavelength λ - The length of one complete wave cycle (cm).

Frequency and wavelength are inversely related (Equation 1):

Figure 2: Electromagnetic spectrum.

3. Electromagnetic Radiation and Spectroscopy

Type of Energy Transfer Region of the Spectroscopic Technique

Figure 3: Relationship between frequency and wavelength.

 Far-Infrared (400-33 cm-1): vibrations of molecules containing heavy atoms, molecular

Information achievable with Infrared spectroscopy includes:

1. The type of atoms within the molecule.

2. The type of bonds between atoms.

Both structures have the molecular formula C2H4O

Figure 4: Infrared spectroscopy regions (oversimplified).

The HCl molecule possesses a permanent dipole moment, so it is infrared active.

Oscillator Wavenumber (cm-1)

Figure 5: Representation of a polyatomic molecule.

In terms of the wavenumber ( ̅ ) (Equation 4):

6. Calculation of Molecular Vibrations

The stretching frequency ̅ (cm-1) of C-H can be calculated as follows:

Force constants (k):

Single bond = 5x105 dyne/cm

Remember higher wavenumber = higher frequency

2) Decrease in reduced mass (μ) (Figure 8).

7. Infrared Active Modes

Figure 9: Modes of vibration (oversimplified).

The transmittance can be calculated as follows (Equation 5):

Absorbance is given by Equation 6:

From the previous expression it follows that (Equation 7):

Absorbance is related to concentration via the Beer-Lambert law (Equation 8):

ε is sometimes referred to as the extinction coefficient.

Figure 11: Sample absorption of IR radiation.

Figure 12: Typical IR spectrum.

The wavenumber is usually expressed as cm-1.

10. Infrared Absorption Signals

11. Dispersive IR Instruments

Figure 13: Basic concept of a single beam IR instrument.

Figure 14: The double beam IR instrument.

12. FTIR Instruments

Common IR sources are:

The fixed mirror is a flat highly reflective surface.

Many instruments employ a Helium-Neon laser as an internal wavelength calibration standard. It is

Thermal detectors: detect changes in temperature of an absorbing material (lithium tantalate

13. FTIR Operation

Figure 17: Interferogram.

The interferogram is converted to a more familiar IR spectrum (wavenumber vs. % transmittance)

FTIR instruments have several advantages over dispersive IR instruments including:

14. Sample Preparation

Figure 18: Classification of IR signals.

The plates are then placed in a holder and a spectrum is taken.

Figure 19: Sodium chloride (NaCl) plates in various conditions.

Transmission Characteristics of Common Solvents.

Figure 22: IR spectrum of pure Nujol.

Material Wavelength Range (µm) Wavelength Range (cm-1) Refractive Index at 2 µm

Reasons for Cloudy Discs

 KBr mixture not properly ground

15. Attenuated Total Reflectance (ATR)

Figure 25: Evanescent wave.

Typical ATR Crystal Materials