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P8M 309
UV VIS Spectroscopy
% SpecLroscopy ls Lhe sLudy of Lhe lnLeracLlon
beLween maLLer and radlaLed energy
Electromagnetic radiation (light) is one form of
energy.
Produced by fluctuation of electric charge and
magnetic field.
Electromagnetic radiation may be defined as the
propagation of the energy in the form of simple
harmonic wave which is produced by oscillation of
electric charge and magnetic field residing on the
atom.
Electromagnetic radiation (light)
Electromagnetic radiation is a selfpropagating wave
in space with electric and magnetic components.
EN radiation has an electric and magnetic field
component which oscillate in phase perpendicular to
each other and to the direction of energy
propagation.
Radio waves have longest wavelength and Camma
rays have shortest!
Since electromagnetic radiation (light) is a wave, it
can be characterized by either a wavelength or
frequency or wave numbers.
'he electromagnetic wave consists of two fluctuating
fields-one electric and the other magnetic. 'he two
vectors are at right angles to one another, and both are
perpendicular to the direction of travel.
'he distance between two consecutive peaks or
troughs in a particular wave.
Liner distance from any point on one wave to the
corresponding point on the adjacent wave.
!ncreasing wavelength
/
1
> /
2
> /
3
&4/63 lambda (/)
5; meter (m), practically
measured in micrometers
(m, a millionth of a meter)
or nanometers (nm, a
billionth of a meter )
Wavelength (/)
'he number of waves which can pass through a
given point in one second.
Decreasing frequency
v
1
< v
2
< v
3
&4/63 nu (v)
5;: cycle/sec, may
also be denoted as hertz
(Hz)
Frequency (v)
Creater the wavelength smaller the frequency.
Frequency, ; = C//
Frequency u
1
Wavelength
where C is the velocity of the light, 3.00x10
8
ms
*
n
n*
n
UJ/JIS
Jacuum UJ or Far UJ
(z<190 nm )
Electron transitions
'his is the earliest method of molecular
spectroscopy.
A phenomenon of interaction of molecules with
ultraviolet and visible lights.
Absorption of ENR results in electronic transition of a
molecule, and electrons are promoted from ground
state to higher electronic states.
Uvv!S spectroscopy
'he radiation which is absorbed has an energy
which exactly matches the energy difference
between the ground state and the excited state.
'hese absorptions correspond to electronic
transitions.
At room temperature, most of the atoms, molecules
and electrons are in the lowest energy orbital called
ground state.
'he electron of atom (molecule) at ground state can
absorb ENR and transit to higher energy orbital
called excited state.
Atom or molecule can absorb the radiation only when
the energy of proton is equal to the energy
difference of the two orbitals.
F * 'ransitions
An electron in a bonding orbital is excited to the
corresponding antibonding orbital. 'he energy
required is large.
For example, methane (which has only CH bonds,
and can only undergo F * transitions) shows
an absorbance maximum at 12S nm.
Absorption maxima due to F * transitions are
not seen in typical Uvv!S spectra.
n F * 'ransitions
Saturated compounds containing atoms with lone
pairs (nonbonding electrons) are capable of
n* transitions.
'hese transitions usually need less energy than
* transitions. 'hey can be initiated by light
whose wavelength is in the range 1S0 2S0 nm.
'he number of organic functional groups with
n* peaks in the Uv region is small.
n F n* and n F n* 'ransitions
Nost absorption spectroscopy of organic
compounds is based on transitions of n or n
electrons to the n* excited state.
'hese transitions fall in an experimentally
convenient region of the spectrum (200 700 nm).
'hese transitions need an unsaturated group in
the molecule to provide the n electrons.
nergy
9
6
9
6
n
Atomic orbital Atomic orbital
Nolecular orbitals
Occupied levels
Unoccupied levels
66
max
(Lambda max) lor a glven subsLance Lhe
wave lengLh aL whlch maxlmum absorbance ln Lhe
elecLromagneLlc specLrum occurs ls known as
max
y
p
o
c
h
r
o
m
i
c
ypsochromic
y
p
e
r
c
h
r
o
m
i
c
Bathochromic
Chromophores:
Functional groups that give electronic transitions.
1. Chromophore containing n electrons: ethylene
2. Chromophore containing n and n electron
Auxochromes:
W Substituents with unshared pair e's like OH, NH
2
,
NHR, NR
2
SH, OR, etc when attached to n
chromophore they generally move the /
max
to longer
/ and also increase the intensity of absorption.
W Combination of chromophore and auxochrome give
rise to another chromophore
W Also known as color enhancing group . e.g. Nitro
group impart yellow color.
W Auxochrome increase conjugation of chromophore
by lone pair of electron.
NH
2
ezene
max
2SS nm
max
203
An|||ne
max
280 nm
max
1430
NH
3
+
rotonated An|||ne
max
200 nm
max
???
|pheny|
max
2S0 nm
max
19000
H
3
2Methy| b|pheny|
max
237 nm
max
102S0
max
2SSnm
max
203
max
280nm
max
1430
max
2S0nm
max
19000
max
237nm
max
102S0
Extending conjugation has a larger effect
on 2
max
; shift is again to longer
wavelengths
2
max
170 nm
H
H
H
H
Ethylene (UPA name: ethene)
2
max
217 nm
(conjugated diene)
H
H
H
H
H
H
H
H
3
H
H
H
3
H
H
H
2
max
263 nm
conjugated triene
ycopene
2
max
505 nm
11 conjugated double bonds
orange-red pigment in tomatoes
2
max
451 nm
11 conjugated double bonds
BeerLambert's Law
'he intensity of emitted electromagnetic radiation
through a sample at a given wavelength decreases
exponentially with the sample thickness and
concentration.
BeerLambert's Law
'he BeerLambert Law is rigorously obeyed when
a single species is present at relatively low
concentrations.
'he BeerLamberts Law is not obeyed:
High concentrations
Solute and solvent form complexes
'hermal equilibria exist between the ground state
and the excited state
Fluorescent compounds are present in solution
A= log 1/'
A= absorbance
'= transmittance
!f sample had a transmitted light through
sample of 80. What is the absorbance of the
sample?
Ans: 0.036
'ransmittance is the percentage of incident light that
passes through a sample of material.
'9.5:4;;.50
onventional
Spectrophotometer
Schematic of a conventional single-beam spectrophotometer
onventional
Spectrophotometer
Optical system of a double-beam spectrophotometer
onventional
Spectrophotometer
Optical system of a split-beam spectrophotometer
Sample cells
Uv Spectrophotometer
Quartz (crystalline silica)
visible Spectrophotometer
Class
Light sources
Uv Spectrophotometer
1. Hydrogen Cas Lamp
2. Nercury Lamp
visible Spectrophotometer
1. 'ungsten Lamp
efine: Iectromagnetic spectrum, Vacuum
UV region
2 $how the infIuence of poIarity of soIvent on
different types of transitions onIy by figure in
UV-Vis sppectroscopy