UV / VISIBLE

SPECTROSCOPY
Mr. Santosh M. Damkondwar
4/21/17
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.
Electromagne
tic 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 ():
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 ():
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=
As photon is subjected to energy, so
E=h=hc/
Electromagnetic Radiation
 Electromagnetic Radiation

Violet 400 - 420 Yellow 570 - 585

nm nm
Indigo 420 - 440 Orange 585 - 620
nm nm
Blue 440 - 490 Red 620 - 780
nm nm
Principles of
Spectroscopy
Principles of Spectroscopy
The principle is based on the
measurement of spectrum of a sample
containing atoms / molecules.

Spectrum is a graph of intensity of

absorbed or emitted radiation by sample
verses frequency () or wavelength ().

Spectrometer is an instrument design to

measure the spectrum of a compound.
Principles of Spectroscopy
1. Absorption Spectroscopy:
. An analytical technique which
concerns with the measurement of
absorption of electromagnetic
radiation.

. e.g. UV (185 - 400 nm) / Visible (400 -

800 nm) Spectroscopy, IR
Spectroscopy (0.76 - 15 m)
Principles of Spectroscopy
2. Emission Spectroscopy:
. An analytical technique in which
emission (of a particle or radiation)
is dispersed according to some
property of the emission & the
amount of dispersion is measured.

. e.g. Mass Spectroscopy

Interaction of EMR
with Matter
 Interaction of EMR with
matter
1. Electronic Energy Levels:
. At room temperature the molecules are
in the lowest energy levels E0.

. When the molecules absorb UV-visible

light from EMR, one of the outermost
bond / lone pair electron is promoted to
higher energy state such as E1, E2, En,
etc is called as electronic transition and
the difference is as:
E = h = En - E0 where (n = 1, 2, 3, etc)
E = 35 to 71 kcal/mole
Interaction of EMR with
matter
2. Vibrational Energy Levels:
. These are less energy level than
electronic energy levels.

. The spacing between energy levels are

relatively small i.e. 0.01 to 10 kcal/mole.

. e.g. when IR radiation is absorbed,

molecules are excited from one
vibrational level to another or it vibrates
with higher amplitude.
Interaction of EMR with
matter
3. Rotational Energy Levels:
. These energy levels are quantized &
discrete.

. The spacing between energy levels

are even smaller than vibrational
energy levels.

Erotational < Evibrational < Eelectronic

Lamberts
Law
Lamberts Law
When a monochromatic radiation is
passed through a solution, the decrease
in the intensity of radiation with thickness
of the solution is directly proportional to
the intensity of the incident light.

Let I be the intensity of incident

radiation.
x be the thickness of the solution.
Then
Lamberts Law
dI
I
dx
dI
So, KI
dx
Integrate equation between limit
I = Io at x = 0 and
I = I at x=l,
We get,
I
ln Kl
I0
Lamberts Law
I
2.303 log Kl
I0
I K
log l
I0 2.303
I0
Where, log A
Absorbance
I
K Absorption
E
2.303 coefficient
Lamberts
A E.l
Law
Beers
Law
Beers Law
When a monochromatic radiation is
passed through a solution, the decrease in
the intensity of radiation with thickness of
the solution is directly proportional to the
intensity of the incident light as well as
concentration of the solution.

Let I be the intensity of incident radiation.

x be the thickness of the solution.
C be the concentration of the solution.
Then
Beers Law
dI
C .I
dx
dI
So, K ' C.I
dx
Integrate equation between limit
I = Io at x = 0 and
I = I at x=l,
We get,
I
ln K ' C.l
I0
Beers Law
I0
2.303 log K .C.l
I
I0 K
log C.l
I 2.303
I 0 Absorbance
Where, log A
I
K Molar
E extinction
2.303 coefficient
A E.C.l Beers Law
Beers Law
A E.C.l
I I
T OR log T log A
I0 I0
From the equation it is seen that the
absorbance which is also called as optical
density (OD) of a solution in a container of
fixed path length is directly proportional to
the concentration of a solution.
PRINCIPLES OF
UV - VISIBLE
SPECTROSCOPY
Principle
The UV radiation region extends
from 10 nm to 400 nm and the
visible radiation region extends from
400 nm to 800 nm.
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
Electronic
Transitions
The possible electronic transitions
can graphically shown as:
The possible electronic transitions
are
 1 * transition

electron from orbital is excited to

corresponding anti-bonding orbital
*.

The energy required is large for this

transition.

e.g. Methane (CH4) has C-H bond

only and can undergo *
 2 * transition

electron in a bonding orbital is

excited to corresponding anti-
bonding orbital *.

Compounds containing multiple

bonds like alkenes, alkynes,
carbonyl, nitriles, aromatic
compounds, etc undergo *
transitions.
 3 n * transition

Saturated compounds containing

atoms with lone pair of electrons like
O, N, S and halogens are capable of
n * transition.

These transitions usually requires

less energy than * transitions.

The number of organic functional

groups with n * peaks in UV
 4 n * transition

An electron from non-bonding orbital

is promoted to anti-bonding *
orbital.

Compounds containing double bond

involving hetero atoms (C=O, CN,
N=O) undergo such transitions.

n * transitions require minimum

energy and show absorption at
 5 * transition
& * transition 6
These electronic transitions are
forbidden transitions & are only
theoretically possible.

Thus, n * & * electronic

transitions show absorption in region
above 200 nm which is accessible to
UV-visible spectrophotometer.

The UV spectrum is of only a few

Terms used in
UV / Visible
Spectroscopy
Chromophore
The part of a molecule responsible for
imparting color, are called as
chromospheres.
OR
The functional groups containing multiple
bonds capable of absorbing radiations
above 200 nm due to n * & *
transitions.

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

Chromophore
To interpretate UV visible spectrum
following points should be noted:

1. Non-conjugated alkenes show an intense

absorption below 200 nm & are therefore
inaccessible to UV spectrophotometer.

2. Non-conjugated carbonyl group

compound give a weak absorption band
in the 200 - 300 nm region.
Chromophore
O
e.g. Acetone which has max = 279 nmO
C
H3C CH3
and that cyclohexane has max = 291 nm.

When double bonds are conjugated in

a compound max is shifted to longer
wavelength.
e.g. 1,5 - hexadiene has max = 178 nm
2,4 - hexadiene has max = 227 nm
CH2 CH3
H2C H3C
Chromophore
3. Conjugation of C=C and carbonyl
group shifts the max of both groups
to longer wavelength.
O
e.g. Ethylene has max = 171 nm
C
Acetone
H2C CH2 has max = 279
H3C nmCH3

Crotonaldehyde
O has max = 290
nm
H2C C
CH3
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 max = 255 nm

OH

Phenol max = 270 nm

NH2

Aniline max = 280 nm

Absorption
& Intensity
Shifts
 Bathochromic Shift (Red Shift)
1
When absorption maxima (max) 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,

 Bathochromic Shift (Red Shift)
1
In alkaline medium, p-nitrophenol
shows red shift. Because negatively
charged oxygen delocalizes more
effectively than the unshared pair of
electron.
O + O
-
O + O
-

N N

-
OH

Alkaline
medium -
OH O

p-nitrophenol
 Hypsochromic Shift (Blue Shift)
2
When absorption maxima (max) 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.
 Hypsochromic Shift (Blue Shift)
2
Aniline shows blue shift in acidic
medium, it loses conjugation.

+ -
NH2 + NH3 Cl
H
Acidic
medium

Aniline
max = 280 nm max = 265 nm
 Hyperchromic Effect
3
When absorption intensity () 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

 4 Hypochromic Effect

When absorption intensity () of a

compound is decreased, it is known
as hypochromic shift.

CH3

Naphthalene 2-methyl
naphthalene
= 19000 = 10250
 Shifts and Effects
Hyperchromic shift

Blue Red
Absorbance ( A )

shift shift

Hypochromic shift

max
Wavelength ( )
APPLICATIONS
OF
UV / VISIBLE
SPECTROSCOPY
Applications
Qualitative & Quantitative Analysis:
It is used for characterizing aromatic
compounds and conjugated olefins.
It can be used to find out molar
concentration of the solute under study.
Detection of impurities:
It is one of the important method to detect
impurities in organic solvents.
Detection of isomers are possible.
Determination of molecular weight using
Beers law.
REFERE
NCES
Reference Books
Introduction to Spectroscopy
Donald A. Pavia

Elementary Organic Spectroscopy

Y. R. Sharma

Physical Chemistry
Puri, Sharma & Pathaniya
Resources
http://www2.chemistry.msu.edu/facul
ty/reusch/VirtTxtJml/Spectrpy/UV-Vi
s/spectrum.htm

http://en.wikipedia.org/wiki/Ultrav
iolet%E2%80%93visible_spectroscopy

http://
teaching.shu.ac.uk/hwb/chemistry/tu
torials/molspec/uvvisab1.htm