SPECTROSCOPY By Ayesha shafi

CONTENTS
Basic definition
UV-visible spectroscopy
IR spectroscopy
Mass spectroscopy
DEFINITIONS
Spectroscopy: It is the branch of science that deals
with the study of interaction of matter with light.
OR
It is the branch of science that deals with the study of
interaction of electromagnetic radiation with matter.
Electromagneti
c Radiation
ELECTROMAGNETIC
RADIATION
Electromagnetic radiation consist of
discrete packages of energy which are
called as photons.

A photon consists of an oscillating

electric field (E) & an oscillating
magnetic field (M) which are
perpendicular to each other.
ELECTROMAGNETIC
RADIATION
FREQUENCY AND
WAVELENGTH
 Frequency :It is defined as the number of
times electrical field radiation oscillates
in one second.
 The unit for frequency is Hertz (Hz).
1 Hz = 1 cycle per second
Wavelength (λ):
 It is the distance between two nearest
parts of the wave in the same phase i.e.
distance between two nearest crest or
troughs.
ELECTROMAGNETIC
RADIATION

The relationship between wavelength

& frequency can be written as:
c=νλ
As photon is subjected to energy, so
E = hν = h c / λ
DEFINITIONS
(CONTINUE..)
Wave Number: It is the no of waves per unit
distance.
SPECTRUM: When electromagnetic radiation is
passed through the prism, it splits into the
regions of different colors. Each region is
characterized by the wavelength, frequency,
wavenumber and energy.
Spectrum is a graph of intensity of absorbed
or emitted radiation by sample verses
frequency (ν) or wavelength (λ).
Spectrometer: is an instrument design to
measure the spectrum of a compound.
TYPES OF SPECTRUM
Emission spectrum
Absorption spectrum
[Link] spectrum: The excited atom or molecule
tends to emit the energy in the form of light. The
dispersion of this resulting light by passing through the
prism/grading give rises a spectrum called as the
emission spectrum.
Types of the emission spectrum:
a. Continuous spectrum: when light of different
wavelength are intermixed with each other. It is known
as the continuous spectrum.
TYPES OF SPECTRUM
b. Line spectrum: when different spectral regions are
separated from each other through dark spaces.
c. Band spectrum: it consist of groups of lines closely
packed together giving the appearance of broad
luminous bands sharply defined at one end and
gradually shading off at the other end.
2. Absorption spectrum: it is the spectrum obtained
due to the absorption of electromagnetic radiations
within certain range of wavelength.
ULTRA-VIOLET VISIBLE
SPECTROSCOPY
UV-Visible spectroscopy deals with the absorption of
electromagnetic radiations in the visible and the U.V
region . It is also known as the electronic spectroscopy.
It involves the gain of energy by a molecule and the
transfer of electrons from lower energy level to higher
energy level.
UV-Visible region extends from 200nm-800nm.
PRINCIPLE OF UV-
VISIBLE SPECTROSCOPY
UV-visible being of higher energy than the infrared
radiations cause the transition of valence electrons
from the ground state to excited state on their
absorption by a sample.
Near UV Region: 200 nm to 400 nm
Far UV Region: below 200 nm
Far UV spectroscopy is studied under vacuum
condition.
The common solvent used for preparing sample to be
analyzed is either ethyl alcohol or hexane.
VACUUM UV REGION
The U.V region below 200nm cannot be studied by
conventional UV spectrometer. Because oxygen of air
absorbs strongly in this region. However if oxygen is
expelled out of the instrument with nitrogen, the range
of instrument can be extended down to about 150nm.
The region below 200nm is therefore usually known as
vacuum UV region.
THEORY OF UV-VISIBLE
SPECTROSCOPY
A molecule possess three types of energy
1. electronic transitional energy (Et)
2. rotational energy (Er)
3. vibrational energy (Ev)
Thus the total energy of the molecule is
E= Et + Er+ Ev
So,
Et is energy due to electronic transitions.
Er is energy due to the rotation of the molecule
Ev is energy due to the vibrations of the atoms within a molecule
CONTINUE
So,
E = Et >Er >Ev
when an UV radiation strikes a molecule, it first
increase its rotational energy, then vibrational and
finally electronic energy.
It means electronic transitions are responsible for the
absorption spectrum in UV–visible region.
Electronic
Transitions
THE POSSIBLE ELECTRONIC
TRANSITIONS CAN GRAPHICALLY
SHOWN AS:
ELECTRONIC
TRANSITION
The absorption of U.V-visible radiations by a sample
results in transfer of electrons from one energy states
to another energy states is known as the electronic
transitions.
THE POSSIBLE ELECTRONIC
TRANSITIONS ARE
1 • σ → σ* transition

2 • π → π* transition

3 • n → σ* transition

4 • n → π* transition

5 • σ → π* transition

6 • π → σ* transition
 1 • σ → σ* transition
• σ electron from orbital is excited
to corresponding anti-bonding
orbital σ*.
• The energy required is large for
this transition.
• e.g. Methane (CH4) has C-H bond
only and can undergo σ → σ*
transition and shows absorbance
maxima at 125 nm.
 2 • π → π* transition

• π electron in a bonding orbital is

excited to corresponding anti-
bonding orbital π*.
• Compounds containing multiple
bonds like alkenes, alkynes,
carbonyl, nitriles, aromatic
compounds, etc undergo π → π*
transitions.
• e.g. Alkenes generally absorb in
the region 170 to 205 nm.
 3 • n → σ* transition

• Saturated compounds containing

atoms with lone pair of electrons
like O, N, S and halogens are
capable of n → σ* transition.
• These transitions usually requires
less energy than σ → σ*
transitions.
• The number of organic functional
groups with n → σ* peaks in UV
region is small (150 – 250 nm).
 4 • n → π* transition
• An electron from non-bonding
orbital is promoted to anti-bonding
π* orbital.
• Compounds containing double
bond involving hetero atoms
(C=O, C≡N, N=O) undergo such
transitions.
• n → π* transitions require
minimum energy and show
absorption at longer wavelength
around 300 nm.
 5 • σ → π* transition
6 • π → σ* transition

• These electronic transitions are

forbidden transitions & are only
theoretically possible.
• Thus, n → π* & π → π* electronic
transitions show absorption in
region above 200 nm which is
accessible to UV-visible
spectrophotometer.
• The UV spectrum shows only a
few band of absorption.
Terms used in
UV / Visible
Spectroscopy
 CHROMOPHORE
The part of a molecule responsible for imparting color,
are called as chromophore.
OR
The functional groups or a molecule containing multiple
bonds capable of absorbing radiations above 200 nm due
to n → π* & π → π* transitions.(isolated functional groups
capable of absorption of UV-Visible radiations).

e.g. NO2, N=O, C=O, C=N, C≡N, C=C, C=S, etc

 AUXOCHROME
The functional groups attached to a chromophore which
modifies the ability of the chromophore to absorb light ,
altering the wavelength or intensity of absorption.
OR
The functional group with non-bonding electrons that
does not absorb radiation in near UV region but when
attached to a chromophore alters the wavelength &
intensity of absorption.
 AUXOCHROME
e.g. Benzene λmax = 255 nm

OH
Phenol λmax = 270 nm

Aniline λmax = 280 nm

NH2
Absorption
& Intensity
Shifts
1 • Bathochromic Shift (Red Shift)

2 • Hypsochromic Shift (Blue Shift)

3 • Hyperchromic Effect

4 • Hypochromic Effect
 1 • Bathochromic Shift (Red Shift)
• When absorption maxima (λmax) of
a compound shifts to longer
wavelength, it is known as
bathochromic shift or red shift.
• The effect is due to presence of an
auxochrome or by the change of
solvent.
• e.g. An auxochrome group like –
OH, -OCH3 causes absorption of
compound at longer wavelength.
 1 • Bathochromic Shift (Red Shift)
• In alkaline medium, p-nitrophenol
shows red shift. Because negatively
charged oxygen delocalizes more
effectively than the unshared pair of
electron.
O +
N
O
-
O
N
O +
-

-
OH
Alkaline
medium -
OH O

p-nitrophenol
λmax = 255 nm λmax = 265 nm
 2 • Hypsochromic Shift (Blue Shift)
• When absorption maxima (λmax) of a
compound shifts to shorter
wavelength, it is known as
hypsochromic shift or blue shift.

• The effect is due to presence of an

group causes removal of conjugation
or by the change of solvent.
 3 • Hyperchromic Effect
• When absorption intensity (ε) of a compound is
increased, it is known as hyperchromic shift.

• If auxochrome introduces to the compound, the

intensity of absorption increases.

N N CH3
Pyridine 2-methyl pyridine
λmax = 257 nm λmax = 260 nm
ε = 2750 ε = 3560
 4 • Hypochromic Effect
• When absorption intensity (ε) of a
compound is decreased, it is known
as hypochromic shift.

CH3

Naphthalene

2methylnaphthalene
ε = 19000 ε = 10250
 SHIFTS AND EFFECTS
Hyperchromic shift

Blue Red
Absorbance ( A )

shift shift

Hypochromic shift

λmax
Wavelength ( λ )