By

What is Spectroscopy?
Spectroscopy is the study of the
interaction of Electromagnetic Radiation
(EMR) with the matter as a function of
wavelength or wave number or
frequency. The matter may be atoms,
molecules or ions.
What is spectrometer and spectrum?

Spectrometer is something which can be used

to measure the presence of particular
compound or molecule in a sample.

Spectrum is the amount of light absorbed by a

compound (intensity) versus the energy of
the light in terms of wavelength, frequency or
wave number.
 Electromagnetic Radiation (EMR)

 EMR is a form of energy that is all around us and

has both electrical and magnetic characteristics.
As per electromagnetism theory, the electrically
charged particles interact with each other and the
magnetic fields.
The electric and magnetic fields in an EMR oscillate
along the direction perpendicular to the propagation
direction of wave.
 Various Terms associated with EMR
 Wavelength (λ): The distance between two successive
waves, measured in nm, cm, m.

 Frequency (ν): Number of waves produced in each second,

measured in Hz.

Amplitude: The maximum distance a wave extends beyond its

middle position.

Wave Number (ῡ): The reciprocal of wave length (1/λ),

measured in cm-1, m-1.
Relationships: Velocity (c) = ν.λ
E = hν = h (c/λ) = hc ῡ
 Different Regions of EMR
EMR can be divided into various regions based on
the frequency (energy) for convenience. It ranges
from very short wavelength (gamma ray) to very
long wavelength (Radio wave)
The energy of a molecule/atom can be written as

E total = E trans + E rot + Evib + Eelectronic

And E trans < E rot < E vib < E electronic

As the translational energy levels are very close for a

molecule, they can be considered continuous.

The other three forms of energy levels are quantised and

the magnitude of their energy level gaps varies
considerable.
The effect of interaction of EMR with the matter
depends on the energy associated with the radiation.
Radiation Used in

Gamma ray Nuclear reaction and treatment of cancer

X-ray XRD for determining the microstrcture of

crystalline compounds, to see inside of bodies
and objects
Ultraviolet and UV-Vis Spectroscopy (electronic Transition)
Visible ray
Infrared ray IR spectroscopy (vibrational-rotational
transition)
Microwave Ray Rotational spectroscopy (Rotational
transition), cooking, Radar & Mobile signal
Radiowaves NMR spectroscopy (for determining the
structure of organic compounds),broadcast
of radio & TV
The interaction between EMR and matter may be

1. Absorption
2. Emission
3. Scattering

So we can have three types of spectroscopies:

1. Absorption Spectroscopy
2. Emission Spectroscopy
3. Scattering Spectroscopy

This help in quantitative and qualitative study of

the matter
 Absorption Spectroscopy

• In absorption spectroscopy, an EMR is

absorbed by a molecule or atom

• By the process, it undergoes transition from

lower energy level to higher energy level or
excited state

• The absorption occurs only when the energy

of EMR matches with the energy gap of two
energy levels.
 Emission Spectroscopy

• Atoms or molecules, that are excited to

higher energy level, can decay to lower level
by emitting radiation, which is studied by
emission spectroscopy.

• The substance first absorbs energy and then

emits this energy as light

• Emission can be induced by sources of

energy such as flame or EMR.
 Scattering Spectroscopy

• Scattering spectroscopy measures certain

physical properties by measuring the amount
of light that a matter scatters at certain
wavelength.

• One of the most useful light scattering

spectroscopy is Raman Spectroscopy
 Microwave
Spectroscopy
 Theory of Microwave Spectroscopy
•Microwave radiation: Microwave is an EMR
with wavelength in the range 3 mm to 1.3 mm
and frequency in the range of 200 MHz to
100GHz.
Theory:
• A molecule with permanent dipole moment
generates fluctuating or oscillating electric
field by its rotation. When the frequency of the
oscillation of the molecules matches with the
frequency of the electric field component of
the incident MW radiation, the resonance
occurs and rotational transition takes place.
MW Active molecules:
Molecules with permanent dipole moments exhibit
microwave (rotational) spectrum, and are called as MW
active molecules.
•Example: HCl, HF, CO, CN etc
MW Inactive molecules:
A molecule with no permanent dipole moment does not
exhibits microwave (rotational) spectrum, and are called
as MW inactive molecules.
•Example: H2, O2, N2, (homonuclear molecules) and
heteronuclear molecules with center of symmetry such as,
CO2 etc.
• Rotational spectra are observed only in case of molecules existing
in the gaseous state, but not in their solid or liquid state as free
rotation is restricted and energy levels are not quantised.
 MW spectra of diatomic molecules
Consider a diatomic molecule (AB) of equilibrium bond length r,
rotating about its axis through center of gravity (C.G.). The distance
between C.G. and the atom A (with mass m1) and atom B (with mass
m2) are r1 and r2 respectively.
As r = r1+r2 and m1r1 = m2r2

As r = r1+r2 and m1r1 = m2r2

 It is customary to express the energy (EJ) in cm-1
(wave number) rather than in J. So (viii) in modified
form obtained by dividing EJ by hc is:
EJ = [ h J (J+1)]/ (8 π2 I c) cm-1
= B J (J+1) cm-1
Where B is rotational constant, B = h / (8 π2 I c) cm-1
 Selection rules for Rotational transition
• A rotating molecule must possess a permanent
dipole moment so as to interact with oscillating
electric field of EMR.

• The rotational transition is governed by the

selection rule: ΔJ = ±1

• ΔJ = +1, absorption takes place

• ΔJ = -1, emission takes place
For a rotational transition taking
place from J  J+1, the energy
(frequency) required can be given by

ΔE JJ+1 = EJ+1 – EJ
= B (J+1)(J+2) – B J(J+1)
= B (J+1) [(J+2) - J)]
= 2B (J+1)
Allowed Rotational energies of a diatomic
molecule as per Selection rules ΔJ = ±1
 Applications of Rotational (Microwave) Spectroscopy
1. Determination of bond lengths of gaseous polar molecules:
The bond length of a molecules with permanent dipole moments in
their gaseous state can be determined using the data obtained from
microwave (rotational) spectrum.

I
2B 2B
n
t 2B
e 2B
n 2B 2B
s 2B 2B
i 2B
t 2B
y

2B 4B 6B 8B 10B 12B 14B 16B 18B 20B 22B

Wave number (cm-1)
2. Determination of dipole moment of a of gaseous polar molecule:

It can be determined by recording microwave (rotational) spectrum

in the presence of a strong external electric field. Under this
condition, the rotational spectral line splits as per stark effect.

If Δν = split of rotational frequency caused by external field

μ = dipole moment of the molecule,
E = Strength of the external electric field
Then,
Δν = (µ x E)2
Δν

Wave number (cm-1)

3. Determination of Isotopic mass of an element:
Different molecules having different isotopes of an element possess
unique moment of inertia depending on the mass of the isotopic
nuclei present in them.
B = h / (8 π2 I c) cm-1 = h / (8 π2 µr2I c) cm-1 ,
That means, B α (1/ I) , B α (1/µ)

So (B / B’ ) = (I’/ I) = (µ’/ µ)
 IR
Spectroscopy
 Principle of IR Spectroscopy

•Infrared region is broadly divided into three parts:

Near IR (0.8- 2.5μm),
IR (2.5-15μm)
and Far IR (15- 200μm)

•The absorption of IR by a compound causes molecular

vibrations.
• This changes in vibrational energy levels are
accompanied by rotational levels as well.
• Thus, each line in vibrational Structure is
accompanied by rotational fine structures.
•IR spectrum is also known as vibrational-rotaional
spectrum
 Principle of IR Spectroscopy

•Infrared region is broadly divided into three parts:

Near IR (0.8- 2.5μm),
IR (2.5-15μm)
and Far IR (15- 200μm)

•The absorption of IR by a compound causes molecular

vibrations.
• This changes in vibrational energy levels are
accompanied by rotational levels as well.
• Thus, each line in vibrational Structure is
accompanied by rotational fine structures.
•IR spectrum is also known as vibrational-rotaional
spectrum
 Requisite for IR Spectra

• The essential requirement for a substance to

absorb IR radiation is that the vibrations in the
molecules must give rise to non-zero dipole
moment. (dμ/dr) ≠ 0

• Homonuclear diatomic molecules (H2, O2, N2, Cl2) do

not show change in dipole moment during vibrations.
So these molecules are IR inactive. They do not exhibit
IR spectra.
• CO2 is MW inactive but some of the vibrations are IR
active.
 Vibrational Energy and zero point energy
As per the quantum chemistry, the allowed vibrational energies (Evib)
are governed by the expression:
Evib = (ν + ½) h ν0

Where ν is the vibrational quantum number, with possible values of

0, 1, 2, 3, etc and ν0 is the fundamental frequency.

Fundamental frequency (ν0 ) It is the frequency of EMR

associated with vibrational transition from ground state (i.e., ν = 0) to
next higher energy level (ν = 1)

At the ground state (i.e., ν = 0), the molecule has the vibrational
energy, Evib = ½ hν0

This is called as Zero point energy (ZPE). It signifies that the

molecule can never be at rest, it must always vibrate.
Why IR Spectra is taken for sample in solid, liquid
or solution form?
• Absorption of IR radiation by a molecule
changes its vibrational as well as rotational
levels.
• Due to presence of rotaional fine structures,
the resulted IR spectrum becomes much
complicated.
• In order to avoid this complication, IR
spectrum of the sample is taken in Solid, liquid
or solution form.
•Under this condition, the rotational fine
structures are buried and a broad peak is
observed for vibrational changes only.
 Types of MolecularVibrations

Stretching vibration Bending vibration

(bond length changed) (bond length unchanged)

In-plane bending Out-of-plane

(i) Symmetric vibrations bending
(ii) Asymmetric vibrations
(i) Rocking
(iii) Wagging
(ii) Scissoring
(iv) Twisting

ν stretching ( asym.) > ν stretching ( sym.) > ν bending

 Number of Vibrational modes or Vibrational degrees of freedom

Theoretically calculated number of vibrational

degrees of freedom or normal modes of vibrations
or fundamental vibrations in a polyatomic
molecule containing ‘n’ no. Of atoms can be
described as:
• For Linear Molecules, it is (3n–5)
Ex: CO, CO2, HCl etc.
• For Non-linear molecules, it is (3n-6)
Ex: H2O, Benzene, CCl4, CH4, BH3 etc.
(No. Of vib. Degrees of freedom) =
(Total degrees of freedom of a molecule) – (translational degrees of
freedom) – ( rotational degrees of freedom) = (3n – 3 – 3)
Various terms associated with vibration bands
Actual number of observed vibration bands for a molecule is,
generally, different from Theoretically calculated number of
vibrational modes, since overtones or combination bands are
involved
OVERTONE Bands: This is resulted due to transition from
ground state to higher energy levels which corresponds to
integral multiples of fundamental frequency (ν0).

Combination Bands: Two vibrational frequencies couple to

give rise to a frequency, known as combination band.

Fermi Resonance: When a fundamental vibration couples

with an overtone or a combination band, the couple vibration
is known as Fermi Resonance.
Finger-print region or Group frequencies region
• IR spectra of polyatomic molecules are quite
complex and it may not be easy to assign mode for
each vibration band.
•However we can get important information by
recognising group frequencies.

• Organic compounds, generally, shows many bands in

the energy range 4000 – 500 cm-1.

• The characteristics peaks for functional groups

appears in the range 1500 – 500 cm-1. This range is
known as Finger-print region or Group frequency
region.
 Hooke’s Law for Stretching Vibration
• The vibrational motion in a molecule is similar to
vibration of simple harmonic oscillator, whose
restoring force (F) is proportional to displacement x
(Hooke’s Law):
F α (-x) or, F = -kx

•The frequency (ν) for simple harmonic motion is given by:

ν = (1/2π) √ (k/μ) s-1
(μ = reduced mass and k = Force constant related to
bond strength)

• In terms of wave number, as it is used in spectroscopy

ῡ = ν/c = (1/2πc) √ (k/μ) cm-1
•Vibrational energy levels are quantised
and energy associated with any
vibrational level can be given by:

E = (ν + ½) h ν0,
• Energy difference between two vibrational
states (i.e., vib. quantum no. ν νʹ),

Δ Evib = Eʹ – E = (νʹ + ½) h ν0 – (ν + ½) h ν0
= (νʹ - ν) h ν0 = Δν. h ν0
 Applications of IR Spectroscopy
1. Identification of Compounds:
• Each group of atoms give rise to a
characteristic absorption bands in the IR
spectrum.
• So by comparing the IR spectrum of a
unknown compound with that of known
compounds, its nature can be established.

•Example: The benzaldehyde and phenyl

methyl ketone shows aborption peak at 1700
cm-1 indicating the presence of C=O group.
IR spectrum of Acetophenone
2. Identification of functional groups:
3. Measurement of Force constant:
By using the equation: ῡ = ν/c = (1/2πc) √ (k/μ) s-1

4. Detection Impurities:
The appearance of extra peak other than the
characteristics IR peaks for the compound indicates
the presence of impurity.

5. Detection of Intra- and Inter-molecular

Hydrogen bonding:
The absorption band due to intermolecular H- bonding
decreases and that due to intra-molecular H- bonding
remains unchanged as dilution increases.
6. Studying the progress of a reaction:
The progress of a reaction can be studied by taking the IR
spectra of the sample collected at different time interval.

7. Distinguishing structural isomers of a compound:

As the isomers have different bondlength and bond
strength, they shows different IRabsorption band

8. Metal-Ligand bonding modes in complex

compounds:

9. Quantitative analysis:
IR spectra may be used for rapid quantitative analysis of a
mixture of compounds, e.g. In milk analysis, in pollution
detection etc.
 UV-Visible
Spectroscopy
 Principle of Absorption Spectroscopy
(UV-Vis spectroscopy)
Lambert’s Law:
It states that when monochromatic light passes through
a transparent medium, the intensity of the transmitted
light decreases exponentially with increase in thickness
of the absorbing materials (path length).
Mathematically,
-(dI/dx) α I or -(dI/dx) = k I
On integration with boundary condition that I =
I0 at x = 0 and I = I at x=x, we get

Absorbance, A = log (I0/I) = ε x

log (I0/I) =Absorbance, (A)
 Principle of Absorption Spectroscopy
(UV-Vis spectroscopy)
Beer’s Law:
It states that when monochromatic light passes
through a transparent medium, the intensity of the
transmitted light decreases exponentially with
increase in concentration of the absorbing materials.

Absorbance, A = log (I0/I) = k’ c

 Beer-Lambert’s Law
It states that the quantity of light absorbed by a substance
dissolved in a fully transmitting solvent, is directly
proportional to concentration of the substance and the path
length of the light through the solution.
Mathematically,
Absorbance, A = log (I0/I) = log (1/T)= ε.C.L
[T is transmittance = (I/I0 ), %T = (I/I0 )x100)]
Where, I0 and I are the intensities of the incident and transmitted light
respectively.
C = conc. Of the solution in mol/L
L = path length (usually 1 cm)
ε = molar extinction coefficient or molar absorptivity
Absorbance “A” is a dimensionless quantity
 Limitations of Beer-Lambert’s Law

Beer-Lambert’s Law is not obeyed:

1. When different forms of absorbing
molecules are in equilibrium as in keto-
enol tautomers
2. When fluoroscent compounds are present
3. When solute and solvents form complexes
through some sort of association.
4. When concentration is very high.
 Instrumentation UV-Vis Spectrometer

316
b
Absorbance (a.u.)

246

469
Tetrathiomolybdate
And
oxythiomolybdate 394

200 300 400 500 600

W avelength (nm )
 Molecular orbital approach to absorption
(Theory of electronic absorption)

• LCAO results in molecular orbitals (MO),

one bonding and anti-bonding MO.
• According to MOT, a molecule is excited
when an electron is transferred from bonding
to antibonding MO. In organic compounds
four kinds of electronic transitions may occur.

• σ σ* transition
• n  σ* transition
• ππ* transition
• nπ* transition
 σ*

Energy π*

n
n  σ* nπ*
π
ππ*
σ
σ σ*
Energy requirement for these transitions follows the order as:

σ σ* transition > n  σ* transition > ππ* transition > nπ*

transition
 σ σ* transition:
• High energy transition as σ bond is very strong and
absorption band appears nearly λ value180-200 nm (uv
region)

• Exhibited by saturated compounds like alkane ( Ethane (135

nm), methane (150 nm)

n  σ* transition:
• Less energitic than σ σ* transition
• Absorption occurs at longer λ value
• Exhibited by the saturated compounds with atleast one
heteroatom containing unshared pair of electron
•Examples: alcohol, amines, ketones, aldehydes etc.
• water absorbs at 167 nm, methanol at 174 nm, methyl
chloride at 169 nm.
 π π* transition:
• Less energitic than σ σ* transition and n σ* transition

• Exhibited by compounds containing double and triple bonds

like alkene and alkyne, aromatic compounds
•
n  π* transition:
• Least energitic transition.
• Absorption occurs at longer λ value
•These are forbidden bands
•As non bonded electrons are loosely held, its transition require
less energy.
• Exhibited by the unsaturated compounds with one heteroatom
containing unshared pair of electron like C=O, C=S etc.
 Transition probability

Depending on the value of extinction

coefficient, transitions are classified as
 Allowed transition (if ε max is more
than 104 )
 arises due to π π* transition

 Forbidden transition (if ε max is below

104 )
 arises due to n π* transition
 Chromophores
• Chromophores imparts color to the
compounds
• It is defined as any isolated covalently
bonded group that shows characteristic
absorption in UV-visible region.
• Examples are Ethylinic linkage (C=C) and
acetylinic linkage (C≡C)
• Those which contain both pi electrons and
nonbonding electrons such keto group,
nitrile group, azo group.
 Auxochromes
An auxochrome is color enhancer.
• It is defined as any group that does not
itself act as a chromophore but whose
presence shifts the absorption band towards
longer wavelength.
• It is saturated group which when attached
to a chromophore changes its intensity as
well as λ max.
• Examples are OH, NH2, Cl groupetc.
•Benzene absorbs at 255 nm with εmax 203,
whereas aniline absorbs at 280 nm with εmax
1430
Absorption and intensity Shifts
 Bathochromic Shift
• It is also known as Red Shift
• λ max shifts towards longer wavelength either
due to presence of an auxochrome or due to
effect of solvent
• Example: due to extend in conjugation
CH2=CH- CH=CH2 (λ max=217 nm)
CH2=CH-CH=CH-CH=CH2 (λ max=254 nm)
• Due to change in polarity of the solvent
 Hypsochromic Shift
• It is also known as Blue Shift
• λ max shifts towards shorter wavelength either
due to removal of conjugation or due to effect of
solvent
• Example: due to removal of conjugation
--NH2 (λ max=280 nm)

--NH3+ (In acidic medium λ max=254 nm)

• λ max value for n π* transition decreases by
changing the non polar solvent to polar solvent.
Hyperchromic Shift (Intensity Up-Shift)
• Due to this effect, ε max increases
• It generally occurs due to introduction
of an auxochrome.

• Example: π π* transition of pyridine

at 257 nm with ε max 2750 is shifted to
262 nm with ε max 3560 for methyl
pyridine.
Hypochromic Shift (Intensity Down-Shift)
• Due to this effect, ε max decreases
• It generally occurs due to introduction group
that distorts the geometry of the molecule

Example:
•Biphenyl absorbs at at 250nm with ε max 19000
•But 2-methyl biphenyl absorbs at 237 nm with
ε max 10250 for methyl pyridine.
•The methyl group distorts the geometry of the
biphenyl leading to breaking the extent of
conjugation.
 Solvent Effect
• The solvent used for preparing the solutions
should not absorb in the region under investigation.
• However, the polarity of the solvent affects the
λmax and ε max .
• Non-polar compounds like, ethylene, butadiene,
aromatic hydrocarbons, conjugated hydrocarbons
exhibit little shift on changing the polarity of the
solvent.
•Polar compounds like α,β-unsaturated carbonyl
compounds exhibit two different shifts on changing
the polarity of the solvent.
• Generally n π* transition n σ* transition
band moves towards shorter wavelength (blue shift)
on increasing the polarity of the solvent.

Reason: The polar solvent stabilizes the n electrons

in the ground state more than the excited state (π*
and σ* ) due to H-bonding. Thus, the energy of n
orbital is lowered increasing the energy gap
between the g.s. and excited state. Thus ,absorption
band move towards shorter wavelength.
• Ex: λmax acetone is 279nm in hexane and 264 nm
in water (polar).
• π π* absorption band moves towards longer
wavelength (red shift) on increasing the polarity of
the solvent.

Reason: The excited state (π*) is more polar than

the g.s. (π). So dipole-dipole interactions with the
polar solvent lower the energy of the excited state
more than g.s. due to H-bonding. Thus, the energy
required for π π* transition is less and, absorption
band move towards longer wavelength.
• Ex: λmax is greater in ethanol than that observed
in hexane (non-polar).
 Applications of UV-Vis Spectroscopy
1. Detection of functional groups:
Ethylenic group λmax is 175 nm
Carbonyl group , λmax is 290 nm

2. Extent of Conjugation:
Conjugation lowers the π π* energy gap. So greater the
extent of conjugation, longer is the absorption wavelength.
Ethylene absorbs at 170 nm, whereas butadiene absorbs at
217 nm.
3. Distinction between conjugated and non conjugated
compounds:
Compound having conjugation absorbs at longer
wavelength than non-conjugated compound.
4. Identification of unknown compounds:
Identification of unknown compounds can be done by
comparing its spectra with the spectra of known
compound.
5. Determination of geometrical isomers (Cis-Trans):
Cis isomer absorb shorter wavelength compared to its
trans-isomer. Bcoz, the steric strain in cis isomer
prevents the coplanarity increasing ππ* energy gap.
Cis-stilbene absorbs at 280 nm, whereas trans-stilbene
absorbs at 295 nm.
6. Detection of impurities:
Ethanol containing benzene as impurity can be identified
from the presence of the absorption peak (λmax) for
benzene at 280 nm.
7. Detection of Hydrogen bonding:
Detection of hydrogen bonding can be done on the basis
of the shift absorbed in polar solvent.
8. Quantitative analysis:
This is based on Beer-Lambert’s law; A = εcl
The concentration of unknown solution can be
determined by using the absorbance values of standard
solution (A1) and unknown solution (A2) .
(A1/A2) = (c1/c2)

Problem: For asolution of camphor in hexane in a 5 cm

cell, the absorbance A was found to be 2.52 with εmax 14.
What is the conc of the camphor? (Ans- 3.6*10-2 mol.L-1)