UV

SPECTROSCOPY

Prof.Harminder Kaur
 INTRODUCTION
⦿ Spectroscopy is the tool for study of the
atomic and molecular structures.
⦿ Ultra-violet spectroscopy involves the
measurement of the absorption of light in
the ultra-violet regions by the compounds
under investigation.
⦿ Since it occurs due to the transition between
the electronic energy levels within a
molecule, it is quite often also known as
electronic spectroscopy.
⦿ The UV region
ranges between
100-400 nm.
⦿ Further it is divided
into 2 separate
regions:
✔ far UV region:
wavelength varies
from 100-200 nm.
✔ near UV region:
wavelength varies
from 200-400 nm.
⦿ NOTE 1: in the region at or below 200 nm,
oxygen (O2) absorbs strongly. Therefore, the
measurements are carried by flushing the
instrument with nitrogen.

⦿ NOTE 2: in a region extending at 150 nm or

below, nitrogen also starts absorbing. Therefore,
the investigations in this region can be done
only by using evacuated instruments. This region
is known as vacuum region.
PRINCIPLE OF UV SPECTROSCOPY
⦿ Absorption of ultra-violet light produces changes
in the electronic states of molecules associated
with the excitation of an electron from a lower
to higher energy level. This energy difference is
given by,
E = hν joules
⦿ Each electronic level in a molecule is associated
with a no. of vibrational sublevels with small
energy separations.
⦿ Similarly each vibrational level is associated
with a no. of rotational sub-levels.
⦿ Total energy of the molecule is the sum of the
electronic, vibrational and rotational energies,
 E= Eelec + Evib + Erot

⦿ If the electronic
excitation in the
molecule had
occurred without any
accompanying
changes in vibrational
and rotational
sub-levels, the
spectrum would have
consisted of sharp
lines with each line
representing a change
in electronic state.
⦿ But due to mixing of vibrational and rotational
changes with electronic changes in the
molecules, there will be a large number of
possible transitions requiring only slightly
different energies. This will require the
absorption of a large no. of wavelengths which
are quite close together resulting in the
formation of broad bands in the spectrum
obtained. Thus UV spectra of organic molecules
consist of bands rather than peaks.
ABSORPTION LAWS
⦿ LAMBERT’S LAW: According to the law,
when a beam of monochromatic light passes through a
homogenous medium, the rate of decrease of intensity of
radiation with the thickness of absorbing medium is
proportional to the intensity of the incident radiation.
Mathematically,
-dI/dx = kI
where
I= intensity of the radiation after passing through
a particular thickness x
-dI/dx= rate of decrease of intensity of the radiation
with the thickness of the absorbing medium
k= absorption constant
⦿ BEER’S LAW: According to the law,
when a beam of monochromatic light is passed through a
substance dispersed in a non-absorbing solvent, the absorption
of light is directly proportional to the concentration of the
absorbing medium.
⦿ The two laws can be combined to get a general
absorption law known as Beer-Lambert’s law.
⦿ BEER-LAMBERT’S LAW: According to the law,
the absorption of light by a substance of particular
wavelength is proportional to the number of molecules in the
path of light.
mathematically,
log10 (I0/I) = Є.c.l
Where,
I0 = intensity of incident light
I = intensity of transmitted light
 c = concentration of absorbing substance in
mol/L
l = path length or thickness of cell containing the
sample in cm
Є = proportionality const known as molar
absorptivity or molar extinction coefficient

⦿ Є is the measure of the intensity of absorption at a

particular wavelength.
⦿ The expression log10(I0/I) is termed optical density
or absorbance (denoted as A).
⦿ Since (I0/I) is the ratio of intensity of the incident
light I0 to the transmitted light I, the percent
transmittance of a sample is:
% transmittance = 100 X (I/I0)
⦿ λmax = wavelength of light absorbed with
maximum absorbance

Єmax

Absorbance

λmax

Wavelength 🡪
⚫ We know that absorption of radiant
energy in UV and visible region occurs
due to excitation of electrons . An electron
excites from ground state to any of higher
state.
There are three types of electrons present
in ground state of molecules
1.Sigma
2.Π (pi)
3.n

Types of Electronics Transitions

6/1/2022 12
6/1/2022 13
⚫ The following electronic transition takes place
in uv and visible region:

⚫ Sigma to sigma*
⚫ π to π*
⚫ n to π*
⚫ n to sigma*
Where sigma*, π* are excited state anti-
bonding orbitals

⚫The increasing order of energy are:

n to π* < π to π* < n to sigma* < sigma to
sigma *

6/1/2022 14
⚫ We shall now study each of these transition.

1. sigma to sigma* : electron is excited from

sigma bonding to sigma* anti-bonding
orbital
⚫ This is high energy process, therefore shorter
wavelength is required.
⚫ They occur only in compound having only
sigma bond. Eg : methane absorb at shorter
wavelength than propane.
⚫ NOTE: cyclopropane absorb at longer
wavelength than propane because it is more
strained.
6/1/2022 15
2. n to sigma*: In these transition a non
bonding electron is excited to sigma*
anti-bonding orbitals.
⚫ Occur in the compound having at least
one hetero atom for example, oxygen,
nitrogen, halogen, sulphur or which have
at least one loan pair of electrons e.g.:
haloalkanes, amines, alcohols, aldehydes,
ketones etc.
⚫ They require lower energy than sigma to
sigma*,hence occur at lower wavelength

6/1/2022 16
⚫ It is noted that lower the electro negativity of
hetero atom ,higher will be the wavelength of
absorption, thus methyl chloride absorb at
173 mµ ,while methyl iodide absorb at 258
mµ
⚫ We know that the molecules in both amines
and alcohols are hydrogen bonded ,therefore
require high energy and shorter wavelength
than expected

3. π to π* : in these transition a π electron is

excited to π* antibonding orbitals
⚫ Usually occur in the unsaturated compound
having atleast one multiple bond. E.g. C=O,
C=N, N=O, C=S and also in aromatic
compound

6/1/2022 17
⚫ They require even lesser energy than n to
sigma*, hence higher wavelength, absorption
generally occur in the region of an ordinary
ultra violet spectrometer.
⚫ In carbonyl compound, absorption occur at
wavelength of 180 mµ while in conjugated
dienes it ranges b/w 170 to 190 mµ
⚫ It may be noted that the induction of an
alkyl group raises the wavelength of
absorption by 3to 5 mµ. This shift is called
bathochromic shift depend upon alkyl group
and also on stereochemistry

6/1/2022 18
4. n to π* : these are transitions in which
electron gets excited from non bonded
orbitals to π* molecular orbital.
⚫ There should be presence of hetero atom like
oxygen, nitrogen joined with carbon as
organic compound.
⚫ Out of all transitions, n to π* require least
energy , thus takes place at maximum
wavelength.
⚫ Note: Aldehyde generally show two types of
transitions:

6/1/2022 19
⚫ π to π*: usually occur at wavelength=290
mµ
⚫ n to sigma*: occur at wavelength of 180 mµ
in case of aldehydes.
⚫ It may also be noted that absorption taking
place at lower wavelength is generally more
intense.

6/1/2022 20
Probability of Electronic
Transition
Based upon whether a particular transition takes place
readily or to a very small extent electronic transition are
divided into two types:
⚫ Allowed Transition
⚫ Forbidden Transition

⚫ Allowed Transition : The transition which

takes place for intensity ranging between
10000 to 1000000 are called allowed
transition. They are generally due to π to π*
transition, e.g. 1,3-butadiene is an allowed
transition for π to π* transition. Actually in
this case, both the orbitals lie in same plane
so overlap to a greater extent, so transition
takes place readily
⚫ Forbidden transition: Transition for which
intensity value is b/w 10 to1000 are called
forbidden transition, e.g. n to π* transition in
case of saturated aldehyde is forbidden
because of less value of intensity i.e. 100.

⚫ Forbidden transition implies that probability

of transition is very low. Actually non bonded
atoms are in a plane perpendicular to the
plane containing π* molecular orbital.
Therefore overlap does not takes place to
greater extent. Therefore the probability is
very low.
QUESTION : The UV spectrum for acetone shows two
peaks at λmax =280 mµ, Єmax =15 and λmax =190 mµ, Єmax
=100. identify the electron transition for each and indicate
which is more intense.

ANSWER: λmax =280 mµ represents n to π* transition.

λmax =190 mµ represents π to π* transition.
As π to π* transition has larger Єmax it has,
therefore, a more intense peak.
ROLE OF SOLVENT IN
ELECTRONIC TRANSITION
◼ The Polarity of solvent influence electronic
transition in uv spectroscopy. The solvent
may cause shift in the position of
absorption band either towards longer
wavelength or shorter wavelength, this
depend on type of electronic transition as
follows:
1> Effect of solvent on n to π* and
n to sigma* transitions :
◼ In case a particular solvent is more polar in
ground state than excited state ,the n electrons
in ground state are more stablised in comparison
with excited state because of hydrogen bonding
or dipole dipole interaction.
◼ As a result the energy difference between the
ground state and excited state increases, thus
absorption towards region of shorter wavelength
◼ Eg; carbonyl group is more polar in ground than
in excited state, thus energy difference b/w
ground state and excited state increases, thus
absorption shift towards shorter wavelength eg
wavelength for acetone is 279 mµ but it becomes
264 mµ In polar solvent
DIAGRAM 1

∏*
E1

E2
Ground state
gets stabilised

n
NON-POLAR SOLVENT

POLAR SOLVENT
2. Effect of solvent in π to π*
transition
◼ If a particular solvent is more polar in excited
state than in ground state , the absorption shift
to a region of longer wavelength. this is hydrogen
bonding in excited state, thus excited state
becomes stable , Thus energy difference
decreases, this cause shift in absorption band to
longer wavelength.
◼ Eg; the molecule of ethylene is more polar in
excited state than in excited state due to π to π*
transition, as a result in polar solvent the
absorption shift to longer wavelength.
DIAGRAM 2

Excited state
* gets stabilised
∏

E1
E2

∏
NON-POLAR SOLVENT POLAR SOLVENT
Choice of solvent

◼ A suitable solvent for ultra violet spectroscopy

should meet the following requirements:
◼ It should not absorb radiation in the region under
investigation.
◼ It should be less polar so as minimum interaction
with the solute molecule.
◼ The most commonly employed solvent is 95%
ethanol. It is cheap and does not absorb radiation
above 210 nm. In other words it is transparent.
Commercial ethanol should not be used as it
contains benzene which undergo absorption at
about 280 nm
◼ Some other solvents which are transparent
above 210 nm are n hexane, cyclohexane ,
methanol , water and ether.
◼ Benzene , chloroform and carbon tetra
chloride can not be used as they absorb in
range of 240-280 nm.
◼ n- hexane and other hydrocarbons are
preferred in polar solvent because of
minimum interaction with solute molecule.
CHROMOPHORES
⦿ The term ‘chromophore’ is applied to a group which
imparts characteristics colour to a compound. In
Greek, Chromophore = colour barrier.
⦿ For example, nitro (NO2) group is a chromophore as
it imparts yellow colour to a compound. Similarly,
diazo (-N=N-) group as it imparts orange or red
colour to the organic azo dyes.
⦿ But at present, the concept of chromophore has
undergone a radical change. A chromophore may be
defined as:
the functional group that absorbs electromagnetic
radiations irrespective of the fact whether it imparts
a particular colour to the compound or not.
⦿ For example, carboxyl group (-COOH) does not give
colour but is still regarded as the chromophore, since
it absorbs at λmax 208 mµ in the UV region and has
Єmax equal to approximately 15.
LIST OF SIMPLE ORGANIC ACTING
AS CHROMOPHORES
Chromophores λmax (mµ) nature of Єmax
transitions
__
Nitrile (-C≡N) 165 π > π* 65
Ethylenic (-C=C-) 170 __ 14000
π > π*
Acetylenic (-C≡C-) 175 __ 10000
π > π*
Carbonyl (>C=O) 160 __ 16000
π > π*
__
185 n > σ* 5000
__
280 n > π* 15
__
Nitro (-NO2) 201 n > σ* 5000
__
274 n > π* 17
__
Carboxylic (-COOH) 208 π > π* 32
__
Ester (-COOR) 211 n > π* 15
__
Azo (-N=N-) 338 n > π* 2
⦿ From the data, it is evident that there are many
chromophores which absorb at wavelength below
220 mµ. This region can’t be studied properly. This
problem may be solved by attaching certain groups
to the basic chromophores.

⦿ Substituent Effects
1. The attachment of substituent groups (other than
H) can shift the energy of the transition.
2. Substituents that increase the intensity and often
wavelength of an absorption are called
auxochromes.
3. Common auxochromes include alkyl, hydroxyl,
alkoxy and amino groups and the halogens.
⦿ Substituents may have four effects on chromophore:
1. Bathochromic shift (red shift) :: a shift to longer λ,
lower energy. It may be achieved by the change of
solvent( solvent effect) or by the presence of an
auxochrome.
2. Hypsochromic shift (blue shift) :: a shift to shorter
λ, higher energy. It can be caused by either
changing the solvent or by removing the
conjugation.
3. Hyperchromic shift :: an increase in intensity and
it is caused by the introduction of an auxochrome.
4. Hypochromic shift :: a decrease in intensity and is
affected by a group which disturbs the geometry
of the molecule.
 +auxochrome

HYPERCHROMIC
(Solvent,
Є HYPSOCHROMIC BATHOCHROMIC +auxochrome)
(Solvent, HYPOCHROMIC
-conjugation)

200 nm (+ group to disturb geometry) 700 nm

WAVELENGTH λ (nm)->
AUXOCHROMES
⦿ An auxochrome is a functional group of atoms with
non-bonded electrons which, when attached to a
chromophore, alters both the wavelength and
intensity of absorption.
⦿ They generally cause bathochromic shift i.e. a shift
to longer wavelength and also increase in intensity
of the absorption band.
⦿ If these groups are in direct conjugation with the
pi-system of the chromophore, they may increase
the wavelength at which the light is absorbed and as
a result intensify the absorption.
⦿ A feature of these auxochromes is the presence of
at least one lone pair of electrons which can be
viewed as extending the conjugated system by
resonance.
⦿ The most important auxochromic groups are OH,
NH2, CH3 and NO2 and their properties are acidic
(phenolic) or basic.
⦿ The actual effect of an auxochrome on a
chromophore depends on the polarity of the
auxochrome, e.g. groups like CH3-, CH3CH2-and Cl-
have very little effect, usually a small red shift of
5-10nm. Other groups such as –NH2 and-NO2 are
very popular and completely alter the spectra of
chromophores such as benzene.
⦿ In general it should be possible to predict the effect
of non-polar or weakly polar auxochromes, but the
effect of strongly polar auxochromes is difficult to
predict.
EXAMPLES OF AUXOCHROMES:
⦿ Benzene and Aniline:

(λmax = 254 mµ and Aniline (λmax = 280 mµ and

Єmax = 230) Єmax = 1430)

⦿ the electron pair on –NH2 group

(auxochromophore) is involved in conjugation
with the pi-electron pair of the ring. Therefore,
there is an increase in the value of both λmax
and Єmax.
⦿ Azobenzene and p-ethoxy azobenzene:

Azobenzene p-ethoxy azobenzene

λmax = 320 mµ and λmax = 385 mµ and Єmax =
Єmax = 21,000 42,000
CONJUGATED CHROMOPHORES
⦿ When two chromophoric groups are in isolated
positions i.e. are linked to each other through
more than one single bond, their effect in the
spectrum is additive.
⦿ For eg: CH2=CH-CH2-CH2-CH=CH2
⦿ However, if they are conjugated like
CH2=CH-CH=CH2
There is large effect on the spectrum as there is
bathochromic shift. Increase in λmax and Єmax.
QUESTIONS:
⦿ Q1. the following three compounds have λmax at 176
nm, 211 nm and 215 nm respectively.
a) CH2 = CHCH2CH = CH2 b) CH3 CH = CH2
C=C
H H

c) CH3 H
C=C
H CH = CH2

ANS: a) represents an isolated diene.

b) and c) are geometrical isomers of each other. B)
is a cis isomer and c) is a trans isomer.
⦿ Q2: which will have a greater λmax ?

CH=CH CH=CH-CH=CH

A) B)

ANS: structure B) is expected to have greater λmax

because greater the extent of conjugation, more
the value of λmax in the UV spectrum.
CYCLIC DIENES:
⦿ Cyclic dienes are of two types:
a) Homoannular diene: it is the one in which the
two double bonds are present in the same ring.
b) Heteroannular diene: it is the one in which the
two double bonds are present in the adjacent
rings.

NOTE: In general, a homoannular diene absorbs at a

greater wavelength than the heteroannular diene.
However, its intensity of absorption is lesser.
⦿ Nature of double
bonds in the cyclic
dienes:
a) Endocyclic double
bonds === it is
present within a
particular ring.

b) Exocyclic double
bonds === it lies
outside a particular
ring.

Note: The presence of the exocyclic double bond

generally increases the wavelength of absorption of a
cyclic conjugated diene.
WOODWARD-FIESER RULES FOR
CALCULATING ΛMAX IN DIENES:
⦿ According to these rules, each diene has a certain
fixed basic value and its actual λmax depends upon
the following points:
1. the no. of alkyl substituents or ring residues
present on the diene system.
2. The no. of double bonds extending conjugation
beyond the diene system.
3. The presence of auxochromes i.e. the polar groups
such as –Cl, -Br, -OR, -SR, -NH2, -OCOCH3, etc.

The contribution by the different substituents or

auxochromes; if present, is added to the λmax
value or the basic value of the diene, these rules
are given in the following table:
 Parent open-chain diene base 217 nm

Parent heteroannular diene base 215 nm

base 253 nm
Parent homoannular diene
ATTACHED GROUP INCREMENT, nm
For each double bond extending conjugation +30

For each alkyl substituent (R) or ring residue +5

For each exocyclic double bond +5

For an O-Acyl (-OAc) 0

For an O-Alkyl (-OR) +30

For a S-Alkyl (-SR) +6

For a Cl or a Br atom +5

For a –NR2 +60

 WOODWARD AND FIESER RULES
FOR CALCULATING ΛMAX IN
ENOMES:
Enomes are generally α,β
unsaturated carbonyl
compounds.
β α
For ketones

(i) open chain or six-membered Base 215 nm

(ii) five-membered Base 202 nm

For aldehydes Base 207 nm

For acids and esters Base 197 nm

 δ γ β α
To the base value, add
these increments
α β γ δ
Extend C=C +30
Add exocyclic C=C +5
Homoannular diene +39
alkyl +10 +12 +18 +18
OH +35 +30 +30 +50
O-Acyl +6 +6 +6 +6
O-alkyl +35 +30 +17 +31
NR2 , NH2, NHR +95
Cl/Br +15/+25 +12/+30
SOME WORKED EXAMPLES:
Parent unit : acyclic diene
Basic value = 217 nm
1 alkyl substituent = +5 nm
Total λmax = 222 nm

Observed value= 220 nm

Parent unit : acyclic diene

Basic value = 217 nm
2 ring residues = 2 x (+5 nm) = +10 nm
1 exocyclic double bond = +5 nm
Total λmax = 232 nm

Observed value= 237 nm

Parent unit : heteroannular diene
Basic value = 215 nm
3 ring residues = 3 x (+5 nm) = +15 nm
1 exocyclic double bond = +5 nm
Total λmax = 235 nm

Observed value= 235 nm

Parent unit : heteroannular diene

Basic value = 215 nm
4 ring residues = 4 x (+5 nm) = +20 nm
1 exocyclic double bond = +5 nm
Total λmax = 240 nm

Observed value= 242 nm

Parent unit : homoannular diene

Basic value = 253 nm
4 ring residues = 4 x (+5 nm) = +20 nm
1 exocyclic double bond = +5 nm
Total λmax = 278 nm

Observed value= 278 nm

Three common errors:

This compound has three exocyclic double

bonds; the indicated bond is exocyclic to
two rings

This is not a heteroannular diene; you

would use the base value for an acyclic
diene

Likewise, this is not a homoannular diene;

you would use the base value for an acyclic
diene
 Parent unit : α,β unsaturated carbonyl
Basic value = 215 nm
2 β-alkyl substituents = 2x (+12) = 24 nm
Total λmax = 239 nm

Observed value = 238 nm

Parent unit : α,β unsaturated carbonyl

Basic value = 215 nm
1 β-alkyl substituent = +12 nm
1 δ-alkyl substituent = +18 nm
1 exocyclic double bond = +5 nm
δ β Extended conjugation = +30 nm
γ α Total λmax = 280 nm

Observed value = 280 nm

APPLICATIONS
⦿ DETECTION OF FUNCTIONAL GROUPS-
FOR E.G- If a compound does not show λmax.
above 220nm , then it indicates the presence
of a carboxyl group, a cyano group, a hydroxyl
group or an amino group and absence of
aldehydic or ketonic group and benzene ring
etc.
• EXTENT OF CONJUGATION -
UV SPECTROSCOPY is extensively used in detecting
and estimating the extent of conjugation. For e.g.- 1,3
butadiene

absorbs at 217nm and it is observed that addition of a

conjugated double bond causes an increase in λmax. by
30 nm . Thus , greater the number of such conjugated
double bonds , longer will be the wavelength of
absorption. It is interesting to note that an organic
compound containing eight or more double bonds , the
absorption may occur even in the visible region.
• DISTINCTION BETWEEN CONJUGATED AND
NON-CONJUGATED COMPOUNDS
For e.g- CH2 C CH2 C CH3
CH3 O
(I)

(II)
NOTE- Because of conjugation , UV absorption band in
compound (ii) is at longer wavelength than that in
compound (i).
 • IDENTIFICATION OF
GEOMETRICAL ISOMERS
In general trans isomer absorbs at a longer wavelength with
larger extinction coefficient than the corresponding cis isomer.
For e.g-

Trans Cis
Λmax=290nm Λmax.=278nm
Єmax.=24,000 Єmax.=9350
REASON- The phenyl groups on either sides of cis isomer cause stearic
hindrance and prevent the effective overlap of π electrons of ring with
the π electrons of C=C double bond( by causing the two types of electrons
to be non-coplanar) during conjugation.
 • DETERMINATION OF
TAUTOMERISM EQUILIBRIUM
UV spectroscopy helps us in determining the
percentages of keto and enol forms in tautomeric
equilibrium.

CH3 C CH2COOC2H5 CH3 C CHCOOC2H5

OH
O (KETO FORM) (ENOL FORM)

ΛMAX.=275 nm Λmax.=244 nm
ЄMAX.=16 Єmax.=16000

From these values one can easily measure the

percentages of keto and enol forms present in the
sample.
 • STRUCTURE OF CHLORAL
The compound (shown) in hydrated form can exist in
two structural forms.

Cl O Cl
OH
Cl C C H.H2O Cl C C
OH
Cl (1) Cl H
THIS FORM(in
hexane solvent) (2)
THIS FORM( in
Aqueous solution)

The compound (1) shows UV absorption at 290 nm in

hexane solvent ( because of the presence of c=o bond in
it). Thus with the help of UV spectrum one can
determine the structure of chloral in different solvents.
END OF UV SPECTROSCOPY