Molecular

Spectroscopy
(Electronic Spectroscopy)

(Optical properties)
Dr. Mohan Singh Mehata
(JSPS & CAS Fellow)

Molecular Vibrations
and
IR Spectroscopy

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.

Nobel Prizes
Nobel Prizes awarded in spectroscopy. These Prizes exemplify both experimental and theoretical contributions to
the growth of spectroscopic investigations as well as the development and discoveries related to spectroscopy.

Experimental Contributions to Spectroscopy

H.A. Lorentz and P. Zeeman (1902) discovery of the splitting of spectral lines in magnetic
J. Stark (1919) discovery of the splitting of spectral lines in electric fields
C.V. Raman (1930), the first to demonstrate spectral line shifts due to inelastic light scattering (Raman effect)
W.E. Lamb, Jr. (1955), who discovered the fine structure splitting in the first excited state of atomic hydrogen (this
work was actually done with microwaves, but its origin and impact have been central to studies of atomic spectra)
R.S. Mulliken (1966) and G. Herzberg (1971), for their contributions to molecular spectroscopy
A.L. Schawlow (1981), for work in the field of laser spectroscopy
A. Zewail (1999), for studies of the transition states of chemical reactions using femtosecond spectroscopy

Theoretical Contributions to Spectroscopy

M. Planck (1918), who discovered the elemental quantum of action
N. Bohr (1922), the first to link the regularities of spectral lines to the quantum structure of atoms
P.A.M. Dirac and E. Schrodinger (1933), for their contributions to the quantum theory of atoms
W. Pauli (1945), who discovered the quantum exclusion principle
Inventions and Discoveries Related to Spectroscopy
A.A. Michelson (1907) invention of the interferometer, a hallmark in spectroscopic instrumentation
C.H. Townes , N.G. Basov and A.M. Prokhorov (1964) development of the maser, a source of coherent microwave
radiation, which led to development of the laser and opened the field of modern spectroscopy
A. Kastler (1966) for the development of optical pumping of atoms
N. Bloembergen (1981) for his contributions to nonlinear optics
H.G. Dehmelt and W. Paul (1989) received this award for invention of the ion trap, an important tool in current
spectroscopic research

Symmetrical
stretching

Asymmetrical
stretching

Scissoring

Rocking

Wagging

Twisting

Introduction: Molecular spectroscopy

About the excitation and de-excitation processes
Absorption
Emission
Excitation
Lifetime
Kashas Rule
Franck Condon Principle
Experimental:
Sample preparation
Steady-state
Time-resolved
Stark effects:
Spectral shift, broadening, quenching and enhancement
Results

Electromagnetic Radiation

Electromagnetic Radiation

Violet

400 - 420 nm

Yellow

570 - 585 nm

Indigo

420 - 440 nm

Orange

585 - 620 nm

Blue

440 - 490 nm

Red

620 - 780 nm

Green

490 - 570 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

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.

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

Beer Lamberts Law:

A=bc
A=absorbance
=molar absorbtivity with units of L /mol.cm
b=path length of the sample (cuvette)
c =Concentration of the compound in solution,
expressed in mol /L

The Quantitative Picture

Transmittance:
Absorbance:

I0
(powerin)

T = I/I0

A = -log10 T = log10 I0/I

I
(powerout)

B(path through sample)

The Beer-Lambert Law (Beers Law):

A = bc
Where the absorbance A has no units, since A = log 10 I0 / I
is the molar absorbtivity with units of L mol -1 cm-1
b is the path length of the sample in cm
c is the concentration of the compound in solution, expressed in mol L -1 (or M, molarity)

The possible electronic transitions are

 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 * transition and shows
absorbance maxima at 125 nm.

 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.
e.g. Alkenes generally absorb in the region
170 to 205 nm.

 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 region is small
(150 250 nm).

 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 longer wavelength
around 300 nm.

 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 broad of
absorption.

The possible electronic

graphically shown as:

transitions

can

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

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

 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, -OCH3
causes absorption of compound at longer
wavelength.

 In alkaline medium, p-nitrophenol shows red

shift. Because negatively charged oxygen
delocalizes more effectively than the
unshared pair of electron.
O

OH

Alkaline
OH

medium

p-nitrophenol
max = 255 nm max = 265 nm

 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.

 Aniline shows blue shift in acidic medium, it

loses conjugation.
+

NH2

NH3 Cl

Acidic
medium

Aniline
max = 280 nm

max = 265 nm

 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

CH3

Pyridine
2methylpyridine
max = 257 nm
max = 260
nm

 When absorption intensity () of a

compound is decreased, it is known as
hypochromic shift.
CH3

Naphthalene
= 19000

2-methyl naphthalene
= 10250

Shifts and Effects

Absorbance ( A )

Hyperchromic shift

Red
shift

Blue
shift

Hypochromic shift
max
Wavelength ( )

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 either ethyl
alcohol or hexane.

Instrumentation
Components of UV-Visible spectrophotometer
Source

Filters & Monochromator

Sample compartment
Detector
Recorder

Five Basic Optical Instrument Components

1) Source A stable source of radiant energy at the

desired wavelength (or range).

2) Wavelength Selector A device that isolates a
restricted region of the EM spectrum used for
measurement (monochromators, prisms & filters).
3) Sample Container A transparent container used to
hold the sample (cells, cuvettes, etc).
4) Detector/Photoelectric Transducer Converts the
radiant energy into a useable signal (usually electrical).
5) Signal Processor & Readout Amplifies or attenuates
the transduced signal and sends it to a readout device as
a meter, digital readout, chart recorder, computer, etc.

How do UV spectrometers work?

Rotates,to
achievescan

Matchedquartzcuvettes
Sampleinsolutionatca.105M.
SystemprotectsPMtubefrom
straylight
D2lampUV
TungstenlampVis
DoubleBeammakesita
differencetechnique

Twophotomultiplier
inputs,differential
voltagedrivesamplifier.

LIGHT SOURCES

Various UV radiation sources are as follows

a. Deuterium lamp
b. Hydrogen lamp
c. Tungsten lamp
d. Xenon discharge lamp
e. Mercury arc lamp
Various Visible radiation sources are as follow
a. Tungsten lamp
b. Mercury vapour lamp
c. Carbonone lamp

Wavelength Selectors
Wavelength selectors output a limited, narrow,

continuous group of wavelengths called a

band.
Two types of wavelength selectors:
A) Filters
B) Monochromators
A)Filters
Two types of filters:
a) Interference Filters
b) Absorption Filters

B. Monochromators
Wavelength selector that can continuously
scan a broad range of wavelengths.
Used in most scanning spectrometers
including UV, visible, and IR instruments.
PRISM TYPE

GRATING TYPE

Refractive type
Reflective type
Diffraction type
Transmission

SUMMARY
Types of source, sample holder and detector

for various EM region

REGION

SOURCE

SAMPLE
HOLDER

DETECTOR

Ultraviolet

Deuterium lamp

Quartz/Fused
silica

Phototube, PM
tube, diode array

Visible

Tungsten lamp

Glass/Quartz

Phototube, PM
tube, diode array

Emission
Fluorescence and phosphoresence

Jablonski Diagram

Jablonski Diagram

More Complex Electronic Processes

Fluorescence: absorption of

radiation to an excited state,

followed by emission of radiation to
a lower state of the same
multiplicity
Phosphorescence: absorption of
radiation to an excited state,
followed by emission of radiation to
a lower state of different multiplicity
Singlet state: spins are paired, no
net angular momentum (and no net
magnetic field)
Triplet state: spins are unpaired, net
angular momentum (and net
magnetic field)

Excited State Processes

ESIPT

Ground State
Ionized species

Energy Transfer
Chromophore II

Ground State

Ground State
Chromophore II

Time Scale of various Processes

FranckCondon principle
which states that an electronic transition takes place so rapidly that a vibrating
molecule does not change its internuclear distance appreciable during the transition

Examples
(molecular systems)

Mirror-image rule and Franck-Condon factors. The absorption

and emission spectra are for anthracene. The numbers 0, 1,
and 2 refer to vibrational energy levels

Excited state reactions:

Fluorescence Lifetimes and Quantum Yields

Quantum yield is the ratio of number of photons emitted to the number of absorbed.
Substances with the largest quantum yields, approaching unity, such as rhodamines, display the
brightest emissions

Lifetime
The lifetime of the excited state is defined by the average time the molecule spends in the excited
state prior to return to the ground state.

The lifetime of the fluorophore in the absence of nonradiative, processes is called the intrinsic
or natural lifetime, and is given by

In principle, the natural lifetime n can be calculated from the absorption spectra, extinction
coefficient, and emission spectra of fluorophores.
where F() is the emission spectrum plotted on the
wavenumber (cm1) scale, () is the absorption spectrum,
and n is the refractive index of the medium. The integrals
are calculated over the S0-S1 absorption and emission
spectra.

The natural lifetime can be calculated from the measured

lifetime ( n ) and quantum yield.

Excitation and Emission Spectra

Stokes Shift

Kashas Rule:
Emission Spectra Are Typically Independent of the Excitation Wavelength.

Samples
A
l

Polymer film

ITO
Quartz
plate
C8H17

C8H17

N
O
Al
N

(n = 60, m = 40)

O
N

O
S

Geometry optimized structure of 6-HQ and

Absorption Spectra as a function of AcOH

Emission & Excitation Spectra of

6-HQ as a function of AcOH

Excited State Dynamics of 6-HQ in

presence of AcOH
TRES

M. S. Mehata, Chemical Physics Letters, 436 (2007) 357

Proton Translocations along H-bonded

molecular wire

M. S. Mehata, Journal of Physical Chemistry B 112 (2008) 8383

M. S. Mehata, Chemical Physics Letters, 436 (2007) 357

CONCLUSIONS
6-Hydroxyquinoline in benzene emits normal fluorescence around
357 nm.
In the presence of acetic acid (HOAc), it exhibits two more bands
at ~419 nm and a large Stokes shifted one at ~583 nm with
decreased intensity of the normal fluorescence.
It appears to form (1:1) and (1:2) complexes- (1:2) 6-HQ/HOAc
undergoes an excited state proton rearrangement (via HOAc wire)
resulting in keto tautomer (emitting at ~583 nm).
This appears to be in line with recent findings where ammonia
wires facilitate proton/hydrogen translocation (Science, 302, 1736,
2003). However, (1:1) 6-HQ/HOAc exhibits intermediate emission
(~419 nm) presumably due to ESIPT

Absorption, emission and decay profiles of 6-MQ+

:
an indicator for Cl ions

Stark Effect
Electric field modulation spectroscopy
(Stark Spectroscopy)

How the E-F spectra varies with excitation

M. S. Mehata et al., Journal of Physical Chemistry C (in press)