Spectroscopy

It is the branch of science that deals with the study of interaction of

electromagnetic radiation with matter.

ELECTROMAGNETIC RADIATION
Electromagnetic radiation consists of discrete packet of energy called as
photons.

A photon consists of an oscillating electric field (E) & an oscillating magnetic

field (M)

which are perpendicular to each other.

i.e.; E = hv

where E =Energy of photon

h= plank's constant

and V = frequency.
ELECTROMAGNETIC SPECTRUM

APPLICATION OF SPECTROSCOPY
(1) Used in structural investigation.

(2) Determine electronic energy level, bond angle and strength of bonds.

(3) Used to monitor changing concentration of reactants or products in

kinetic studies.

ADVANTAGES OF SPECTROSCOPY
(1) Requires less time and less amount of sample.

(2) Results obtained are reliable and more accurate.

(3) Sample recovery is possible after spectroscopic analysis because

Sample is not chemically affected.

Energy Level Diagram

Electronic Energy Levels

At room temperature the molecules are in the lowest energy levels Eo.

When the molecules absorb UV-visible light from EMR, one of the outermost

bond/lone pair electron is promoted to higher energy state such as E1,

E2,…...,En etc. Is called as electronic transition and the difference is as:

* Vibrational Energy Levels

These are less energy level than electronic energy levels.

The spacing between energy levels are relatively small i.e., 0.01 to 10
kcal/mole.

E.g., when IR radiation is absorbed, molecules are excited from one

vibrational level to another or it vibrates with higher amplitude.

Rotational Energy Levels

These energy levels are quantized & discrete.

The spacing between energy levels is even smaller than vibrational energy
levels.

ΔE = ΔErotational + ΔEvibrational + ΔEeletronic + ΔEtranslational

ΔEtranslational < ΔErotational < ΔEvibrational < ΔEeletronic

ABSORPTION LAW

Beer-Lambert’s Law

When a monochromatic radiation is passed through a solution, the

absorbance done

is directly proportional to product of concentration of the solution and the

path length of the solution.

OR

The intensity of the emitted light decreases exponentially as the thickness

and concentration of the absorbing medium increases.

UV-VISIBLE SPECTROSCOPY
> In U.V. spectroscopy when a molecule absorbs EMR of UV range its
electron undergoes transition.

> When a sample absorbs UV light, valance electrons are promoted to

higher energy levels in accordance with the frequency of absorbed light.
> The UV spectrum of a compound gives information about its electronic

structure. In other words, a sample with pi electrons, with pi bonds, with lone

pairs, with conjugation, will show characteristic absorption of UV light.

When a molecule is subjected to UV range, a spectrum is obtained in

spectrophotometer. This spectrum consists of bands which are the plots

between absorbed light and wavelength.

> Through the study of these bands we can elucidate the molecular
structure.

UV region is divided into

a) Near UV region (wavelength region nearer to the visible region, 𝜆 ~ 250

nm-400nm)

b) Far UV region (wavelength region farther to the visible region, 𝜆 ~ 190 nm-
250 nm)

c) Vacuum UV region (𝜆 < 190nm)

Electronic Transitions
The absorption of UV and visible light is through the transition of an electron
in the molecule from lower to a higher energy molecular orbital. The various
electronic transitions observed in organic compound.

The different types of electrons found in molecules are: sigma electrons, pi

electrons and n electrons.

𝛿 Electrons: These are found in saturated compounds like alkanes. They are
highly stable are very difficult to move.

π electrons: These are found in unsaturated compounds like alkenes and

alkynes. Less stable than sigma electrons as they have more energy in
comparison to them.
n electrons: These are non-bonding electrons which do not participate in
bonding. They are found as lone pairs. They have highest amount of energy.

1. 𝜎 — 𝜎 * transition

• 𝜎 electron from 𝜎 bonding orbital is excited to corresponding anti-

bonding orbital 𝜎 *.
• The energy required is large for this transition. These transitions can be
found in alkanes like- Methane (CH4) has C-H bond only and can
undergo 𝜎 — 𝜎 * transition and shows absorbance maxima at 125 nm.

2. 𝜋 — 𝜋* transition

• 𝜋electron in a bonding orbital is excited to corresponding anti-bonding

orbital 𝜋*.
• Compounds containing multiple bonds like alkenes, alkynes, carbonyl,
nitriles, aromatic compounds, etc. undergo 𝜋 — 𝜋* transitions. e.g.,
Alkenes generally absorb in the region 170 to 205 nm.

3. n — 𝜎 * transition

• Saturated compounds containing atoms with lone pair of electrons like

O, N, S and halogens are capable of n ~ 𝜎 * transition.
• These transitions usually require less energy than 𝜎 — 𝜎 * transitions.

4. n — 𝜋* transition
 • An electron from non-bonding orbital is promoted to anti-bonding 𝜋*
orbital.
• Compounds containing double bond involving hetero atoms (C=O,
C=N, N=O) undergo such transitions.
• n — 𝜋* transitions require minimum energy and show absorption at
longer wavelength around 300 nm.

TERMS USED IN UV/ VISIBLE SPECTROSCOPY

CHROMOPHORE
The part of a molecule that is responsible for absorption of EMR in UV regions
known as chromophore. Chromophores are responsible to produce colour to
the compound.

These are the functional groups containing multiple bonds capable of

absorbing radiations above 200 nm due to n --> 𝜋* & 𝜋 — 𝜋* transitions. e.g.,
NO, N=O, C=O, C=N, C=C, C=S, etc.

AUXOCHROME
The functional group with non-bonding electrons that does not absorb
radiation in near UV region but when attached to a chromophore alters the
wavelength & intensity of absorption.

e.g., Benzene 𝜆 max = 255 nm

Phenol 𝜆 max = 270 nm

Aniline 𝜆 max = 280 nm

ABSORPTION AND INTENSITY SHIFTS

1. Bathochromic Shift (Red Shift)
• When absorption maxima (Amax) of a compound shift to longer
wavelength, it is known as bathochromic shift or red shift.
• The effect is due to presence of an auxochrome or by the change of
solvent. E.g. An auxochrome group like -OH, -OCH; causes absorption
of compound at longer wavelength.
• In alkaline medium, p-nitrophenol shows red shift. Because negatively
charged oxygen delocalizes more effectively than the unshared pair of
electrons.

2. Hypsochromic Shift (Blue Shift)

 • When absorption maxima (𝜆 max) of a compound shift to shorter
wavelength, it is known as hypsochromic shift or blue shift. The effect is
due to removal of conjugation or auxochrome or by the change of
solvent.
• Aniline shows blue shift in acidic medium; it loses conjugation.

3. Hyperchromic Effect
• When absorption intensity (𝜖 ) of a compound Is increased, it 1s known
as hyperchromic shift.
• If auxochrome is introduced to the compound, the intensity of
absorption increases.

4. Hypochromic Effect
• When absorption intensity (𝜖 ) of a compound Is decreased, it is known
as hypochromic shift.
• It occurs when the symmetry of the compound is disturbed.
Factors causing shifts in UV-VIS spectroscopy
1. Conjugation
a) Conjugation brings about a bathochromic shift (shift towards higher
wavelength) in the absorption bands.

b) The higher the extent of conjugation. the more is the bathochromic shift.

c) Increase in Conjugation decreases the energy differences between the

highest occupied molecular orbital (HOMO) and the lowest unoccupied
molecular orbital (LUMO. Therefore, the electronic transition becomes
possible even at lower energy (higher wavelength).

2. Auxochrome
• Auxochromes are the chemical groups that result in a bathochromic
shift when attached to a chromophore.
• They bring about bathochromic shift in absorption by extending
conjugation through resonance.

3. Solvents
The solvents used in any spectroscopic method should be non-absorbing to
the electromagnetic radiation being used. Water and many organic solvents
such as ethanol, acetonitrile, cyclohexane, dimethylformamide (DMF),
dimethyl sulfoxide (DMSO) etc.

APPLICATIONS OF UV / VISIBLE SPECTROSCOPY

• Qualitative & Quantitative Analysis.
• It is used for characterizing aromatic compound.
• Detection of impurities: It is one of the important methods to detect
impurities in organic solvents.
 • Detection of geometrical isomers is possible: Trans isomers exhibit 𝜆 max,
at slightly longer wavelength than the cis-isomer.
• Determination of molecular weight using Beer’s law.

INFRA RED SPECTROSCOPY

IR spectroscopy is concerned with the study of absorption of infrared
radiation, which causes vibrational transition in the molecule. Hence, IR
spectroscopy also known as Vibrational spectroscopy.

IR spectra gives valuable information about

a) Functional Groups.

b) Bonds present in a compound.

IR SPECTROSCOPY PRINCIPLE
At ordinary temperature the molecules are generally in their lowest
vibrational energy levels and are vibrating with its natural frequency. When
sufficient amount of energy in the form of infrared radiations (equal to
difference between two vibrational levels) is supplied to the molecule and
when the natural frequency of molecule becomes equal to the radiant
frequency (IR radiations) then the absorption takes place and vibrational
transition occurs.

Criteria for a compound to absorb IR radiation

The two essential criteria for a molecule to be IR active are:

1. Correct wavelength of radiation.

2. Change in dipole moment.

1. Correct wavelength of radiation.

For a molecule to absorb IR radiation, the natural frequency of vibrations of

some part of a molecule is the same as the frequency of incident radiation.

2. Change in dipole moment.

A molecule can only absorb IR radiation when its absorption causes a

change in its electric dipole.
A molecule is said to have an electric dipole when there is a slight positive
and a slight negative charge on its component atoms.

Example:

> All the homo-nuclear diatomic molecules such as H, N, O, etc. are IR

inactive.

> Vibration of a bond involving two atoms that have large electro-negativity
difference is usually IR active.

> Vibration of carbon monoxide (C = O) causes a change in dipole moment

and is therefore IR active

MOLECULAR VIBRATIONS
Due to increase in vibrational and rotational energies of the molecule, two
types of fundamental vibrations are observed.

1. Stretching vibration

2. Bending vibrations

1. Stretching vibration
These are the vibration or oscillation along the line of bond. In stretching
vibrations there occurs a change in bond length.

They occur at higher energy: 4000-1250 cm”.

It is of 2 types:

a) Symmetrical stretching

b) Asymmetrical stretching

a) Symmetrical stretching:

The atoms of a molecule either move away or towards the central atom but
in same direction.

b) Asymmetrical stretching:
One atom approach toward the central atom while other departs from it.

2. Bending vibrations
Vibration or oscillation are not along the line of bond.

• These are also called as deformations.

• In this, bond angle/bond axis is altered.
• Occurs at low energy: 1400-666 cm-1

These are of 2 types:

a) In plane bending

• Scissoring
• Rocking

b) Out plane bending

• Wagging
• Twisting

Scissoring: This is in plane blending in which two atoms approach each other
and bond angles decrease.

Rocking: Movement of atoms take place in the same direction.

Wagging: Two atoms move to one side of the plane. They move up and
down the plane.
Twisting: One atom moves above the plane and another atom moves below
the plane.

Number of fundamental vibrations

For a linear molecule = 3n -5

For a non-linear molecule = 3n -6

Where n is the number of atoms in a molecule.

e.g., CO, (linear molecule); n = 3

No. of fundamental vibrations = 3n -5 = (3 x 3)-5=4

H2O (non-linear molecule); n=3

So, no. of fundamental vibrations will be: 3n -6 = (3 x 3) - 6 =3

FUNCTIONAL GROUP REGION (4000-1500 cm-1)

Region from 4000-1500 cm-1 in an IR spectrum is useful for identification of
functional groups. This region shows the absorption due to stretching mode.

FINGER PRINT REGION

The region below 1500 cm-1 (1500-600 cm-1) is known as finger print region. It
contains number of vibrations caused by bending and stretching vibrations.

The molecules containing same functional groups show similar absorption

above 1500 cm-1, but if the module is different, then the spectra differ in
finger print region. On the other hand, if along with high frequency region
(above 1500 cm-1), the finger print region also matches, the two molecules
are of same compound. This region thus provides confirmation of identity of
molecules.

APPLICATION OF IR SPECTROSCOPY
1. Qualitative Analysis

Main application of IR spectroscopy is compound identification.

2. Quantitative Analysis:

From the equation A = ECL we can determine the concentration of a

sample.

3. Determination of Purity of a sample.

4. To identify cis and trans isomers

IR absorption of a trans isomer is very weak and of cis is stronger.

5. Distinguish between inter and intra molecular H bonding

IR spectra of the compound at different concentration are recorded with

increase in concentration, the absorption band due to inter molecular H-
bonding increases while that due to intra molecular H-bonding remains
unchanged.

6. Structural information

IR spectra provides valuable information regarding molecular symmetry,

dipole moments, bond strength, characteristic absorption etc.

NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY

Nuclear magnetic resonance (NMR) spectroscopy is the study of interaction
of radio Frequency electromagnetic radiations with the nuclei of molecules
placed in a strong magnetic field.

Principle

The nuclei carry a charge, this charge spin on the nuclear axis and this

circulation of nuclear charge generates a magnetic dipole along the axis.

STUDY OF NMR SPECTRUM
1. The number of signals.
2. The position of the signal.
3. The intensities of the signal.
4. The splitting of the signal.

THE NUMBER OF SIGNALS

THE POSITION OF THE SIGNAL

Chemical shift
It refers to the difference between the resonant frequency of spinning
protons and the signal of the reference molecule.

Shielding shifts the absorption upfield and deshielded shifts to the downfield

• Shielded protons have lower resonance frequency, and smaller

chemical shift values.
• Deshielded protons have higher resonance frequency, and larger
chemical shift values.

THE SPLITTING OF SIGNAL

OPTICAL ISOMERISM
• Optical isomers refer to those stereoisomers that have the same
molecular formula and even exhibit identical physical and chemical
properties.
• However, they have a different spatial arrangement, and because of
this, they react differently to the plane-polarised light.
• They are non-superimposable mirror images of each other and are
found in molecules having asymmetric carbon.
• One of the isomers rotates the plane-polarised light when passed
through its solution in the clockwise direction. It is referred to as (+) or D-
isomer (dextrorotatory).
• On the other hand, the other one rotates the plane-polarised light
when passed through its solution in the anti-clockwise direction and is
referred to as (-) or L-isomer (laevorotatory).

Compounds with No Chiral Carbon

It is widely known that optical isomer compound must have a chiral carbon.
However, there are a few compounds that do not have chiral carbon yet
exhibit optical activity.

These compounds are non-planar and give rise to non-superimposable mirror

images. To find the chirality of these compounds, we look for symmetry
elements that are the centre of symmetry and plane of symmetry.

> A plane of symmetry is an imaginary plane that bisects the molecule in

such a way that it gives identical mirror images.

> The centre of symmetry is an imaginary point at the centre of a molecule

(Can be on an atom, on a bond, or in space) from where if we move
equidistantly in the opposite directions, it will lead us to the same atoms or
groups.

If any of these symmetry elements are present, the molecule will be optically
inactive. Three of the molecules that bear no chiral carbon but are optically
active molecules are

1. Allenes,

2. Spiro compounds, and

3. Biphenyls

OPTICAL ISOMERISM IN ALLENE

A successive diene or a cumulated diene is known as an allene.

The 3 carbon atoms in allene are doubly bonded, so the central carbon
atom Is sp hybridised while the 2 carbon atoms on either side are sp2
hybridised.

This way, no plane of symmetry or centre of symmetry will exist and will make
them optically active.
OPTICAL ISOMERISM IN SPIRO COMPOUNDS
Spiro compounds are made up of two rings attached to a single carbon
atom (Spiro carbon atom), which is sp3 hybridised.

This results in a tetrahedral geometry which makes the 2 rings perpendicular

to each other (non-planar in 3-D). Spiro compounds thus give non-
superimposable mirror images.

OPTICAL ISOMERISM IN BIPHENYL

Biphenyls are formed by 2 benzene rings attached via a single sigma bond.

When biphenyl rings are substituted over ortho position, the carbon atoms in
benzene are sp2 hybridised, thus the rotation across the sigma bond is
restricted.

Optical activity in biphenyls is observed when ortho substitution is present

over both the rings and the groups are not identical. Otherwise, a plane of
symmetry will exist and it will become optically inactive.

TYPES OF STEREOISOMERISM
a) Conformational isomerism: Also termed as rotational isomers as they have
free rotation of groups about C - C bond axis as they have different
arrangements of atoms in space. This type of isomerism is found in alkanes,
cycloalkenes and their derivatives.

b) Configurational isomerism: They differ in the arrangement of atoms and

have different properties. It is of two types geometrical isomerism and optical
isomerism. These isomers do not have free rotation around the C-C single
bond.

GEOMETRICAL ISOMERISM
Geometrical isomers are stereoisomers that have a different arrangement of
groups or atoms around double bonds.

Cis isomers of compounds are less stable than trans-isomers.

Cis isomers: If the functional groups (substituents) are on the same side of the
plane.

Trans isomers: If functional groups are on the opposing sides.

Necessary conditions for geometrical isomerism:

1. Restricted rotation around the C — C double bond.

2. Two atoms or groups attached to the molecule with the doubly bonded
carbon atom must be different. However, it doesn’t matter whether the left
side groups are the same or different on the right side.

E/Z SYSTEM OF NOMENCLATURE OF GEOMETRICAL ISOMERS

If the two group of highest priority are on the opposite side of the double
bond.

The alphabet E (Entagagaen: German word meaning opposite) is used for

the structure.

If the groups of highest priority are on the same side the alphabet Z
(Zusammen: German word meaning together) is used. Thus, E stands for
opposite side and Z for the same side
Determination of the configuration of geometrical isomers
(I) Physical methods: - Generally trans-isomer had high m.p. and low b.p.
than cis isomers.
(II) Solubility: Solubility of cis isomer is more. For example, solubility of maleic
acid is 3.0 g/100 mL of water at 293 K and solubility of fumaric acid is 0.7
g in 100 mL of water at 293 K