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uv vlS SpecLroscopy

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

All types of radiations travel with the same velocity


and no medium is required for their propagation. 'hey
can travel through vacuum.
Wave number is defined as the total number of
waves passing through a space of unit length (1cm
or 1m).
We can calculate the wave number by taking the
reciprocal value of wave length of a radiation. i.e.
Wave number
Wave number =
1
Wavelength
Energy of a particular wave of the particular
radiation can also be calculated by applying the
relation:
Energy
E=h.; = h. (c//)
Here
h = Planck's constant = 6.626x10
27
erg sec
; = Frequency of radiation
c = Velocity of EM radiation
/ = Wavelength
'he arrangement of all types of electromagnetic
radiations in order of their increasing wavelength or
decreasing frequencies is known as complete
electromagnetic spectrum.
'he full electromagnetic radiation spectrum is
continuous and each region merges slowly into the
next.
For convenience of reference, definitions of the
various spectral regions have been set by the ]oint
Committee on Nomenclature in Applied
Spectroscopy:
Electromagnetic spectrum
Fig: Electromagnetic spectrum
Region WaveIength (nm)
Far ultraviolet 10-200
Near ultraviolet 200-380
Visible 380-780
Near infrared 780-3000
Middle infrared 3000-30,000
Far infrared 30,000-300,000
Microwave 300,000-1,000,000,000
!t may be defined as the analysis of chemical
substance by the measurement of the amount of
radiation absorbed by this substance.
Absorption spectroscopy refers to a range of
techniques employing the interaction of
electromagnetic radiation (ENR) with matter.
!n absorption spectroscopy, the intensity of a beam
of light measured before and after interaction with a
sample is compared.
Absorption spectroscopy
!nfrared spectroscopy
Near infrared spectroscopy
Nicrowave spectroscopy
Xray spectroscopy (emission spectroscopy)
Uvvisible spectroscopy
NNR spectroscopy
Examples of absorption spectroscopy
Absorption spectroscopy is widely used for both
qualitative (is a chemical present?) and
quantitative (how much?) and structural (is it
degraded?) work in a wide range of fields.
'he Uv light below 200nm is relatively uninformative
and difficult to measure. Since air also absorb
radiation in this region.
!n order to measure absorption of Uv radiation below
200nm, air must be removed from the instrument.
'his region is called vacuum Uv region.
!t can excites sigma bond electrons.
'he Uv light above 200nm excites pi bond electrons
and the region give informative spectra and can be
easily measured.
vacuum ultraviolet region
3 kinds of electrons in organic molecules are involved
Uv or visible spectroscopy
Bonding electron (ground level)
Nonbonding electron (ground level)
Antibonding electron (excited level)
Electrons of ground states are in outer most shell or
valence electrons.
Kinds of electron involved
While two atoms form chemical bond, their atomic
orbital combine together to form molecular orbital.
Bonding orbital and antibonding orbital
Bonding orbital energy level is always lower than that
of the original atomic orbital
Antibonding orbital energy higher
Electrons which are involved in bond formation.
'hey are present in the lower energy level in the
molecule.
'hey are of two types:
bond electrons (electrons in the single bond,
electrons in saturated compounds)
nbond electrons (electrons in double and
triple bond, electrons in unsaturated compounds)
Bonding electron
Paired electrons in outer most shell which are not
involved in bond formation between atoms in
molecule.
'hey are present in the higher energy level in the
molecule than bonding electrons.
'hey are represented by n.
Organic compounds contains nitrogen, oxygen,
halogen, sulphur.
Nonbonding electron
Electrons which are excited to a higher energy level
by absorbing energy (ENR) are called antibonding
electron.
'hey are present in the higher energy level.
'hey are represented by:
Antibonding sigma (*) and
Antibonding pi (n*)
Antibonding electron
E
n
e
r
g
y

*
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

SolvenL selecLlon crlLerla


ulluLed soluLlon ls used for analysls so
solvenLs has greaL lnfluence on Lhe process
1he crlLerla are
% Should noL absorb radlaLlon (1100)
% Should be less polar Lo avold lnLeracLlon
% SolvenLs do noL conLaln double bonds are
mosL sulLable
% Should be free from lmpurlLles
SolvenL selecLlon crlLerla
MosL commonly used solvenL ls 93 eLhanol
Commerclal grade solvenL conLaln benzene as
lmpurlLy
So|vent At wh|ch or above
wh|ch rema|n
transparent
JaLer 203 nm
MeLhanol LLhanol LLher
Cyclohexane nhexane
210 nm
Chloroform 243 nm
8enzene 280 nm
'ransition of electron of acetone (n n*)
Polarity of solvent decrease
Solvents H
2
O CH
3
OH C
2
H
S
OH CHCl
3
C
6
H
14
/
max
(nm) 264.S 270 272 277 273
CH
3
CH
3
C
=
O
Polarity
!n chemistry, polarity refers to the dipoledipole
intermolecular forces between the slightly positively
charged end of one molecule to the negative end of
another or the same molecule.
Nolecular polarity is dependent on the difference in
electronegativity between atoms in a compound and
the asymmetry of the compound's structure.
For example, a molecule of water is polar because of
the unequal sharing of its electrons in a "bent"
structure, whereas methane is considered nonpolar
because the carbon shares the hydrogen atoms
uniformly.
Bathochromic shift (red shift) a shift to longer /
lower energy
Hypsochromic shift (blue shift) shift to shorter /
higher energy
Hyperchromic effect an increase in intensity
Hypochromic effect a decrease in intensity
Chromophores: functional groups that give electronic
transitions.
Auxochromes: substituents with unshared pair e's
like OH, NH, SH ..., when attached to n chromophore
they generally move the absorption max. to longer /.
Bathochromic shift: shift to longer /, also called red
shift.
Hysochromic shift: shift to shorter /, also called blue
shift.
Hyperchromism: increase in s of a band.
Hypochromism: decrease in s of a band.
200 nm 700 nm
1

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