SPECTROSCOPY: Spectroscopy may be defined as the interaction between

the matter and electromagnetic radiations.

There are two types of spectrums:

Emission spectra: Molecules give emission spectra when subjected to intense heat or electric
discharge. The molecules obtain the necessary energy to become excited. On returning to their
lower energy state, molecules may emit radiation, which is the result of a transition of a
molecule from an excited state to one of lower energy, usually the ground state. This excess
energy is emitted as photon and the corresponding frequency is recorded as the emission
spectrum.

Absorption spectra: When a substance is irradiated with electromagnetic radiation, the

energy of the incident photons may be transferred to the molecules raising them from the
ground state to an excited state. This process is known as absorption and the resultant spectrum
is known as absorption spectrum. Energy absorption occurs only when the energy difference
between the ground state and higher energy level is exactly matched by the energy of the
incident electromagnetic radiation.

Representation of absorption spectrum: When electromagnetic radiation interacts with the

molecules, then some of wavelengths (or frequency or wavenumbers) of radiation are absorbed
while the rest of its is transmitted. Consequently, we can plot either
1) Absorbance (A) versus λ or frequency or wave number
2) Percent transmittance (%T) versus λ or frequency or wave number.

Principal of IR spectroscopy: The IR spectroscopy theory utilizes the concept that molecules
tend to absorb specific frequencies of light that are characteristic of the corresponding structure
of the molecules. The energies are reliant on the shape of the molecular surfaces, the associated
vibronic coupling, and the mass corresponding to the atoms.
For instance, the molecule can absorb the energy contained in the incident light and the result
is a faster rotation or a more pronounced vibration.
 INFRARED (IR) SPECTROSCOPY

OR

VIBRATIONAL SPECTROSCOPY

 Infrared spectroscopy is used in identification of functional groups in pure compounds.

 Infra-red (IR) does not have sufficient energy to induce electronic transition as seen in UV
spectroscopy. When molecule absorbed electromagnetic radiation in IR region, undergoes
vibrational or a rotational transition whichcauses net change in the dipole moment in the
molecule (IR active, for example HCl, CO etc), if dipole moment does not change in
molecules, then they are IR inactive (for example: O2, H2, N2 etc.) means they does not
absorb IR radiation. IR region ranges from 4000-400 cm-1.
 If the frequency of IR radiation matched with the vibrational frequency of molecule, then
molecule absorb radiation.
 IR spectroscopy based on Hooke’s law, suppose two atoms or masses are connected
through spring (bond), then frequency of vibration can be represented by following
equation:

1 𝗆 or ῡ = 1 𝗆
ν= √
2π μ 2πc √μ

Where, 𝜅 is force constant of the bond, ῡ is wave-number (cm-1), ν is the frequency, c is

speed of light and 𝜇 is reduce mass (m1 and m2 are the masses of atoms)

m1 × m2
μ =
(m1 + m2)

 Stronger the bond, greater the value of force constant (𝜅), higher the frequency
vibration or wave-number (cm-1). For example,
C−C C=C C≡C
dynes
force constant 5 x 105 10 x 105 15 x 105 ( )
cm
wavenumber (cm−1) 1200 1650 2100
 IR spectrum is divided mainly into two region as follows;
 FUNDAMENTAL VIBRATION

These vibrations are arising when molecule promoted from ground state to lower excited state.
The fundamental vibrations for linear and non-linear molecules are determined by following
way:

Molecule Degree of freedom

Linear 3n-5
Non-linear 3n-6
Where, ‘n’ is number of atom present in the moleculeThe

vibrations discuss below are fundamental vibrations.

a) Stretching vibration: Distance between two atoms increase and decrease butbond angle
remains constant.

Types of stretching vibrations

i) Symmetric stretching vibration: In this case both the atoms stretchedor
compressed in same direction.

ii) Asymmetric stretching vibration: In this vibration one atom undergoesstretching

and other atom undergoes compression and vice versa.

b) Bending vibrations: Distance between two atom remains constant but bondangle
changes. These vibrations can occur either in plane or out of plane.
Types of bending vibrations
1) In plane bending vibrations:
i) Scissoring: both the atom move towards each other just likescissor.

ii) Rocking: both the atoms move in same direction, either in left sideor right
side.

2) Out of plane bending vibrations:

i) Wagging: both the atom move up and down with respect to centralatom.
ii) Twisting: one atom move up and other atom move down with r espect to

central atom.
 OVERTONES AND COMBINATION BANDS

When molecule absorbed electromagnetic radiation in IR region, and then molecule promoted
from ground state to second, third or even fourth vibrational excited state. These bands are known
as Overtones. The intensity of these bands is very weak. It is helpful in characterization of aromatic
compounds.

When two fundamental vibrational frequencies (ν 1 + ν2) in a molecule couple to give rise to a new
vibrational frequency within the molecule, it is known ascombination band.

COUPLED VIBRATIONS

The coupled vibrations are observed in group like –CH2, NH2 etc. In these groups same atoms are
attached to the central atom. When –CH2 undergoes vibration by the absorption of IR radiation,
due to internal perturbation, energy of one C-H bond is transfer to neighboring C-H bond which
enhance its vibrational frequency. Therefore, two stretching vibrational frequencies for –CH2
group is observed at 2950 cm-1 (asymmetric stretching) and 2860 cm-1 (symmetric stretching).

FERMI RESONANCE

When fundamental vibration coupled with overtones or combination band, the coupled vibration
is called Fermi resonance or when molecule absorb IR radiation then it transfers its energy or
intensity from fundamental vibration to overtones, then Fermi resonance is observed. As we know
that the intensity of overtones band is very weak as compare to fundamental vibrations. But, due to
transfer of energy, the strong band is observed for overtones along with the fundamental frequency.
Fermi resonance is generally observed in carbonyl groups. For example, in benzoyl chloride –
C=O stretching vibration observed at 1790 cm-1 and 1745 cm-1. The lower frequency band at 1745
cm-1 is observed due to combination of
overtones of CH bending vibration at 875 cm-1 with the fundamental vibration of C=O stretching.

FINGERPRINT REGION

The region from 1500-600 cm-1 in IR spectrum is known as Fingerprint region. In this region
number of bending vibration is more than the number of stretching vibration.

Number of molecules contains same functional group & show similar peak above 1500 cm-1 but
they show different peak in finger print region. Therefore, we can saythat each and every molecule
have unique peak or band which is observed in finger print region, it is just like the finger print of
human.
IR correlation chart

Types of vibration Frequency (cm-1)

Alkane C-C stretching 1200
C-H stretching 3000-2840
-CH2 bending 1465
-CH3 bending 1375
CH2 rocking 720
Alkenes C=C stretching 1650
=C-H stretching 3095-3010
=C-H bending 1000-650
Alkynes C≡C stretching 2100
≡C-H stretching 3300
≡C-H bending 700-600
Aromatic C=C stretching 1600, 1500, 1450
=C-H stretching 3040-3010
=C-H bending Below 900
C=O stretching Amide 1680
Carboxylic acid 1710
Ketone 1715
Aldehyde 1725
Ester 1735
Acid chloride 1800
Anhydride 1760 (I), 1810 (II)
C-O Stretching 1300-1000
O-H Alcohol, phenol
Free 3600
H-bonded 3400-3200
Carboxylic acid 3400-2400
Amine -N-H stretching 3440 (as), 3350 (s)
-N-H bending 1650-1580
C-N stretching 1350-1000
Amide -N-H stretching 3370 (as), 3150 (s)
-N-H bending 1650-1560
Nitriles C≡N 2250
Imines C=N 1690-1640
Nitro group (NO2) N=O 1550 (as), 1350 (s)
Alkyl halide (C-X) C-F 1350
C-Cl 750
C-Br, C-I <660
Factor affecting vibrational frequency

a) Conjugation: As the conjugation increase, stretching frequency decreases,because force

content decreases due to conjugation.

b) Inductive effect and resonance effect:

Oxygen is more electronegative than nitrogen, therefore nitrogen easily donate electron or
lone pair of nitrogen undergoes delocalization with C=O bond. Due to delocalization
double bond of C=O changes into partial double bond therefore force constant decreases
which decrease the C=O stretching frequency.

c) Hydrogen bonding:
Intermolecular hydrogen bonding weakens the O-H bond, thereby shifting the band to
lower frequency. For example, in neat solution O-H stretching
 vibration of phenol observed in the range from 3400-3300 cm-1. Whensolution is dilute
then O-H frequency shifted towards higher frequency at 3600 cm-1.
Whereas in case of methyl salisilate, intramolecular hydrogen bonding lower down the

stretching frequency of O-H at 3200 cm-1. Intramolecular hydrogen bonding does not
change its frequency even in very dilute solution because upon dilution structure of
compound does not change.

d) Ring strain: As the size of the ring decrease, vibrational frequency of C=Oincrease.
For example.

Application of IR Spectroscopy

a) Identification of different functional group.

b) Distinction between intermolecular and intra-molecular hydrogen bonding.
c) Identification of purity of the compound, if compound is impure thenadditional
peaks are observed in the IR spectrum.
d) Study of chemical reaction:

e) Identification of geometrical isomers (cis-trans).

 UV-VISIBLE SPECTROSCOPY

 Ultraviolet and visible spectroscopy also known as electronic spectroscopy is

used to measure the number of conjugations of double bond and aromatic
conjugation within the molecule.
 It involves the promotion of electrons from HOMO to LUMO (HOMO means
Highest Occupied Molecular Orbital whereas LUMO means Lowest
Unoccupied Molecular Orbital).
 The HOMO-LUMO gap decrease as the conjugation increase.

 The ultraviolet region corresponds to 400-200 nm and visible region to 800-

400 nm.
 UV-visible spectroscopy based on Beer-Lambert law.
I
log o⁄I = ε. 𝑙. 𝑐 or ϵ = 𝐴/𝑐 𝑙

Where,

Io = intensity of incident light

I = intensity of transmitted light

ε = molar absorptivity

l = path length of sample

c = concentration of sample

A = Absorbance

 Absorbance (A): It is reciprocal of Transmittance Io

1 or log
Absobance (A) = Optical density (D) = log I
T

 Transmittance (T): The fraction of incident light transmitted is known as

Transmittance.
 I
Transmittance (T) =
Io

Electronic transitions
When molecule is getting excited by the absorption of electromagnetic radiation in
UV-visible region then its electrons are promoted from ground state to excited
state or from bonding orbital to anti-bonding orbital.

Types of electronic transitions

a) 𝜎 − 𝜎* Transition: transition of an electron from bonding sigma orbital (𝜎) to

anti-bonding sigma orbital ( 𝜎 *), is represented by 𝜎 − 𝜎 * transition. For
example, alkanes because in alkane all the atoms are held together with sigma
bond.
b) 𝑛 − 𝜎* & 𝑛 − 𝜋* Transition: Transitions from non-bonding molecular (𝑛)
orbital to anti-bonding sigma orbital or anti-bonding pi orbital (𝜋* ), are
represented by 𝑛 − 𝜎 * or 𝑛 − 𝜋 * transition respectively. This transition
required less energy than 𝜎 − 𝜎 * transition. For example, alkyl halide,
aldehydes, ketones etc.
c) 𝜋 − 𝜋 * Transition: This type of transition generally shows in unsaturated
molecules like alkenes, alkynes, aromatics, carbonyl compounds etc. This
transition required less energy as compare to 𝑛 − 𝜎* transition.
 
n-    n-  
* *

Energy

Absorption, intensity shift & UV spectrum

a) Bathochromic shift: It is also known as Red shift, in this case absorption

shift towards longer wavelength (𝜆max).
b) Hypsochromic shift: It is also known as Blue shift, in this case absorption
shift towards shorter wavelength (𝜆max).
c) Hyperchromic shift: Intensity of absorption maximum (𝜀max) increases.
d) Hypochromic shift: Intensity of absorption maximum (𝜀max) decrease.

Absorption & intensity shift

UV spectrum of unknown compound

 Chromophores & Auxochromes

a) Chromophores: Chromophore is a covalently unsaturated group absorbed

electromagnetic radiation in UV-visible region and impart color to the
compound. For example, C=C, C≡C, benzene, NO2 etc.
b) Auxochromes: Auxochrome is a saturated group (containing lone pair of
electron), when auxochrome attach to the Chromophore absorption shift
towards longer wavelength. For example, -OH, -SH, -NH etc.

Factor affecting absorbance & intensity

a) Conjugation

Absorption shift towards longer wavelength, if double bonds (chromophore)

present in the molecule are in conjugation. For example,

In compound ‘A’, double bonds are in conjugation therefore ‘A’ possessing higher
wavelength (𝜆max) as compare to compound ‘B’ (non-conjugated derivative).
We know that,
ℎ𝑐
E=
𝜆

As the conjugation increase transition energy (E) between the orbitals is decrease
and therefore wavelength (𝜆max) increase.

b) Effect of solvent:
In polar solvent, 𝜋 − 𝜋* transitions shift towards longer wavelength (Red
shift), because dipole interactions with polar solvent molecules lower the
energy of excited state (𝜋*) than that of the ground state (𝜋* orbital’s are
stabilized by hydrogen bonding with polar solvent).

Whereas in polar solvent, 𝑛 − 𝜋* transitions shift towards lower wavelength

(Blue shift), because dipole interactions with polar solvent molecules lower
the energy of ground state (𝑛) than that of the excited state (𝑛 orbital’s are
stabilized by hydrogen bonding with polar solvent).

The Woodward-Fieser rules (Calculation of 𝝀max (nm) in

conjugated dienes)

𝜆max (nm)
Base value:
Acyclic or heteroannular dienes 214
Homoannular dienes 253
Increments for:
Double bond extending conjugation 30
R alkyl substituent or ring residue 5
Exocyclic double bond 5
Polar groupings:
-OCOCH3 0
-OR 6
-Cl, -Br 5
-NR2 60

Explanation of terminology used in Woodward-Fieser rules

 The Woodward-Fieser rules (Calculation of 𝝀 max (nm)
in α,β-unsaturatedcarbonyl compounds)

𝜆max (nm)
Base value:
Acyclic or 6-membered cyclic ketone 215
5-membered cyclic ketone 202
Aldehyde 210
Increments for:
Homocyclic diene component 39
Double bond extending conjugation 30
R alkyl substituent or ring residue 𝛼 = 10, 𝛽 = 12, 𝛾 & ℎ𝑖𝑔ℎ𝑒𝑟 = 18
Exocyclic double bond 5
Polar groupings:
-OH 𝛼 = 35, 𝛽 = 30, 𝛾 = 30
-OR 𝛼 = 35, 𝛽 = 30, 𝛾 = 17
-Cl, 𝛼 = 15, 𝛽 = 12, 𝛾 = 12
-Br 𝛼 = 25, 𝛽 = 30, 𝛾 = 25
-NR2 𝛽 = 95
-OCOCH3 𝛼 = 6, 𝛽 = 6, 𝛾 = 6
Application of UV spectroscopy

i) Extent of conjugation: As the double bond increase, absorption shift towards

longer wavelength.
ii) Distinction between conjugated and non conjugated compounds:

Both the above compound A & B are isomer of each other. Due to 𝑛 − 𝜋*
transition, compound A appear at longer wavelength than compound B.
iii) Study of geometric isomerism:

In isomer A, due to steric hindrance, absorption shift towards lower

wavelength. We know that, steric hindrance destabilizes the compound
means energy of the molecule increase which decreases the wavelength.
iv) Used in the detection of impurities: Due to presence of impurities in the
compound, give additional peaks are observed in UV spectrum.
INSTRUMENTATION:

Source: Supplied light in the UV range (200-400 nm) hydrogen or deuterium lamp
and Visible range (400-800 nm) Tungsten lamp.
Monochromator: Allows only the chosen wave length and absorb all the other
wave length and this happen for the complete chosen range of wave length. Eg
Prism, diffraction etc.
Beam splitter: Splits the beam of light of each wavelength into two halves of equal
intensities. One goes to the test sample and other to the blank /reference sample.
Cuvettes: One cell is filled with the test sample and the other with the reference
sample.
Detector: It detects the intensities of the transmittance light coming out of the cells
and compares for each wave length and generates currents proportional to the
differences in the intensities of the sample and blank.
Recorder: It records an Uv/ visible spectrograph with absorbance against the wave
length.
 ATOMIC ABSORPTION SPECTROSCOPY

 Atomic absorption spectroscopy is a very common technique for detecting

metals and metalloids in samples.
 It is very reliable and simple to use.
 It also measures the concentration of metals in samples.

Principal: The basic principles of AAS can be expressed as follows. Firstly, all atoms or ions
can absorb light at specific, unique wavelengths. When a sample containing copper (Cu) and
nickel (Ni), for example, is exposed to light at the characteristic wavelength of Cu, then only
the Cu atoms or ions will absorb this light. The amount of light absorbed at this wavelength is
directly proportional to the concentration of the absorbing ions or atoms.

The electrons within an atom exist at various energy levels. When the atom is exposed to its
own unique wavelength, it can absorb the energy (photons) and electrons move from a ground
state to excited states. The radiant energy absorbed by the electrons is directly related to the
transition that occurs during this process. Furthermore, since the electronic structure of every
element is unique, the radiation absorbed represents a unique property of each individual
element and it can be measured.

Instrumentation: An atomic absorption spectrometer uses these basic principles and applies
them in practical quantitative analysis. A typical atomic absorption spectrometer consists of
four main components: the light source, the atomization system, the monochromator and the
detection system.
Uv