UNIT III: Spectroscopy

Basic Principle and Applications of UV-

Visible, FT-IR, Raman, Microwave and
Nuclear Magnetic Resonance (NMR)
Spectroscopy.
 WHAT IS SPECTROSCOPY?
➢ Atoms and molecules interact with electromagnetic radiation (EMR) in a
wide variety of ways.
➢ Atoms and molecules may absorb and/or emit EMR.
➢ Absorption of EMR stimulates different types of motion in atoms and/or
molecules.
➢ The patterns of absorption (wavelengths absorbed and to what extent)
and/or emission (wavelengths emitted and their respective intensities) are
called ‘spectra’.
➢ The field of spectroscopy is concerned with the interpretation of spectra in
terms of atomic and molecular structure (and environment).
➢ Spectroscopy is the study of the interaction of light with matter. The
fundamental measurement obtained in spectroscopy is a spectrum, which
is a plot of measured light intensity versus some property of light.
 Electromagnetic radiation
Electromagnetic radiation (EM radiation or EMR) refers to the
waves (or their quanta, photons) of the electromagnetic field,
propagating (radiating) through space carrying electromagnetic
radiant energy.

3
4
5
 Regions of Electromagnetic Radiation
Frequency ()

Wavelength ()

Radio-waves Microwaves Infra-red Visible Ultra-violet X-ray -ray

Region Region Region Region Region Region Region
Frequency 106 - 1010 1010 - 1012 1012 - 1014 1014 - 1015 1015 - 1016 1016 - 1018 1018- 1020
(HZ)
Wavelength 10 m – 1 cm 1 cm – 100 µm 100 µm – 1 µm 700 – 400 nm 400 - 10 nm 10 nm – 100 pm –
100 pm 1 pm
NMR, ESR Rotational Vibrational Electronic Electronic
Spectroscopy spectroscopy Spectroscopy Spec.

Energy 0.001 – 10 Order of some Some 104 Some 100 Some 100s 107 - 109 109 - 1011
J/mole 100 J/mole J/mole kJ/mole kJ/mole J/mole J/mole

UHF TV, cellular, Table lamp,

telephones. Tube light (400
(300 MHz and 3 GHz) nm -800 nm or
400–790 THz)
FM Radio, VHF TV. Microwave ovens, Sun Lamp X-ray
AM Radio Police radar, satellite
stations-- (3 to 30 GHz)
 Regions of Electromagnetic Radiation
The figure illustrate in pictorial fashion the various regions of electromagnetic radiation
and its molecular processes, which are quite different.
NMR

ESR
REGION PROCESS
X-ray Bond-breaking
UV/Visible Electronic
Infrared Vibrational
Microwave Rotational
Radio Frequency (nmr) Nuclear and
Electronic Spin
The interaction of radiation with matter

http://hyperphysics.phy-astr.gsu.edu/hbase/mod3.html
Planck suggests that radiation (light, energy) can
only come in quantized packets that are of size, hν.
 Energy is quantized
Planck suggests that radiation (light, energy) can
only come in quantized packets that are of size hν.

E = h
Planck’s constant
h = 6.626 × 10-34 J·s
Energy (J)
Planck, 1900

Frequency (s-1)
Note that we can specify the energy by specifying any one of the following:

1. The frequency, n (units: Hz or s-1): E = h

2. The wavelength, λ, (units: m or cm or mm): hc
 = c
E=
Recall: 
3. The wavenumber, ~ (units: cm-1 or m-1)
1 E = hc~
Recall: ~ =

THE ELECTROMAGNETIC SPECTRUM
 3.1. UV-Vis Spectroscopy
Electronic absorption spectroscopy
Ultraviolet-visible spectroscopy refers to absorption spectroscopy in
the ultraviolet-visible spectral region. In this region of the
electromagnetic spectrum (10-800 nm), atoms and molecules undergo
electronic transitions.
“ultra” means “beyond”
UV-Visible Spectroscopy-Basics

• Ultraviolet radiation stimulates

molecular vibrations and electronic
transitions.
• Absorption spectroscopy from 160 nm
to 780 nm
• Measurement absorption or
transmittance
• Identification of inorganic and organic
species.
 Electronic transitions
When an atom or molecule absorbs
energy, electrons are promoted from their
ground state to an excited state. In a
molecule, the atoms can rotate and
vibrate with respect to each other. These
vibrations and rotations also have
discrete energy levels, which can be
considered as being packed on top of
each electronic level.
Electronic transitions:
The absorption of UV or Visible
radiation corresponds to the excitation of
outer electrons. There are three types of
electronic transition which can be
considered:
• Transitions involving , , and n
electrons
• Transitions involving charge-
transfer electrons
• Transitions involving d and f
electrons
Fig. The Excitation Process.
Bonding and antibonding MO Orbitals (Sigma and Pi Bond)
 Absorbing species containing , , and n electrons

All molecules will undergo electronic excitation following absorption of light, but for most
molecules very high energy radiation (in the vacuum ultraviolet, <200 nm) is required.
Consequently, absorption of light in the UV-visible region will only result in the following
transitions: n-* and - *.
UV-Visible Spectroscopy-Principle
➢ In order to absorb light
in the region from 200
- 800 nm (which is
where the spectra are
measured), the
molecule must contain
either pi bonds or
atoms with non-
bonding orbitals.

➢ A non-bonding orbital
is a lone pair on, say,
oxygen, nitrogen or a
halogen.

➢ Molecules or parts of
molecules that absorb
light strongly in the
UV-vis region are
called chromophores
(Color loving).
Chromophores and Auxochromes

C=C, C=O, −C≡N, N=N,

NO
OH, NH2, NHR
Electronic Transitions-Conjugated π Systems
➢ If the molecule is exposed to light
of a wavelength with energy equal
to ΔE, the HOMO-LUMO energy
gap, this wavelength will be
absorbed and the energy used to
excite one of the electrons from
the HOMO to the LUMO – in other
words, from the σ to the σ*
orbital. This is referred to as a σ -
σ* transition.
➢ For H2 molecules ΔE for this
electronic transition is 258
kcal/mol, corresponding to light
with a wavelength of 111 nm.
 Electronic Transitions-Conjugated π Systems
➢ When a double-bonded molecule such as ethylene absorbs light, it undergoes
a π - π* transition. Because π- π* energy gaps are narrower than σ - σ* gaps,
ethylene absorbs light at 165 nm - a longer wavelength than molecular
hydrogen.

Electronic transitions of both molecular hydrogen and

ethylene are too energetic to be accurately recorded by
standard UV spectrophotometers, which have a range of
220 – 700 nm.

➢ It is a powerful tool in the study of molecules

with conjugated π systems. In these groups,
the energy gap for π - π* transitions is smaller
than for isolated double bonds, and thus the
wavelength absorbed is longer.
Electronic Transitions-Conjugated π Systems
1,3-butadiene is an example of
conjugated double bond system
absorbs UV light with a wavelength
of 217 nm.

As conjugated pi systems become

larger, the energy gap for a π - π*
transition becomes increasingly
narrow, and the wavelength of light
absorbed correspondingly
becomes longer.
 Electronic Transitions-Conjugated π Systems
The absorbance due to the π - π*
transition in 1,3,5-hexatriene occurs at
258 nm, corresponding to a ΔE of 111
kcal/mol.
In molecules with extended  systems,
the HOMO-LUMO energy gap becomes
so small that absorption occurs in the
visible rather then the UV region of the
electromagnetic spectrum.

The conjugated  system in 4-

4-methylpent-3en-2-one
methylpent-3en-2-one gives rise to a
strong UV absorbance at 236 nm due
to a π - π* transition. However, this
molecule also absorbs at 314 nm. This
second absorbance is due to the
transition of a non-bonding (lone pair)
electron on the oxygen up to a π* anti-
bonding MO. This is referred to as an n
- π* transition.
The nonbonding (n) MO’s are higher in energy than the highest bonding π
orbitals, so the energy gap for an n→π∗ transition is smaller that that of a π - π*
transition – and thus the n - π* peak is at a longer wavelength.
In general, n - π* transitions are weaker (less light absorbed) than those due to π -
π* transitions.

Molecule Wavelength at Maximum ∆E Electronic Transitions

Absorbance (λMax) (Kcal/mol)
Hydrogen 111nm (σ→σ*) 258
Ethylene 165nm (π→π*) 173
1,3-Butadiene 217nm ((π→π*) 132
1,3,5-Hexatriene 258nm (π→π*) 111
4,methylpent-3-en-2-one 236nm (π→π*) 121
314nm (n→π*) 91

β-carotene 470nm (π→π*) ?

Erythrosine (Red # 3) 524nm (π→π*) ?
Indigotine (Blue #2) 630nm (π→π*) ?
 UV-Visible Spectroscopy of β-Carotene

β-Carotene

β-Carotene, with its system of 11 conjugated double

bonds, absorbs light with wavelengths in the blue region of the
visible spectrum while allowing other visible wavelengths -
mainly those in the red-yellow region - to be transmitted. This is
why carrots or papaya are orange.

Problem: What is the energy of the photons (in kcal/mol) of light with wavelength of 470 nm,
the λmax of -carotene?
 Problem: 1.2

What is the energy of the photons (in kcal/mol) of light with wavelength
of 470 nm, the λmax of -carotene?

Solution:
Given, λ=470nm, h=6.626x10-34Js; velocity of light c=3x108ms-1

𝒉𝒄 (𝟔. 𝟔𝟐𝟔 × 𝟏𝟎−𝟑𝟒 𝑱. 𝒔)(𝟑 × 𝟏𝟎𝟖 𝒎𝒔−𝟏 )

𝑬= =
λ 𝟒𝟕𝟎 × 𝟏𝟎−𝟗 𝒎
= 𝟒. 𝟐𝟐𝟗𝟒 × 𝟏𝟎−𝟏𝟗 𝐉/𝐩𝐡𝐨𝐭𝐨𝐧

Energy of 1 mole of photons,

𝟒.𝟐𝟐𝟗𝟒×𝟏𝟎−𝟏𝟗 𝐉 𝟔.𝟎𝟐𝟑×𝟏𝟎𝟐𝟑 𝒑𝒉𝒐𝒕𝒐𝒏𝒔
𝐄= × = 𝟐𝟓𝟒𝟕𝟑𝟒 𝑱/𝒎𝒐𝒍
𝑷𝒉𝒐𝒕𝒐𝒏 𝒎𝒐𝒍
= 𝟐𝟓𝟒. 𝟕𝟑𝟒𝒌𝑱/𝒎𝒐𝒍

WKT, 1 Cal =4.184 J

𝟐𝟓𝟒.𝟕𝟑𝟒𝒌𝑱 𝟏 𝑪𝒂𝒍
𝑬= × = 𝟔𝟎. 𝟖𝟖𝟑𝒌𝑪𝒂𝒍/𝒎𝒐𝒍
𝒎𝒐𝒍 𝟒.𝟏𝟑𝟒𝑱
 Problem:1.2
Calculate the energy required to induce the (σ→σ*) transition in
Hydrogen if the λmax for this transition is 111nm.
Solution:
Given, λ = 111 nm, h = 6.626x10-34 Js; velocity of light,c = 3x108 ms-1; E=h

𝒉𝒄 (𝟔. 𝟔𝟐𝟔 × 𝟏𝟎−𝟑𝟒 𝑱. 𝒔)(𝟑 × 𝟏𝟎𝟖 𝒎𝒔−𝟏 )

𝑬= =
λ 𝟏𝟏𝟏 × 𝟏𝟎−𝟗 𝒎
= 𝟏. 𝟕𝟗𝟎𝟖 × 𝟏𝟎−𝟏𝟖 𝐉/𝐩𝐡𝐨𝐭𝐨𝐧
Energy of 1 mole of photons,
𝟏.𝟕𝟗𝟎𝟖×𝟏𝟎−𝟏𝟖 𝐉 𝟔.𝟎𝟐𝟑×𝟏𝟎𝟐𝟑 𝒑𝒉𝒐𝒕𝒐𝒏𝒔
𝐄= × = 𝟏. 𝟎𝟕𝟗 × 𝟏𝟎𝟔 𝑱/𝒎𝒐𝒍
𝑷𝒉𝒐𝒕𝒐𝒏 𝒎𝒐𝒍
= 𝟏𝟎𝟕𝟗 𝒌 𝑱/𝒎𝒐𝒍

WKT, 1 Cal = 4.184 J

𝟏𝟎𝟕𝟗 𝒌𝑱 𝟏 𝑪𝒂𝒍
𝑬= × = 𝟐𝟓𝟕. 𝟕𝟗 𝒌𝑪𝒂𝒍/𝒎𝒐𝒍
𝒎𝒐𝒍 𝟒.𝟏𝟑𝟒 𝑱

Problem: Caffeine molecules absorb infrared radiation at 1656 cm−1. Calculate the
following:
(i) wavelength of this radiation; ~ = 1 1 cm = 107 nm

(ii) frequency of this radiation; 
(iii) energy change associated with this absorption.
 UV-Visible Spectroscopy
• THE BEER-LAMBERT
LAW
I  It 
• For a light absorbing T= t %T =   x 100
Io Io  Io 
medium, the light

light intensity (I)

intensity falls
exponentially with
sample depth. Io It

• For a light absorbing l

medium, the light
It cuvette
intensity falls
exponentially with
increasing sample l Sample depth
concentration.
 UV-Visible Spectroscopy
A =   cl A = − log10 T
Absorbance

The negative logarithm of T is called

the absorbance (A) and this is directly
proportional to sample depth (called
pathlength, l in cm) and sample
concentration (c in M). The equation is
called the Beer-Lambert law.

 is called the molar absorption coefficient

and has units of (M-1 cm-1) dm3 mol-1 cm-1

Concentration
Beer-Lambert’s law states that when a beam of monochromatic light is passed through
a solution, some of the radiations were getting absorbed (A), which decreases the
intensity of the transmitted radiation (I) compared to the intensity of incident radiation
(I0). The decrease in the intensity of incident radiation is directly proportional to the
concentration (c) as well as thickness (path length, l) of the solution (however this
law does not hold good at higher and lower concentrations of the solutions).
log I0 /I = A = εlc
 Problems UV-Visible Spectroscopy
1. Guanosine has a maximum absorbance of 275 nm. ϵ275 = 8400 M−1cm−1 and
the path length is 1 cm. Using a spectrophotometer, you find the that A275
= 0.70. What is the concentration of Guanosine?
2. The literature value of ε for 1,3-pentadiene in hexane is 26,000 (L.mol-1.cm-
1) at its λ
max of 224 nm with a path length of 2 cm. You prepare a sample
and take a UV spectrum, finding that A224 = 0.850. What is the
concentration of your sample?
3. A substance is dissolved and made into a solution (4 g/Liter). The length
of cuvette is 2 cm and only 50% of the certain light beam is transmitted.
(a). What is the extinction coefficient? (b). what is the molar absorption
coefficient, if the molecular weight is 100?

𝐈𝟎 𝟏𝟎𝟎
Solution: (a). 𝐀 = 𝐥𝐨𝐠 = 𝐥𝐨𝐠
= 𝟎. 𝟑𝟎𝟏 −− −(𝟏)
𝑰𝒕 𝟓𝟎
𝟒𝐠 𝟖𝛆 𝐠
𝐀 = 𝛆𝐜𝐥 = 𝛆 × × 𝟐 𝐜𝐦 = . 𝐜𝐦 −− − 𝟐
𝐋𝐢𝐭𝐞𝐫 𝐥𝐢𝐭𝐞𝐫
Equating 1 & 2, 𝟖𝛆 𝐠. 𝐋𝐢𝐭𝐞𝐫 −𝟏 . 𝐜𝐦 = 𝟎. 𝟑𝟎𝟏
𝟖𝛆 𝐠. 𝐋𝐢𝐭𝐞𝐫 −𝟏 . 𝐜𝐦 = 𝟎. 𝟑𝟎𝟏
𝟎. 𝟑𝟎𝟏
𝛆= −𝟏
= 𝟎. 𝟎𝟑𝟕𝟔 𝐋𝐢𝐭𝐞𝐫 𝐠 −𝟏 𝐜𝐦−𝟏
𝟖𝐠. 𝐥𝐢𝐭𝐞𝐫 . 𝐜𝐦
𝟎.𝟎𝟑𝟕𝟔 𝐋𝐢𝐭𝐞𝐫 𝟏𝟎𝟎 𝐠
(b) × = 𝟑. 𝟕𝟔 𝐋𝐢𝐭𝐞𝐫. 𝐦𝐨𝐥−𝟏 𝐜𝐦−𝟏 𝐨𝐫 𝟑.
𝐠.𝐜𝐦 𝐦𝐨𝐥
𝟕𝟔 𝐌 −𝟏 𝐜𝐦−𝟏
 Applications of UV-Visible Spectroscopy
1. To study the kinetics of a reaction: rate of reaction, rate equation and
mechanism of a reaction.
2. UV spectroscopy is used extensively in teaching, research and analytical
laboratories for the quantitative analysis of all molecules that absorb
ultraviolet and visible electromagnetic radiation.
3. To study enzyme kinetics: In clinical diagnostics for estimation of glucose,
tissue damage.
4. UV-visible spectroscopy is used for dissolution testing of tablets and
products in the pharmaceutical industry.
5. In-vitro and In-vivo drug release kinetics can also be studied.
6. In environmental and agricultural fields the quantification of organic
materials and heavy metals in fresh water can be carried out using UV-
visible spectroscopy.
7. In the dye, ink and paint industries UV-visible spectroscopy is used in the
quality control in the development and production of dyeing reagents, inks
and paints and the analysis of intermediate dyeing reagents.
8. In the biochemical and genetic fields, UV-visible spectroscopy is used in
the quantification of DNA and protein/enzyme activity as well as the
thermal denaturation of DNA.
 3.2. Infrared Spectroscopy
• IR spectra produced by the absorption of energy by a molecule in
the infrared region and the transition occur between vibrational
levels. This is also known as vibrational spectroscopy.

• IR radiation lies in the wavelength range of 0.7-400 m.

• An IR spectrum is a plot of per cent transmittance (or absorbance)

against wave-number (frequency or wavelength).

• A 100 per cent transmittance in the spectrum implies no absorption

of IR radiation. When a compound absorbs IR radiation, the intensity
of transmitted radiation decreases. This results in a decrease of
percent transmittance and hence a dip in the spectrum. The dip is
often called an absorption peak or absorption band.

• Different types of groups of atoms (C-H, O-H, N-H, etc.) absorb

infrared radiation at different characteristic wave numbers.
 IR Spectroscopy-Principles
A molecule can only absorb radiation when the incoming infrared radiation is of the
same frequency as one of the fundamental modes of vibration of the molecule.

A molecule to absorb infrared radiations it must possess a specific feature, i.e. an

electric dipole moment of the molecule must change during the vibration. This is
the selection rule for infrared spectroscopy.

A dipole moment is a quantity that describes two opposite charges separated by a

distance.

The dipole moment of a hetero nuclear

diatomic molecule (HF, HCl) changes
as the bond expands and contracts. By
comparison, an example of an ‘infrared-
inactive’ molecule is a homonuclear
diatomic molecule (H2, O2, N2, Cl2)
because its dipole moment remains
zero no matter how long the bond.
The interactions of infrared
radiation with matter may be
understood in terms of
changes in molecular
dipoles associated with
vibrations and rotations.

symmetric stretching of the C–H bonds of

bromomethane

In order to begin with a basic model, a molecule can be

looked upon as a system of masses joined by bonds
with spring-like properties.
Taking first the simple case of diatomic molecules,
such molecules have three degrees of translational
freedom and two degrees of rotational freedom. The
atoms in the molecules can also move relative to one
other, that is, bond lengths can vary or one atom can
move out of its present plane. This is a description of
stretching and bending movements that are collectively
referred to as vibrations.
 Molecular vibrations in IR
A molecular vibration 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 and the typical frequencies of
molecular vibrations range from less than 1013 to approximately 1014 Hz, corresponding
to wavenumbers of approximately 300 to 3000 cm−1.
Stretching Vibrations: a change in the length of a bond, such as C-H or C-C;
In stretching vibration, the distance between two atom decrease or increase, but
bond angle unaltered.

Symmetrical Asymmetrical
stretching stretching
 Bending Vibrations
In plane bending or scissoring: a change in the angle between two bonds, such as
the HCH angle in a methylene group
In plane bending or scissoring

In plane bending or rocking: a change in angle between a group of atoms, such as

a methylene group and the rest of the molecule.
In plane bending or rocking
Out of plane bending or wagging: a change in angle between the plane of a group
of atoms, such as a methylene group and a plane through the rest of the
molecule. Out of plane bending or wagging

Out of plane bending or twisting: a change in the angle between the planes of two
groups of atoms, such as a change in the angle between the two methylene
groups. Out of plane bending or twisting

In bending or deformation vibration, the bond angle decrease or increase, but

bond distance remain same.
 Molecular vibrations
 For linear molecule
 linear molecule containing “N” atoms has (3N-5) fundamental
vibration modes.
E.g., CO2 – (3 x 3 – 5) = 4 fundamental vibration modes.
1. Symmetric stretching,
2. Asymmetric stretching,
3. In-plane bending (scissoring, rocking)
4. Out of plane bending (twisting, waging)
E.g. HCl, BeH2, diatomic molecules (N2, H2, etc.,)
 Molecular vibrations
 For non – linear molecule

 non – linear molecule containing “N” atoms has (3N-6)

fundamental vibration modes.
E.g., CH4 – (3 x 5 – 6) = 9 fundamental vibration modes.
E.g. C6H6 – (3 x 12 – 6) = 30 fundamental vibration modes.
E.g., H2O– (3 x 3 – 6) = 3 fundamental vibration modes.
1. Symmetric stretching,
2. Asymmetric stretching,
3. Bending stretching
 INFRARED SPECTROSCOPY

• Methane H H H H H H

C C C

H H H H H H

Asymmetrical Symmetrical Bending or

stretching stretching scissoring

H H H H H H

C C C

H H H H H H

Rocking or in Twisting or out-of- Wagging or out-

plane bending plane bending of-plane bending
 IR Spectrum
• An IR spectrometer measures the
percent transmittance as a
function of frequency. This plot is
called an absorption spectrum.
The location of each signal on the
spectrum can be specified either
by the corresponding wavelength
or by the corresponding
frequency of radiation that was
absorbed.

• IR signals more often reported in Wavenumber is proportional to frequency,

terms of a frequency-related unit,
called wavenumber ( ഥ 𝝂 ).The
wavenumber corresponds to the
frequency of light divided by a
constant (the speed of light, c).
The units of wavenumber are
inverse centimeters (cm−1), and
the values range from 400 to 4000
cm−1.
and therefore, a larger wavenumber
represents higher energy. Signals that
appear on the left side of the spectrum
correspond with higher energy radiation,
while signals on the right side of the
spectrum correspond with lower energy
radiation. Every signal in an IR spectrum
has three characteristics: wavenumber,
intensity, and shape.
 Hooke's law

Hooke's law: the force is

proportional to the extension
Hooke’s Law:
FT-IR Principle
• For every bond, the wavenumber of absorption associated with bond stretching is
dependent on two factors: (1) bond strength and (2) masses of the atoms sharing the
bond. The impact of these two factors can be rationalized when we treat a bond as if it
were a vibrating spring connecting two weights.

• Using vibrating spring analogy following equation is constructed, which is derived from
Hooke’s law, which enables us to approximate the frequency of vibration for a bond
between two atoms of mass m1 and m2:
𝟏 𝒇 𝒎 .𝒎
• ෥=
𝒗 and 𝝁𝒓𝒆𝒅 = 𝒎 𝟏+𝒎𝟐 The equation shows that the frequency of the vibration,
𝟐𝝅𝒄 𝝁𝒓𝒆𝒅 𝟏 𝟐
෥ is proportional to the (root of) a force constant f-more or less the bond strength and
𝒗
inversely proportional to the (root of) a reduced mass μred, that is, the product of the
masses of the two atoms forming the bond divided by their sum. It is evident from the
above equations that stronger bonds vibrate faster and so do lighter atoms.

For each and every bond in a molecule, the

energy gap between vibrational states is very
much dependent on the nature of the bond. For
example, the energy gap for a C−H bond is much
larger than the energy gap for a C−O bond. Both
bonds will absorb IR radiation but the C−H bond
will absorb a higher energy photon.
 FT-IR Absorption Band Positions
𝟏 𝒇
෥=
𝒗
𝟐𝝅𝒄 𝝁𝒓𝒆𝒅

Use of the reduced mass in this equation allows us to treat the two atoms as one
system. Since that μred appears in the denominator the smaller atoms give bonds that
vibrate at higher frequencies, thereby corresponding to a higher wavenumber of
absorption.
A comparison of absorption wavenumbers for different groups where different
atoms bonded with carbon is given. The C−H bond involves the smallest atom (H)
and therefore appears at the highest wavenumber.

C-H C-D C-C C-O C-Cl

3000 cm-1 2200 cm-1 1200 cm-1 1100 cm-1 700 cm-1

While the force constant (f ) which is a measure of bond strength appears in the
numerator. This means that stronger bonds will vibrate at higher frequencies, thereby
corresponding to a higher wavenumber of absorption.
For example, compare the following bonds. The C≡N bond is the strongest of the
three bonds and therefore appears at the highest wavenumber.
𝑪≡𝑵 𝑪=𝑵 𝑪−𝑵
2200cm-1 1600cm-1 1100cm-1
 FT-IR Absorption Band Positions
• Single bonds (except for X−H bonds) appear on the right side of the spectrum
(below 1500 cm−1) because single bonds are generally the weakest bonds.

• Double bonds appear at higher wavenumber (1600-1850 cm−1) because they are
stronger than single bonds, while triple bonds appear at even higher wavenumber
(2100-2300 cm−1) because they are even stronger than double bonds.

• And finally, the left side of the spectrum contains signals produced by X−H bonds
(such as C−H, O−H, or N−H), all of which stretch at a high wavenumber because
hydrogen has the smallest mass.
FT-IR Absorption Band Positions

Bond → C-H C-D C-O C-Cl

/cm-1 → 3000 2200 1100 700
Bond → CO C=O C-O
/cm-1 → 2143 1715 1100
 FT-IR Absorption Band Positions

4000-3000 3000-2000 2000-1500 1500-1000

cm-1 cm-1 cm-1 cm-1

O-H CC C=C C-O

N-H CN C=O C-F
C-H C-Cl
deformations

Increasing energy

Increasing frequency
 FT-IR Absorption Band Positions
The reduced masses (μred) of the C–H, N–H, and O–H combinations are all
almost have same value. Any difference between the positions of the IR
bands of these bonds must then be due to bond strength. In practice, C–H
stretches occur at around 3000 cm–1 (though they are of little use as
virtually all organic compounds have C–H bonds), N–H stretches occur at
about 3300 cm–1, and O–H stretches higher still. We can immediately
deduce that the O-H bond is stronger than N–H which is stronger than C–
H. IR is a good way to measure such bond strengths.

Due to higher electronegativity difference between the atoms, O-H has a higher
ionic character than N-H & C-H.
 FT-IR Absorption bands of –NH and -NH2
On comparison in the IR spectra (A & B), the
(A)
IR peak of an NH group is different from that
of an NH2 group. A group gives an
independent vibration only if both bond
strength and reduced mass are different from
those of neighboring bonds.

This is true in the case of N-H, which reflects

in sharp peak at about 3300 cm–1.

The NH2 group is also independent of the rest

of the molecule, but the two NH bonds inside
the NH2 group have identical force constants
(B) and reduced masses and so vibrate as a
single unit.

Two equally strong bands appear, one for the

two N-H bonds vibrating in phase
(symmetric) at 3360 cm-1 and one for the two
Anti-Symmetric N-H bonds vibrating in opposition (anti-
symmetric). The anti-symmetric vibration
requires more energy and is at slightly higher
frequency, 3442 cm-1.
 Infrared spectra of O-H Group

O-H stretch: The free O-H stretch is a small sharp peak at 3650-3600 cm-1.

The hydrogen-bonded O-H stretch is a broad strong peak at 3400-3300 cm-1.

For the neat liquid, all the alcohol is considered hydrogen bonding, so the broad peak is
the only one of the two visible. As the alcohol is dissolved in a non-hydrogen bonding
solvent, the free O-H peak becomes more visible:
FT-IR Absorption bands of –OH and -COOH

(C)

The O–H bands occur at higher

frequency, sometimes as a sharp
absorption at about 3600 cm–1 which
is non-hydrogen bonded. Broad
signal of phenol (Spec. (C), 3500-
2900 cm-1) is due to strong H-
bonding. In benzoic acid, H-bonded
(D) dimers with 2 strong H bond between
the carbonyl oxygen atom of one
molecule and the acidic hydrogen
results in very broad V-shaped
absorbance (3500-2500 cm-1).
 IR Spectrum of 2-Butanol and 2-Propanol

IR spectra is divided into two main regions namely the diagnostic or Functional group
region (above 1500 cm−1) generally has fewer peaks and provides the clearest information.
This region contains all signals that arise from double bonds, triple bonds, and X−H bonds.

The fingerprint region (below 1500 cm−1) contains signals resulting from the vibrational
excitation of most single bonds (stretching and bending). This region is called the
fingerprint region because each compound has a unique pattern of signals in this region,
much the way each person has a unique fingerprint.

To illustrate this point, compare the spectra for 2-butanol and 2-propanol.The diagnostic
regions of these compounds are virtually indistinguishable (they both contain characteristic
C−H and O−H signals), but the fingerprint regions of these compounds are very different.
Effect of Hybridization States on Wavenumber of Absorption

• sp orbitals have more s character than the other hybridized atomic orbitals, and
therefore sp orbitals more closely resemble s orbitals.

• Compare the shapes of the hybridized atomic orbitals and note that the electron
density of an sp orbital is closest to the nucleus (much like an s orbital).

• As a result, a Csp−H bond will be shorter than other C−H bonds and these shorter
bonds due to increased force constant (f) vibrates at higher wavenumber.
Therefore the peak of appears at the highest wavenumber.
 INFRARED SPECTROSCOPY

• Human Breath

100

80 O O
H H H H
60
%T

40 O C O

20
O C O O C O
0
4000 3500 3000 2500 2000 1500 1000 500
wavenumber/cm -1
 O

C
O O CH3

C
HO

Wavenumber = 1/ wavelength
INFRARED SPECTROSCOPY

Ethane Chloroethane
INTERPRETATION OF INFRARED SPECTRA

Ethanoic acid
Infrared Experimental

Benzoic acid
 Application
➢ Identity of the compound
• Resemblance of two spectra gives the nature of the
compound.
• E.g.: C=O in aldehyde and ketone
➢ Detection of functional group
• C=O in Acetone and diethyl ketone give absorption peak in
same place
• But C=O in acetic acid will show peak in different place.
➢ Testing purity of the sample
• Pure sample give sharp and well resolved bands
• But impure sample give broad and poor resolved bands
 Application
➢ Study progress of chemical reaction
• IR spectrum of test solution at different time interval give the
progress of the chemical reaction
➢ Determination of shape or symmetry of the molecule
• Linear or non linear nature can be determined
• Eg; NO2 gives 3 peaks at 750, 1323, 1616 cm-1
For non linear molecule = (3n – 6) = 3 peaks, so NO2 is non-linear
➢ To study tautomerism
• These shows the IR band for C=O, - OH, and - NH

HN CH2 HC NH N CH2
C C C

O OH
OH
 Application
➢ Industrial applications
a. Determination of structure of chemical compounds
b. Determination of molecular weight
c. Crystallinity
E.g.; Crystalline Nylon 6,6 – at 943 cm-1
Amorphous Nylon 6,6 – at 1238 cm-1
➢ Determination of isomers
• Isomers show similar spectra in group frequency region, but
show different peaks in finger print region
➢ Determination of hydrogen bonding
• Inter and intra molecular hydrogen bonding determined using
solutions of different dilution
Raman spectroscopy (named after Indian physicist Sir C.
V. Raman) is a spectroscopic technique used to observe
vibrational, rotational, and other low-frequency
modes in a system. Raman spectroscopy is commonly
used in chemistry to provide a structural fingerprint by
which molecules can be identified.
 Raman Spectroscopy
Introduction
 Raman Spectroscopy-Introduction
❖ Raman spectroscopy is a spectroscopic technique based on inelastic
scattering of monochromatic light, usually from a laser source.

❖ Frequent inelastic scattering means that the frequency of photons in

monochromatic light changes upon interaction with a sample.

❖ Photons of the laser light are absorbed by the sample and then re-
emitted.
❖ Frequency of the re-emitted photons
is shifted up or down in comparison
with original monochromatic
frequency, which is called the Raman
effect.
❖ This shift provides information about
vibrational, rotational and other low
frequency transitions in molecules.
❖ Raman spectroscopy can be used to
study solid, liquid and gaseous
samples.
 Raman Spectroscopy- Principle
➢ Raman effect studies molecular
deformations in electric field (E)
determined by molecular polarizability (α).
➢ The laser beam can be considered as an
oscillating electromagnetic wave with
electrical vector (E). Upon interaction with
the sample it induces electric dipole
moment,  = αE which deforms molecules.
➢ Because of periodical deformation,
molecules start vibrating with
characteristic frequency m.

➢ IR and Raman are the most common vibrational spectroscopies for assessing
molecular motion and fingerprinting species.
- Based on inelastic scattering of a monochromatic excitation source
- Routine energy range: 200 - 4000 cm–1
➢ Complementary selection rules to IR spectroscopy
- Selection rules dictate which molecular vibrations are probed.
- Some vibrational modes are both IR and Raman active.
Fig. The chloroform molecule, CHCl3, and its
polarizability ellipsoid from across and along
the symmetry axis.
 Raman Spectroscopy - Principle
When a Monochromatic radiation is
incident upon a sample, then this
light will interact with the sample in a
unique fashion. The light may be
absorbed, reflected or scatted.
While most of the radiation is
elastically scattered called Rayleigh
scattering. A small proportion is
inelastically scattered (Raman
scatter, composed of Stokes and
anti-Stokes portions). The resultant
scattering of the radiation gives the
information of molecular Sample
polarizability and hence structure.

Spontaneous Raman scattering is typically very weak, and as a result the main
difficulty of Raman spectroscopy is separating the weak inelastically
scattered light from the intense Rayleigh scattered laser light.
 Raman Spectroscopy-Selection Rules
Amplitude of vibration is called a nuclear displacement. In other words,
monochromatic laser light with frequency, 0 excites molecules and transforms
them into oscillating dipoles. Such oscillating dipoles emit light of three different
frequencies when:
1. A molecule with no Raman-active modes absorbs a photon with the incident
frequency, vo. If the excited molecule returns back to the same basic vibrational
state and emits light with the same frequency, vo as an excitation source. This
type if interaction is called an elastic Rayleigh scattering.
2. A photon with frequency v0 is absorbed by Raman-active molecule which at the
time of interaction is in the basic vibrational state. Part of the photon’s energy is
transferred to the Raman-active mode with frequency, vm and the resulting
frequency of scattered light is reduced to v0-vm. This Raman frequency is called
Stokes frequency, or just “Stokes”.
3. A photon with frequency, v0 is absorbed by a Raman-active molecule, which, at
the time of interaction, is already in the excited vibrational state. Excessive energy
of excited Raman active mode is released, molecule returns to the basic
vibrational state and the resulting frequency of scattered light goes up to v0 + vm.
This Raman frequency is called Anti-Stokes frequency, or just “Anti-Stokes”.
Basic Principals of Raman Spectroscopy: Light is scattered by the sample at various
angles by momentary absorption to virtual state and re-emission.
A molecule with no Raman-active modes absorbs a photon with the incident
frequency, vo. If the excited molecule returns back to the same basic vibrational state
and emits light with the same frequency, o as an excitation source. This type of
interaction is called an elastic Rayleigh scattering.

Elastic collision: Collision between photon and molecule results in no change in energy
 Two Types of Raman Scattering

Inelastic: Collision between photon and

molecule results in a net change in energy
Anti-Stokes: E = h + DE

Stokes: E = h - DE

DE – the energy of the first vibration level of the ground state – IR vibration absorbance
Raman frequency shift and IR absorption peak frequency are identical.

A photon with frequency, v0 is absorbed by Raman-active molecule, which, at the time of interaction is in
the basic vibrational state. Part of the photon’s energy is transferred to the Raman-active mode with
frequency, vm and the resulting frequency of scattered light is reduced to v0-vm (E= h - DE). This Raman
frequency is called Stokes frequency, or just “Stokes”.

A photon with frequency, v0 is absorbed by a Raman-active molecule, which, at the time of interaction, is
already in the excited vibrational state. Excessive energy of excited Raman active mode is released,
molecule returns to the basic vibrational state and the resulting frequency of scattered light goes up to v0
+ vm (E = h + DE). This Raman frequency is called Anti-Stokes frequency, or just “Anti-Stokes”.
 - Resulting Raman Spectrum

Lower energy higher energy

Probability of Emission Observed Intensity

Raleigh scattering >> Stokes >> anti-Stokes
difference in population of energy levels of vibrational transitions

Intensity of Raman lines are 0.001% intensity of the source

Active Raman Vibrations:
- Polarizability of Molecules need to change in during vibration
- Polarizability causes deformation of electron cloud distribution

example:
O=C=O IR inactive
Raman active

O=C=O IR active
Raman inactive
IR & Raman are complimentary. Can be cases where vibration is both IR & Raman
active (e.g. SO2 – non-linear molecule)
In general:
IR tends to emphasize polar
O

functional groups (R-OH, C

, etc.)

Raman emphasizes aromatic & carbon

backbone (C=C, -CH2-, etc.)

- Raman does not “see” many common polar

solvents can use with aqueous samples –
advantage over IR.

Raman frequency range: 4000-50 cm-1

(Stokes and anti-stokes)
 Raman Spectroscopy: Classical Treatment
• Number of peaks related to degrees of freedom
DoF = 3N - 6 (bent) or 3N - 5 (linear) for N atoms
• Energy related to harmonic oscillator,  or D = c k(m1 + m2 )
2 m1m2

• Selection rules related to symmetry


Rule of thumb: If the molecule is symmetric it will be Raman active, if
it is a asymmetric molecule then it must be IR active

CO2 H2O

Raman: 1335 cm–1

Raman + IR: 3657 cm–1
IR: 2349 cm–1
Raman + IR: 3756 cm–1

IR: 667 cm–1

Raman + IR: 1594 cm–1
 Raman Spectroscopy
A) Introduction

1.) Raman spectroscopy: complementary to IR spectroscopy.

- radiation of certain frequency is scattered by the molecule.
with shifts in the wavelength of the incident beam.
- observed frequency shifts are related to vibrational changes in the
molecule and also associated with IR absorbance.
- Raman Scattering Spectrum Resembles IR absorbance spectrum.
- but the basic mechanisms of Raman & IR differ.

a) Comparison of Raman & IR:

IR Raman
i. Vibrational modes due to absorption Vibrational modes due to scattering

ii. Change in dipole Change in polarizability

extend compress

d- 2d+ d-
iii. excitation of molecule to momentary distortion of the
excited vibrational state electrons distributed around the bond

iv. asymmetric vibrations (active) symmetric vibrations (active)

IR and Raman Spectra of Benzene
Mutually exclusive principle
Surface Enhanced Raman
Spectra (SERS)
 Applications of Raman Spectroscopy
1. Used in identification of functional group of unknown samples, structural elucidation
and identification of polymorphs in pharmaceutical industries.
2. RS is useful in studies of carbon polymorphs CNT and diamond manufacturing.
3. Raman spectroscopy is an appropriate method to analyze hydrogen bonding in aqueous
solution. Intramolecular interactions caused by hydrogen bond formation, or very weak
intermolecular forces indicated by very low frequency vibrations (down to 10 cm-1) can
be investigated directly.
4. In semiconductor industries RS is used for composition determination, analysis of
crystal structure, surface and interface analysis, phase determination, doping, point
defects, temperature influence and mechanical stress.
5. Used in biochemical and biomedical investigations such as proteins, nucleic acids and
bio-membranes. The dynamics of biological systems such as living cells or to study
biological interfaces. Tissue differentiation has a great potential for clinical use in the
near future.
6. Used in medical diagnosis like endoscopy, tissue diagnosis and malignant
transformation. Raman microscopy is used in medical microscopic disease diagnostics
such as tumour characterization
7. In the food industry, the degree of hydrogenation of fats (number of C-C bonds in the
skeletal backbone of fat molecules) can be measured in seconds.
8. RS is used In weather forecasting.
9. Remote sensing and measurement of temperatures in satellites from earth is possible
due to Raman Spectroscopy.
 3.4.Microwave (or) Rotational Spectroscopy

Microwave spectroscopy began in 1934 when Cleeton and Williams observed absorption at
microwave frequencies for NH3 contained in a rubber bag. The second molecule to have its
microwave absorption observed was H2O, the "work" being done by the U. S. Navy; during
World War II, the Navy was developing radar and observed greater attenuation of signals
than expected. Eventually it was realized that the radar frequency being used was one for
which atmospheric water vapor absorbed. The development of radar provided the
technology needed for microwave spectroscopy, and the field flourished after the war.
 Microwave Spectroscopy - Introduction
❖ Microwave photon excites the molecules interacting with its electric dipole
moment causes molecule to undergo rotational transition resulting in
microwave rotational spectrum. Microwave rotational spectroscopy uses
microwave radiation to measure the energies of rotational transitions for
molecules in the gas phase.

• H-Cl and C=O give rotational

spectrum (microwave active).
• H-H and Cl-Cl don't give rotational
spectrum (microwave inactive).

❖ Photons that have energy in MW region causes transitions between the

quantum rotational energy levels of a gas molecule.
❖ In liquid or solid state rotations of molecules are hindered due to enhanced
interactions for MW spectroscopy, hence the sample must be in the gas phase.
❖ Rotational spectroscopy is only really practical in the gas phase where the
rotational motion is quantized. In solids or liquids the rotational motion is
usually quenched due to collisions between their molecules.
Which of the following molecules would show rotational spectrum: Br2, HBr and CS2? Why?
❖ Molecules can be broken down into 5
categories based on their shape and
the moment of inertia around their 3
orthogonal rotational axes.
❖ These 5 categories include diatomic
molecules, linear molecules, spherical
tops, symmetric tops and asymmetric
tops.
Moment of inertia can be
measured using a simple
pendulum, because it is the
resistance to the rotation
caused by gravity.
 RIGID ROTOR (diatomic molecule)
For simplicity, we can consider only rotational motion of rigid diatomic molecule,

A diatomic molecule can rotate around a vertical

axis. The rotational energy is quantized.

Assume a rigid (not elastic) bond

r0 = r1 + r2 (3.1)
For rotation about center of gravity, C :
m1r1 = m2r2
m1r1 = m2 (r0 - r1)
m2r2 = m1 (r0 – r2)
0
m1r1 = m2 (r0 - r1) m2r2 = m1 (r0 – r2)
m1r1 = m2r0-m2r1 m2r2 = m1r0-m1r1
M1r1 + m2r1 = m2r0 m2r2 + m1r1 = m1r0

m2r0 m1r0
r1 = r2 =
m1 + m2 m1 + m2
 RIGID ROTOR m1r1 = m2r2

Moment of inertia about C, Centre of gravity: m1 = m2r2/r1 m2 = m1r1/r2

IC = m1r12 + m2r22 = m2r2r1 + m1r1r2 = r1r2 (m1 + m2) (3.2.) sub r1,r2
mm 2 (3.3)  = reduced mass, 1 = 1 + 1
 I = 1 2 r0 = μr0
2

m1 + m2 μ m1 m2

I = Moment of inertia = mr2;  = reduced mass = m1m2/(m1 + m2); r = inter-nuclear distance

A diatomic molecule can rotate around a vertical axis. The rotational energy is quantized. By
the using Schrödinger equation, the rotational energy levels allowed to the rigid diatomic
molecule are given by,

𝒉𝟐
𝑬𝑱 = 𝟖𝝅𝟐 𝑰 𝑱 𝑱 + 𝟏 𝒋𝒐𝒖𝒍𝒆𝒔 −−− − (3.4) where, J = 0,1,2,3…

Where, J is a rotational quantum number and h is the Planck's constant, I is the moment of
inertia, either IB or IC.

Equation 3.4 expresses the allowed energies in joules, we, however, are interested in
differences between these energies, or, more particularly, in the corresponding frequency, 
= DE/h, or wavenumber,  = DE/hc cm-1, of the radiation emitted or absorbed as a
consequence of changes between energy levels.
 Rotational Spectra of Rigid Diatomic molecule
Rotational Energy Levels for rigid rotor:
𝑬𝑱 𝒉
𝜺𝑱 =
𝒉𝒄
=
𝟖𝝅𝟐 𝑰𝒄
𝑱 𝑱 + 𝟏 𝒄𝒎−𝟏 ----- (3.5) E = hc~
𝒉 𝐸𝑗
Since, Rotational constant, 𝑩 = ------ (3.6) 𝜀𝑗 = = ത
𝟖𝝅𝟐 𝑰𝒄 ℎ𝑐

𝜺𝑱 = 𝑩𝑱 𝑱 + 𝟏 𝒄𝒎−𝟏 −−− − (3.7) Where, J = 0,1,2,3…

Where,
m1m2 2 2
I= r0 = μr0 𝜺𝑱 = ത = 𝑩𝑱 𝑱 + 𝟏 𝒄𝒎−𝟏
m1 + m2
Where, J = J+1
For rigid rotor, J → J + 1,
ഥ𝑱→𝑱+𝟏 = B (J+1) (J+2) – BJ (J+1) 𝒄𝒎−𝟏
𝒗
ഥ𝑱→𝑱+𝟏 = 2B(J+1) 𝒄𝒎−𝟏 ---(3.8)
𝒗 ഥ𝑱→𝑱+𝟏 = B (J+1) ((J+2) – J)) 𝒄𝒎−𝟏
𝒗

Selection rule
1. The molecule must have a permanent dipole moment (μ≠0).
2. ∆𝐽 = ∓1. (∆𝐽 =+1 and ∆𝐽 =-1 refer to absorption and emission, respectively: in fact
∆𝐽 = ∓1 applies to both.)
3. ∆MJ= 0, ±1, a rule which is important only if the molecule is in an electric or magnetic
field
 Rotational Spectra of Rigid Diatomic molecule
For rigid rotor, J → J + 1,

The allowed rotational energy Allowed transitions between the energy levels
levels of a rigid diatomic molecule of a rigid diatomic molecule and the spectrum
Considering the transition energy between two energy levels, the difference is a multiple of
2. That is, from J=0 to J=1, the ΔE0→1 is 2B and from J = 1 to J = 2, the ΔE1→2 is 4B.

Energy levels and line positions calculated in the rigid rotor approximation. This
diagram illustrates how transitions between the rotational energy levels of
molecules map onto the energies at which these transitions are observed during
laboratory experiments.
 Rotational Spectra of Rigid Rotor
Selection Rule: Apart from Specific rule, DJ= 1, Gross rule-the molecule should have a
permanent electric dipole moment,  . Thus, homonuclear diatomic molecules do not have a
pure rotational spectrum. Heteronuclear diatomic molecules do have rotational spectra.

Dj = 1
Dj = +1 (absorption)
Dj = −1 (emission)
MW spectroscopy of Oxygen
 The rotation spectrum of 12C16O at 40 K.

Far-infrared spectrum of CO showing transitions with J”=3 to 9

The lines are nearly equally spaced and vary in intensity.

We also will learn why the lines are nearly equally spaced and vary in intensity. Such
spectra can be used to determine bond lengths, and even bond angles in polyatomic
molecules.
Microwave Spectroscopy in Space Research- Detection of Interstellar Molecules
(Cyanodiacetylene)

A part of the microwave spectrum of cyanodiacetylene (H-C≡C-C≡C-C≡N) which

has such a small B value (1331.331 MHz) that six transitions with J”= 9 to 14 lie in
the 26.5 to 40.0 GHz region.

The many ‘satellite’ transitions in each group are due to the molecule being in not
only the zero-point vibrational state but also a multitude of excited vibrational
states.
 The central part of the galaxy M77,
also known as NGC 1068,

This graphic illustrates how scientists think

“solid state” chemistry may be taking place
in ice particles that form clouds in the
atmosphere of Saturn’s moon Titan. Image
credit: NASA/JPL-Caltech/GSFC.

The first step in the proposed process is the

formation of ice particles made from the related
chemical cyanoacetylene (HC3N). As these tiny bits
of ice move downward through Titan’s stratosphere,
they get coated by hydrogen cyanide (HCN). At this
stage, the ice particle has a core and a shell
comprised of two different chemicals. Occasionally,
a photon of ultraviolet light tunnels into the frozen
shell and triggers a series of chemical reactions in
the ice. These reactions could begin either in the
core or within the shell. Both pathways can yield
dicyanoacteylene ice and hydrogen as products.
Cyanoacetylene in IC 342
 MW Spectroscopy-Applications

1. Used to precisely measure bond-length, bond angle and dipole moment of a

molecule.
2. Microwave spectroscopy is used for the measurement of dipole moments of
molecules.
3. Used in space research for the detection of Interstellar molecules (130
molecules/ions detected.
4. To Study Stark Effect using which dipole moment (μ) can be calculated.
(Under normal conditions these (2J+1) components of each J level remain
degenerate but, in the presence of an electric field, the degeneracy is partly
removed: each level is split into (J+1) components according to the value of J.
This splitting in an electric field is known as the Stark effect and is small in a
linear molecule.