Module -II

By
In spectroscopy, transitions between different energy levels
within atoms and molecules are recorded and then used to give
information on chemical structure.
UV Spectroscopy

UV Spectroscopy

• UV & electronic transitions

• Usable ranges & observations
• Selection rules
• Band Structure
• Instrumentation & Spectra
• Beer-Lambert Law
• Application of UV-spec
UV Spectroscopy

I. Introduction
A. UV radiation and Electronic Excitations
1. The difference in energy between molecular bonding, non-bonding and
anti-bonding orbital's ranges from 125-650 kJ/mole

2. This energy corresponds to EM radiation in the ultraviolet (UV) region,

100-400 nm, and visible (VIS) regions400-800 nm of the spectrum

3. For comparison, recall the EMR spectrum:

-rays X-rays UV IR Microwave Radio

4. Using IR we observed vibrational transitions with energies of 8-40

kJ/mol at wavelengths of 2500-15,000 nm

5. For purposes of our discussion, we will refer to UV and VIS

spectroscopy as UV
UV Spectroscopy
I. Introduction
B. The Spectroscopic Process
1. In UV spectroscopy, the sample is irradiated with the broad spectrum
of the UV radiation.

Where has the energy

that was within the
photons gone to ?

2. If a particular electronic transition matches the energy of a certain band of

UV, it will be absorbed.
3. The remaining UV light passes through the sample and is observed.
4. From this residual radiation a spectrum is obtained with “gaps” at these
discrete energies – this is called an absorption spectrum.
UV Spectroscopy

In a typical experiment, the molecules or atoms start at lower

energy and go to a higher energy level upon absorption of radiation
of appropriate wavelength.
After

Before E
Energy
Energy
UV Spectroscopy

Electronic Transitions





LUMO


HOMO
 UV Spectroscopy

I. Introduction
Selection Rules
1. Not all transitions that are possible are observed.

2. For an electron to transition, certain quantum mechanical constraints

apply – these are called “selection rules”.

3. For example, an electron can change its spin quantum number during
a transition – these are “forbidden”
Other examples include:
• the number of electrons that can be excited at one time
• symmetry properties of the molecule
• symmetry of the electronic states

4. To further complicate matters, “forbidden” transitions are sometimes

observed.
UV Spectroscopy

Band Structure

1. UV tends to give wide i.e. broad, overlapping bands; as the electronic

excitation is associated with vibrational and rotational excitation which
cannot be resolved.

2. It would seem that since the electronic energy levels of a pure sample of
molecules would be quantized, fine, discrete bands would be observed –
for atomic spectra, this is the case.
3. This effect will impact the wavelength at which a transition is observed –
very similar to the effect of H-bonding on the O-H vibrational energy
levels in neat samples.
UV Spectroscopy

Instrumentation and Spectra

1. The construction of a traditional UV-VIS spectrometer is very similar to an
IR, as similar functions – sample handling, irradiation, detection and output
are required

2. Here is a simple schematic that covers most modern UV spectrometers:

log(I0/I) = A
UV-VIS sources
I0 I

sample
200 , nm 700

detector
monochromator/
beam splitter optics I0 I0
reference

10
UV Spectroscopy

3. Two sources are required to scan the entire UV-VIS band:

• Deuterium lamp – covers the UV – 200-330
• Tungsten lamp – covers 330-700

4. As with the dispersive IR, the lamps illuminate the entire band of UV or visible
light; the monochromator (grating or prism) gradually changes the small bands of
radiation sent to the beam splitter .

5. The beam splitter sends a separate band to a cell containing the sample solution
and a reference solution.

6. The detector measures the difference between the transmitted light through the
sample (I) vs. the incident light (I0) and sends this information to the recorder.

7. As with dispersive IR, time is required to cover the entire UV-VIS

band due to the mechanism of changing wavelengths.
UV Spectroscopy

B. Instrumentation – Sample Handling

1. Virtually all UV spectra are recorded solution-phase.

2. Cells can be made of plastic, glass or quartz.

3. Only quartz is transparent in the full 200-700 nm range; plastic and glass are only
suitable for visible spectra.

4. Concentration is empirically determined.

A typical sample cell (commonly called a cuvet):

12
UV Spectroscopy

5. Solvents must be transparent in the region to be observed; the wavelength where

a solvent is no longer transparent is referred to as the cutoff.

6. Since spectra are only obtained up to 200 nm, solvents typically only need to
lack conjugated π systems or carbonyls .

Common solvents and cutoffs:

acetonitrile 190 nm
chloroform 240 nm
cyclohexane 195 nm
1,4-dioxane 215 nm
95% ethanol 205 nm
n-hexane 201 nm
methanol 205 nm
isooctane 195 nm
water 190 nm
UV Spectroscopy

7. Additionally solvents must preserve the fine structure (where it is actually

observed in UV)
UV Spectroscopy

The Spectrum

1. The x-axis of the spectrum is in wavelength; 200-400 nm for UV, 200-800 nm for
UV-VIS determination.

2. Due to the lack of any fine structure, spectra are rarely shown in their raw form,
rather, the peak maxima are simply reported as a numerical list of “ λ max”
values.

NH2

λ max = 206 nm
O O 252 nm
317 nm
376 nm
UV Spectroscopy

A . Introduction
Observed electronic transitions

1. The lowest energy transition (and most often obs. by UV) is typically
that of an electron in the Highest Occupied Molecular Orbital (HOMO)
to the Lowest Unoccupied Molecular Orbital (LUMO).

2. There are three kinds of electrons present in molecules as

a) ϭ-electrons – present in ϭ-bonds.
b) π-electrons- present in π-bonds.
c) “n”-nonbonding electrons- present as unshared electrons.

3. Sigma and pi electrons have corresponding higher energy antibonding

molecular orbitals (σ* π*), but there is no higher energy antibonding
molecular orbital for non bonding electrons.
UV Spectroscopy

4. From the molecular orbital diagram, there are several possible electronic
transitions that can occur, each of a different relative energy:


  alkanes

  carbonyls

  unsaturated cmpds.
Energy
n
n  O, N, S, halogens

n  carbonyls



UV Spectroscopy

5. Although the UV spectrum extends below 100 nm (high energy),

oxygen in the atmosphere is not transparent below 200 nm.

6. Special equipment to study vacuum or far UV is required.

7. Routine organic UV spectra are typically collected from 200-800 nm.

8. This limits the transitions that can be observed:

  alkanes 150 nm

  carbonyls 170 nm

  unsaturated cmpds. 180 nm √ - if conjugated!

n  O, N, S, halogens 190 nm

 carbonyls 300 nm √
n
UV Spectroscopy

.
B Instrumentation and Spectra

The Spectrum

1. The y-axis of the spectrum is absorbance, A.

2. From the spectrometers point of view, absorbance is the inverse of transmittance:

A = log10 (I0/I)

3. From an experimental point of view, three other considerations must be made:

i. a longer path length, l through the sample will cause more UV light
to be absorbed – linear effect
ii. the greater the concentration, c of the sample, the more UV light
will be absorbed – linear effect
iii. some electronic transitions are more effective at the absorption
of photon than others – molar absorptivity, e this may vary by
orders of magnitude
UV Spectroscopy

4. These effects are combined into the

Beer-Lambert Law: A = ε C l

i. For most UV spectrometers, l would remain constant (standard cells are

typically 1 cm in path length).

ii. Concentration is typically varied depending on the strength of absorption

observed or expected – typically dilute – sub .001 M.

iii. molar absorptivities vary by orders of magnitude:

• values of 104-106 are termed high intensity absorptions
• values of 103-104 are termed low intensity absorptions
• values of 0 to 103 are the absorptions of forbidden transitions
A is unit less, so the units for e are cm-1 · M-1 and are rarely expressed
UV Spectroscopy

C. Practical application of UV spectroscopy

1. UV was the first organic spectral method, however, it is rarely used as a primary
method for structure determination.

2. It is most useful in combination with NMR and IR data to elucidate unique

electronic features.
3. Determination of sterochemistry (cis and trans): A trans isomer usually has a
greater λ max and ԑ max value than the corresponding cis isomer. Thus UV
spectra is useful to distinguish between cis and trans isomer.
4. The strength of Hydrogen bond can be calculated by running the UV spectrum in
different solvents like polar and non-polar.
UV Spectroscopy

Chromophores

Definition:
1. Remember the electrons present in organic molecules are involved in covalent
bonds or lone pairs of electrons on atoms such as O or N.

2. Since similar functional groups will have electrons capable of discrete classes
of transitions, the characteristic energy of these energies is more representative
of the functional group than the electrons.

3. A functional group capable of having characteristic electronic

transitions is called a chromophore (color loving).

4. Structural or electronic changes in the chromophore can be quantified and used

to predict shifts in the observed electronic transitions.
5. Chromophore are compounds containing unsaturation i.e. Alkenes, Alkynes,
Azo compounds, Nitriles, Nitro compounds etc.
UV Spectroscopy

Organic Chromophores

1. Alkanes – only posses s-bonds and no lone pairs of electrons, so only the
high energy σ → σ * transition is observed in the far UV.

This transition is destructive to the molecule, causing cleavage of the σ –

bond.

 C C

C C

UV Spectroscopy

2. Alcohols, ethers, amines and sulfur compounds –

in the cases of simple, aliphatic examples of these compounds the n →
σ * is the most often observed transition; like the alkane σ → σ * it
is most often at shorter wavelength than 200 nm.

Note how this transition occurs from the HOMO to the LUMO

C N
CN

C N

nN sp 3 C N anitbonding
orbital

CN C N
UV Spectroscopy

3. Alkenes and Alkynes – in the case of isolated examples of these

compounds the π → π * is observed at 175 and 170 nm, respectively.

Even though this transition is of lower energy than σ → σ *, it is still in

the far UV – however, the transition energy is sensitive to substitution.




UV Spectroscopy

4.Carbonyls – unsaturated systems incorporating N or O can undergo

n → π* transitions (~285 nm) in addition to π → π * Despite the fact this
transition is forbidden by the selection rules (e = 15), it is the most often observed
and studied transition for carbonyls.

This transition is also sensitive to substituents on the carbonyl

Similar to alkenes and alkynes, non-substituted carbonyls undergo the π → π *

transition in the vacuum UV (190 nm, e = 900); sensitive to substitution effects.
UV Spectroscopy

4. Carbonyls – n → π * transitions (~285 nm); π → π * (188 nm)


It has been determined
from spectral studies,
n that carbonyl oxygen
C O

more approximates sp
rather than sp2
 O

CO transitions omitted for clarity

UV Spectroscopy

General – from our brief study of these general chromophores, only the weak n →π*
transition occurs in the routinely observed UV region.

The attachment of substituent groups (other than H) can shift the energy
of the transition.

Substituent's that increase the intensity and often wavelength of

an absorption are called Auxochrome.

Common auxochromes include alkyl, hydroxyl, alkoxy and amino groups

and the halogens
UV Spectroscopy

General – Substituents may have any of four effects on a chromophore

i. Bathochromic shift (red shift) – a shift to longer wavelength.
ii. Hypsochromic shift (blue shift) – shift to shorter wavelength.
iii. Hyperchromic effect – an increase in intensity.
iv. Hypochromic effect – a decrease in intensity.

Hyperchromic


Hypsochromic Bathochromic
Hypochromic

200 nm 800 nm
UV Spectroscopy

Substituent Effects
1. Conjugation – most efficient means of bringing about a bathochromic and
hyperchromic shift of an unsaturated chromophore:
max nm 
175 15,000

217 21,000

258 35,000

465 125,000

-carotene

O
n → * 280 12
 → * 189 900
n → * 280 27
O
 → * 213 7,100
UV Spectroscopy

Conjugation – Alkenes
The observed shifts from conjugation imply that an increase in conjugation
decreases the energy required for electronic excitation.
From molecular orbital (MO) theory two atomic p orbitals, f1 and f2 from
two sp2 hybrid carbons combine to form two MOs Y1 and Y2* in
ethylene



 

 
UV Spectroscopy

Conjugation – Alkenes
When we consider butadiene, we are now mixing 4 p orbitals giving 4 MOs
of an energetically symmetrical distribution compared to ethylene


 





 


E for the HOMO → LUMO transition is reduced

UV Spectroscopy

Chromophores
Conjugation – Alkenes
Extending this effect out to longer conjugated systems the energy gap becomes
progressively smaller:

Energy Lower energy =

Longer wavelengths

ethylene
butadiene
hexatriene
octatetraene
UV Spectroscopy

Conjugation – Alkenes
Methyl groups also cause a bathochromic shift, even though they are
devoid of π- or n-electrons.
This effect is thought to be through what is termed
“Hyperconjugation” or sigma bond resonance.

C
H
C C H
UV Spectroscopy

The effect of substituent groups can be reliably quantified

from empirical observation of known conjugated structures and applied to new
systems.
This quantification is referred to as the Woodward-Fieser Rules which
we will apply to two specific chromophores:
1. Conjugated dienes
2. Conjugated dienones

max = 239 nm
UV Spectroscopy

Structure Determination
Dienes
Woodward-Fieser Rules
Woodward and the Fiesers performed extensive studies of terpene and
steroidal alkenes and noted similar substituents and structural features
would predictably lead to an empirical prediction of the wavelength for
the lowest energy π → π * electronic transition.

The general approach for calculating λ max value is to begin with an

absorption wavelength for the parent chromophore and add a value of
each substituent attached to the conjugated system.
UV Spectroscopy

Woodward-Fieser Rules - Dienes

The rules begin with a base value for λ max of the chromophore being
observed:

acyclic butadiene = 217 nm

The incremental contribution of substituents is added to this base value

from the group tables:
Group Increment (nm)
Extended conjugation +30
Homoannular component +39
Each exo-cyclic C=C +5
Alkyl +5
-OCOCH3 +0
-OR +6
-SR +30
-Cl, -Br +5
-NR2 +60
UV Spectroscopy

Woodward-Fieser Rules – Dienes

For example:

Isoprene - acyclic butadiene = 217 nm

one alkyl subs. + 5 nm
222 nm
Experimental value 220 nm

Allylidenecyclohexane
- acyclic butadiene = 217 nm
one exocyclic C=C + 5 nm
2 alkyl subs. +10 nm
232 nm
Experimental value 237 nm
UV Spectroscopy

Woodward-Fieser Rules – Cyclic Dienes

There are two major types of cyclic dienes, with two different base values:
Heteroannular diene is the cyclic diene in which the two double bonds in
conjugation are present in two different rings.
Homoannular diene is the cyclic diene in which both the conjugated double
bonds are present in same ring .
Heteroannular (transoid): Homoannular (cisoid):

e = 5,000 – 15,000 e = 12,000-28,000

base λ max = 215 nm base λ max = 253 nm

The increment table is the same as for acyclic butadienes with a couple additions:
Group Increment (nm)
Additional homoannular +39
Where both types of
diene are present, the
one with the longer max
becomes the base
UV Spectroscopy

Woodward-Fieser Rules – Cyclic Dienes

For example:
1,2,3,7,8,8a-hexahydro-8a-methylnaphthalene
heteroannular diene = 215 nm

3 alkyl subs. (3 x 5) +15 nm

1 exo C=C + 5 nm
235 nm

Experimental value 235 nm

UV Spectroscopy

Woodward-Fieser Rules – Cyclic Dienes

heteroannular diene = 215 nm

4 alkyl subs. (4 x 5) +20 nm
1 exo C=C + 5 nm
240 nm
C OH
O

homoannular diene = 253 nm

4 alkyl subs. (4 x 5) +20 nm
1 exo C=C + 5 nm
278 nm
C OH
O
UV Spectroscopy

Woodward-Fieser Rules – Cyclic Dienes

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 homooannular diene; you

would use the base value for an acyclic diene.
UV Spectroscopy

Enones
1. General Features

Carbonyls, as we have discussed have two primary electronic transitions:

 Remember, the π → π * transition is

allowed and gives a high intensity, but lies
outside the routine range of UV observation.
n
The n → π * transition is forbidden and
gives a very low intensity, but can routinely
be observed.

UV Spectroscopy

IV. Structure Determination

B. Enones
1. General Features
For auxochromic substitution on the carbonyl, pronounced hypsochromic shifts
are observed for the n → π * transition (λmax):
O

H
293 nm This is explained by the inductive withdrawal of
electrons by O, N or halogen from the carbonyl
O carbon – this causes the n-electrons on the
CH3 279 nm carbonyl oxygen to be held more firmly

O
It is important to note this is different from the
235 nm
Cl auxochromic effect on π → π * which extends
conjugation and causes a bathochromic shift
O

NH2
214 nm
In most cases, this bathochromic shift is not
O
enough to bring the π → π * transition into the
204nm observed range
O

O
204 nm
OH
44
UV Spectroscopy

IV. Structure Determination

B. Enones
1. General Features
Conversely, if the C=O system is conjugated both the n → π * and π → π *
bands are bathochromically shifted.

Here, several effects must be noted:

i. the effect is more pronounced for π → π *

ii. if the conjugated chain is long enough, the much higher intensity
π → π * band will overlap and drawn out the n → π * band.

i. the shift of the n → π * transition is not as predictable.

For these reasons, empirical Woodward-Fieser rules for conjugated enones are
for the higher intensity, allowed π → π * transition.

45
UV Spectroscopy

IV. Structure Determination

B. Enones
1. General Features
These effects are apparent from the MO diagram for a conjugated enone:



 


n n



 

O O

46
UV Spectroscopy

IV. Structure Determination

     
B. Enones
 C C C  C C C C C
2. Woodward-Fieser Rules - Enones O O

Group Increment (nm)

6-membered ring or acyclic enone Base 215 nm
5-membered ring parent enone Base 202 nm
Aldehyde Base 207 nm

Double bond extending conjugation 30

Alkyl group or ring residue α, β,γ, and higher 10, 12, 18
-OH α, β,γ, and higher 35, 30, 18
-OR α, β,γ,δ 35, 30, 17, 31
-O(C=O)R α, β,γ, 6
-Cl α, β 15, 12
-Br α, β 25, 30
-NR2 β 95
Exocyclic double bond 5
Homocyclic diene component 39

47
UV Spectroscopy

IV. Structure Determination

B. Enones
2. Woodward-Fieser Rules - Enones
Unlike conjugated alkenes, solvent does have an effect on λmax
These effects are also described by the Woodward-Fieser rules.

Solvent correction Increment (nm)

Water +8
Ethanol, methanol 0
Chloroform -1
Dioxane -5
Ether -7
Hydrocarbon -11

48
UV Spectroscopy

IV. Structure Determination

B. Enones
2. Woodward-Fieser Rules - Enones
Some examples – keep in mind these are more complex than dienes
cyclic enone = 215 nm
O 2 x β- alkyl subs. (2 x 12) +24 nm
239 nm

Experimental value 238 nm

R
cyclic enone = 215 nm
extended conj. +30 nm
β-ring residue +12 nm
δ-ring residue +18 nm
O exocyclic double bond + 5 nm
280 nm

Experimental 280 nm 49
Thank you…..