Ultraviolet-Visible Spectroscopy

# What do you mean by Ultraviolet-Visible Spectroscopy ?

# What is electromagnetic spectrum?

# What is absorption Spectroscopy ?

Electromagnetic Spectrum
Violet:   400 - 420 nm

Indigo:   420 - 440 nm

Blue:   440 - 490 nm c

Green:   490 - 570 nm

Yellow:   570 - 585 nm

Orange:   585 - 620 nm

Red:   620 - 780 nm

What happens when a beam of electromagnetic
radiation is passed through a substance?

When a beam of electromagnetic radiation is passed

through a transparent medium of a substance, the
radiation can be either absorbed or transmitted
depending upon its wavelength and the structure of the
substance. The radiation not absorbed i.e. the transmitted
light can be measured by means of a suitable instrument,
known as spectrometer and the record is known as the
spectrum of the substance.
 UV-visible
spectrophotometer

UV-visible spectra
Electromagnetic radiation may be described by its
wavelength(λ) or by its frequency ()

Define Wavelength

Define frequency
• The frequency and wavelength are
related by the expression

λ
Or cλ Where, c is the velocity of light
= 3 x 1010 cm/sec
What happens when a molecule absorb energy?

Electromagnetic radiation is energy.

Therefore, when a substance absorb radiation it gains energy.

The higher the frequency the greater is the gain in energy.

The absorbed energy brings about changes within the

molecule.
# The absorbed energy may break bonds within the
molecule

# It may raise electrons to the higher energy level

# It may bring about increased vibration or rotation of the

atoms

# It may change nuclear or electronic spin.

The UV and visible light which ranges from 10 nm to 780

nm causes electronic transitions within the molecules.

The commonly used UV-visible spectrometers can

measure absorption of light from 190 to 780 nm.
• Vacuum ultraviolet region
• The UV light below 200nm is relatively uninformative and difficult to
measure. Since oxygen in the air also absorb UV light in this region.

• Therefore, if we want to measure absorption of UV light below 200 nm,

air must be removed from the instrument. The region is known as
Vacuum ultraviolet and it ranges from 120 nm to 200 nm. This region
excites the σ-bond electrons.

• The UV light above 200 nm, causes excitation of electrons from π-

orbitals, particularly π-conjugated system. Absorption in this region can
be readily measured and it gives informative spectra.
• Laws of spectrometry
• If a substance is placed in the path of light , a number of events
may takes place.

• Absorption of light by a sample

There are two laws which govern the absorption of light by the
molecules.

These are Lambert’s law and Beer’s law.

Lambert’s law : It states that the fraction of the incident light absorbed
is independent of the intensity of the light source.

According to this law each layer of equal thickness absorb an equal

fraction of radiant light which passes through it.

Beer’s law : It states that the absorption is proportional to the number

of absorbing molecules.
Equation of absorbance:
Combining Beer’s law and Lambert’s Law the equation of absorbance is
obtained.

log10 I0 / I = A = a. b. c

Where , I0 is the intensity of the incident light

I is the intensity of the transmitted light

a is the proportionality constant and called absorptivity.

A is the absorbance

b is the path length of the sample or absorbing solution

in cm

c is the concentration of the absorbing solute.

When c is expressed in moles / litre,

the absorptivity is called

molar absorptivity or molar extinction coefficient

and is written by ε.

When c is expressed as percent w/v,

the absorptivity is called A1%1cm.

Types of electronic transitions.
According to molecular orbital theory when a molecule is excited by the

absorption of UV or visible light its electrons are changed from one

orbital to another orbital of higher energy. The electrons can be σ, π

or an n electrons.

The various transitions involved in ultraviolet-visible spectroscopy are

i) σ  σ*

ii) n  σ*

iii) π  π*

iv) n  π*
i) σ  σ* transition
This type of transition takes place in saturated centres of the molecules.

Here an electron from a stable σ orbital goes to an unstable or anti

bonding σ orbital.

A σ electron is held tightly and a high amount of energy is required to

excite it. Hence, the saturated compounds do not show absorption in

the normal ultraviolet and visible region i.e. 190nm to 780nm.

Therefore to study σ  σ* transition, air must be removed from the

instrument. This region is less informative.

.
C C
 
C .C
 ii) n σ* transition
This type of transition takes place in saturated compounds containing a

heteroatom with unshared pair of electrons (n electrons).

Here an electron from an unshared pair goes to an unstable or anti

bonding σ orbital.

n σ* transition takes place in halogen, oxygen, nitrogen and sulfur

compounds.

These are also high energy transitions and most of the absorbtion

occur at shorter wavelength and can not be measured by the usual

UV-visible spectrophotometers.
n σ* transition

.. n  *
.
C X* * C X **
* *
* *

.. n  . .
O O

.. n  . .
N N

.. n  . .
S S
 iii) π  π* transition
This type of transition takes place in unsaturated compounds containing

double or triple bonds.

Here an electron from stable or π orbital goes to an unstable or antibonding

π orbital.

Alkenes, alkynes, carbonyl compounds, azo compounds etc. show π  π*

transition.

..   *
*
..
C O ** C O *
*
Excitation of π electrons requires less energy.

Conjugation of double bonds further lowers the energy required for

transition and absorption moves to longer wavelength where it can

be more conveniently measured.

If there is enough double bonds in conjugation absorption will move to

visible region and the compound will be colored.

β-carotene, for example, contain 11 c-c double bonds in conjugation.

It is a colored compound and absorb at 451 nm.

iv) n  π* transition
This type of transition takes place in unsaturated compounds containing a
heteroatom with unshared pair of electrons (n electrons).

Here an electron from an unshared pair goes to an unstable π orbital.

... n  . ..
C O *
C O *

n  π* transition takes place in aldehydes, carbonyl compds, cyanides,

etc.

These are low energy transitions and most of the absorption occur at longer
wavelength where they can be measured by the UV-visible
spectrophotometers.
• Choice of solvents:
Solvents Minimum wavelength for I cm
cell, nm
Acetonitrile 190
Water 191
Hexane 201
Methanol 203
Ethanol 204
Ether 215
Chloroform 237
Carbon tetrachloride 257
Benzene 280

• Commercial absolute ethanol should not be used as it

contain benzene which absorb strongly in the UV region.
• Effect of solvents on different transitions

i) n  π*


... n  . ..
C O* C O x More energy
*

y Shorter
More polar less polar n wavelength

Dipole- dipole interactions with solvent molecules will lower the

energy of the ground state more than that of the excited state.
ii) π  π*


..   * ..
O **
*
Less energy
C C O ** x
less polar y Longer
More polar wavelength

iii) n  σ*


.. n  . . More energy
C O C O x
Shorter
y
wavelength
More polar less polar n
UV terminology:
• Chromophore

• Auxochrome

• OH, OR, NH2, NHR, NR2 SH etc.

CH CH λmax below 190 nm

CH CH NR2
λmax 230 nm
Enamine
• Bathochromic shift (Red shift)

• Hypsochromic shift (Blue shift)

Hexane
aniline max 230 nm
A

aq. acid
max 203 nm

• Hyperchromic effect


• Hypochromic effect


Instrumentation:
Spectrometer / spectrophotometer
Colorimeter
The ultraviolet visible spectrometer consists of the following
parts
i) Radiation source
ii) Monochromator
iii)Sample compartment
iv)Detector
v) Recorder
Radiation source
Hydrogen (or deuterium) discharge lamp emits radiant
energy from 185 to 365 nm

A 6 or 12 v tungsten lamp emits radiation from 350 to

800nm.

Monochromator
• The monochromator disperse the polychromatic light by
means of a prism or grating.
Sample compartment
Optical windows of cuvettes are made
of quartz or glass

Detector
Radiant light electric energy

Photomultiplier tube: a cathode, an

anode and dynodes
Anode
III
Dynode I

light

II Cathode

Photomultiplier tube
Recorder
Double beam spectrophotometer
The necessity for making two separate measurement has
been eliminated with double beam spectrophotometer.

With this instrument the monochromatic ray is split

into two identical beams; one beam is passed
through the reference cell and the other is passed
through the sample cell in order to cancel the
solvent absorption.
Electronic transitions in polyenes
C C λ max below 190 nm
an isolated double bond
excited state

C C C C

two double bonds in conjugation

ground state

Ethylene absorbs at 175 nm

1,3-butadiene absorbs at 217 nm
 Energy

Ethylene Butadiene Hexatriene Octetraene

Fig. Energy gap in polyene

The longer the conjugated system the longer the

wavelength of the absorption maximum. β-Carotrne
contain eleven C-C double bonds in conjugation and
shows λ max at 451 nm
 C C C C

Unoccupied
C C

C C C C LUMO

 

C C C C HOMO

C C

C C C C

From 2p orbitals From 4p orbitals

2 molecular orbitals 4 molecular orbitals

As the number of p orbitals increased the transition from the highest

occupied molecular orbitals (HOMO) to the lowest unoccupied molecular
orbitals (LUMO) need progressively lower energy i.e. energy gap
becomes progressively smaller with increasing conjugation.
• The two atomic orbitals can overlap in two ways depending on their phase relationship. The phase of an orbital is a direct consequence of
the wave-like properties of electrons. In graphical representations of orbitals, orbital phase is depicted either by a plus or minus sign
(confusing because there is no relationship to electrical charge) or simply by shading one lobe. The sign of the phase itself does not have
physical meaning except when mixing orbitals to form molecular orbitals.
• Then two same-sign orbitals have a constructive overlap forming a molecular orbital with the bulk of electron density located between the
two nuclei. This MO is called the bonding orbital and its energy is lower than that of the original atomic orbitals. A bond involving molecular
orbitals which are symmetrical with respect to rotation around the bond axis (no change) and is called a sigma bond (σ-bond). In the event of
a phase change, the bond becomes a pi bond (π-bond). Symmetry labels are further defined by whether the orbital maintains its original
character after an inversion about its center; if the orbital does retain its original character it is defined gerade, g, or if the orbital does not
maintain its original character, ungerade, u.
• Atomic orbitals can also interact with each other out-of-phase which leads to destructive cancellation and no electron density between the
two nuclei at the so-called nodal plane depicted as a perpendicular dashed line. In this anti-bonding MO with energy much higher than the
original AO's, any electrons present are located in lobes pointing away from the central internuclear axis. For a corresponding σ-bonding
orbital, such an orbital would be symmetrical but differentiated from it by an asterisk as in σ*. For a π-bond, corresponding bonding and
antibonding orbitals would not have such symmetry around the bond axis and be designated π and π*, respectively.
• The next step in constructing an MO diagram is filling the newly formed molecular orbitals with electrons. Three general rules apply:
• The Aufbau principle states that orbitals are filled starting with the lowest energy
• The Pauli exclusion principle states that the maximum number of electrons occupying an orbital is two having opposite spins
• Hund's rule states that when there are several MO's with equal energy, the electrons fill into the MO's one at a time before filling two
electrons into any.
• The filled MO highest in energy is called the Highest Occupied Molecular Orbital or HOMO and the empty MO just above it is then the
Lowest Unoccupied Molecular Orbital or LUMO. The electrons in the bonding MO's are called bonding electrons and any electrons in the
antibonding orbital would be called antibonding electrons. The reduction in energy of these electrons is the driving force for chemical bond
formation. Whenever mixing for an atomic orbital is not possible for reasons of symmetry or energy, a so-called non-bonding MO is created,
which is often quite similar to and has energy level equal or close to its constituent AO, thus not contributing to bonding energetics. The
resulting electron configuration can be described in terms of bond type, parity and occupancy for example dihydrogen 1σg2. Alternatively it
can be written as a molecular term symbol e.g. 1Σg+ for dihydrogen. Sometimes, the letter n is used to designate a non-bonding orbital.
• WiKI